Heat preservation vacuum sampling tube for measuring number of platelets

Abstract

The invention provides a heat preservation vacuum sampling tube for detecting the number of platelets, which comprises a special-shaped test tube body, the special-shaped test tube body is composed of an upper test tube body and a lower test tube body, the diameter of the upper test tube body is larger than that of the lower test tube body, the extension line of the lower test tube body is internally tangent to the upper test tube body, and the extension line of the lower test tube body is internally tangent to the lower test tube body. A sealing cover is sleeved at a port of the upper test tube body, a thermal insulation sleeve is sleeved outside the lower test tube body, the outer side surface of the thermal insulation sleeve is flush with the upper test tube body, and a longitudinal notch is reserved in the thermal insulation sleeve at the longitudinal tangent line of the upper test tube body and the lower test tube body. The special-shaped test tube body is filled with an anti-coagulation solution, and the anti-coagulation solution is applied to a full-automatic blood cell analyzer and comprises 1.5 to 2.2 mg / ml of EDTA-K2 (Ethylene Diamine Tetraacetic Acid) and 9.5 to 19 umol / L of P2Y12 receptor antagonist ticagrelor.

Description

Technical Field

[0001] The invention belongs to the technical field of medical analysis, and in particular relates to a heat-insulating vacuum sampling tube for measuring the number of platelets. Background Art

[0002] After blood leaves the body, it loses the regulation of internal environmental factors such as temperature, endothelial cell secretory factors, and flow shear force. The physiological status of blood cells changes rapidly with the changes in the environment, and fragmentation, dissolution, coagulation, and other phenomena may occur at any time, affecting the detection of various blood cell indicators. How to mimic the internal environment of the cardiovascular system? How to keep blood cells in a good physiological state for a longer time? How to obtain the "true value" when counting blood cells? Preventing or reducing changes in the morphology and quantity of platelets will help medical technicians observe the dynamic physiological changes of blood cells in the static (ex vivo) mode, and can present various data to doctors in a timely and accurate manner, which is beneficial for doctors to diagnose and treat patients' diseases.

[0003] Blood cell analysis is a routine test item in clinical application. Although the probability of EDTA-dependent pseudothrombocytopenia (EDTA-PTCP) is low, EDTA-K 2 Anticoagulants cause platelet aggregation, which makes it impossible for blood cell analyzers to correctly identify platelets, accounting for more than 90% of pseudothrombocytopenia. This can easily lead to misdiagnosis of patients and cause trouble for patients and clinical diagnosis and treatment.

[0004] Platelet-neutrophil aggregates in the body are formed under the action of platelets and neutrophils, which in turn provide favorable conditions for thrombosis and inflammation, thereby affecting the occurrence and development of the disease, causing intravascular plaques to rupture, resulting in thrombosis and embolism, and triggering myocardial infarction, cerebral infarction and other diseases. The neutrophil / lymphocyte ratio (NLR) and platelet / lymphocyte ratio (PLR) have attracted much attention in recent years. Acute myocardial infarction (AMI) is the most critical emergency event among cardiovascular diseases. With the continuous advancement of percutaneous coronary intervention (PCI) technology and the continuous improvement of stent materials. Studies have shown that an early feature of cerebral infarction is a decrease in lymphocytes. In the prognosis prediction of patients with acute cerebral infarction, the clinical value of the neutrophil-to-lymphocyte ratio is significantly higher than that of the platelet-to-lymphocyte ratio, indicating that among the factors affecting the poor prognosis of patients with acute cerebral infarction after thrombolytic therapy, increased NLR and PLR are independent risk factors, which can provide important clinical basis. Platelets, neutrophils and lymphocytes are easily affected by the state of the body and are not stable. In contrast, PLR and NLR are more reliable and can effectively reflect the inflammatory state of the body. Many studies have shown that both NLR and PLR are related to risk stratification and poor prognosis in patients with acute coronary syndrome (ACS). Compared with the prediction of a single indicator, combined prediction can improve its sensitivity and specificity to a certain extent. Compared with the use of NLR and PLR alone, NLR+PLR has a higher ability to predict the risk of in-hospital death in patients with AMI. In addition, compared with NSTEMI patients, NLR+PLR is better in predicting the risk of in-hospital mortality in STEMI patients. Combined predictive indicators can be used to risk stratify AMI patients during hospitalization to improve their clinical prognosis.

