OBJECTIVE: To investigate the role of platelet-rich plasma (PRP) in inducing the M2 macrophage polarization via regulating AMPK singling pathway. METHODS: The expressions of M1 marker CD11c and M2 marker CD206 in macrophages of blank control group, LPS group, LPS+PRP group, and LPS+PRP+Compound C group were detected by flow cytometry. Western blot was used to observe the effects of PRP on the expression of AMPK-mTOR signaling pathway-related proteins at different times (12 h, 18 h and 24 h) after LPS treatment. RNA interference technology was used to silence the expression of AMPK in macrophages, and the expression of TGF-β protein was subsequently examined by Western blot. RESULTS: LPS significantly reduced the expression of CD206 and increased the expression of CD11c (P <0.05). After the addition of PRP, the expression of CD206 was significantly increased (P <0.05), while the expression of CD11c was significantly decreased (P <0.05). Compared with LPS group, PRP treatment significantly increased the expressions of p-AMPK and p-ULK1 proteins at 12 h, 18 h and 24 h, while significantly decreased the expression of p-mTOR protein (P <0.05). After the addition of AMPK inhibitor Compound C, the expression of CD206 was significantly reduced (P <0.05) and the expression of CD11c was significantly increased compared with LPS+PRP group (P <0.05). After silencing the expression of AMPK in macrophages, the promotion effect of PRP on TGF-β was significantly reduced (P <0.05). CONCLUSION PRP can stimulate the transformation of macrophages to M2 type via AMPK signalling pathway.
Humans ; AMP-Activated Protein Kinases/pharmacology* ; Lipopolysaccharides/pharmacology* ; Macrophages/metabolism* ; Transforming Growth Factor beta/metabolism* ; Platelet-Rich Plasma/metabolism*

【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 analyze the clinical features of an outbreak of extensive drug resistant typhoid fever, and to provide experience for the diagnosis and treatment of drug resistant typhoid fever.Methods:Seven patients with confirmed diagnosis of extensive drug resistant typhoid fever who visited Beijing You′an Hospital, Capital Medical University, from January 27 to February 15, 2022 were included. The clinical characteristics, drug sensitivity tests, consultation and treatment history and prognosis of the patients were analyzed through descriptive study.Results:Of the seven extensive drug resistant typhoid fever patients, three were male and four were female, one of whom was pregnant (at 32-week gestation), aged (29.8±6.8) years, with a range of 22 to 42 years. There were seven cases with fever, and the course of fever ranged from six to 20 days. There were five cases with diarrhea and lack of typhoid-specific manifestations such as rose spot, apathetic facial expression and relatively slow pulse. Four cases were complicated with intestinal bleeding and six cases developed liver function injury. Six cases had loss or decrease in eosinophil ratio and two cases had decreased white blood cell count. The results of drug susceptibility tests showed that seven strains of Salmonella typhi were resistant to chloramphenicol, ampicillin, sulfamethoxazole-trimethoprim, quinolones, ceftriaxone, cefepime, ceftazidime, cefuroxime, and sensitive to carbapenem antibiotics, tigecycline and piperacillin/tazobactam. All seven cases had a history of antimicrobial use before admission. One case was administered with intravenous ceftizoxime for seven days after admission. After discharge, cefixime was administered orally for seven days. Six patients were given intravenous piperacillin sodium/tazobactam sodium for 14 days. All blood/fecal cultures were negative and the patients were cured and discharged. During the follow-up, one patient developed splenic abscess. All the seven patients were residents of the same apartment in Beijing City, and there were water cuts and turbid odors in the incubation period, which were considered as typhoid fever outbreak caused by waterborne transmission. Conclusions:With the use of antimicrobial agents, the typical clinical manifestations of typhoid fever are absent, and the drug resistance rates to quinolone and third-generation cephalosporins increase. Appropriate antimicrobial agents should be selected and the anti-infection course should be prolonged.

