Pseudo-ginseng polysaccharide carbon quantum dot polymer as well as preparation method and application thereof

Abstract

The invention relates to the technical field of medical nano materials, and particularly discloses a panax notoginseng polysaccharide carbon quantum dot polymer as well as a preparation method and application thereof. The preparation method of the panax notoginseng polysaccharide carbon quantum dot polymer comprises the following steps: S1, extracting panax notoginseng polysaccharide from decoction dregs after total saponins are extracted from panax notoginseng; s2, dispersing panax notoginseng polysaccharide in water according to a ratio of 1g: (10-40) mL, carrying out hydrothermal reaction at 200-300 DEG C for 1-3 hours, and cooling to room temperature; s3, performing ultrasonic treatment, centrifuging to take supernate, filtering, dialyzing and purifying, and dialyzing by using a dialysis bag with the molecular weight cutoff of 500-800D; and S4, freeze-drying the dialyzed solution to obtain the panax notoginseng polysaccharide carbon quantum dot polymer. The medical dressing prepared from the pseudo-ginseng polysaccharide carbon quantum dot polymer provided by the invention is spongy and has a remarkable hemostatic effect, and a mouse liver injury model result shows that the hemostatic time is only about 56s.

Description

Technical Field

[0001] The present invention belongs to the technical field of medical nanomaterials, and particularly relates to a panax notoginseng polysaccharide carbon quantum dot polymer, a preparation method thereof, and an application thereof. Background Art

[0002] Panax notoginseng, as a traditional Chinese medicine, its active ingredient, total saponins of Panax notoginseng, has been widely used in the treatment of cardiovascular and cerebrovascular diseases. However, the medicinal residues generated after the industrial extraction of Panax notoginseng saponins account for 60% - 70% of the total raw materials. Due to the lack of high-value utilization methods, they are often directly discarded. Although existing technologies have tried to extract Panax notoginseng polysaccharides from the residues, there are still the following technical defects: purity and ash content problems. The polysaccharides obtained by the traditional water extraction and alcohol precipitation method have a high ash content (>4%) and low purity (<70%), seriously affecting their bioavailability and subsequent applications; process complexity and high cost. The existing processes require multiple steps of enzymatic hydrolysis, resin purification, and repeated centrifugation, with cumbersome operations and difficulty in large-scale production; limited functional development. Although Panax notoginseng polysaccharides have activities such as anti-fatigue and immunomodulation, their applications in the field of nanomaterials have not been fully explored.

[0003] Carbon Quantum Dots (CQDs), as a new type of nanomaterial, show potential in the fields of drug delivery and bioimaging due to their fluorescence properties, biocompatibility, and easy surface functionalization. However, traditional chemical synthesis methods rely on non-renewable raw materials (such as graphene) and are prone to produce toxic by-products. Although natural polysaccharides as carbon sources have biodegradability, low toxicity, and abundant functional groups (such as hydroxyl groups, carboxyl groups), existing research mainly focuses on common polysaccharides such as cellulose and chitosan, and the research on the carbon quantum dotization of Panax notoginseng polysaccharides is still blank. Existing patents (such as CN106832037A) have disclosed methods for extracting polysaccharides from Panax notoginseng residues, but do not involve the functional modification of polysaccharides.

[0004] How to convert waste medicinal residues into carbon quantum dot polymers meets the necessity and innovation of technology integration. It is urgent to develop a technical path to convert Panax notoginseng saponin extraction residues into high-value-added nanomaterials, providing a new path for the development of the entire industrial chain of Panax notoginseng. Summary of the Invention

[0005] The purpose of the present invention is to provide a panax notoginseng polysaccharide carbon quantum dot polymer, a preparation method thereof, and an application thereof, aiming at the deficiencies in the existing technology. The panax notoginseng polysaccharide carbon quantum dot polymer is a hemostatic material that can achieve rapid hemostasis, wound repair, and in vivo absorption.