[0005] In the actual clinical testing work, it is quite difficult to complete a routine blood test within 15 minutes after the patient's blood is drawn (except in emergency cases). However, if the blood cell analysis is done more than 30 minutes after the EDTA-K2 anticoagulant is used, the platelet count of DTA-PTCP patients is likely to be reduced, but the number of white blood cells will increase relatively due to the increase in platelet aggregates.

[0006] At present, the methods for calibrating EDTA-PTCP include: dilution mode method, anticoagulant replacement method, microcentrifugation method, drug stabilization method, and pre-dilution vacuum blood collection tube detection method. However, the dilution mode method, microcentrifugation method, and drug stabilization method are inconvenient to operate in actual applications. Although the anticoagulant replacement method is simple to operate, it has the defect of incomplete calibration. Summary of the invention

[0007] The invention provides a heat-insulating vacuum sampling tube for measuring the number of platelets, which solves the problem that the sample temperature affects the measurement result when measuring the number of platelets in the prior art.

[0008] In order to solve the above technical problems, the technical solution of the present invention is:

[0009] A heat-insulating vacuum sampling tube for detecting the number of platelets comprises a special-shaped test tube body, the special-shaped test tube body is composed of an upper test tube body and a lower test tube body, the diameter of the upper test tube body is larger than the diameter of the lower test tube body, the extension line of the lower test tube body is inscribed with the inner part of the upper test tube body, a sealing cover is provided at the port of the upper test tube body, a heat-insulating sleeve is provided on the outer surface of the heat-insulating sleeve, the outer side of the heat-insulating sleeve is flush with the upper test tube body, and the heat-insulating sleeve has a longitudinal notch at the longitudinal tangent of the upper test tube body and the lower test tube body, and the longitudinal notch with a spacing is conducive to observing the liquid level in the test tube from the outside. The special-shaped test tube body can be made of glass or PE material.

[0010] Among them, the insulation sleeve is filled with self-heating material or phase change material, and the outer membrane of the moisturizing sleeve is made of breathable and water-impermeable material, which is conducive to heat dissipation. Self-heating material or phase change material is a special material that can automatically release heat energy under certain conditions. Self-heating material is usually produced by contacting chemical heating agents with oxygen, and phase change material uses the phase change of matter to release or absorb latent heat to adjust the temperature. It is often used in cushions, knee pads and other products that require insulation function. The self-heating material and phase change material of the present invention are composed of common materials on the market. The breathable and water-impermeable materials are common commercially available materials such as expanded polytetrafluoroethylene and expanded polytetrafluoroethylene. The insulation sleeve of the present invention is made of "self-heating" material or phase change material. After the seal is opened and properly handled, it can quickly enter the heating and heat preservation state within 10 minutes. An inner lining temperature control film is provided between the moisturizing sleeve and the lower test tube body to ensure uniform heating. The temperature generated by the self-heating material can make the liquid in the tube reach 26°C, and the duration is >120min.

[0011] Among them, the upper test tube body is 20mm, the lower test tube body has a diameter of 7-8mm, the width of the longitudinal notch is 2-3mm, the height is 50-55mm, and the total length D of the test tube body and the insulation sleeve can be greater than 75mm.

[0012] The test tube is vacuum-filled and sealed with a rubber stopper, and the special-shaped test tube can hold more than 3 ml of blood sample.

[0013] The special-shaped test tube contains an anticoagulant solution, which is used in a fully automatic blood cell analyzer and contains 1.5-2.2 mg / ml EDTA-K 2 and 9.5-19umol / L P2Y 12 Receptor antagonist ticagrelor.

