Objective:To investigate effects of human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSCs-Exo)on bisphenol AF(BPAF)-induced oxidative damage and inflammatory factor release from endometrial stromal cells(hESCs).Methods:hESCs were divided into Control group,BPAF group(25 μmol/L BPAF treatment),BPAF+Exo group(25 μmol/L BPAF+hUCMSCs-Exo treatment),BPAF+Exo+LY group(25 μmol/L BPAF+hUCMSCs-Exo+10 μmol/L LY294002 treatment).Cell prolifera-tion was detected by MTT assay;apoptosis,intracellular ROS level,and mitochondrial membrane potential level were detected by flow cytometry;protein expressions of Bcl-2,Bax,Cleaved-caspase-3 and PI3K/AKT signaling pathway were detected by Western blot;mRNA expressions of inflammatory factors TNF-α,IL-6 and IL-1β were detected by RT-qPCR.Results:Compared with Control group,hESCs survival rate was gradually decreased(P<0.01),apoptosis rate was gradually increased with the increased concentration of BPAF(≥25 μmol/L).Compared with Control group,BPAF group showed increased ROS level,decreased mitochondrial membrane potential level,increased Bax and Cleaved-caspase-3 protein expressions,and decreased Bcl-2,p-PI3K and p-AKT protein expressions.Compared with BPAF group,cell survival rate of BPAF+Exo group was increased(P<0.01),ROS level decreased,mitochondrial membrane potential level increased,expressions of Bax and Cleaved-caspase-3 proteins decreased,and expressions of Bcl-2,p-PI3K and p-AKT increased.Compared with BPAF+Exo group,expressions of Bax and Cleaved-caspase-3 protein in cells of BPAF+Exo+LY group were increased,while expressions of Bcl-2,p-PI3K and p-AKT protein were decreased.Expressions of inflammatory factors TNF-α,IL-6 and IL-1β mRNA were significantly up-regulated in BPAF group compared with Control group(P<0.01),and expressions of inflammatory factors mRNA were significantly down-regulated in BPAF+Exo group compared with BPAF group(P<0.05).Conclu-sion:BPAF(≥25 μmol/L)inhibits proliferation of hESCs and promoted apoptosis.hUCMSCs-Exo inhibits BPAF-induced oxidative dam-age and inflammatory factors expressions in hESCs through PI3K/AKT signaling pathway.

Objective:To investigate effects of human umbilical cord mesenchymal stem cell-derived exosomes(hUCMSCs-Exo)on bisphenol AF(BPAF)-induced oxidative damage and inflammatory factor release from endometrial stromal cells(hESCs).Methods:hESCs were divided into Control group,BPAF group(25 μmol/L BPAF treatment),BPAF+Exo group(25 μmol/L BPAF+hUCMSCs-Exo treatment),BPAF+Exo+LY group(25 μmol/L BPAF+hUCMSCs-Exo+10 μmol/L LY294002 treatment).Cell prolifera-tion was detected by MTT assay;apoptosis,intracellular ROS level,and mitochondrial membrane potential level were detected by flow cytometry;protein expressions of Bcl-2,Bax,Cleaved-caspase-3 and PI3K/AKT signaling pathway were detected by Western blot;mRNA expressions of inflammatory factors TNF-α,IL-6 and IL-1β were detected by RT-qPCR.Results:Compared with Control group,hESCs survival rate was gradually decreased(P<0.01),apoptosis rate was gradually increased with the increased concentration of BPAF(≥25 μmol/L).Compared with Control group,BPAF group showed increased ROS level,decreased mitochondrial membrane potential level,increased Bax and Cleaved-caspase-3 protein expressions,and decreased Bcl-2,p-PI3K and p-AKT protein expressions.Compared with BPAF group,cell survival rate of BPAF+Exo group was increased(P<0.01),ROS level decreased,mitochondrial membrane potential level increased,expressions of Bax and Cleaved-caspase-3 proteins decreased,and expressions of Bcl-2,p-PI3K and p-AKT increased.Compared with BPAF+Exo group,expressions of Bax and Cleaved-caspase-3 protein in cells of BPAF+Exo+LY group were increased,while expressions of Bcl-2,p-PI3K and p-AKT protein were decreased.Expressions of inflammatory factors TNF-α,IL-6 and IL-1β mRNA were significantly up-regulated in BPAF group compared with Control group(P<0.01),and expressions of inflammatory factors mRNA were significantly down-regulated in BPAF+Exo group compared with BPAF group(P<0.05).Conclu-sion:BPAF(≥25 μmol/L)inhibits proliferation of hESCs and promoted apoptosis.hUCMSCs-Exo inhibits BPAF-induced oxidative dam-age and inflammatory factors expressions in hESCs through PI3K/AKT signaling pathway.