[0006] To achieve the above purpose, the present invention adopts the following technical solutions: The first aspect of the present invention is to provide a preparation method of a panax notoginseng polysaccharide carbon quantum dot polymer, comprising the following steps: S1. Extract notoginseng polysaccharide from the residue after extracting total saponins from notoginseng; S2. Disperse notoginseng polysaccharide in water at a ratio of 1 g:(10 - 40) mL, carry out hydrothermal reaction at 200 - 300 °C for 1 - 3 h, and then cool to room temperature; S3. Perform ultrasonic treatment, centrifuge to obtain the supernatant, filter and purify by dialysis. Use a dialysis bag with a cut-off molecular weight of 500 - 800 D for dialysis; S4. Freeze-dry the dialyzed solution to obtain the carbon quantum dot polymerized notoginseng polysaccharide Further, in step S3, the ultrasonic treatment time is 15 - 30 min, the ultrasonic power is 100 - 500 W, and the frequency is 10 - 50 kHz.

[0007] Further, in step S3, the centrifugation speed is 3000 - 10000 r / min, and the centrifugation time is 5 - 30 min.

[0008] Further, in step S3, the filtration uses a 0.22 - 0.45 μm microporous filter membrane, the dialysis time is 6 - 24 h, and the dialysis solution is changed every 2 - 6 h.

[0009] Further, the extraction process of the notoginseng polysaccharide is as follows: dry and crush the residue after extracting total saponins from notoginseng, sieve it, extract with alkali, then concentrate and cool, add an equal volume of protein removal sevag solution, remove the precipitate, add absolute ethanol to the filtrate until precipitation occurs, freeze-dry the filter cake, and sieve it to obtain.

[0010] The second aspect of the present invention is to provide a carbon quantum dot polymerized notoginseng polysaccharide prepared by the above method.

[0011] Further, the particle size of the carbon quantum dot polymerized notoginseng polysaccharide is not greater than 5 nm.

[0012] Further, the fluorescence spectrum of the carbon quantum dot polymerized notoginseng polysaccharide shows that the maximum excitation wavelength is 370 nm and the maximum emission wavelength is 468 nm.

[0013] The third aspect of the present invention is to provide the application of the above carbon quantum dot polymerized notoginseng polysaccharide in medicine, health products or cosmetics.

[0014] The fourth aspect of the present invention is to provide a medical dressing, which is obtained by injecting the above carbon quantum dot polymerized notoginseng polysaccharide solution into a prefabricated mold and freeze-drying.

[0015] Compared with the prior art, the beneficial effects brought by the technical solution provided by the present invention are: (1)The polysaccharide carbon quantum dot polymer of Panax notoginseng provided by the present invention has a particle size not greater than 5 nm. The fluorescence spectrum shows that the maximum excitation wavelength is 370 nm and the maximum emission wavelength is 468 nm.

[0016] (2)The medical dressing is prepared from the polysaccharide carbon quantum dot polymer of Panax notoginseng provided by the present invention. It is light yellowish-white or grayish-white, flat and spongy, and feels soft to the touch. Compared with the existing Surgicel (regenerated oxidized fiber), its hemostatic effect is more significant. The results of the mouse liver injury model show that the hemostasis time is only about 56 s.

[0017] (3)The medical dressing provided by the present invention can be widely used for traumatic hemostasis, intraoperative hemostasis, and adjuvant treatment of burns and scalds, etc., and has broad application potential. Description of the Drawings

[0018] Figure 1 It is a transmission electron microscope image of the polysaccharide carbon quantum dot polymer of Panax notoginseng prepared in Example 1; Figure 2 It is a particle size distribution diagram of the polysaccharide carbon quantum dot polymer of Panax notoginseng prepared in Example 1; Figure 3 It is a fluorescence spectrum diagram of the polysaccharide carbon quantum dot polymer of Panax notoginseng prepared in Example 1; Figure 4 It is an optical image of the medical dressing prepared from the polysaccharide carbon quantum dot polymer of Panax notoginseng; Figure 5 It is a longitudinal scanning electron microscope image of the medical dressing provided by the present invention prepared from the polysaccharide carbon quantum dot polymer of Panax notoginseng; Figure 6 It is a cross-sectional scanning electron microscope image of the medical dressing provided by the present invention prepared from the polysaccharide carbon quantum dot polymer of Panax notoginseng; Figure 7 It is a microscopic image of a tissue section of the negative control group; Figure 8 It is a microscopic image of a tissue section of the positive control group; Figure 9 It is a microscopic image of a tissue section of the medical dressing provided by the present invention prepared from the polysaccharide carbon quantum dot polymer of Panax notoginseng. Detailed Embodiments

