Method and equipment for quantifying oxygen release capacity of HBOC, medium and program product

Abstract

The invention provides a method, a device, a medium and a program product for quantifying the oxygen release capacity of HBOC, further discloses a method, a device, a medium and a program product for evaluating the oxygen release speed of HBOC based on the oxygen release speed, further discloses a method, a device, a medium and a program product for predicting the Boer effect of HBOC, and relates to the field of intelligent medical treatment. The in-vivo oxygen release capacity of the HBOC is evaluated by utilizing a high-flux oxygen dissociation analysis technology, and the method has the advantages that a plurality of HBOC samples can be detected at the same time, the oxygen release ratio and the oxygen release rate of the HBOC samples can be obtained, prescription screening can be performed more quickly, meanwhile, the Boer effect of the HBOC can be semi-quantitatively represented, the acidic Boer effect and the alkaline Boer effect can be distinguished, and the method has a good application prospect. Therefore, technical support is provided for research and development of HBOC.

Description

Technical Field

[0001] The present invention relates to the field of intelligent medical treatment, and more specifically, to a method, device, medium and program product for quantifying the oxygen release capacity of HBOC. Background Art

[0002] Blood transfusion is an intervention measure with standardized procedures, which can supplement missing blood components in critical situations and save patients' lives. Therefore, the demand for blood substitutes - hemoglobin-based oxygen carriers (HBOCs) is becoming more and more urgent. Hemoglobin oxygen carriers are based on human or animal hemoglobin, obtained by polymerization, modification or encapsulation. Because of their good biocompatibility, safety and stability, as well as oxygen carrying / release capacity similar to natural red blood cells, they have gradually become the main research direction of blood substitutes. The main function of HBOC is to improve tissue oxygen supply by relying on its oxygen carrying / release capacity. It can be used to treat hypoxic diseases and can also be used for perfusion and preservation of transplanted organs. Evaluating the oxygen release capacity in vivo is particularly important for the development of effective HBOCs. Summary of the invention

[0003] The present invention aims to solve at least one of the technical problems existing in the prior art. To this end, the present invention provides a method, device, medium and program product for quantifying the oxygen release capacity of HBOC; the method of the present invention proposes the concept of absolute oxygen release capacity, so that the test results between samples are more comparable; at the same time, the method also proposes an oxygen release ratio and introduces a standardization process to reduce the error generated in each test process, so that the test results of different batches are comparable.

[0004] In a first aspect, the present application discloses a method for quantifying the oxygen release capacity of HBOC, the method comprising: 101: Obtain OxyHb%max-HBOC sample to be tested, OxyHb%min-HBOC sample to be tested, OxyHb% Max-bHb standard and OxyHb%-MetbHb in the detection process; 102: Based on the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested, the absolute oxygen release capacity of the sample is calculated and recorded as δOxyHb%; 103: Based on the δOxyHb%, OxyHb% Max-bHb standard, and OxyHb%-MetbHb, calculate the oxygen release ratio of the HBOC sample to be tested, recorded as △OxyHb%; 104: Outputting the oxygen release capacity of the HBOC sample to be tested based on the ΔOxyHb%.

[0005] In some embodiments, the step 103 is specifically: calculating the difference between the OxyHb% Max-bHb standard and the OxyHb%-MetbHb; calculating the ratio of the δOxyHb% to the difference, the ratio being the oxygen release ratio; Optionally, the oxygen hemoglobin content in 101 is calculated by the following method: Based on the characteristic spectral data, the concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are respectively calculated; The concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are added together to obtain a sum value; and the ratio between the concentration of oxyhemoglobin and the sum value is calculated to obtain the oxyhemoglobin content.

[0006] In some embodiments, the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested in 101 are calculated by characteristic spectrum data of the HBOC sample to be tested; OxyHb%max-HBOC sample to be tested is the maximum value of the oxygenated hemoglobin content of the HBOC sample to be tested, and OxyHb%min-HBOC sample to be tested is the minimum value of the oxygenated hemoglobin content of the HBOC sample to be tested; Optionally, the OxyHb% Max-bHb standard in 101 is calculated by characteristic spectrum data of a bHb standard; the OxyHb% Max-bHb standard is the maximum value of the oxygenated hemoglobin content of the bHb standard; Optionally, the OxyHb%-MetbHb in 101 is calculated by characteristic spectrum data of MetHb standard; OxyHb%-MetbHb is the average value of the oxygenated hemoglobin content of the MetHb sample; Optionally, the characteristic spectral data is obtained by: determining a sample including any one or more of the HBOC sample to be tested, the bHb standard, and the MetHb sample; and detecting the sample based on an ELISA instrument to obtain its characteristic spectral data.

[0007] The second aspect of the present application discloses a method for evaluating the oxygen release rate of HBOC based on the oxygen release rate, the method comprising: According to the method disclosed in the first aspect of the present application, the absolute oxygen release capacity of the HBOC sample to be tested is calculated; 50% of the absolute oxygen release capacity is calculated and recorded as 1 / 2δOxy-Hb%; the time corresponding to the acquisition of 1 / 2δOxy-Hb% is recorded as t; the ratio of the 1 / 2δOxy-Hb% and t is calculated to obtain the oxygen release rate V-deoxy; The oxygen release rate of the HBOC sample to be tested is evaluated based on the oxygen release rate V-deoxy; if the oxygen release rate V-deoxy is high, the oxygen release rate of the HBOC sample to be tested is fast; if the oxygen release rate V-deoxy is low, the oxygen release rate of the HBOC sample to be tested is slow.

[0008] In some embodiments, the method further comprises: The oxygenated hemoglobin content of the HBOC sample to be tested is calculated according to the method disclosed in the first aspect of the present application; if the oxygenated hemoglobin content of the HBOC sample to be tested and that of HbA are within the same range, the result that the oxygen carrying capacity of the HBOC sample to be tested and that of HbA in the body are the same is obtained; Optionally, the method further includes: if the oxygen release ratio of the HBOC sample to be tested is lower than that of HbA, and the oxygen release rate of the HBOC sample to be tested is lower than that of HbA, the result is obtained that the oxygen release ratio and oxygen release rate of the HBOC sample to be tested in the body are lower than those of HbA, and oxygen can be carried to peripheral tissues and oxygen is supplied to hypoxic tissues.

[0009] The third aspect of the present application discloses a method for predicting the HBOC Bohr effect, the method comprising: Adjust the pH of the DPBS buffer to at least 2 pH values. When it is 2 pH values, the pH values ​​are recorded as the first pH value (<7.4) and the second pH value (>7.4); Calculate the absolute oxygen release capacity of each pH group of the HBOC sample to be tested according to the method disclosed in the first aspect of the present application, and calculate the oxygen release ratio at different pH values ​​as needed; The Bohr effect BE is calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the second pH value: △OxyHb% first pH value - △OxyHb% second pH value; Based on the Bohr effect BE=△OxyHb%first pH value-△OxyHb%second pH value, the strength of the Bohr effect of the HBOC sample to be tested within the interval between the first pH value and the second pH value is obtained.