Objective:To investigate damaging effects of clomifene citrate(CC)on endometrial stromal cells(hEndoSCs),and to study effects of spermidine on autophagy and inflammatory cytokine expression in damaged endometrial stromal cells.Methods:Groups were firstly divided into control group,spermidine group,clomiphene group(CC group),CC+Spermidine group.MTT assay was used to detect cell survival rate of hEndoSCs after co-incubation with different concentrations of CC or Spermidine for 24 h.Con-tent of intracellular reactive oxygen species(ROS)and level of apoptosis in cells of the 4 groups were detected by flow cytometry tech-nique.Western blot was used to detect expressions of autophagy pathway-related proteins ULK1,p-ULK1,LC-3Ⅱ,and apoptosis-re-lated proteins Bax,Bcl-2,Cleaved-caspase 3.RT-qPCR was used to detect mRNA expressions of IL-6,IL-1β and TNF-α.Results:Compared with control group,CC group showed decreased cell survival,increased apoptosis rate,ROS content,Bax,Cleaved-cas-pase 3 expressions,decreased Bcl-2 expression,decreased levels of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ,and elevated expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA(P<0.01).There was no significant changes viability of cells in spermidine group compared with control group(P>0.05).Compared with CC group,cell survival rate in CC+spermidine group was sig-nificantly increased,apoptosis rate,ROS content,Bax and Cleaved-caspase 3 expressions were decreased,Bcl-2 expression was in-creased,expressions of autophagy-related proteins p-ULK1 and LC-3Ⅱ/Ⅰ were elevated,while expressions of inflammatory factors IL-6,IL-1β and TNF-α mRNA were decreased(P<0.01).Conclusion:CC can inhibit endometrial stromal cell proliferation,promote apoptosis,and increase the transcript levels of inflammatory factors IL-6,IL-1β and TNF-α.Spermidine can reduce intracellular ROS in clomiphene-injured endometrial stromal cells by activating cellular autophagy,increase cell survival,and inhibit the expressions of inflammatory factors IL-6,IL-1β and TNF-α.

Objective:To discuss the effect of Wnt/β-catenin signaling pathway inhibitor methyl 3-｛[(4-methyl-phenyl)sulfonyl]amino ｝ benzoate(MS AB)on the fibrogenic response of the human endometrial stromal cells(HESCs),and to provide the foundation for the application of MSAB in the target therapy of intrauteriue adhesion(JUA).Methods:The normal HESCs were cultured in vitro and divided into two groups:control group and transforming growth factor β1(TGF-β1)group;the HESCs from the adhesion part of the IUA patients were cultured in vitro,regarded as IUA group.Western blotting method was used to detect the expression levels of fibrotic marker protein type Ⅰ collagen α1(COL1A1)in the cells in various groups at different time points(0,12,24,48,and 60 h)after treated with TGF-β1.MTT assay was used to detect the proliferation activities of the cells in various groups.Western blotting method was used to detect the expression levels of the fibrotic marker protein COL1A1,stromal marker proteins such as N-cadherin and α-smooth muscle actin(α-SMA),and Wnt/β-catenin signaling pathway-related protein β-catenin in the cells in control and IUA groups.Based on the MSAB concentrations,the normal HESCs were divided into 0(control),0.25,0.50,0.75,and 1.00 μmol·L-1 MSAB groups,and MTT assay was used to detect the survival rates of the cells in various groups.After treated with MSAB,the normal HESCs were divided into control group(normal HESCs),TGF-β1 group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with complete culture medium,and the cells continued to be cultured for 24 h),and MSAB group(10 μg·L-1 TGF-β1 induced normal HESCs for 24 h then the drug was withdrawn,replaced with a complete medium containing 0.75 μmol·L-1 MSAB and the cells continued to be cultured for 24 h).Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of epithelial-mesenchymal transition(EMT)-related transcription factors Snail,Slug,Smuc,ZEB1,and ZEB2,and COL1A1 mRNA in the cells in various groups.Western blotting method was used to detect the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in various groups.Results:Compared with control group(after treated with TGF-β1 for 0 h),the expression levels of COL1A1 proteins in the HESCs after treated with TGF-β1 for 12,24,48,and 60 h in TGF-β1 group were increased(P＜0.05 or P＜0.01).Compared with control group,there was no significant difference in the proliferation activity of the HESCs in IUA and TGF-β1 groups(P＞0.05).Compared with control group,the expression levels of COL1A1,β-catenin,N-cadherin,and α-SMA proteins in the cells in IUA group were increased(P＜0.05 or P＜0.01).Compared with control group,the survival rates of the cells in 0.75 and 1.00 μmol·L-1 MSAB groups were decreased(P＜0.05 or P＜0.01).Compared with control group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in TGF-β1 group were increased(P＜0.05 or P＜0.01);compared with TGF-β1 group,the expression levels of Snail,Slug,and COL1A1 mRNA in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Compared with control group,after treated with TGF-β1 for 24 h,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in TGF-β1 group were increased(P＜0.01);compared with TGF-β1 group,the expression levels of COL1A1,N-cadherin,α-SMA,β-catenin,and c-myc proteins in the cells in MSAB group were decreased(P＜0.05 or P＜0.01).Conclusion:MSAB can inhibit the fibrogenic responses of the HESCs in vitro,and the results provide the theoretical basis for the application of MSAB in the target therapy of IUA.

Objective:To investigate the effects of bisphenol A(BPA)on the proliferation activity and stemness characteristics of endometrial mesenchymal stem/stromal cells(eMSCs),and to elucidate the improvement effect of human umbilical cord mesenchymal stem cell-derived supernatant(hUCMSC-Sup)on the cell injury.Methods:The eMSCs were cultured in vitro and treated with different concentrations of BPA(0,200,250,300,350,and 400 μmol·L-1).The eMSCs were divided into control group(only cultured with culture solution),BPA group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA),BPA+hUCMSC-Sup group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 50%volumetric ratio of hUCMSC-Sup),and BPA+CHIR-99021 group(cultured with isovolumetric culture solution including 200 μmol·L-1 BPA and 10 μmol·L-1 CHIR-99021).The survival rates of eMSCs in various groups were detected by methyl thiazolyl tetrazolium(MTT)assay.The numbers and diameters of the spheroids in various groups were detected by spheroids formation assay,the proliferation activities of the cells in eMSCs stem cell spheroids in various groups were detected by CCK-8 assay;the percentage of CD73+cells in eMSCs in various groups were detected by flow cytometry;the expression levels of sex determining region Y-box 2(Sox2),octamer-binding transcription factor 4(Oct4),and Nanog mRNA in the eMSCs in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method,the expression levels of β-catenin protein in the eMSCs in various groups were detected by Western blotting method.Results:The MTT results showed that after treated with BPA for 24 and 48 h,compared with 0 μmol·L-1 BPA group,the survival rates of eMSCs in 200,250,300,350,and 400 μmol·L-1 BPA groups were significantly decreased(P<0.01).At 24 and 48 h after treatment,compared with control group,the survival rate of the eMSCs in BPA group was significantly decreased(P<0.01);at 48 h after treatment,compared with BPA group,the survival rate of the eMSCs in BPA+hUCMSC-Sup group was significantly inereased(P<0.05).The spheroids formation assay results showed that compared with culture 3 d group,the numbers and diameters of stem cell spheroids of the eMSCs in culture 4 d group and culture 5 d group were significantly increased(P<0.05 or P<0.01);compared with control group,after 48 h of culture,the number and diameter of the cells in eMSCs stem cell spheroids in BPA group were significantly decreased(P<0.05 or P<0.01).The CCK-8 results showed that after 24 and 48 h of treatment,compared