[0014] The automatic blood cell analyzer uses blood cell counting to analyze samples 60 minutes to 120 minutes after the anticoagulant and venous blood are mixed.

[0015] The invention also contains 10-20umol / L of an adenylate cyclase activator, wherein the adenylate cyclase activator is forskolin.

[0016] The automatic blood cell analyzer uses blood cell counting to analyze samples within 120 minutes after the anticoagulant and venous blood are mixed.

[0017] The samples for testing include 30-40ul of blood cell counting analysis anticoagulant and 2ml of venous blood.

[0018] Compared with the prior art, the technical solution provided by the present invention has the following beneficial effects:

[0019] The insulated vacuum sampling tube in the present invention simulates the internal environment of the human body, and combines the in vitro anti-platelet aggregation effects of ticagrelor and forskolin under insulated conditions, and significantly inhibits the pseudoplatelet aggregation phenomenon under multiple activation mechanisms, thereby reducing the impact on other blood cell analysis effects and the number of blood cells, and may provide timely and accurate basis for the diagnosis of clinical laboratory platelet counts, platelet dysfunction-related diseases, and cardiovascular and cerebrovascular diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 60min blood smear (10×10)

[0021] Figure 2 60min blood smear microscopy (100×10)

[0022] Figure 3 Satellite phenomenon in 60-minute blood smear microscopy (100×10)

[0023] Figure 4 This is a schematic diagram of the structure of the heat-insulating vacuum sampling tube of the present invention;

[0024] Figure 5 It is a cross-sectional view of the heat-insulating vacuum sampling tube of the present invention along the AA direction (without the heat-insulating sleeve);

[0025] Figure 6 It is a cross-sectional view of the heat-insulating vacuum sampling tube of the present invention along the BB direction.

[0026] In the figure, 1. special-shaped test tube body, 1-1. upper test tube body, 1-2. lower test tube body, 2. sealing cover, 3. insulation sleeve, 4. longitudinal notch, 5. lining temperature control film. DETAILED DESCRIPTION

[0027] The following embodiments are provided for a better understanding of the present invention, but are not limited to the best mode of implementation, and do not limit the content and protection scope of the present invention. Any product identical or similar to the present invention obtained under the inspiration of the present invention or by combining the features of the present invention with other prior arts shall fall within the protection scope of the present invention.

[0028] Example 1

[0029] like Figure 4-Figure 6 As shown, this embodiment provides a heat preservation vacuum sampling tube for detecting the number of platelets, including a special-shaped test tube body 1, the special-shaped test tube body 1 is composed of an upper test tube body 1-1 and a lower test tube body 1-2, the diameter of the upper test tube body 1-1 is larger than the diameter of the lower test tube body 1-2, the extension line of the lower test tube body is inscribed with the upper test tube body 1-1, a sealing cover 2 is sleeved at the port of the upper test tube body 1-1, and a heat preservation sleeve 3 is sleeved on the lower test tube body 1-2, the outer side of the heat preservation sleeve 3 is flush with the upper test tube body 1-1, and the heat preservation sleeve 3 has a longitudinal notch 4 at the longitudinal tangent of the upper test tube body 1-1 and the lower test tube body 1-2, and the longitudinal notch 4 with a spacing is conducive to observing the liquid level in the test tube from the outside. The special-shaped test tube body 1 is longitudinally provided with a scale corresponding to the longitudinal notch 4, and the special-shaped test tube body 1 can be made of glass or PE material.

[0030] Among them, the insulation sleeve 3 is filled with self-heating material or phase change material, the outer membrane of the moisturizing sleeve is made of breathable and water-impermeable material, and an inner lining temperature control membrane 5 is provided between the moisturizing sleeve and the lower test tube body 1-2. The temperature generated by the self-heating material can make the liquid in the tube reach 26°C, and the duration is >120min.

[0031] Among them, the upper test tube body 1-1 is 20mm, the lower test tube body 1-2 has a diameter of 7.5mm, the width of the longitudinal notch 4 is 2.5mm, the height is 52mm, and the total length D of the test tube body and the insulation sleeve 3 is 75mm.