[0010] In some embodiments, the method further comprises: Adjust the pH of the DPBS buffer to at least 1 third pH value. When it is 1 pH value, the pH value is recorded as the third pH value (pH=7.4); The Bohr effect BE was calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the third pH value. 酸 =△OxyHb% first pH value-△OxyHb% third pH value; The Bohr effect BE was calculated based on the oxygen release ratio at the second pH value and the oxygen release ratio at the third pH value. 碱=△OxyHb% third pH value-△OxyHb% second pH value; Calculate the Bohr effect BE 酸 and Bohr effect BE 碱 The Bohr effect index EI is calculated by the ratio of The acidic or alkaline Bohr effect is determined based on the Bohr effect index EI; if EI>1, it is mainly the acidic Bohr effect; if EI<1, it is mainly the alkaline Bohr effect; the stronger the acidic Bohr effect, the stronger the oxygen release capacity; the stronger the alkaline Bohr effect, the stronger the oxygen carrying capacity.

[0011] The fourth aspect of the present application discloses a computer device, which includes: a memory and a processor; the memory is used to store a computer program; and the processor executes the computer program to implement the steps of the above method.

[0012] A fifth aspect of the present application discloses a computer-readable storage medium having a computer program stored thereon, which implements the steps of the above method when the computer program is executed by a processor.

[0013] A sixth aspect of the present application discloses a computer program product, including a computer program, which implements the steps of the above method when executed by a processor.

[0014] This application has the following beneficial effects: 1. This application innovatively discloses a method for quantifying the oxygen release capacity of HBOC, which determines the time parameters of the oxygenation process and the deoxygenation detection process in the ODA detection process. Due to the different oxygen carrying / release capacity of HBOC, the oxygenated hemoglobin content in the air is also different. Increasing the oxygenation process can make different test samples start deoxygenation in the oxygen saturated state, so that the test results between samples are more comparable. By comparison, it is clear that the oxygenation time of the sample test is 10 cycles and the deoxygenation time is 50 cycles, which avoids excessive gas use during the experiment and saves time costs. Subsequently, this study proposed a method for quantifying the oxygen release capacity, that is, the oxygen release ratio △OxyHb%, and introduced a standardization process, using the theoretical oxygen release range of the standard in each test as the standard, so as to reduce the error generated in each test process and make the test results of different batches comparable, so that the oxygen release capacity of different varieties of samples can be compared during the HBOC research process, and better prescription screening can be carried out.

[0015] 2. This application innovatively discloses a method for evaluating the oxygen release rate of HBOC based on the oxygen release rate, and proposes that V-deoxy be used to characterize the difference in oxygen release rate. When conducting research on the oxygen carrying / release capacity of HBOC, the two can be compared and verified with each other; this well solves the current problem that the oxygen carrying / release capacity is mainly calculated by analyzing the oxygen dissociation curve. 50, can only characterize the affinity of hemoglobin to oxygen under a single condition (P 50 It reflects the relationship between hemoglobin oxygen saturation and PO2), but cannot reflect the difference in oxygen release rate (no time dimension). Bovine hemoglobin (bHb) and adult hemoglobin (HbA) were introduced as control groups in the study. 50 The P of PEG-bHb was 30.87±0.77 mmHg. 50 was 6.35±0.18 mmHg, HbA 50 Therefore, theoretically, when deoxygenated, at the same oxygen partial pressure, the oxyhemoglobin content of bHb is the lowest, followed by HbA, and the highest is PEG-bHb, which is consistent with the results of this study. Fig.14 B. In the deoxygenation stage, PEG-bHb released the slowest oxygen (0.92±0.05). This oxygen release rate is consistent with that of sample P 50 The values ​​are positively correlated, so this method can be used to obtain the oxygen release rate and to characterize the oxygen affinity of HBOC to a certain extent. In addition, based on the comparison of parameters between samples, it can be determined whether the HBOC sample to be tested can be further optimized according to the specific application scenario.

[0016] 3. The present application innovatively discloses a method for predicting the Bohr effect of HBOC. The method uses the difference in oxygen release ratios at different pH values ​​to semi-quantitatively characterize the Bohr effect. The method utilizes high-throughput oxygen dissociation analysis (ODA) technology to quickly and intuitively obtain the Bohr effect of HBOC in different pH ranges. In addition, the method can simultaneously obtain the test results of multiple samples with high throughput, and can also examine the changes in the Bohr effect of samples in different pH ranges. At the same time, the method can further distinguish between the acidic Bohr effect and the alkaline Bohr effect. Currently, the Bohr factor is often used in the literature to characterize the Bohr effect, that is, pH is used as the horizontal coordinate, logP 50 As the vertical axis, the pH and the corresponding logP 50 Perform linear fitting and the slope of the straight line (Φ = ΔlogP 50 / ΔpH) is the Bohr factor. This method is not only cumbersome to operate, but can only obtain an average Bohr factor. In fact, the Bohr effect of samples in different pH ranges is not the same. This method can solve the above problem well.

[0017] In summary, this study used high-throughput oxygen dissociation analysis (ODA) technology to evaluate the in vivo oxygen release capacity of HBOC. Its advantage is that it can detect multiple HBOC samples at the same time to obtain their oxygen release ratio and oxygen release rate, which can be used for faster prescription screening. It can also semi-quantitatively characterize the Bohr effect of HBOC and distinguish between acidic Bohr effect and alkaline Bohr effect, thereby providing technical support for the research and development of HBOC. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative work.

[0019] Figure 1 is a schematic diagram of a method flow chart provided by the first aspect of an embodiment of the present invention; Figure 2 is a schematic diagram of a method flow chart provided by the second aspect of an embodiment of the present invention; Figure 3 is a schematic diagram of a method flow chart provided by the third aspect of an embodiment of the present invention; Figure 4 is a schematic diagram of a system for quantifying the oxygen release capacity of HBOC provided by an embodiment of the present invention; Figure 5 Schematic diagram of a system for evaluating HBOC oxygen release rate based on oxygen release rate provided by an embodiment of the present invention; Figure 6 is a schematic diagram of a system for predicting the HBOC Bohr effect provided by an embodiment of the present invention; Figure 7 is a schematic diagram of a computer device provided by an embodiment of the present invention; Figure 8 is a schematic diagram of the architecture of an exemplary computing device provided by an embodiment of the present invention; Fig. 9 is a schematic diagram of a storage medium provided by an embodiment of the present invention; Fig.10 It is a connection diagram of a computer system, an ELISA device, a nitrogen bottle, and an air bottle provided in an embodiment of the present invention; Fig.11 is a schematic diagram of changes in oxygenated hemoglobin content during the oxygenation process provided by an embodiment of the present invention; Fig.12 is a schematic diagram of changes in the oxygenated hemoglobin content during the deoxygenation process provided by an embodiment of the present invention; Fig.13 Schematic diagram of calculating △Oxy-Hb% by different methods provided by embodiments of the present invention, wherein: Fig.13 A is the calculation method without standardization. Fig.13 B is the standardized calculation method; Fig.14Schematic diagram of V-deoxy calculation provided by an embodiment of the present invention, wherein A. 1 / 2δOxy-Hb% interval; B. ODA test results of different samples; C. V-deoxy calculation results of different samples; D. P of different samples 50 Value test results; EV-deoxy and P 50 Correlation analysis of values; Fig.15 Schematic diagram of ODA detection of bHb at different pH values ​​provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to better understand the solutions of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention.