[0019] To make the objectives, technical solutions, and advantages of the present invention clearer, the following further describes the detailed embodiments of the present invention in conjunction with specific examples and drawings. For those not specified in the examples regarding specific test methods, instrument equipment, or conditions, they are all carried out according to the techniques or conditions described in the literature in this field or according to the product specifications. For reagents or instruments without indicating the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0020] Example 1 The residue of notoginseng after extracting total ginsenosides was dried, pulverized, passed through a 200-mesh sieve, added with ultrapure water, and the solid-liquid weight ratio was maintained at 1:40. The pH was adjusted to pH≈13 with NaOH, magnetically stirred and soaked at 60°C for 60 min, ultrasonically extracted for 8 min, filtered under negative pressure, the filtrate was concentrated at 60°C with a rotary evaporator, after cooling, an equal volume of protein-removing sevag solution was added, shaken at room temperature for 15 min, allowed to stand for layering, and the precipitate (i.e., impurity protein) was filtered off. Absolute ethanol was added to the clear liquid until precipitation occurred, refrigerated and allowed to stand at 4°C for 12 h, then filtered under negative pressure. The filter cake was redissolved with ultrapure water (heated appropriately if necessary, preferably not exceeding 60°C), decolorized with activated carbon, and the insoluble matter was filtered off. Absolute ethanol was added to the filtrate until precipitation occurred, refrigerated and allowed to stand at 4°C for 12 h, then filtered under negative pressure. The filter cake was freeze-dried, passed through a 100-mesh standard sieve, weighed and the yield was calculated, and stored at -18°C for standby.

[0021] According to the ratio, 1.5 g of notoginseng polysaccharide was dispersed in 30 mL of ultrapure water, placed in a 50 mL polytetrafluoroethylene-lined reaction kettle, reacted in a muffle furnace at 240°C for 2 h, then cooled to room temperature, ultrasonically treated for 20 min, centrifuged at 6000 r / min for 10 min, and the supernatant was taken. The supernatant was filtered through a 0.22 μm microporous filter membrane. The filtrate was dialyzed for 12 h (the dialysis solution was changed every 4 h) in a purified water system using a dialysis bag with a molecular weight cut-off of 700D with the aid of a magnetic stirrer. The dialyzed solution was collected and freeze-dried to obtain notoginseng polysaccharide carbon quantum dot polymer, and the yield was about 25%.

[0022] Example 2 The residue of notoginseng after extracting total ginsenosides was dried, pulverized, passed through a 200-mesh sieve, added with ultrapure water, and the solid-liquid weight ratio was maintained at 1:40. The pH was adjusted to pH≈13 with NaOH, magnetically stirred and soaked at 60°C for 60 min, ultrasonically extracted for 8 min, filtered under negative pressure, the filtrate was concentrated at 60°C with a rotary evaporator, after cooling, an equal volume of protein-removing sevag solution was added, shaken at room temperature for 15 min, allowed to stand for layering, and the precipitate (i.e., impurity protein) was filtered off. Absolute ethanol was added to the clear liquid until precipitation occurred, refrigerated and allowed to stand at 4°C for 12 h, then filtered under negative pressure. The filter cake was redissolved with ultrapure water (heated appropriately if necessary, preferably not exceeding 60°C), decolorized with activated carbon, and the insoluble matter was filtered off. Absolute ethanol was added to the filtrate until precipitation occurred, refrigerated and allowed to stand at 4°C for 12 h, then filtered under negative pressure. The filter cake was freeze-dried, passed through a 100-mesh standard sieve, weighed and the yield was calculated, and stored at -18°C for standby.