[0032] The test tube is vacuumed and sealed with a rubber stopper, and the special-shaped test tube body 1 can accommodate more than 3 ml of blood sample.

[0033] The special-shaped test tube body 1 contains an anticoagulant solution, which is used in a fully automatic blood cell analyzer and contains 2.0 mg / ml EDTA-K 2 and 9.5umol / L P2Y 12 Receptor antagonist ticagrelor.

[0034] The automatic blood cell analyzer uses blood cell counting to analyze samples 60 minutes to 120 minutes after the anticoagulant and venous blood are mixed.

[0035] The samples for testing included 30ul of blood cell counting analysis anticoagulant and 2ml of venous blood.

[0036] The above-mentioned method for correcting the number of false platelets by blood cell counting and analyzing anticoagulant solution comprises the following steps:

[0037] Step 1: Add the blood cell counting and analysis anticoagulant solution into the vacuum blood collection tube, then pass 2 ml of venous blood into it, and then quickly invert it 5 times to mix well;

[0038] Step 2: Use a fully automatic blood cell analyzer to test the blood sample obtained in step 1 to obtain a platelet detection value, where the platelet number is the true value.

[0039] Example 2

[0040] like Figure 4-Figure 6 As shown, this embodiment provides a heat preservation vacuum sampling tube for detecting the number of platelets, including a special-shaped test tube body 1, the special-shaped test tube body 1 is composed of an upper test tube body 1-1 and a lower test tube body 1-2, the diameter of the upper test tube body 1-1 is larger than the diameter of the lower test tube body 1-2, the extension line of the lower test tube body is inscribed in the upper test tube body 1-1, a sealing cover 2 is sleeved at the port of the upper test tube body 1-1, the lower test tube body 1-2 is covered with a heat preservation sleeve 3, the outer side of the heat preservation sleeve 3 is flush with the upper test tube body 1-1, and the heat preservation sleeve 3 has a longitudinal notch 4 at the longitudinal tangent of the upper test tube body 1-1 and the lower test tube body 1-2, and the longitudinal notch 4 with a spacing is conducive to observing the liquid level in the test tube from the outside. The special-shaped test tube body 1 can be made of glass or PE material.

[0041] Among them, the insulation sleeve 3 is filled with self-heating material or phase change material, the outer membrane of the moisturizing sleeve is made of breathable and water-impermeable material, and an inner lining temperature control membrane 5 is provided between the moisturizing sleeve and the lower test tube body 1-2. The temperature generated by the self-heating material can make the liquid in the tube reach 26°C, and the duration is >120min.

[0042] Among them, the upper test tube body 1-1 is 20mm, the lower test tube body 1-2 has a diameter of 7mm, the width of the longitudinal notch 4 is 3mm, the height is 50mm, and the total length D of the test tube body and the insulation sleeve 3 is 80mm.

[0043] The test tube is vacuumed and sealed with a rubber stopper, and the special-shaped test tube body 1 can accommodate more than 3 ml of blood sample.

[0044] The special-shaped test tube body 1 contains an anticoagulant solution, which is used in a fully automatic blood cell analyzer and contains 1.5 mg / ml EDTA-K 2 and 19umol / L P2Y 12Receptor antagonist ticagrelor.

[0045] The automatic blood cell analyzer uses blood cell counting to analyze samples 60 minutes to 120 minutes after the anticoagulant and venous blood are mixed.

[0046] The samples for testing included 40ul of blood cell counting analysis anticoagulant and 2ml of venous blood.

[0047] The above-mentioned method for correcting the number of false platelets by blood cell counting and analyzing anticoagulant solution comprises the following steps:

[0048] Step 1: Add the blood cell counting and analysis anticoagulant solution into the vacuum blood collection tube, then pass 2 ml of venous blood into it, and then quickly invert 4-6 times to mix;

[0049] Step 2: Use a fully automatic blood cell analyzer to test the blood sample obtained in step 1 to obtain a platelet detection value, where the platelet number is the true value.