[0021] In some of the processes described in the specification and claims of the present invention and the above-mentioned figures, multiple operations that appear in a specific order are included, but it should be clearly understood that these operations may not be executed in the order in which they appear in this article or executed in parallel. The serial numbers of the operations, such as 101, 102, etc., are only used to distinguish different operations, and the serial numbers themselves do not represent any execution order. In addition, these processes may include more or fewer operations, and these operations may be executed in sequence or in parallel. It should be noted that the descriptions of "first", "second", etc. in this article are used to distinguish different messages, devices, modules, etc., do not represent the order of precedence, and do not limit the "first" and "second" to be different types.

[0022] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0023] definition: The Bohr effect refers to the effect of pH on the oxygen affinity of hemoglobin. That is, as the H+ concentration increases, hemoglobin maintains in the T state and reduces its affinity for oxygen, which is the acidic Bohr effect. + As the concentration decreases, hemoglobin tends to the R state and its affinity for oxygen increases, which is the alkaline Bohr effect. The Bohr effect is of great significance to life activities. When blood flows through tissues and organs, the accumulation of metabolites such as lactic acid and carbon dioxide reduces the pH of tissues, promotes hemoglobin to release oxygen, and allows tissues to obtain more oxygen. Conversely, when blood flows through the lungs, it promotes H +The release of carbon dioxide is beneficial to the combination of hemoglobin and oxygen. In special environments (such as plateaus), red blood cells with low oxygen affinity and strong Bohr effect will be more beneficial, which can promote both oxygen uptake and oxygen release.

[0024] P 50 , which is the oxygen partial pressure corresponding to an oxygen saturation of 50%, is mainly used to characterize hemoglobin oxygen affinity and is also used to represent oxygen carrying / release capacity. 50 value [8] , such as Hemopure P 50 The value is about 40 mmHg, while Sanguinate is 7~16 mmHg, which is a big difference. 50 This means that HBOC has a strong oxygen-carrying capacity and can release oxygen to peripheral tissues with low oxygen partial pressure, which is less likely to cause vasoactive side effects. 50 HBOC is easy to release oxygen and is often used to enhance the sensitivity of tumor tissue chemotherapy. 50 It can be detected by the hemoglobin oxygen carrying / release analyzer developed by the Military Medical Research Institute. Based on the different light absorption characteristics of Hb and HbO2 (at 560nm, the light absorption values ​​of the two are significantly different, and 570nm is the isosbestic point of the two), the dual-wavelength spectrophotometry method is used to detect the light absorption value of hemoglobin at different oxygen partial pressures, and the oxygen binding / dissociation curve is drawn after conversion to oxygen saturation, and the P is directly output. 50 value.

[0025] ODA is the abbreviation of high-throughput oxygen dissociation analysis technology, which can monitor the oxygen release process of hemoglobin based on the characteristic spectral changes during the deoxygenation process of hemoglobin. This method has the advantages of easy operation and high throughput. The oxygenated hemoglobin content is calculated by the five-wavelength method. Therefore, it is suitable for high-throughput detection of HBOC's in vivo oxygen release capacity. However, there is still room for improvement in the detection of HBOC using ODA. First, HBOC is different from natural hemoglobin, and the oxygen release process is related to its characteristics, and the details of the detection process need to be explored. Second, the changes in the ODA curve can only be used as a qualitative basis, and the test results need to be quantitatively compared. Third, ODA can only characterize the oxygen release process, and it is necessary to supplement the hemoglobin oxygen release capacity parameter characterization method including the Bohr effect. The solution provided in this application can optimize the above-mentioned defects.

[0026] Figure 1 1 is a flow chart of a method for quantifying the oxygen release capacity of HBOC provided by an embodiment of the present invention. Specifically, the method comprises the following steps: 101: Obtain OxyHb%max-HBOC sample to be tested, OxyHb%min-HBOC sample to be tested, OxyHb% Max-bHb standard and OxyHb%-MetbHb in the detection process; In some embodiments, the oxygen hemoglobin content in 101 is calculated by the following method: Based on the characteristic spectral data, the concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are respectively calculated; The concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are added together to obtain a sum value; and the ratio between the concentration of oxyhemoglobin and the sum value is calculated to obtain the oxyhemoglobin content.

[0027] In some embodiments, the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested in 101 are calculated by characteristic spectrum data of the HBOC sample to be tested; OxyHb%max-HBOC sample to be tested is the maximum value of the oxygenated hemoglobin content of the HBOC sample to be tested, and OxyHb%min-HBOC sample to be tested is the minimum value of the oxygenated hemoglobin content of the HBOC sample to be tested; Optionally, the OxyHb% Max-bHb standard in 101 is calculated by characteristic spectrum data of a bHb standard; the OxyHb% Max-bHb standard is the maximum value of the oxygenated hemoglobin content of the bHb standard; Optionally, the OxyHb%-MetbHb in 101 is calculated by characteristic spectrum data of MetHb standard; OxyHb%-MetbHb is the average value of the oxygenated hemoglobin content of the MetHb sample; Optionally, the characteristic spectrum data is obtained by: determining a sample including any one or more of the HBOC sample to be tested, the bHb standard, and the MetHb sample; and detecting the sample based on an ELISA instrument to obtain its characteristic spectrum data. The maximum value is for the experimental sample and the standard, the minimum value is for the experimental sample, the average value is for methemoglobin, and OxyHb% is calculated at each detection point; 102: Based on the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested, the absolute oxygen release capacity of the sample is calculated and recorded as δOxyHb%; 103: Based on the δOxyHb%, OxyHb% Max-bHb standard, and OxyHb%-MetbHb, calculate the oxygen release ratio of the HBOC sample to be tested, recorded as △OxyHb%; In some embodiments, the step 103 is specifically: calculating the difference between the OxyHb% Max-bHb standard and the OxyHb%-MetbHb; calculating the ratio of the δOxyHb% to the difference, the ratio being the oxygen release ratio; 104: Outputting the oxygen release capacity of the HBOC sample to be tested based on the ΔOxyHb%.