[0023] Disperse 1.5 g of notoginseng polysaccharide proportionally into 10 mL of ultrapure water, place it in a 50 mL polytetrafluoroethylene-lined reaction kettle, react in a muffle furnace at 200 °C for 1 h, then cool to room temperature, ultrasonicate for 15 min, centrifuge at 3000 r / min for 30 min, and take the supernatant. Filter the supernatant through a 0.22 μm microporous membrane. Dialyze the filtrate using a dialysis bag with a molecular weight cut-off of 800 D in a purified water system with the aid of a magnetic stirrer for 6 h (change the dialysis solution every 2 h). Collect the dialyzed solution and perform freeze-drying to obtain the carbon quantum dot-modified polymer of notoginseng polysaccharide.

[0024] Example 3 The residue after extracting total saponins from notoginseng is dried, pulverized, and passed through a 200-mesh sieve. Add ultrapure water to keep the solid-liquid weight ratio at 1:40, adjust the pH to pH≈13 with NaOH, soak with magnetic stirring at 60 °C for 60 min, perform ultrasonic extraction for 8 min, carry out negative pressure filtration, concentrate the filtrate with a rotary evaporator at 60 °C, add an equal volume of protein-removing sevag solution after cooling, shake at room temperature for 15 min, let it stand for layer separation, filter out the precipitate (i.e., impurity proteins), add absolute ethanol to the clear liquid until precipitation occurs, refrigerate and stand at 4 °C for 12 h, perform negative pressure filtration, take the filter cake, redissolve it with ultrapure water (heat appropriately if necessary, preferably not exceeding 60 °C), decolorize with activated carbon and filter out the insoluble substances, add absolute ethanol to the filtrate until precipitation occurs, refrigerate and stand at 4 °C for 12 h, perform negative pressure filtration, freeze-dry the filter cake, pass through a 100-mesh standard sieve, weigh and calculate the yield, and store it for later use at -18 °C.

[0025] Disperse 1.5 g of notoginseng polysaccharide proportionally into 40 mL of ultrapure water, place it in a 50 mL polytetrafluoroethylene-lined reaction kettle, react in a muffle furnace at 300 °C for 3 h, then cool to room temperature, ultrasonicate for 30 min, centrifuge at 10000 r / min for 5 min, and take the supernatant. Filter the supernatant through a 0.22 μm microporous membrane. Dialyze the filtrate using a dialysis bag with a molecular weight cut-off of 500 D in a purified water system with the aid of a magnetic stirrer for 24 h (change the dialysis solution every 6 h). Collect the dialyzed solution and perform freeze-drying to obtain the carbon quantum dot-modified polymer of notoginseng polysaccharide.

[0026] Comparative Example 1 It is basically the same as Example 1, except that in step S3, the filtrate uses a dialysis bag with a molecular weight cut-off of 1000 D. Allowing substances with a larger molecular weight to dialyze out results in surface defects on the CQDs, weak fluorescence performance, and low quantum yield, leading to poor fiber orientation of the prepared medical dressing and affecting material demolding.

[0027] Comparative Example 2 It is basically the same as Example 1, except that in step S1, the temperature of the hydrothermal reaction is 350 °C. If the temperature is too high, excessive carbonization will lead to an increase in surface defects of the carbon quantum dots, triggering non-radiative energy transfer, and the fluorescence quantum yield will instead decrease. Moreover, particle aggregation occurs, and the particle size is greater than 10 nm.

[0028] Comparative Example 3 It is basically the same as Example 1, except that in step S1, the temperature of the hydrothermal reaction is 180 °C. When the temperature is low, the particle size of the prepared carbon quantum dots is 5-10 nm.

[0029] The pseudo-ginseng polysaccharide carbon quantum dot polymers prepared in Examples 1-3 all have the same microscopic morphology and fluorescence characteristics. The following takes Example 1 as an example for detailed description: Reference Figure 1 and Figure 2 , it can be seen that the diameter of the prepared carbon quantum dots <5 nm, which are uniformly dispersed spherical particles with good dispersibility. The statistical analysis of the particle size shows that the average particle size is about 2.15 nm.

[0030] Reference Figure 3 , the prepared carbon quantum dots have inherent fluorescence characteristics. As shown in the figure, they are light yellow-brown in the environment, while under ultraviolet light with a wavelength of 373 nm, they show obvious blue-green fluorescence. At the same time, the fluorescence spectrum shows that the maximum excitation wavelength is 370 nm and the maximum emission wavelength is 468 nm.