[0050] Example 3

[0051] like Figure 4-Figure 6 As shown, this embodiment provides a heat preservation vacuum sampling tube for detecting the number of platelets, including a special-shaped test tube body 1, the special-shaped test tube body 1 is composed of an upper test tube body 1-1 and a lower test tube body 1-2, the diameter of the upper test tube body 1-1 is larger than the diameter of the lower test tube body 1-2, the extension line of the lower test tube body is inscribed in the upper test tube body 1-1, a sealing cover 2 is sleeved at the port of the upper test tube body 1-1, the lower test tube body 1-2 is covered with a heat preservation sleeve 3, the outer side of the heat preservation sleeve 3 is flush with the upper test tube body 1-1, and the heat preservation sleeve 3 has a longitudinal notch 4 at the longitudinal tangent of the upper test tube body 1-1 and the lower test tube body 1-2, and the longitudinal notch 4 with a spacing is conducive to observing the liquid level in the test tube from the outside. The special-shaped test tube body 1 can be made of glass or PE material.

[0052] Among them, the insulation sleeve 3 is filled with self-heating material or phase change material, the outer membrane of the moisturizing sleeve is made of breathable and water-impermeable material, and an inner lining temperature control membrane 5 is provided between the moisturizing sleeve and the lower test tube body 1-2. The temperature generated by the self-heating material can make the liquid in the tube reach 26°C, and the duration is >120min.

[0053] Among them, the upper test tube body 1-1 is 20mm, the lower test tube body 1-2 has a diameter of 8mm, the width of the longitudinal notch 4 is 2mm, the height is 55mm, and the total length D of the test tube body and the insulation sleeve 3 is 80mm.

[0054] The test tube is vacuumed and sealed with a rubber stopper, and the special-shaped test tube body 1 can accommodate more than 3 ml of blood sample.

[0055] The special-shaped test tube body 1 contains an anticoagulant solution, which is used in a fully automatic blood cell analyzer and contains 2.0 mg / ml EDTA-K 2 and 9.5umol / L P2Y 12 Receptor antagonist ticagrelor and 10umol / L adenylate cyclase activator forskolin.

[0056] The automatic blood cell analyzer uses blood cell counting to analyze samples within 120 minutes after the anticoagulant and venous blood are mixed.

[0057] The samples for testing included 30ul of blood cell counting analysis anticoagulant and 2ml of venous blood.

[0058] The above-mentioned method for correcting the number of false platelets by blood cell counting and analyzing anticoagulant solution comprises the following steps:

[0059] Step 1: Add the blood cell counting and analysis anticoagulant solution into the vacuum blood collection tube, then pass 2 ml of venous blood into it, and then quickly invert it 5 times to mix well;

[0060] Step 2: Use a fully automatic blood cell analyzer to test the blood sample obtained in step 1 to obtain a platelet detection value, where the platelet number is the true value.

[0061] Comparative Example 1

[0062] The anticoagulant solution in this embodiment contains 2.0 mg / ml EDTA-K 2 The rest are the same as in Example 1.

[0063] This embodiment provides a method mainly using P2Y 12 The method of correcting the false platelet count with receptor antagonists comprises the following steps:

[0064] Step 1: Add 30ul of blood cell counting and analysis anticoagulant solution into a vacuum blood collection tube, then pass 2ml of venous blood, and then quickly invert 6 times to mix; the blood cell counting and analysis anticoagulant solution contains 2.0mg / ml EDTA-K 2 ;

[0065] Step 2: Use a fully automatic blood cell analyzer to test the blood sample obtained in step 1 to obtain a platelet detection value, where the platelet number is the true value.