[0028] In some embodiments, the ΔOxyHb% and the oxygen release capacity of the HBOC sample to be tested in 104 are regression results. The high and low ΔOxyHb% are relative to the standard. If the ΔOxyHb% is high (the calculated value of the oxygen release ratio is high), the result of the oxygen release capacity of the HBOC sample to be tested is output as strong; if the ΔOxyHb% is low (the calculated value of the oxygen release ratio is low), the result of the oxygen release capacity of the HBOC sample to be tested is output as weak. The second aspect of the present application discloses a method for evaluating the oxygen release rate of HBOC based on the oxygen release rate. Figure 2 As shown, the method includes: 201, according to the method disclosed in the first aspect of the present application, the absolute oxygen release capacity of the HBOC sample to be tested is calculated; 50% of the absolute oxygen release capacity is calculated and recorded as 1 / 2δOxy-Hb%; the time corresponding to the time of obtaining 1 / 2δOxy-Hb% is recorded as t; the ratio of the 1 / 2δOxy-Hb% and t is calculated to obtain the oxygen release rate V-deoxy; 202. Evaluate the oxygen release rate of the HBOC sample to be tested based on the oxygen release rate V-deoxy; if the oxygen release rate V-deoxy is high, the oxygen release rate of the HBOC sample to be tested is fast; if the oxygen release rate V-deoxy is low, the oxygen carrying rate of the HBOC sample to be tested is slow.

[0029] In some embodiments, the method further comprises: The oxygenated hemoglobin content of the HBOC sample to be tested is calculated according to the method disclosed in the first aspect of the present application; if the oxygenated hemoglobin content of the HBOC sample to be tested and that of HbA are within the same range, the result that the oxygen carrying capacity of the HBOC sample to be tested and that of HbA in the body are the same is obtained; Optionally, the method further includes: if the oxygen release ratio of the HBOC sample to be tested is lower than that of HbA, and the oxygen release rate of the HBOC sample to be tested is lower than that of HbA, the result is obtained that the oxygen release ratio and oxygen release rate of the HBOC sample to be tested in the body are lower than those of HbA, and oxygen can be carried to peripheral tissues and supplied to hypoxic tissues.

[0030] The third aspect of the present application discloses a method for predicting the HBOC Bohr effect, specifically: Figure 3 As shown, the method includes: 301, adjusting the pH of the DPBS buffer to at least 2 pH values, when it is 2 pH values, the pH values ​​are recorded as the first pH value (<7.4) and the second pH value (>7.4); 302, calculating the absolute oxygen release capacity of each pH group of the HBOC sample to be tested according to the method disclosed in the first aspect of the present application, and calculating the oxygen release ratio at different pH values ​​as needed; 303, based on the oxygen release ratio of the first pH value and the oxygen release ratio of the second pH value, the Bohr effect BE is calculated as: △OxyHb%first pH value-△OxyHb%second pH value; 304, based on the Bohr effect BE=△OxyHb%first pH value-△OxyHb%second pH value, obtain the strength of the Bohr effect of the HBOC sample to be tested within the first pH value and the second pH value interval. Specifically, this judgment belongs to the regression result, and the numerical value is compared with the standard. The larger the numerical value, the stronger the Bohr effect. If BE is greater than the standard, output the result that the Bohr effect of the HBOC sample to be tested is stronger than that of the standard; if BE is less than the standard, output the result that the Bohr effect of the HBOC sample to be tested is weaker than that of the standard; Among them, when calculating the Bohr effect and the Bohr effect index, the requirements for taking different pH values ​​are: with 7.4 as the center, taking values ​​on both sides, the oxygen release ratio of the low pH value - the oxygen release ratio of the high pH value, the low pH value is the first pH value, and the high pH value is the second pH value.

[0031] In some embodiments, the method further comprises: Adjust the pH of the DPBS buffer to at least 1 third pH value. When it is 1 pH value, the pH value is recorded as the third pH value (=7.4); The Bohr effect BE was calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the third pH value. 酸 =△OxyHb% first pH value-△OxyHb% third pH value; The Bohr effect BE was calculated based on the oxygen release ratio at the second pH value and the oxygen release ratio at the third pH value. 碱 =△OxyHb% third pH value-△OxyHb% second pH value; Calculate the Bohr effect BE 酸 and Bohr effect BE 碱 The Bohr effect index EI is calculated by the ratio of The acidic or alkaline Bohr effect is determined based on the Bohr effect index EI; if EI>1, it is mainly the acidic Bohr effect; if EI<1, it is mainly the alkaline Bohr effect; the stronger the acidic Bohr effect, the stronger the oxygen release capacity; the stronger the alkaline Bohr effect, the stronger the oxygen carrying capacity.

[0032] Figure 7is a schematic diagram of a computer device provided by an embodiment of the present invention, such as Figure 7 As shown, the device may include: one or more processors, and one or more memories; wherein the memories store computer-readable codes, and when the computer-readable codes are run by the one or more processors, the method described above may be executed.

[0033] The processor in this embodiment can be an integrated circuit chip with signal processing capabilities. The above processor can be a general-purpose processor, a digital signal processor (DSP), an application-specific integrated circuit (ASIC), a field-programmable gate array (FPGA) or other programmable logic devices, discrete gates or transistor logic devices, discrete hardware components. The disclosed methods, operations and logic block diagrams in the embodiments of the present disclosure can be implemented or executed. The general-purpose processor can be a microprocessor or the processor can also be any conventional processor, etc., which can be an X86 architecture or an ARM architecture.

[0034] In general, various example embodiments of the present disclosure may be implemented in hardware or dedicated circuits, software, firmware, logic, or any combination thereof. Certain aspects may be implemented in hardware, while other aspects may be implemented in firmware or software that may be executed by a controller, microprocessor, or other computing device. When various aspects of the disclosed embodiments are illustrated or described as block diagrams, flow charts, or using some other graphical representation, it will be understood that the blocks, devices, systems, techniques, or methods described herein may be implemented in hardware, software, firmware, dedicated circuits or logic, general purpose hardware or controllers or other computing devices, or some combination thereof as non-limiting examples.

[0035] For example, the method or device according to the embodiment of the present disclosure may also be implemented by Figure 8 The architecture of the computing device 3000 shown in FIG. Figure 8 As shown, the computing device 3000 may include a bus 3010, one or more CPUs 3020, a read-only memory (ROM) 3030, a random access memory (RAM) 3040, a communication port 3050 connected to a network, an input / output component 3060, a hard disk 3070, etc. The storage device in the computing device 3000, such as ROM 3030 or hard disk 3070, may store various data or files used for processing and / or communication of the method provided by the present disclosure and program instructions executed by the CPU. The computing device 3000 may also include a user interface 3080. Of course, Figure 8 The architecture shown is only exemplary and can be omitted according to actual needs when implementing different devices. Figure 8 One or more components of a computing device are shown.

