Umbilical cord mesenchymal stem cells (UC-MSCs) have been widely used in regenerative medicine, but there is limited research on the stability of UC-MSCs formulation during production. This study aims to assess the stability of the cell stock solution and intermediate product throughout the production process, as well as the final product following reconstitution, in order to offer guidance for the manufacturing process and serve as a reference for formulation reconstitution methods. Three batches of cell formulation were produced and stored under low temperature (2-8 ℃) and room temperature (20-26 ℃) during cell stock solution and intermediate product stages. The storage time intervals for cell stock solution were 0, 2, 4, and 6 h, while for intermediate products, the intervals were 0, 1, 2, and 3 h. The evaluation items included visual inspection, viable cell concentration, cell viability, cell surface markers, lymphocyte proliferation inhibition rate, and sterility. Additionally, dilution and culture stability studies were performed after reconstitution of the cell product. The reconstitution diluents included 0.9% sodium chloride injection, 0.9% sodium chloride injection + 1% human serum albumin, and 0.9% sodium chloride injection + 2% human serum albumin, with dilution ratios of 10-fold and 40-fold. The storage time intervals after dilution were 0, 1, 2, 3, and 4 h. The reconstitution culture media included DMEM medium, DMEM + 2% platelet lysate, 0.9% sodium chloride injection, and 0.9% sodium chloride injection + 1% human serum albumin, and the culture duration was 24 h. The evaluation items were viable cell concentration and cell viability. The results showed that the cell stock solution remained stable for up to 6 h under both low temperature (2-8 ℃) and room temperature (20-26 ℃) conditions, while the intermediate product remained stable for up to 3 h under the same conditions. After formulation reconstitution, using sodium chloride injection diluted with 1% or 2% human serum albumin maintained a viability of over 80% within 4 h. It was observed that different dilution factors had an impact on cell viability. After formulation reconstitution, cultivation in medium with 2% platelet lysate resulted in a cell viability of over 80% after 24 h. In conclusion, the stability of cell stock solution within 6 h and intermediate product within 3 h meets the requirements. The addition of 1% or 2% human serum albumin in the reconstitution diluent can better protect the post-reconstitution cell viability.

Objective To explore the correlation between sarcopenia and body fat percentage(BF%)in elderly male patients with type 2 diabetes mellitus(T2DM).Methods A total of 291 elderly male T2DM patients who were hospitalized in the Endocrinology Department of Xuanwu Hospital,Capital Medical University were enrolled in this study from December 2018 to September 2019.All the patients were divided into sarcopenia group(n=35)and non sarcopenia group(n=256).BF%,skeletal muscle mass index(SMI),grip strength,gait speed,HbA1c,hemoglobin(Hb),blood uric acid(SUA),albumin and vitamin D(Vit D)were measured in all the participants.Results Compared with the non sarcopenia group,age,HbA1c and BF%were increased,while BMI,Vit D,SMI,Hb,SUA,ALB,grip strength,and gait speedwere decreased in sarcopenia group(P＜0.05 or P＜0.01).Pearson correlation analysis showed that SMI was positively correlated with BMI,Vit D,Hb,and SUA(P＜0.05 or P＜0.01),and negatively correlated with HbA1c(P＜0.01).Grip strength and gait speed are positively correlated with Vit D and Hb(P＜0.01),and negatively correlated with BF%(P＜0.01).Logistic regression analysis showed that age and BF%were risk factors for sarcopenia,while BMI and Vit D were protective factors for sarcopenia.Conclusions High BF%significantly increases the risk of sarcopenia in elderly male T2DM patients.

Clostridioides difficile/genetics* ; Clostridioides ; Real-Time Polymerase Chain Reaction ; Feces ; Bacterial Proteins/genetics*

From 2015 to 2019, the annual average incidence rate of scarlet fever was 7.80/100 000 in Yantai City, which showed an increasing trend since 2017 (χ²_trend=233.59, P<0.001). The peak period of this disease was from April to July and November to January of the next year. The ratio of male to female was 1.49∶1, with a higher prevalence among cases aged 3 to 9 years (2 357/2 552, 92.36%). Children in kindergartens, primary and middle school students, and scattered children were the high risk population, with the incidence rate of 159.86/100 000, 25.57/100 000 and 26.77/100 000, respectively. The global spatial auto-correlation analysis showed that the global Moran's I index of the reported incidence rate of scarlet fever in Yantai from 2015 to 2019 was 0.28, 0.29, 0.44, 0.48, and 0.22, respectively (all P values<0.05), suggesting that the incidence rate of scarlet fever in Yantai from 2015 to 2019 was spatial clustering. The local spatial auto-correlation analysis showed that the "high-high" clustering areas were mainly located in Laizhou City, Zhifu District, Haiyang City, Fushan District and Kaifa District, while the "low-high" clustering areas were mainly located in Haiyang City and Fushan District.
Child ; Humans ; Male ; Female ; Scarlet Fever/epidemiology* ; Spatial Analysis ; Cities/epidemiology* ; Seasons ; Risk Factors ; Incidence ; Cluster Analysis ; China/epidemiology*