[0066] Effect verification

[0067] 1. Instruments and reagents: Beijing Jiuchunjian JCJ500 intelligent temperature inspection instrument, Shenzhen Mindray BC-6900 blood cell analyzer (optical method + impedance method) and supporting reagents, Japan Olympus (Olymplus) BX43 binocular microscope and graphic system, VP-10L oil-free vacuum filtration pump (Wenling Shenba Air Compressor Manufacturing Co., Ltd.), Finnpipette pipette (10-100ul), purple head tube for blood cell analysis (containing 30ul 2.0mg / mLEDTA-K2), purchased from Shanshan Medical Supplies Co., Ltd., Zhengzhou City, Henan Province, negative pressure 3ml, made of glass. Platelet antagonist Ticagrelor API T822289 was purchased from Shanghai MacLean Biochemical Technology Co., Ltd., protein kinase A activator forskolin API S1612 was purchased from Shanghai Biyuntian Biotechnology Co., Ltd., and blood smear stain was purchased from Zhuhai Beso Company. The instrument is calibrated and compared according to the operating procedures before use, and daily quality control is carried out, and the results are under control.

[0068] 2. Whole blood mode detection: The blood cells of the samples were counted by the electrical impedance method (CD) and the fluorescent dye laser scattering method (CDR) of the Mindray BC-6900 blood cell analyzer in whole blood mode; the platelet and other blood cell values ​​were recorded at 10min, 60min, and 120min by time resolution. The three 30ul anticoagulant containing different types and concentrations of whole blood samples were numbered as T: (2.0mg / mL EDTA-K2), T1: (2.0mg / mLEDTA-K2+9.5umol / L ticagrelor), T1F: (2.0mg / mL EDTA-K2+9.5umol / L ticagrelor+10umol / L forskolin).

[0069] 3. Statistical analysis Statistical analysis was performed using Graph pad Prism 8 software. Measurement data were presented as mean ± standard deviation. One-way analysis of variance (ANOVA) was used for comparison among multiple groups, and paired T test was used for analysis between two groups. P < 0.05 was considered statistically significant.

[0070] result:

[0071] From November 2023 to March 2024, a total of 12 samples of pseudo-thrombocytopenia (including 1 case of satellite phenomenon) were collected. Because the instrument impedance method cannot fully and accurately count platelets in in vitro activated aggregation, platelet aggregates of varying sizes cause most T sample test results to gradually decrease at different time periods. The data are shown in Tables 1, 2, and 3.

[0072] Table 1. Platelet CD mode detection data of Mindray 6900 blood cell analyzer at different time periods

[0073]

[0074]

[0075] Table 2. Platelet CDR mode detection data of Mindray 6900 blood cell analyzer at different time periods

[0076] 10min 60min 120min T T1 T1F T T1 T1F T T1 T1F 1 189 188 189 208 214 188 218 220 181 2 261 243 263 174 150 259 188 170 232 3 341 345 326 254 369 350 131 353 351 4 333 333 284 298 311 336 279 283 330 5 258 242 250 261 245 246 229 243 246 6 145 163 204 148 141 194 165 151 203 7 218 167 207 192 129 211 150 130 202 8 107 122 113 116 115 113 118 121 115 9 17 18 17 15 16 17 16 18 17 10 227 222 226 212 220 224 207 234 227 11 162 170 137 153 158 121 142 146 157 12 114 112 111 110 112 112 108 111 116

[0077] Table 3 Statistical differences between platelet data of each group and group T (p value)

[0078] T T1 T1F 10min 0.5548 0.2594 CD Mode 60min 0.9724 0.0557 120min 0.545 0.0434 10min 0.4724 0.6117 CDR Mode 60min 0.7874 0.1267 120min 0.3327 0.0788

[0079] 1. The results of statistical analysis are shown in Table 3. Under the whole blood mode electrical impedance method (CD mode) of the BC-6900 blood cell analyzer, at 60 minutes of sampling, there was no statistical difference in the platelet data of the T1 and T1F groups with those of the T group (P>0.05); while there was a statistical difference between the 120-minute data of the T1F group and the T group (P<0.05), indicating that ticagrelor alone could not effectively inhibit or dissociate aggregated platelets, while ticagrelor combined with forskolin had a significant inhibitory effect on in vitro platelet aggregation, and both of them had a time- and concentration-dependent inhibition of in vitro platelet aggregation. Under the whole blood mode fluorescent staining laser scattering method (CDR mode) of the Mindray BC-6900 blood cell analyzer, whether the sampling was 60 minutes or 120 minutes, there was no statistical difference between the T1, T1F groups and the T group (P>0.05), indicating that the combined drug T1F group could inhibit or disaggregate platelets.