[0031] Zeta potential detection shows that the surface of the carbon quantum dots contains hydroxyl groups (-OH) and carboxyl groups (-COOH), so they carry a weak negative charge. The surface charge detected by a nano-particle size potentiometer is -5.30 ± 0.12 mV.

[0032] Example 4 In order to investigate the hemostatic effect of the pseudo-ginseng polysaccharide carbon quantum dot polymer provided by the present invention, the pseudo-ginseng polysaccharide carbon quantum dot polymer prepared in Example 1 was used to prepare a medical dressing. The specific process is as follows: Weigh an appropriate amount of dry pseudo-ginseng polysaccharide carbon quantum dot polymer, add it to ultrapure water, with a mass fraction of 1% - 3%, stir and shake for 2 hours to obtain a polysaccharide solution, and adjust its pH value to 7 - 8 (adjusted by 1 mol / L sodium hydroxide solution).

[0033] Inject the pseudo-ginseng polysaccharide carbon quantum dot polymer solution into a prefabricated mold, pour liquid nitrogen for pre-freezing for 15 min, place it in a -40 °C ultra-low temperature refrigerator, freeze and store for 12 h to form, then place it in a freeze dryer for freeze drying for 24 h (pressure <5 Pa, temperature -50 °C), and demold in a super-clean workbench to obtain a medical dressing. The demolded test dressing is sheared and then internally packaged with a paper-plastic packaging bag and sterilized with ethylene oxide.

[0034] The prepared medical dressing has no stains on its appearance, is well-sealed, and has no peculiar smell. Refer to Figure 4 , the medical dressing is light yellowish-white or grayish-white, flat and spongy, and feels soft to the touch. The appearance shows fibrous stripes.

[0035] Refer to Figure 5 and Figure 6 , which are the SEM images of the longitudinal and cross-sectional surfaces of the medical dressing respectively. It has a three-dimensional structure with loose pores, a large specific surface area, and a thickness of 8 ± 1 (mm).

[0036] The medical dressing was also tested for acidity, alkalinity, and heavy metals using the 2020 edition of the Pharmacopoeia. The pH value of the dissolution solution of the medical dressing is 6 - 8, and the heavy metal content should not be greater than 20 mg / kg.

[0037] Example 5 Systemic acute toxicity test and muscle implantation test.

[0038] (1) Test materials Positive control product: Surgicel, a regenerated oxidized fiber, purchased from Johnson & Johnson (Shanghai) Medical Devices Co., Ltd.

[0039] Sample group: Example 4.

[0040] All test materials were cut into pieces of 5 mm × 3 mm size under a laminar flow hood environment before the operation for test implantation.

[0041] Animals: Adult New Zealand long-eared rabbits for testing.

[0042] Animal operating table, sterile gauze, surgical knives, hemostats, disposable sterile syringes, stainless steel boxes, autoclaves, and disposable surgical gowns, gloves, and masks. Sodium pentobarbital (Guangdong Bangmin Pharmaceutical Factory Co., Ltd.).

[0043] (2) Test methods and procedures Six New Zealand long-eared rabbits were randomly divided into 3 groups by gender, with 2 animals in each group, 1 male and 1 female. Group A: Negative control group; Group B: Positive control group (Surgicel), Group C: Sample group.

[0044] Before the operation, each New Zealand long-eared rabbit was weighed, recorded, and marked, and then anesthetized and tested one by one. The materials of each group were implanted under the muscles on the ventral side of the right hind leg of the New Zealand long-eared rabbit. The sham operation was used as the negative control, and Surgicel was used as the positive control. The reactions and survival rates of the animals were observed within 72 hours.

[0045] The present invention adopts the implantation method of planting, that is, using a minimally invasive surgical forceps to pick up small pieces of test materials, piercing the corresponding muscle tissue and then taking out the minimally invasive surgical forceps, suturing the skin, disinfecting and dressing the local area to complete the implantation. Disinfect and dress the wound every day, one animal per cage, and raise them normally. This method forms a smaller wound at the muscle part of the surgical implantation, and the muscle wound does not need to be sutured to reduce the interference of the suture on the absorption of the test material. The negative group only forms the same wound without implanting materials.

