【Objective】 To investigate the effects of different anticoagulants on platelet-rich plasma(PRP) release content of growth factor and injection pain. 【Methods】 A total of 15 voluntary blood donors were selected, with each blood donor using four kinds of anticoagulant tubes with EDTA-K2 anticoagulation, EDTA-NA2 anticoagulation, citrate anticoagulation, ACD-A anticoagulation respectively as group A, B, C and D. PRP was isolated and prepared by the rich plasma method, and the contents of PDGF-AA, TGF-β, IGF-1, VEGF, and PF-4 were detected by enzyme-linked immunosorbent assay. Meanwhile, SD rats (20, 4 / group) were injected subcutaneously or intradermally with the supernatant of PRP and PG gel prepared in the 4 groups and normal saline in the control group. The pain status of SD rats during the injection was observed and recorded. The pain status of the 5 groups of experimental animals was evaluated according to the American Laboratory Animal Pain Guide. 【Results】 The platelet counts in PRP in group D was the highest [(1 294.53±277.37) × 10⁹/L], which was significantly higher than that in group A [ (789.13±377.13) ×10⁹/L] and group C [ (990.94±493.12) ×10⁹/L] (P<0.05). The OD value of PDGF-AA in group A, B, C, and D were 1.51± 0.18, 1.69±0.21, 0.66±0.19and 1.72±0.13, respectively, with statistically significant difference between groups (P<0.05 ) and group D better than the other three groups. The OD value of PF-4 was 1.18±0.24, 1.61±0.14, 0.65±0.26 and 1.72±0.10 respectively, with statistically significant difference between groups (P<0.05) and group D better than other three groups. The OD value of IGF-1 was 1.02±0.08, 0.98±0.11, 1.06±0.11 and 1.32±0.65 respectively, with no significant difference between groups (P>0.05). The OD value of VEGF was 0.13±0.04, 0.21±0.14, 0.08±0.02 and 0.13±0.04 respectively, with statistically significant difference between group B and C (P<0.05). The OD value of TGF-β was 0.14±0.01, 0.15±0.01, 0.28±0.17 and 1.10±0.37 respectively, with statistically significant difference between groups (P<0.05) and group D better than other three groups. Comparison of injection pain: when the supernatant of PRP and PG gel was injected, there were significant differences between group A, B, C and D, and the control group (P<0.05) . The median pain scores of PRP injection of group A, B, C, and D were 6 (1.5), 5 (0.75), 4.5 (2.5), and 3(3) respectively, with group D lower than other three groups, and no statistically significant difference was noticed (P>0.05) . The median pain scores of the PG supernatant injection of group A, B, C, and D were 4 (2.25), 3 (2.75), 4 (3), 1 (1.5), and the difference was not statistically significant (P>0.05). There was no significant difference between the PRP injection group and the PG supernatant group (P> 0.05). 【Conclusion】 PRP prepared by two-step centrifugation with ACD-A anticoagulant can obtain the higher platelet counts and the maximum release of PDGF-AA, PF-4, IGF-1, and TGF-β. In terms of pain, ACD-A anticoagulant injection has the lowest pain with the animals.

【Objective】 To develop a spray-on membrane dressing for wound repair containing platelet rich plasma (PRP) sodium alginate (SA)/agarose(AG)/carboxymethyl chitosan (CMCS). 【Methods】 SA/AG/ CMCS were mixed in different proportions to prepare biodegradable quick setting spray (BQSS) by blending film method, and the film-forming time, moisture retention and compression resistance of the prepared BQSS were tested. Then PRP and BQSS were mixed in the proportion of 3∶7, 4∶6, 5∶5, 6∶4 and 7∶3 to prepare PRP-BQSS spray film dressings. The film-forming time, moisture retention, compressive strength, porosity and slow-release effect of growth factors of PRP-BQSS spray film dressings were studied. 【Results】 In the preparation of BQSS compound spray film solution, when SA, AG, CMCS and sterile distilled water were 0.6∶0.6∶0.6∶98.2g, the film-forming time (7.73±0.31) s, moisture retention (75. 54±3.03) % and compression resistance (791.00±68.02) g of the spray-film dressing were the best. The basic properties of PRP-BQSS spray-on film dressings and the release of growth factors show that PRP-BQSS spray-on film dressings can exist in different forms, and with the decrease of PRP concentration percentage, its film-forming time, moisturizing performance and compressive strength showed an upward trend. When the PRP content is 30%, the porosity of the dressing is the highest, about(84.34±0.90)%. The release of platelet-derived growth factor-AA(PDGF-AA), platelet factor-4(PF-4) and transforming growth factor beta (TGF-β) was in a slow upward trend, and the release of the three growth factors was higher than that of PRP group in 48 hours. 【Conclusion】 The preparation method of PRP-BQSS spray film dressing designed in this study is simple and mild, and can form a film quickly, with good biological properties and better growth factor inhibition and sustained-release effect.