[0080] 2. Using the 10-minute data of CDR mode as the standard result, there was no statistical difference in the 10-minute data of platelets in the CD mode T and T1F groups (P>0.05), and there were statistical differences in the 60-minute and 120-minute data (P<0.05); there was no difference in the data of the T1F group in each time period of the CDR mode (P>0.05). This shows that the SF-Cube technology of the BC-6900 blood cell analyzer can assist in correcting some pseudo-thrombocytopenia, and the use of combined drugs has no drug effect on the platelet count. See Table 4.

[0081] Table 4 Statistical differences (p values) between the two counting modes of platelet T1F groups, based on the 10-min data of the CDR mode

[0082]

[0083]

[0084] 3. Using the 10-min data of CDR mode as the standard result, there was no statistical difference in the white blood cell and lymphocyte data of the CD mode T and T1F groups (P>0.05), see Table 5. Table 6.

[0085] Table 5 Statistical differences (p value) between the two counting modes of white blood cell data in the T1F group, based on the 10-minute data of the CDR mode

[0086] T T1F 10min 0.9725 CD Mode 60min 0.8780 120min 0.9739 10min 0.9803 CDR Mode 60min 0.9335 120min 0.9622

[0087] Table 6 Statistical differences (p value) between the two lymphocyte counting modes in the T1F group, based on the 10-min data of the CDR mode

[0088]

[0089]

[0090] 4 Results of artificial smears of the three samples. Artificial smears of the three samples T, T1, and T1F were observed at 60 minutes: There were fewer platelet aggregates in the head and body of the T sample smear, and the platelets were aggregated in clusters and arranged closely in the tail. There were fewer platelet aggregates in the head and body of the T1 sample smear, and the platelets were aggregated in clusters at the tail. Compared with the T sample, the platelet aggregates in various parts were slightly smaller in size, the platelet aggregates were looser, and the gaps were slightly larger than those of the T sample. No obvious platelet aggregation was observed in the head, body, and tail of the T1F sample smear. The platelets were in small clusters (ranging from 2 to 9) or scattered distribution with obvious gaps. In the T1F smear with satellite phenomenon, it can be seen that the aggregation of "satellite" platelets is significantly reduced, and the platelets tend to be scattered and independent. The results of low-power and oil-immersion smears are shown in Figure 1 , Figure 2 , satellite phenomena Figure 3 .

[0091] 5 Results of Beijing Jiuchunjian JCJ500 Intelligent Temperature Patrol Meter: The probe was inserted into a test tube containing 3 ml of physiological saline for 15 minutes. The temperature generated by 5 grams of "self-heating" material can make the liquid in the tube reach 26°C, and the duration is >120 minutes. See Table 7.

[0092] Table 7. Test results of intelligent temperature inspection instrument

[0093]

[0094]

[0095] discuss

[0096] After blood samples were anticoagulated with EDTA, Ca 2+Although chelated, platelet counts still show a "pseudo-reduction" phenomenon, proving that multiple activation and inhibition mechanisms affect the high reactivity and low threshold of platelets in vitro, as well as the complexity of the mechanism. In 2000, the International Committee for Standardization in Hematology and the International Society of Experimental Hematology established a reference method for platelet counting based on flow cytometry. The comparative use of the electrical impedance method and the fluorescence laser scattering method based on flow cytometry may be helpful in observing the effect of antagonists combined with activators against "pseudo-platelet aggregation" in vitro.