ObjectiveTo examine the current status of preschoolers' eating behaviors and investigate its correlation with family cohesion and adaptability. MethodsA cross-sectional study was conducted involving 21，954 preschoolers and their families from Pingshan District， Shenzhen， between September 2021 and December 2021. A general demographic questionnaire， the Chinese version of Family Adaptability and Cohesion Evaluation Scale Ⅱ （FACESⅡ-CV） and Chinese Preschoolers’ Eating Behavior Questionnaire （CPEBQ） were used to collect the relevant information. Multiple linear regression was used to analyze the association of family cohesion and adaptability with eating behaviors of preschoolers. ResultsTypes of family cohesion and adaptability were significantly correlated with all the 7 dimensions of preschoolers' eating behaviors， including food fussiness （R²=0.252， F=114.457， P<0.001）， food responsiveness （R² = 0.111， F =24.973， P<0.001）， eating habit （R²= 0.304， F =139.658， P<0.001）， satiety responsiveness （R² = 0.259， F =105.332， P<0.001）， external eating （R² = 0.182， F =50.150， P<0.001）， emotional eating （R² = 0.234， F =91.084， P<0.001） and initiative eating （R² = 0.349， F =168.608， P<0.001）. After adjusting for confounding factors， our study showed that types of family cohesion and adaptability were independent predictors of preschoolers' eating behaviors （P<0.05）. ConclusionsTypes of family cohesion and adaptability have a significant predictive effect on the 7 dimensions of preschoolers' eating behaviors. Higher scores of family cohesion and adaptability imply stronger initiative eating ability and less poor dietary behaviors in preschoolers.

【Objective】 To discuss the effect of temperature on the quality of platelet-rich plasma (PRP) prepared manually. 【Methods】 A total of 120 peripheral blood samples (60 mL/ person) were collected from healthy voluntary blood donors in the Blood Center of General Hospital of Southern Theater Command from May 10, 2010 to September 10, 2010. Each whole blood sample (60mL/ person) was randomly divided into 3 aliquots(20 mL each), totaling 360 aliquots, then divided into 9 groups (A1, A2, A3, B1, B2, B3, C1, C2, and C3), with 40 aliquots in each group. In group A: the ambient temperature for PRP preparation was set to (4±2)℃, and the temperature for PRP preparation and centrifugal bin was (4±2)℃, (24±2)℃ and (30±2)℃ for group A1, A2, and A3, respectively. In group B: the ambient temperature for PRP preparation was set to (24±2)℃, and the temperature for PRP preparation and centrifugal bin was (4±2)℃, (24±2)℃ and (30±2)℃ for group B1, B2 and B3 respectively. In group C: the ambient temperature for PRP preparation was set to (30±2)℃, and the temperature for PRP preparation and centrifugal bin was (4±2)℃, (24±2)℃ and (30±2)℃ for group C1, C2 and C3 respectively. Platelet concentrates were separated and prepared by rich slurry method, and the platelet recovery rate was calculated for each group, the platelet morphology was observed under the microscope, and the concentrations of platelet-derived growth factor (PDGF-BB), transforming growth factor β (TGF-β) and vascular endothelial growth factor (VEGF) were quantitatively determined using enzyme-linked immunosorbent assay (ELISA). 【Results】 The platelet count and platelet recovery rate were compared as follows: among group A1, A2, A3, B1, B2, B3, C1, C2 and C3, the results of A3, B3 and C3were (489.2±21.47) × 10⁹/L vs (495.7±23.2) ×10⁹/L vs (489.4±17.1) ×10⁹/L and (57.4±2.3)% vs (54.9±1.3)% vs (50.7±2.3)%, respectively, all showed platelet aggregation, and the differences, relative to A1 and A2, B1 and B2, and C1 and C2, were statistically significant (P<0.05). Microscopic observation was as follows: in the PRPs collected from group A1, A2, B1, B2, C1 and C2, the platelet cells were uniform in size and without aggregation; in the PRPs collected from group A3, B3 and C3, some platelets showed aggregation. Determination of growth factor content was as follows: the content of TGF-β(ng/L) and PDGF-BB(ng/L) was 375.0±119.1 vs 183.67±106.2 and 933.0±273.0 vs 656±113.0 in the non-aggregation group and the aggregation group, respectively (P<0.05), while the content of VEGF(ng/L) was 217.5±93.5 vs 155.3±103.4 (P>0.05), with no statistically difference. 【Conclusion】 The ambient temperature had little effect on the preparation of PRP, and the temperature of the centrifuge bin was better to be maintained at (24±2)℃.

【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.