[0046] One week later, the skin sutures were removed. Three weeks later, the animals were euthanized painlessly, and the test results were observed. The leg muscle tissues except the part containing the implanted material were taken out, fixed in 10% formalin solution for 24 hours, embedded in paraffin, sectioned, stained with HE, and pathological sections were made to observe the inflammatory reaction under an optical microscope.

[0047] (3)Test results Continuously observe the reaction of New Zealand long-eared rabbits for 72 hours. None of the animals in groups A, B, and C died, had no adverse reactions, maintained normal body temperature, had no obvious change in body weight, and showed normal animal activities. The main evaluation index of microscopic observation is the grade of inflammation occurrence, and the inflammation judgment grade is shown in Table 1.

[0048] Table 1. Inflammatory reaction grading criteria

[0049] Three weeks after the test, the animals were anesthetized, and the skin at the previously implanted material site was cut open to observe the morphological changes of the muscle tissue at the implanted site. Visible to the naked eye: The muscle tissues of groups A, B, and C were all normal, the wounds healed well, there was no bleeding or nodules, Surgicel and the test samples had been completely absorbed, and no residues were found with the naked eye; a little scar tissue could be seen at the wound site, which was the scar produced by autologous healing. The muscle tissue at the surgical site was taken out, tissue sections were prepared, and microscopic observation was carried out.

[0050] The experimental results are as Figures 7 - 9 shown. The results of fiber observation of HE sections at the test site three weeks after the test showed that: the muscle fibers of groups A, B, and C had good continuity, no obvious inflammatory reaction, only a small amount of inflammatory cell infiltration in the tissue stroma, and a small amount of fibrous tissue hyperplasia at the edge of the embedded tissue. The tissue structure was normal, and all were judged as grade 0 of inflammatory reaction.

[0051] The HE sections showed that the test results of the three groups were similar. The biocompatibility of the medical dressing provided by the present invention is equivalent to that of the currently sold products, and the biocompatibility is good.

[0052] Example 6 The evaluation of hemostatic performance is that in vitro coagulation tests and the establishment of in vivo hemostatic models are conventional means for comprehensively evaluating the hemostatic performance of hemostatic materials. In vitro coagulation tests evaluate their procoagulant performance by detecting the whole blood clotting time, activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT), and preliminarily infer their hemostatic mechanisms.

[0053] Test materials: Grouping: Control group: Surgicel, a regenerated oxidized fiber, purchased from Johnson & Johnson (Shanghai) Medical Devices Co., Ltd.; Blank group: Ordinary medical gauze; Notoginsenoside polysaccharide group: A hemostatic dressing prepared from notoginsenoside polysaccharide without carbon quantum dot preparation.

[0054] Animal operating table, sterile gauze, surgical knives, hemostats, disposable sterile syringes, stopwatches, stainless steel boxes, autoclaves, and disposable surgical gowns, gloves, and masks. Sodium pentobarbital (Guangdong Bangmin Pharmaceutical Co., Ltd.), sodium citrate (Zhongshan Xinning Pharmaceutical Co., Ltd.), APTT, PT, and TT kits (Wuhan Sevier Biotechnology Co., Ltd.). New Zealand long-eared rabbits.

[0055] (1) Determination of whole blood clotting time The test was set up with experimental groups, control groups, and blank groups, and the notoginsenoside polysaccharide group, n = 10, using disposable sterile plastic test tubes. The self-made multifunctional absorbable hemostatic wound dressings 500D and 1000D semi-permeable membrane products in the laboratory were respectively prepared into solutions with concentrations of 5%, 2.5%, and 1%, for a total of 6 test groups to be measured; Surgicel was prepared into a 5% solution with normal saline as the control solution, and the notoginsenoside polysaccharide group prepared a 5% solution of notoginsenoside polysaccharide without carbon quantum dot preparation for standby. 20 μl of the sample solution was placed in each test tube of the experimental group, 20 μl of the control solution was placed in each test tube of the control group, 20 μl of normal saline was added to the blank group, and 20 μl of the corresponding notoginsenoside polysaccharide solution was added to the notoginsenoside polysaccharide group. After all the test tubes of each group were sealed, they were pre-warmed at 37 °C for standby.