[0036] The embodiment of the present invention also provides a computer-readable storage medium, such as Fig. 9 As shown, it is a schematic diagram of a storage medium provided by an embodiment of the present invention, and a computer readable instruction 4010 is stored on the computer storage medium 4020. When the computer readable instruction 4010 is executed by the processor, the method according to the embodiment of the present disclosure described with reference to the above figures can be executed. The computer readable storage medium in the embodiment of the present disclosure can be a volatile memory or a non-volatile memory, or can include both volatile and non-volatile memories. The non-volatile memory can be a read-only memory (ROM), a programmable read-only memory (PROM), an erasable programmable read-only memory (EPROM), an electrically erasable programmable read-only memory (EEPROM) or a flash memory. The volatile memory can be a random access memory (RAM), which is used as an external cache. By way of example and not limitation, many forms of RAM are available, such as static random access memory (SRAM), dynamic random access memory (DRAM), synchronous dynamic random access memory (SDRAM), double data rate synchronous dynamic random access memory (DDRSDRAM), enhanced synchronous dynamic random access memory (ESDRAM), synchronous link dynamic random access memory (SLDRAM), and direct memory bus random access memory (DR RAM). It should be noted that the memory of the methods described herein is intended to include, but is not limited to, these and any other suitable types of memory. It should be noted that the memory of the methods described herein is intended to include, but is not limited to, these and any other suitable types of memory.

[0037] The embodiments of the present disclosure also provide a computer program product or system, including a computer program, which implements the steps of the above method when executed by a processor.

[0038] In some embodiments, the present embodiment also discloses a system for quantifying the oxygen release capacity of HBOC, such as Figure 4 As shown, the system comprises: The first acquisition module 401 is used or configured to acquire OxyHb%max-HBOC sample to be tested, OxyHb%min-HBOC sample to be tested, OxyHb% Max-bHb standard and OxyHb%-MetbHb in the detection process; The first calculation module 402 is used or configured to calculate the absolute oxygen release capacity of the sample based on the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested, which is recorded as δOxyHb%; The second calculation module 403 is used or configured to calculate the oxygen release ratio of the HBOC sample to be tested based on the δOxyHb%, OxyHb% Max-bHb standard and OxyHb%-MetbHb, which is recorded as △OxyHb%; The oxygen release capacity quantification module 404 is used for or configured to output the oxygen release capacity of the HBOC sample to be tested based on the ΔOxyHb%.

[0039] In some embodiments, this embodiment also discloses a system for evaluating the oxygen release rate of HBOC based on the oxygen release rate, such as Figure 5 As shown, the system comprises: The third calculation module 501 is used or configured to calculate the absolute oxygen release capacity of the HBOC sample to be tested according to the method disclosed in the first aspect of the present application; 50% of the absolute oxygen release capacity is calculated as 1 / 2δOxy-Hb%; the time corresponding to the acquisition of 1 / 2δOxy-Hb% is recorded as t; the ratio of the 1 / 2δOxy-Hb% to t is calculated to obtain the oxygen release rate V-deoxy; The oxygen release rate prediction module 502 is used for or configured to evaluate the oxygen release rate of the HBOC sample to be tested based on the oxygen release rate V-deoxy; if the oxygen release rate V-deoxy is high, the oxygen release rate of the HBOC sample to be tested is fast; if the oxygen release rate V-deoxy is low, the oxygen release rate of the HBOC sample to be tested is slow.

[0040] In some embodiments, this embodiment also discloses a system for predicting the HBOC Bohr effect, such as Figure 6 As shown, the system comprises: A first pH value adjustment module 601 is used or configured to adjust the pH of the DPBS buffer to at least 2 pH values. When the pH values ​​are 2, the pH values ​​are recorded as a first pH value (<7.4) and a second pH value (>7.4). The fourth calculation module 602 is used or configured to calculate the absolute oxygen release capacity of each pH group of the HBOC sample to be tested according to the method disclosed in the first aspect of the present application, and calculate the oxygen release ratio at different pH values ​​as needed.

[0041] A fifth calculation module 603 is used or configured to calculate the Bohr effect BE based on the oxygen release ratio of the first pH value and the oxygen release ratio of the second pH value: ΔOxyHb% first pH value-ΔOxyHb% second pH value; The Bohr effect calculation module 604 is used for or configured to obtain the strength of the Bohr effect of the HBOC sample to be tested within the range of the first pH value and the second pH value based on the Bohr effect BE=ΔOxyHb%first pH value-ΔOxyHb%second pH value.

[0042] In some embodiments, the system further comprises: A second pH value adjustment module, used for or configured to adjust the pH of the DPBS buffer to at least a third pH value, when the pH value is 1, the pH value is recorded as the third pH value (=7.4); The Bohr effect BE was calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the third pH value. 酸 =△OxyHb% first pH value-△OxyHb% third pH value; The sixth calculation module is used or configured to calculate the Bohr effect BE based on the oxygen release ratio of the second pH value and the oxygen release ratio of the third pH value. 碱 =△OxyHb% third pH value-△OxyHb% second pH value; A seventh calculation module is used or configured to calculate the Bohr effect BE 酸 and Bohr effect BE 碱 The ratio of is the Bohr effect index EI; The Bohr effect judgment module is used for or configured to judge the acidic or alkaline Bohr effect based on the Bohr effect index EI; if EI>1, it is mainly the acidic Bohr effect; if EI<1, it is mainly the alkaline Bohr effect; the stronger the acidic Bohr effect, the stronger the oxygen release capacity; the stronger the alkaline Bohr effect, the stronger the oxygen carrying capacity.

[0043] Currently, at the user end, when determining the oxygen-release capacity of the sample to be tested, the bHb sample as the standard, the HBOC sample to be tested, the HbA and MetHb samples need to be placed in the microplate reader at the same time to start measuring and calculating the oxygen-release capacity of the sample to be tested; with the continuous deepening of research, each time the measurement is performed, it is only necessary to place the HBOC sample to be tested in the microplate reader, and the relevant values ​​of the other three samples can be used as fixed values ​​during the research process and used directly later.