Objective:To investigate the psychological resilience level of Chinese children and adolescents with type 1 diabetes mellitus (T1DM) and to explore its influence on glucose sugar control.Methods:Using the convenient sampling method, a total of 200 children and adolescents with T1DM in the First Affiliated Hospital of Nanjing Medical University and Nanjing Children's Hospital were selected as research obejcts from January 2019 to January 2021. The general information questionnaire, Diabetes Strengths and Resilience Measure for Adolescents (DSTAR-Teen) and Diabetes Behavior Rating Scale (DBRS) were used to investigate the level of psychological resilience and self-management behavior. And the glycated hemoglobin (HbA1c) value was collected, and analyzed the effect of mental toughness level on it.Results:A total of 200 questionnaires were distributed in this survey, and 196 valid questionnaires were recovered. The total score of DSTAR-Teen for 196 children and adolescents with T1DM was (43.60±8.55) and the total score of DBRS was (0.64±0.16) , the HbA1c value was 7.2% (6.5%, 8.3%) . The results of multiple linear regression analysis showed that the level of mental toughness was an influencing factor of HbA1c level in children and adolescents with T1DM ( P<0.05) . Conclusions:In this study, children and adolescents with T1DM have a higher level of psychological resilience, which may affect their blood glucose control. In the future, interventions that increase the level of mental resilience are suggested to be implemented to promote the improvement of health outcomes of patients.

【Objective】 To analyze the influencing factors of clinical efficacy of autologous platelet-rich plasma in the treatment of androgenetic alopecia. 【Methods】 From January, 2019 to December, 2021, 151 outpatients with androgenetic alopecia were treated in our hospital. All patients were treated with autologous PRP injection once a month and planned to received injection for 4-6 occasions. According to the hair growth before and after treatment, the treatment results were evaluated. The influencing factors in the treatment results were grouped and analyzed, including the treatment numbers (divided into 1-6 occasions), the grade of alopecia(three grades: mild, moderate and severe alopecia) and the Plt concentration in PRP[five grades(×10⁹/L): extremely low concentration (<800), low concentration (801-1 000) and medium concentration (1 001-1 200), high concentration(1 201-1 500)and extremely high concentration(>1 501)]. 【Results】 The correlation coefficient between the number of treatments and the effective rate was pearson=0.986, P<0.001, showing a positive correlation. There was a statistical difference in the total number of treatments between groups (P<0.01). As to the severity of alopecia: The effective rates of mild and moderate alopecia patients (66.67%, 56.95%) were higher than those of severe alopecia patients (35%). The difference between groups was statistically significant (P<0.01). As to the concentration of Plt in PRP: The concentration of Plt directly affected the therapeutic effect. When the Plt count was (1 200-1 500)×10⁹/L, the effect was optimal, reaching 68.12%, which was significantly higher than the other four groups(P<0.01). The treatment numbers, alopecia grade and Plt concentration in PRP were all related to the effectiveness of treatment (P<0.05). When the OR value of treatment numbers and Plt concentration was >1, the effective rate increased by 2.619 times and 2.033 times, respectively, as treatment numbers and Plt concentration increased. When the OR value of the grade of alopecia was 0.338 (0<0.338<1), the effective rate decreased with the increase of alopecia severity; For each grade increased, the effective rate would be compromised by 0.338. 【Conclusion】 When PRP is applied to treat AGA patients, the clinical therapeutic effect will be better in patients who received more treatments, PRP with higher Plt count, and be in the early degree of hair loss. Whether the higher the concentration of Plt in PRP, the better the therapeutic effect will be, remains to be further verified.