[0056] After taking blood from the heart of New Zealand long-eared rabbits, it was mixed evenly with sodium citrate at a ratio of 9:1 to obtain anticoagulated whole blood. The anticoagulated whole blood was respectively added to the test tubes of the experimental group, control group, and blank group, 1 mL per tube, incubated at 37 °C for 3 min, and after adding 25 mmol / L CaCl2 solution, the stopwatch was immediately started. The test tubes were tilted every 30 s to observe whether the blood was flowing until the blood coagulated (the inverted test tube showed no blood flow). The time from adding the CaCl2 solution to the solidification of the liquid surface without flow was recorded, which was the whole blood clotting time.

[0057] (2) Determination of in vitro coagulation indexes APTT, PT, and TT Centrifuge the anticoagulated whole blood obtained in the previous step at 3000 rpm and low temperature for 15 minutes, and collect the upper platelet-poor plasma for later use.

[0058] For the determination of the three in vitro coagulation indexes APTT, PT, and TT, experimental groups, control groups, and blank groups were set up, with n = 10. In each test tube of the experimental group, a small piece of multifunctional absorbable hemostatic wound dressing (2 mg) was placed; in each test tube of the control group, a small piece of Surgicel (20 mg) was placed; nothing was placed in the blank group; in the panax notoginseng polysaccharide group, a dressing prepared with 2 mg of the corresponding panax notoginseng polysaccharide was added. All test tubes were preheated at 37 °C for 5 minutes for later use.

[0059] APTT determination: Add 0.5 mL of plasma to each test tube of the experimental group, control group, blank group, and panax notoginseng polysaccharide group, incubate at 37 °C for 5 minutes, add 0.5 mL of APTT reagent pre-warmed at 37 °C and mix well, continue to incubate at 37 °C for 5 minutes, add 0.5 mL of 25 mmol / L CaCl2 solution pre-warmed at 37 °C, immediately start the stopwatch, and stop the watch until plasma coagulation occurs, and record the plasma coagulation time (in seconds).

[0060] PT and TT determination: Use PT and TT reagent kits respectively, and the test method is the same as above.

[0061] (3) Test results The determination results of whole blood coagulation time, activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) are shown in Table 2. It can be seen from Table 1 that the whole blood coagulation time of the multifunctional absorbable hemostatic wound dressing sample group is about 160 s, that of the commercially available Surgicel in the control group is about 206 s, that of the blank group is about 227 s, and that of the panax notoginseng polysaccharide group is about 190 s.

[0062] Table 2. Determination results of whole blood coagulation time, APTT, PT, and TT (n = 10)

[0063] Note: The data in this table are analyzed by one-way ANOVA with the blank group as the control group, and the analysis results are represented by ##; the data in this table are analyzed by one-way ANOVA with the control group (5%) as the control, and the analysis results are represented by **.

[0064] Analyzed by ANOVA of SPSS-19.0 software, the experimental groups (500D), experimental groups (1000D), control groups, blank groups, and panax notoginseng polysaccharide groups were compared pairwise with the blank group and the control group (5%) as the controls respectively.

[0065] The determination results of whole blood clotting time, APTT, and PT kits showed that P < 0.001 between experimental group 500D (5%) and experimental group 1000D (5%) and the blank group, control group, and panax notoginseng polysaccharide group, with significant differences, indicated by **. The determination results of the TT kit showed that P < 0.01 between experimental group 500D (5%) and experimental group 1000D (5%) and the control group, blank group, and panax notoginseng polysaccharide group, also with significant differences, indicated by *.

[0066] The comparison results between experimental group 500D (2.5%), experimental group 500D (1%), experimental group 1000D (2.5%), experimental group 1000D (1%) and the control group, blank group, and panax notoginseng polysaccharide group for whole blood clotting time and PT showed that P < 0.01, with significant differences, indicated by *.

[0067] The comparison results between experimental group 500D (1%) and experimental group 1000D (1%) and the control group, blank group, and panax notoginseng polysaccharide group for whole blood clotting time showed that P < 0.01, with significant differences, indicated by *.