[0044] Example

[0045] Objects and methods: 1. Materials and instruments: 1.1 Materials: DPBS buffer (Sigma, USA), 1% NaOH (analytical grade, Sinopharm Chemical Reagent Co., Ltd.), hydrochloric acid (analytical grade, Sinopharm Chemical Reagent Co., Ltd.), nitrogen (purity ≥ 99.9%, Beijing Huanyu Jinghui Jingcheng Gas Technology Co., Ltd.), air (Beijing Huanyu Jinghui Jingcheng Gas Technology Co., Ltd.), 96-well plate (Beijing blabgic Technology vloted), 2-iminothiolane (IT, Sigma), 5kDa PEG maleimide (PEG5K-mal, Jenkem Technology, Beijing), potassium ferrocyanide (analytical grade, Sinopharm Chemical Reagent Co., Ltd.); 1.2 Instruments: Blood gas analyzer (ABL90, Radiometer, Denmark); pH meter (METTLER TOLEDO, Switzerland); Bloodox-2018 oxygen carrying / release analyzer (Beijing Ruanlong Biotechnology Co., Ltd.); microplate reader FLUOstar Omega (BMG, LRBTECH).

[0046] 2. Methods: 2.1 Preparation of bHb, PEG-bHb, HbA and methemoglobin (MetHb): Bovine whole blood was obtained from Beijing Kerunweide Biotechnology Co., Ltd. (China), and bovine hemoglobin (bHb) was purified from bovine blood and used as a standard.

[0047] bHb was reacted with 2-iminothiolane and PEG maleimide in PBS buffer (pH 7.4) overnight to obtain PEG-modified bovine hemoglobin (PEG-bHb) as an HBOC experimental product.

[0048] The anticoagulated blood of the volunteers was purified to obtain an adult hemoglobin solution (HbA) with a concentration of 5 g / dL as a control.

[0049] MetHb was prepared by mixing bHb with potassium ferrocyanide as a negative control.

[0050] 2.2ODA detection and analysis process: bHb, PEG-bHb, and MetHb were diluted to 0.25 mg / ml with DPBS, placed in a 96-well plate, and placed in a microplate reader FLUOstar Omega for detection. The connections of the computer system, microplate reader, nitrogen cylinder, and air cylinder are as follows: Fig.10As shown, connect an external nitrogen bottle and an air bottle, switch as needed, scan range 350-700nm, scan time <1s per well. Perform a spectral scan every 5 minutes (1 cycle) (spectral resolution 1 nm), and record the characteristic spectrum of the sample. In between each measurement, shake the 96-well plate in the form of a graph "8" for 1 minute (300 rpm / min). Export the characteristic spectrum data and calculate according to the following formula.

[0051] Methemoglobin concentration: C MetHb =2.985*(OD 630 -OD 700 ) +0.194*(OD 576 -OD 700 )-0.4023*(OD 560 -OD 700 ) Deoxyhemoglobin concentration: C DeoxyHb =1.373*(OD 560 -OD 700 )-0.747*(OD 576 -OD 700 )-0.737*(OD 630 -OD 700 ) Oxyhemoglobin concentration: C OxyHb =1.013*(OD 576 -OD 700 )-0.3269*(OD 630 -OD 700 )-0.7353*(OD 560 -OD 700 ) Oxyhemoglobin content: OxyHb%=

C OxyHb / ( C MetHb + C DeoxyHb +C OxyHb )

[0052] 2.2.2 Determine the deoxygenation time: After the sample to be tested reaches saturation, stop the air flow, open the nitrogen passage, and pass nitrogen into the detection chamber at a rate of 20 L / min. Record the changes in the characteristic spectrum and determine the deoxygenation time.

[0053] 2.3 Data Analysis: 2.3.1 Oxygen release ratio △Oxy-Hb%: The maximum oxygen release capacity of the standard was used as the standard, and the absolute oxygen release capacity of the experimental group was compared with it to calculate the oxygen release ratio of the HBOC sample to characterize its actual oxygen release capacity.

[0054] Absolute oxygen release capacity of sample: δOxyHb% (experimental group) = OxyHb% Max (Experimental group) - OxyHb% min (Experimental group) Oxygen release ratio: △OxyHb% (experimental group) =

[0055] Among them, OxyHb% Max (experimental group) represents the maximum value of oxygenated hemoglobin content in the experimental group during the detection process; OxyHb% min (experimental group) represents the minimum value of oxygenated hemoglobin content in the experimental group during the detection process; OxyHb% Max (standard product) represents the maximum value of oxygenated hemoglobin content of the standard product during the detection process; OxyHb% (MetbHb) represents the average value of oxygenated hemoglobin content in the MetHb group.

[0056] 2.3.2 Oxygen release rate V-deoxy: The oxygen release rate refers to the rate at which the HBOC sample releases oxygen during the deoxygenation process. The calculation formula for the oxygen release rate defined in this study is as follows: V-deoxy = δOxyHb%÷t in δOxy-Hb% is 50% of the δOxyHb% of the sample group, and t is The time corresponding to δOxy-Hb% is as follows: Fig.14 As shown in A. According to this method, V-deoxy was detected and statistically analyzed for bHb, PEG-bHb and HbA.

[0057] 2.3.3 Semi-quantitative analysis of the Bohr effect: Use dilute hydrochloric acid and 1% sodium hydroxide solution to adjust the pH of DPBS buffer to 6.8, 7.2, 7.6, and 8.0, and repeat the steps in 2.2 for bHb to perform ODA detection.

[0058] The semi-quantitative analysis method of the Bohr effect is introduced as follows: (1) Calculate δOxyHb% for each pH group.

[0059] (2) Calculate △OxyHb% at different pH values ​​as needed.

[0060] (3) Calculate the Bohr effect (BE). Set the pH interval to calculate the △OxyHb% difference as the Bohr effect. 6.8-8.0 =△OxyHb%(pH=6.8) - △OxyHb%(pH=8.0).

[0061] (4) Calculate the Bohr effect index (EI). Calculate the acidic Bohr effect (BE 酸 ) and the alkaline Bohr effect (BE 碱 ), B.E. 酸 =△OxyHb%(pH=6.8) - OxyHb%(pH=7.4), BE 碱 =△OxyHb%(pH=7.4) - OxyHb%(pH=8.0). EI=BE 酸 / BE 碱 If EI>1, it is mainly acidic Bohr effect; if EI<1, it is mainly alkaline Bohr effect.

[0062] 2.3.4 P 50 Detection: The Bloodox-2018 red blood cell oxygen carrying / release function evaluation device was used to measure the P 50 3 mg hemoglobin sample and 4 ml P 50 Buffer solution, 20 µl 20% BSA solution, and 20 µl defoamer were mixed evenly and added to the sample pool. Air was introduced to saturate the hemoglobin with oxygen. At 37°C, nitrogen was introduced to start drawing the oxygen dissociation curve and read the P 50 value.

[0063] result: 3.1 Determine the starting point of deoxygenation: The initial oxygenated hemoglobin content of the standard is 89.44% ± 1.29%. Fig.11 It can be seen that the oxygenated hemoglobin content of the standard gradually increases with the number of cycles, and increases significantly before the 10th cycle, and then no longer increases significantly, reaching an approximate stable value of 97.27%±0.48%. Therefore, the oxygenation time is determined to be 10 cycles, *, p<0.05; **, p<0.01.