【Objective】 To discuss the repair effect of lyophilized platelet lysate (PL) products on articular cartilage injury model of rats. 【Methods】 A total of 25 SD rats were injected with typeⅡcollagenase at the right knuckle articular cavity respectively on day 1, 3 and 5 of experiment, and the modeling conditions were observed 14 days after the last injection of collagenase. The SD rats with successful modeling were randomly divided into three groups, and were injected with lyophilized PL [Group A, 1 mL/(mouse·time)], PL [Group B, 1 mL/(mouse·time)], and normal saline[Group C, 1 mL/(mouse·time)]. The above three substances were injected with corresponding drugs on day 0, 7, 14 and 21 of experiment based on the grouping conditions, and the changes of knee joint diameters of the rats from the three groups were observed and compared. On day 14 and 28, one rat in each group was randomly killed and two knuckle articular cavities of each were taken for tissue sampling, using hematoxylin-eosin staining (HE) and immunohistochemistry. 【Results】 After 14 days of modeling by injection of type Ⅱ collagenase, the proportion of successful modeling in rats was 84% (21/25), with the knee joint diameter (mm) before and after modeling at 12.84±1.14 vs 14.11±1.17(P<0.01). On day 14, 21 and 28, groups A and B were superior to group C in the knee joint diameter and activity improvement (P<0.05), with 13.33±1.16 vs 13.37±1.08 vs 14.21±1.08, 13.10±1.09 vs 13.01±1.04 vs 14.09±1.09 and 12.38±1.08 vs 12.51±1.03 vs 14.01±1.07, respectively. Histological observation showed that group A and B were superior to group C in the production and arrangement of chondrocytes and the positive expression of type Ⅱ collagen, and there was no significant difference between group A and group B. 【Conclusion】 The lyophilized PL has similar therapeutic effect to PL in the treatment and repair of articular cartilage injury, and is worthy of clinical application.

【Objective】 To explore the clinical effects of autologous platelet-rich plasma (PRP) in the treatment of postoperative bone nonunion for long bone shaft fractures during operation and after operation. 【Methods】 A total of 34 patients with postoperative bone nonunion for long bone shaft fracture, who were admitted and cured in the Orthopedics Department of our hospital from January 2019 to June 2020, were selected. They were randomly divided into the experimental group(n=17) and control group(n=17). Individuals in the control were treated with autologous iliac bone alone., while the experimental group were treated with autologous iliac bone graft plus autologous PRP during and after operation. After surgery, the autogenous PRP was accurately injected with ultrasound guidance every 7 days (5 mL/person, 4 occasions in total). The clinical healing time of the fracture, the grading of callus formation and the functional rehabilitation level of limbs on the affected side at different time were observed in both groups. 【Results】 All 34 patients were followed up. In the experimental group and the control group, the clinical healing time (month) of the fracture was (5.03±1.24) vs (6.91±1.41), P<0.05. The healing rate of the fracture for 6 months and 9 months was 94%(16/17)vs 59%(10/17)and 94%(16/17)vs 82%(14/17), respectively (P<0.05). The grading of callus formation within 3, 6 and 9 months was (2.11±0.69) vs (1.53±0.80), (3.06±0.90) vs (2.59±0.87) and (3.82±0.73) vs (3.35±0.86), respectively (P<0.05). The acceptance rate of functional rehabilitation of limbs on the affected side was 82.35%(14/17)vs 76.47%(13/17), P<0.05. 【Conclusion】 The application of autologous PRP in the treatment of postoperative bone nonunion for long bone shaft fractures during operation and after operation can achieve good clinical treatment.

【Objective】 To study the therapeutic effect of autologous platelet-rich plasma(PRP) on joint injury. 【Methods】 Selected patients with joint injury treated in the Department of Transfusion Medicine of General Hospital of Southern Theatre Command of PLA from 2019 to 2020 were enrolled as the research objects, including 5 patients with shoulder joint injury, 34 patients with knee joint injury and 9 patients with ankle joint injury. All patients were treated with PRP injection at the injury site. The functional score and VAS score before and after treatment were compared. 【Results】 After 6 months of treatment, the CMS score and VAS of 5 patients with shoulder joint injury after treatment were (83.00±5.39) and (1.60±0.40), better than those before treatment (60.00±7.58)and (4.20±0.49)(P<0.05); The Lysholm knee score and VAS of 34 patients with knee joint injury after treatment were (80.73±2.43) and (2.07±0.24), better than those before treatment(50.30±2.96) and (4.28±0.33) (P<0.05); The AOFAS Ankle Hindfoot Scale and VAS of 9 patients with ankle joint injury after treatment were (68.44±4.59) and (2.56±0.53), better than those before treatment (42.67±4.57) and (4.89±0.63) (P<0.05). 【Conclusion】 For common joint injury sites, the clinical effect of using PRP injection is significant, which can effectively relieve pain and improve motor function, which is worthy of clinical application.