[0097] Platelet activation in the anticoagulant environment of EDTA makes the regulation of a series of signaling pathways more complicated. In the blood flow, a delicate balance between activation and inhibition mechanisms can prevent spontaneous platelet aggregation and vascular occlusion. 12 Receptor antagonists and cAMP-dependent inhibition are a coordinated effort involving multiple signaling pathways, regulated by at least 16 kinases and 7 phosphatases, and involve extensive crosstalk with activated signaling pathways.

[0098] Preliminary analysis based on statistical results:

[0099] 1. Although ticagrelor combined with forskolin inhibited most of the platelet activation in vitro, which was better than ticagrelor alone in inhibiting platelet activation, the concentration needed to be controlled and there was also forskolin dose dependence. There was no statistical difference between the 10-minute data of the T1F group in the CD mode and the 10-minute data in the CDR mode, which was the same as the results of the impedance dilution mode. However, in actual work, difficulties such as sample transportation, inability to load samples for testing in time after blood collection, and temperature influence were encountered, which made it impossible to correct "pseudo-thrombocytopenia" in time.

[0100] 2. In CDR mode, the data from samples 2 and 7 show that the application of ticagrelor combined with forskolin in platelet count is better than that of SF-Cube technology alone.

[0101] 3. In CDR mode, there was no statistical difference in the white blood cell and lymphocyte data between the T and T1F groups in CD mode (P>0.05). The combination of ticagrelor and forskolin was relatively accurate in counting white blood cells and lymphocytes, and the PLR ​​and NLR results can be used clinically.

[0102] 4. 5 grams of "self-heating" material can maintain the required time and temperature of the liquid in the test tube for 120 minutes, meeting the needs of clinical trials.

[0103] Although the embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and cannot be understood as limiting the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention without departing from the principles and purpose of the present invention. The protection scope of the present invention is defined by the claims and their equivalent technical solutions.

Claims

1. A heat-insulating vacuum sampling tube for detecting the number of platelets, characterized in that: The invention comprises a special-shaped test tube body, which is composed of an upper test tube body and a lower test tube body. The diameter of the upper test tube body is larger than that of the lower test tube body. The extension line of the lower test tube body is inscribed with the upper test tube body. A sealing cover is provided at the port of the upper test tube body. The outer surface of the insulation cover is flush with the upper test tube body. The insulation cover has a longitudinal notch at the longitudinal tangent of the upper test tube body and the lower test tube body.

2. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 1, characterized in that: The heat preservation sleeve contains a self-heating material or a phase change material, and the outer membrane of the moisture-retaining sleeve is made of an air-permeable and water-impermeable material.

3. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 2, characterized in that: An inner lining temperature control film is provided between the moisturizing sleeve and the lower test tube body.

4. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 1, characterized in that: The upper test tube body is 20 mm, the lower test tube body has a diameter of 7-8 mm, the width of the longitudinal notch is 2-3 mm, and the height is 50-55 mm.

5. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 1, characterized in that: The special-shaped test tube body can accommodate a blood sample larger than 3 ml.

6. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 1, characterized in that: The special-shaped test tube contains an anticoagulant, which is used in a fully automatic blood cell analyzer and contains 1.5-2.2 mg / ml EDTA-K2 and 9.5-19 umol / L P2Y 12 Receptor antagonist ticagrelor.

7. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 6, characterized in that: The automatic blood cell analyzer uses blood cell counting to analyze samples 60 minutes to 120 minutes after the anticoagulant solution and venous blood are mixed.

8. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 6, characterized in that: The invention also contains 10-20umol / L of an adenylate cyclase activator, wherein the adenylate cyclase activator is forskolin.

9. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 6, characterized in that: The automatic blood cell analyzer uses blood cell counting to analyze samples within 120 minutes after the anticoagulant solution and venous blood are mixed.

10. The heat-insulating vacuum sampling tube for detecting the number of platelets according to claim 7 or 9, characterized in that: The sample for testing contains 30-40ul of blood cell counting anticoagulant and 2ml of venous blood.

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