[0064] 3.2 Determine the detection time: The initial oxygenated hemoglobin content of the standard is 91.18% ± 2.1%. After N2 is introduced for deoxygenation, Fig.12 It can be seen that the oxygenated hemoglobin content of the sample began to decrease, and decreased significantly with the increase in the number of cycles. After the 50th cycle, it no longer decreased significantly and reached an approximate stable value of 6.13%±0.49%. Therefore, the oxygenation time was determined to be 50 cycles, *, p<0.05; **, p<0.01.

[0065] 3.3 Calculation of △Oxy-Hb%: In order to quantify the oxygen-releasing capacity of HBOC samples and reduce the systematic error, two calculation methods were used for the same PEG-bHb sample. Fig.13 A is the calculation method without standardization, that is, directly calculating δOxyHb%. Fig.13 B is the standardized calculation method, that is, the absolute oxygen release capacity of bHb measured as a control substance is used as the standard for standardization to calculate △OxyHb%, *, p<0.05; **, p<0.01. At the same time, the non-standardized measurement value is compared and analyzed with the standardized measurement value to calculate the coefficient of variation. The results are shown in Table 1. The coefficient of variation of the non-standardized measurement value is 0.1, and the coefficient of variation of the standardized measurement value is 0.03. The coefficient of variation of the standardized method is significantly reduced, which also indicates that the PEG-bHb sample retains 58.35% of the oxygen release capacity of natural bHb.

[0066] Table 1. Comparison of unstandardized and standardized measurements

[0067] 3.4 Oxygen release rate V-deoxy: like Fig.14 As shown in B, ODA detection of bHb, PEG-bHb, and HbA was performed, and V-deoxy ( Fig.14 C). The oxygen release rate of bHb (2.59±0.12) was significantly greater than that of HbA (1.36±0.14), and the oxygen release rate of the modified PEG-bHb (0.92±0.05) was significantly lower than that of bHb and HbA.

[0068] like Fig.14 As shown in D, P of bHb 50 The P of PEG-bHb was 30.87±0.77 mmHg. 50 was 6.35±0.18 mmHg, HbA 50 The V-deoxy was compared with the measured P 50 Linear regression analysis was performed using R 2 =0.98( Fig.14 E), *, p<0.05; **, p<0.01, Table 2 shows the calculated values ​​of V-deoxy for different samples.

[0069] Table 2. Calculated V-deoxy values ​​of different samples

[0070] 3.5 Calculation of Bohr effect: One of the main characteristics of hemoglobin is the Bohr effect. The Bohr effect makes hemoglobin oxygen affinity decrease in low pH environments, such as tissue cells, making it easier to release oxygen. Conversely, in high pH environments, such as lungs, hemoglobin oxygen affinity increases, making it easier to bind oxygen. Fig.15 The ODA test results of bHb at different pH values ​​are shown. As the pH value increases, the deoxygenation rate of the sample is slower. Table 3 shows the calculated values ​​of the Bohr effect in different pH ranges. It can be seen that the Bohr effect in the pH 7.2-7.6 range is 0.0281±0.0086, which is significantly lower than 0.2485±0.0283 in the pH 6.8-8.0 range, which is consistent with the theory.

[0071] In addition, this study also introduced the calculation of the Bohr effect index EI. As shown in Table 3, the standard deviation of the EI calculation results in the pH 7.2-7.6 range is large, indicating that the calculation has a large variation and the results are inaccurate. The I calculation result in the pH 6.8-8.0 range is 0.1134±0.0798, showing a more obvious alkaline Bohr effect.

[0072] Table 3. Calculated values ​​of Bohr effect in different pH ranges

[0073] It should be noted that the flowcharts and block diagrams in the accompanying drawings illustrate the possible architecture, functions and operations of the systems, methods and computer program products according to various embodiments of the present disclosure. In this regard, each box in the flowchart or block diagram can represent a module, a program segment, or a part of a code, and the module, program segment, or a part of the code contains one or more executable instructions for implementing the specified logical function. It should also be noted that in some alternative implementations, the functions marked in the box can also occur in a different order from the order marked in the accompanying drawings. For example, two boxes represented in succession can actually be executed substantially in parallel, and they can sometimes be executed in the opposite order, depending on the functions involved. It should also be noted that each box in the block diagram and / or flowchart, and the combination of boxes in the block diagram and / or flowchart can be implemented with a dedicated hardware-based system that performs a specified function or operation, or can be implemented with a combination of dedicated hardware and computer instructions.

[0074] In general, various example embodiments of the present disclosure may be implemented in hardware or dedicated circuits, software, firmware, logic, or any combination thereof. Certain aspects may be implemented in hardware, while other aspects may be implemented in firmware or software that may be executed by a controller, microprocessor, or other computing device. When various aspects of the disclosed embodiments are illustrated or described as block diagrams, flow charts, or using some other graphical representation, it will be understood that the blocks, devices, systems, techniques, or methods described herein may be implemented in hardware, software, firmware, dedicated circuits or logic, general purpose hardware or controllers or other computing devices, or some combination thereof as non-limiting examples.

[0075] Those skilled in the art can clearly understand that, for the convenience and brevity of description, the specific working processes of the systems, devices and units described above can refer to the corresponding processes in the aforementioned method embodiments and will not be repeated here.

[0076] In the several embodiments provided in the present application, it should be understood that the disclosed systems, devices and methods can be implemented in other ways. For example, the device embodiments described above are only schematic. For example, the division of the units is only a logical function division. There may be other division methods in actual implementation, such as multiple units or components can be combined or integrated into another system, or some features can be ignored or not executed. Another point is that the mutual coupling or direct coupling or communication connection shown or discussed can be an indirect coupling or communication connection through some interfaces, devices or units, which can be electrical, mechanical or other forms.

[0077] The units described as separate components may or may not be physically separated, and the components shown as units may or may not be physical units, that is, they may be located in one place or distributed on multiple network units. Some or all of the units may be selected according to actual needs to achieve the purpose of the solution of this embodiment.

[0078] In addition, each functional unit in each embodiment of the present invention may be integrated into one processing unit, or each unit may exist physically separately, or two or more units may be integrated into one unit. The above-mentioned integrated unit may be implemented in the form of hardware or in the form of software functional units.

[0079] The exemplary embodiments of the present disclosure described in detail above are merely illustrative and not restrictive. It should be understood by those skilled in the art that various modifications and combinations may be made to these embodiments or their features without departing from the principles and spirit of the present disclosure, and such modifications should fall within the scope of the present disclosure.