[0068] For the control group, panax notoginseng polysaccharide group, and blank group, with the blank group as the comparison object, the statistical analysis of the determination results of whole blood clotting time showed that P < 0.001, with significant differences, indicated by ##; the statistical analysis of the determination results of APTT, PT, and TT kits showed that P < 0.01, with significant differences, indicated by #.

[0069] Example 6 Mouse liver injury model.

[0070] In vivo liver hemostasis experiments were carried out on New Zealand short-haired experimental white rabbits. When the experimental animals met the experimental conditions, the liver was cut with surgical scissors in a clean environment to form a 1-cm-long wound. Hemostatic materials were applied to the wound and pressed moderately, and the hemostasis time and subsequent oozing time were observed. The hemostasis time was recorded when no oozing was observed. The average data after statistics are shown in Table 3 below.

[0071] Experimental group: The medical dressing prepared in Example 4; Control group: Surgicel, a regenerated oxidized fiber, purchased from Johnson & Johnson (Shanghai) Medical Devices Co., Ltd.; Blank group: Ordinary medical gauze; Panax notoginseng polysaccharide group: Hemostatic dressing prepared from panax notoginseng polysaccharide without carbon quantum dots.

[0072] Table 3. Hemostasis time.

[0073]

[0074] Note: The time data in the table are statistical data.

[0075] In the case of no conflict, the above-mentioned embodiments and the features in the embodiments in this article may be combined with each other.

[0076] The above are only the preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a notoginseng polysaccharide carbon quantum dot polymer, characterized in that: The following steps are involved: S1. extracting Panax notoginseng polysaccharide from the residue after extracting total saponins from Panax notoginseng; S2, disperse Panax notoginseng polysaccharide in water at a ratio of 1 g: (10-40) mL, perform hydrothermal reaction at 200-300°C for 1-3 hours, and then cool to room temperature; S3, ultrasonic treatment, centrifugation to obtain the supernatant, filtration and dialyzation purification, dialysis using a dialysis bag with a molecular weight cutoff of 500-800D; S4. Freeze-dry the dialyzed solution to obtain Panax notoginseng polysaccharide carbon quantum dot polymer.

2. The method according to claim 1, characterized in that In step S3, the ultrasonic treatment time is 15-40 min, the ultrasonic power is 100-500 W, and the frequency is 10-50 kHz.

3. The method according to claim 1, characterized in that In step S3, the centrifugal speed is 3000-10000 r / min, and the centrifugal time is 5-30 min.

4. The method according to claim 1, characterized in that: In step S3, the filtration uses a 0.22-0.45 μm microporous filter membrane, the dialysis time is 6-24 hours, and the dialysate is replaced every 2-6 hours.

5. The method according to claim 1, characterized in that The extraction process of Panax notoginseng polysaccharide is as follows: drying and crushing the residue after extracting total saponins from Panax notoginseng, sieving, alkali extraction, and then concentrating and cooling, adding an equal volume of protein-removing sevag solution, removing precipitates, adding anhydrous ethanol to the filtrate until precipitates are precipitated, freeze-drying the filter cake, and sieving to obtain the polysaccharide.

6. A Panax notoginseng polysaccharide carbon quantum dot polymer prepared by the method described in any one of claims 1 to 5.

7. The notoginseng polysaccharide carbon quantum dot polymer according to claim 6, characterized in that: The particle size of the notoginseng polysaccharide carbon quantum dot polymer is no more than 5 nm.

8. The notoginseng polysaccharide carbon quantum dot polymer according to claim 6, characterized in that: The fluorescence spectrum of the Panax notoginseng polysaccharide carbon quantum dot polymer shows that the maximum excitation wavelength is 370nm and the maximum emission wavelength is 468nm.

9. Use of the Panax notoginseng polysaccharide carbon quantum dot polymer as described in any one of claims 6 to 8 in the preparation of medicines, health products or cosmetics.

10. A medical dressing, characterized in that: The notoginseng polysaccharide carbon quantum dot polymer solution as described in any one of claims 6 to 8 is injected into a prefabricated mold and freeze-dried to obtain the product.

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