The present study investigated the mechanism of total flavonoids from Ampelopsis grossedentata(AGTF) against gouty arthritis(GA) by network pharmacology and experimental validation. The main active ingredients and targets of AGTF, as well as disease targets, were screened out using relevant databases and literature data. The "protein-protein interaction"(PPI) network and "drug-ingredient-target-pathway" network were constructed, and the potential targets and mechanism of AGTF against GA were predicted. The hyperuricemia(HUA) combined with GA model was induced in rats. The gait behaviors of rats were scored, and ankle swelling degree was observed. The uric acid(UA) level and xanthine oxidase(XOD) activity in the rat serum were detected, and the levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) were measured. The protein expression of toll-like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), and nuclear factor-kappa B(NF-κB) in the synovial tissues of the rat ankle joint was determined by immunohistochemistry. Ten active ingredients of AGTF and 73 candidate targets of AGTF against GA were screened out by network pharmacology. Eighty-six signaling pathways were enriched, including TNF signaling pathway, NF-κB signaling pathway, TLR signaling pathway, Nod-like receptor signaling pathway, and purine metabolism signaling pathway, which were closely related to AGTF against GA. Animal experimental results showed that AGTF could effectively improve the abnormal gait behaviors of GA rats, relieve ankle inflammation, and reduce ankle joint swelling. In addition, AGTF could significantly reduce UA level, inhibit XOD activity, decrease TNF-α, IL-6, and IL-1β content, and down-regulate the expression of TLR4, MyD88, and NF-κB in ankle synovial tissues(P<0.05, P<0.01). The results of network pharmacology and experimental validation are consistent, indicating that AGTF exerts its therapeutic effect on GA by regulating UA metabolism, improving abnormal UA level, reducing the release of inflammatory factors, and regulating immunity and the TLR4/MyD88/NF-κB inflammatory pathway.
Ampelopsis/chemistry* ; Animals ; Arthritis, Gouty/drug therapy* ; Flavonoids/therapeutic use* ; Interleukin-1beta/metabolism* ; Interleukin-6/metabolism* ; Myeloid Differentiation Factor 88/metabolism* ; NF-kappa B/metabolism* ; NLR Proteins/metabolism* ; Rats ; Toll-Like Receptor 4/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Uric Acid ; Xanthine Oxidase

Background and Objectives: The goal of this study was to investigate the mechanism of mesenchymal stem cell (MSC)-derived microRNA (miR)-150-5p-expressing exosomes in promoting skin wound healing through activating PI3K/AKT pathway by PTEN. Methods: and Results: Human umbilical cord (HUC)-MSCs were infected with miR-150-5p overexpression and its con-trol lentivirus, and HUC-MSCs-derived exosomes (MSCs-Exos) with stable expression of miR-150-5p were obtained. HaCaT cells were induced by H2O2 to establish a cellular model of skin injury, in which the expression of miR-150-5p and PTEN and the phosphorylation of PI3K and AKT were evaluated. HaCaT cells were transfected with pcDNA3.1-PTEN or pcDNA3.1 and then cultured with normal exosomes or exosomes stably expressing miR-150-5p. Cell proliferation was inspected by CCK-8. Cell migration was detected by scratch test and cell apoptosis by flow cytometry. The starBase tool was used to predict the binding site of miR-150-5p to PTEN. Dual-luciferase reporter assay and RIP assay were applied to assess the interaction between miR-150-5p and PTEN. In H2O2 -induced HaCaT cells, the miR-150-5p expression decreased, and PTEN expression increased in a concentration-dependent manner. MSCs-Exos promoted the growth and migration of H2O2 -induced HaCaT cells and inhibited their apoptosis. In addition, overexpression of exosomal miR-150-5p enhanced the protective effect of MSCs-Exos on H2O2 -induced HaCaT cells; PTEN overexpression in HaCaT cells partially restrained miR-150-5p-mediated inhibition on H2O2 -induced injury in HaCaT cells. PTEN was a target gene of miR-150-5p. MiR-150-5p regulated PI3K/AKT pathway through PTEN. Conclusions MSCs-derived miR-150-5p-expressing exosomes promote skin wound healing by activating PI3K/AKTpathway through PTEN.