Objective: To develop a method of quantitative analysis of multi-components by single marker (QAMS) for six active ingredients in Gualoupi injection. Methods: An HPLC method was used with a Waters Sunfire ODS C18column (250 mm×4. 6mm, 2. 5 μm), the mobile phase was methanol(A)-0. 2% glacial acetic acid solution(B) with gradient elution, the flow rate was 1. 0 ml· min-1,the detection wavelength was 254 nm,the column temperature was 30℃ and the injection volume was 20 μl. Using 3, 29-dibenzoyl rarounitriol as a reference, the relative correction factors among karounidiol, vanillic acid, adenosine, quercitrin and cynaro-side were detected by QAMS and their contents were calculated, and the results were compared with those of the external standard method. Results: The differences were not statistically significant between the calculated values of karounidiol, vanillic acid, adeno-sine, quercitrin and cynaroside and those measured by the external standard method (P>0. 05). Conclusion: QAMS can be used for the determination of 6 effective components in Gualoupi injection, and the result is accurate, simple and effective.

OBJECTIVE:To investigate the correlation of blood concentrations of tamoxifen (TAM) and its metabolites with endometrial hyperplasia in estrogen receptor(ER)-positive breast cancer patients. METHODS:A total of 69 patients with ER-posi-tive breast cancer selected from our hospital during Mar. 2015-Apr. 2016 received TAM (twice a day,one tablet each time) for more than 6 months. According to endometrial thickness,they were divided into abnormal hyperplasia group(40 cases)and normal group(29 cases). The steady state concentrations of TAM and its metabolites [4-OH-tamoxifen(OHT),N-demethylation tamoxifen (DMT),Endoxifen] were determined by HPLC-FLU. The correlation of blood concentration and other factors with endometrial thickness were investigated by Pearson test and multiple regression analysis. RESULTS:The medication time and Endoxifen steady state concentration in abnormal hyperplasia group were both significantly longer or higher than normal group,with statistical signifi-cance (P<0.05). There was no statistical significance in age,BMI,complication and steady state concentrations of TAM,OHT and DMT between 2 groups(P>0.05). The endometrial thickness was positively correlated with Endoxifen steady state concentra-tion and medication time(r=0.447,0.460,P<0.05). Using endometrial thickness(y)as dependent variable,medication time(x1) and Endoxifen steady state concentration(x2)as independent variable,multiple regression analysis was conducted. Multiple regres-sion equation was calculated as follows:y=2.436+0.123x1+0.082x2(F=12.610,r=0.526,P<0.05). CONCLUSIONS:Medication time and Endoxifen steady state concentration may be related to endometrial hyperplasia,which can provide reference for predicting TAM induced endometrial abnormal hyperplasia in ER-positive breast cancer patients.

OBJECTIVE:To discuss the correlation between Interntion Normalized Ratio(INR)with concentration of warfarin enantiomer and dose density after cardiac valve replacement. METHODS:The plasma concentration of R-warfarin or S-warfarin, dose of unit weight and INR of 176 patients with anticoagulation after cardiac valve replacement in each time point were moni-tored,and the correlation of INR and warfarin plasma concentration or dose was observed. RESULTS:Doses of all patients in 24-288 h were significantly higher than 0 h,the differences were statistically significant(P<0.001),the dose was fluctuated within a certain range after 96 h,and compared with 96 h,there were no significant differences(P>0.05). Average plasma concentration of R-warfarin or S-warfarin had consistent trend,plasma concentration of enantiomers was fluctuated within a certain range after 108 h,there were no significant differences(P>0.05). Compared with 0 h,there was no significant difference when INR was 12 h (P>0.05),it increased significantly in 36 h and had been increasing,then fluctuated within a certain range after 108 h,compared with 108 h,there were no significant differences(P>0.05). The correlation coefficient between INR and unit weight dose of warfa-rin in 60 h>36 h>12 h,it showed correlation(P<0.001). There was certain correlation between plasma concentration of warfarin and dose density in 12,36 and 60 h. CONCLUSIONS:The correlation between plasma concentration of warfarin and unit weight dose is stronger than the correlation between INR and unit weight dose. When combined with other coagulation,it may help to man-age warfarin dose.