Claims

1. A method for quantifying the oxygen release capacity of HBOC, characterized in that: The method comprises: 101: Obtain OxyHb%max-HBOC sample to be tested, OxyHb%min-HBOC sample to be tested, OxyHb%Max-bHb standard and OxyHb%-MetbHb in the detection process; 102: Based on the OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested, the absolute oxygen release capacity of the sample is calculated and recorded as δOxyHb%; 103: Based on the δOxyHb%, OxyHb% Max-bHb standard, and OxyHb%-MetbHb, calculate the oxygen release ratio of the HBOC sample to be tested, recorded as △OxyHb%; 104: Outputting the oxygen release capacity of the HBOC sample to be tested based on the ΔOxyHb%.

2. The method for quantifying the oxygen release capacity of HBOC according to claim 1, characterized in that: The step 103 specifically includes: calculating the difference between the OxyHb% Max-bHb standard and the OxyHb%-MetbHb; calculating the ratio of the δOxyHb% to the difference, wherein the ratio is the oxygen release ratio; Optionally, the oxygen hemoglobin content in 101 is calculated by the following method: Based on the characteristic spectral data, the concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are respectively calculated; The concentration of methemoglobin, the concentration of deoxyhemoglobin, and the concentration of oxyhemoglobin are added together to obtain a sum value; and the ratio between the concentration of oxyhemoglobin and the sum value is calculated to obtain the oxyhemoglobin content.

3. The method for quantifying the oxygen release capacity of HBOC according to claim 1, characterized in that: The OxyHb%max-HBOC sample to be tested and the OxyHb%min-HBOC sample to be tested in 101 are calculated by the characteristic spectrum data of the HBOC sample to be tested; OxyHb%max-HBOC sample to be tested is the maximum value of the oxygenated hemoglobin content of the HBOC sample to be tested, and OxyHb%min-HBOC sample to be tested is the minimum value of the oxygenated hemoglobin content of the HBOC sample to be tested; Optionally, the OxyHb% Max-bHb standard in 101 is calculated by characteristic spectrum data of a bHb standard; the OxyHb% Max-bHb standard is the maximum value of the oxygenated hemoglobin content of the bHb standard; Optionally, the OxyHb%-MetbHb in 101 is calculated by characteristic spectrum data of MetHb standard; OxyHb%-MetbHb is the average value of the oxygenated hemoglobin content of the MetHb sample; Optionally, the characteristic spectrum data is obtained by: determining a sample including any one or more of the HBOC sample to be tested, the bHb standard, and the MetHb sample; and detecting the sample based on an ELISA instrument to obtain its characteristic spectrum data; Optionally, the characteristic spectral data is obtained by: determining a sample including any one or more of the HBOC sample to be tested, the bHb standard, and the MetHb sample; and detecting the sample based on an ELISA instrument to obtain its characteristic spectral data.

4. A method for evaluating the oxygen release rate of HBOC based on the oxygen release rate, characterized in that: The method comprises: The absolute oxygen release capacity of the HBOC sample to be tested is calculated according to the method described in any one of claims 1 to 3; 50% of the absolute oxygen release capacity is calculated and recorded as 1 / 2δOxy-Hb; the time corresponding to the time of obtaining 1 / 2δOxy-Hb% is recorded as t; the oxygen release rate V-deoxy is calculated by calculating the ratio of the 1 / 2δOxy-Hb% and t; The oxygen release rate of the HBOC sample to be tested is evaluated based on the oxygen release rate V-deoxy; if the oxygen release rate V-deoxy is high, the oxygen release rate of the HBOC sample to be tested is fast; if the oxygen release rate V-deoxy is low, the oxygen release rate of the HBOC sample to be tested is slow.

5. The method for evaluating HBOC oxygen release rate based on oxygen release rate according to claim 4, characterized in that: The method further comprises: The oxygenated hemoglobin content of the HBOC sample to be tested is calculated according to the method described in any one of claims 1 to 3; if the oxygenated hemoglobin content of the HBOC sample to be tested and that of HbA are within the same range, the result that the oxygen carrying capacity of the HBOC sample to be tested and that of HbA in the body are the same is obtained; Optionally, the method further includes: if the oxygen release ratio of the HBOC sample to be tested is lower than that of HbA, and the oxygen release rate of the HBOC sample to be tested is lower than that of HbA, the result is obtained that the oxygen release ratio and oxygen release rate of the HBOC sample to be tested in the body are lower than those of HbA, and oxygen can be carried to peripheral tissues and oxygen is supplied to hypoxic tissues.

6. A method for predicting the HBOC Bohr effect, characterized in that: The method comprises: The pH of the DPBS buffer solution is adjusted to at least 2 pH values. When the pH values ​​are 2, the pH values ​​are recorded as the first pH value and the second pH value. Calculate the absolute oxygen release capacity of each pH group of the HBOC sample to be tested according to the method described in any one of claims 1 to 3, and calculate the oxygen release ratio at different pH values ​​as needed; The Bohr effect BE is calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the second pH value: △OxyHb% first pH value - △OxyHb% second pH value; Based on the Bohr effect BE=△OxyHb%first pH value-△OxyHb%second pH value, the strength of the Bohr effect of the HBOC sample to be tested within the interval of the first pH value and the second pH value is obtained.

7. The method for predicting the HBOC Bohr effect according to claim 6, characterized in that: The method further comprises: The pH of the DPBS buffer solution is adjusted to at least 1 third pH value. When it is 1 pH value, the pH value is recorded as the third pH value; The Bohr effect BE was calculated based on the oxygen release ratio of the first pH value and the oxygen release ratio of the third pH value. 酸 =△OxyHb% first pH value-△OxyHb% third pH value; The Bohr effect BE was calculated based on the oxygen release ratio at the second pH value and the oxygen release ratio at the third pH value. 碱 =△OxyHb% third pH value-△OxyHb% second pH value; Calculate the Bohr effect BE 酸 and Bohr effect BE 碱 The Bohr effect index EI is calculated by the ratio of Optionally, the acidic or alkaline Bohr effect is judged based on the Bohr effect index EI; if EI>1, it is mainly the acidic Bohr effect; if EI<1, it is mainly the alkaline Bohr effect; the stronger the acidic Bohr effect, the stronger the oxygen release capacity; the stronger the alkaline Bohr effect, the stronger the oxygen carrying capacity.

8. A computer device, characterized in that: The device comprises: a memory and a processor; the memory is used to store a computer program; the processor executes the computer program to implement the steps of the method according to any one of claims 1 to 7.

9. A computer-readable storage medium, characterized in that: A computer program is stored thereon, and when the computer program is executed by a processor, the steps of the method according to any one of claims 1 to 7 are implemented.

10. A computer program product, comprising a computer program, characterized in that When the computer program is executed by a processor, the steps of the method described in any one of claims 1 to 7 are implemented.

Images