Objective:To analyze the material basis, target and pathway of Chebulae fructus and Margarita in Mongolian medicine Garidi-13 Pill on stroke with the method of network pharmacology and molecular docking technology, so as to better understand its "detoxification" mechanism. Methods:TCMIP and BATMAN-TCM were used to predict the target of Chebulae fructus and Margarita composition, and GeneCards were used to search for the target of stroke. The overlapping targets of the two platforms were imported into the Metascape software for GO biological analysis and KEGG pathway analysis. The molecular docking of key molecules and targets was carried out with LEDOCK. Results:A total of 22 active components were collected and 217 targets related to stroke were predicted. Among them, 1-O-galloyl-glucose, cuprum, ellagicacid, arjungenin and corilagin were the key substances playing the role of "detoxification" of the Chebulae fructus and Margarita; IL6, TNF, HSP90AA1, PTGS2, CASP3, NR1I2, VKORC1 and ATP1A1 were the key targets playing the role of "detoxification" . These targets were significantly enriched in cell response, humoral level regulation, hemostasis, response to steroid hormones, steroid metabolism, coagulation and other biological processes, as well as nitrogen metabolism, IL-17 signaling pathway and other pathways. Molecular docking verified the accuracy of previous prediction results from computer simulation level. Conclusion:The process of Chebulae fructus and Margarita intervening strok is closely related to the elimination of harmful metabolites and calmingthe inflammatory reaction, which was not only consistent with the modern medicine on the pathological process of stroke, but also consistent with the interpretation of "evil and poison" with Mongolian medicine theory.

Objective:To investigate the application effect of reverse abdominal wall advancement flap in repairing chest wall soft tissue defect after breast tumor resection.Methods:From October 2020 to April 2021, the Department of Plastic Surgery and Breast Surgery of Hainan Cancer Hospital cooperated to repair the chest wall wounds of 4 female patients with unilateral giant breast tumors after primary lesion resection. Patients aged 40-63 years old, with an average of 51.5 years old. The size of the tumor estimated by physical examination was 7 cm × 6 cm-15 cm × 20 cm. The flaps were closely monitored after surgery, and complications were recorded. Local recurrence was followed-up.Results:The wound size of 4 patients after mastectomy was 16 cm×14 cm-20 cm×18 cm. Abdomen separation range reached anterior axillary line laterally and contralateral clavicle midline medially. Inferiorly, 1 case reached umbilical level, 1 case reached 2 cm below the umbilicus, and 2 cases reached 3 cm below the umbilicus. Three cases were diagnosed as breast lobulated tumors, and 1 case invasive lobular carcinoma. Among the 4 cases, 3 flaps survived completely and healed by first intention, while another flap healed under blister scab. There was no necrosis, infection, hematoma, seroma, or vascular crisis of the flap. All patients were satisfied. Three patients received radiotherapy and one received radiotherapy combined with oral chemotherapy. All patients were followed up for 3-6 months. No local recurrence was discovered. The abdomen was tighter than before operation, but no stiffness was found. There was no abdominal pain, abdominal wall bulging or abdominal hernia.Conclusions:Reverse abdominal wall advancedment flap was used to repair soft tissue defect of the chest wall after breast tumor resection. The operation was simple and fast, with no need for additional donor site incision. Quick recovery allowed shorter interval between surgery and next scheduled treatments.

Objective:To investigate the application effect of reverse abdominal wall advancement flap in repairing chest wall soft tissue defect after breast tumor resection.Methods:From October 2020 to April 2021, the Department of Plastic Surgery and Breast Surgery of Hainan Cancer Hospital cooperated to repair the chest wall wounds of 4 female patients with unilateral giant breast tumors after primary lesion resection. Patients aged 40-63 years old, with an average of 51.5 years old. The size of the tumor estimated by physical examination was 7 cm × 6 cm-15 cm × 20 cm. The flaps were closely monitored after surgery, and complications were recorded. Local recurrence was followed-up.Results:The wound size of 4 patients after mastectomy was 16 cm×14 cm-20 cm×18 cm. Abdomen separation range reached anterior axillary line laterally and contralateral clavicle midline medially. Inferiorly, 1 case reached umbilical level, 1 case reached 2 cm below the umbilicus, and 2 cases reached 3 cm below the umbilicus. Three cases were diagnosed as breast lobulated tumors, and 1 case invasive lobular carcinoma. Among the 4 cases, 3 flaps survived completely and healed by first intention, while another flap healed under blister scab. There was no necrosis, infection, hematoma, seroma, or vascular crisis of the flap. All patients were satisfied. Three patients received radiotherapy and one received radiotherapy combined with oral chemotherapy. All patients were followed up for 3-6 months. No local recurrence was discovered. The abdomen was tighter than before operation, but no stiffness was found. There was no abdominal pain, abdominal wall bulging or abdominal hernia.Conclusions:Reverse abdominal wall advancedment flap was used to repair soft tissue defect of the chest wall after breast tumor resection. The operation was simple and fast, with no need for additional donor site incision. Quick recovery allowed shorter interval between surgery and next scheduled treatments.