with control group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA group was significantly decreased(P<0.01);compared with BPA group,the proliferation activity of the cells in eMSCs stem cell spheroids in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The flow cytometry results showed that compared with control group,the percentage of the CD73+cells in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the percentage of the CD73+cells in eMSCs in BPA+hUCMSC-Sup group was significantly increased(P<0.01).The RT-qPCR results showed that compared with control group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA group were significantly decreased(P<0.01);compared with BPA group,the expression levels of Sox2,Oct4,and Nanog mRNA in the cells in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were significantly increased(P<0.01).The Western blotting results showed that compared with control group,the expression level of β-catenin protein in the eMSCs in BPA group was significantly decreased(P<0.01);compared with BPA group,the expression levels of β-catenin protein in the eMSCs in BPA+hUCMSC-Sup group and BPA+CHIR-99021 group were signifrcantly inereased(P<0.01).Conclusion:BPA can inhibit the stemness characteristics of the eMSCs,and injury the self-renewal and repair of endometrium;its mechanism may be related to down-regulating the activity of Wnt/β-catenin signal pathway in the cells.hUCMSC-Sup can promote the proliferation of injured eMSCs,and has improvement effect on the stemness injury induced by BPA.

Objective:To investigate the clinical efficacy of botulinum toxin A in the treatment of hand hyperhidrosis.Methods:One hundred patients with hand hyperhidrosis were treated with botulinum toxin A (BTXA, Lanzhou Biotechnology Development Co., Ltd., Botulinum Toxin Type A for Injection Hengli) injection, a total of 200 U. Each hand was injected with 100 U respectively. The curative effect was evaluated by self-made questionnaire. The scores of the two were added. The subjective and objective evaluation were carried out before and after injection, and the patients were rechecked 2 weeks, and 1, 4 and 6 months after injection. Efficacy, patient satisfaction and adverse reactions were evaluated.Results:Compared with before treatment, the effective rate increased 2 weeks after injection, 1 month after injection, 4 months after injection and 6 months after injection, and the difference was statistically significant (Chi-square value was 31.54, 36.33, 28.34, 25.23, respectively, P<0.05). After 6 months of follow-up, the effect gradually decreased, and the curative effect could be maintained for about 10 months. After recurrence, the symptoms of hand sweating were still reduced. Satisfaction 96%; Adverse reactions were mild, subcutaneous blood stasis, 27% hand muscles were slightly weak, and returned to normal after 2-3 weeks. Conclusions:Botulinum toxin A injection has certain curative effect, high safety and less adverse reactions. It is an ideal method for the treatment of hand hyperhidrosis.

Objective:To explore the mechanism of miR-205-5p/E2F1 signal axis in regulating the glioma U251, U87 radiotherapy resistance.Methods:X-ray gradual ascending and intermittent induction method was used to irradiate the glioma U251 cells to establish U251/TR, U87/TR radiation-resistant cell lines. Then, the morphology, migration, invasion and proliferation abilities of cells (U251/TR, U87/TR radiation-resistant cells and U251, U87 radiation-sensitive cells) were analyzed. Luciferase gene detection system and point mutation technique were employed to analyze the mechanism of miR-205-5p and E2F1 gene activity on U251 and U87 radiation-resistant cell lines.Results:Compared with the radiation-sensitive U251 cells, the radiation-resistant cells U251/TR, U87/TR showed increased proliferation activity, enhanced migration and invasion abilities and decreased apoptosis under X-ray irradiation. miR-205-5p mimics transfection could down-regulate the expression of E2F1 factor in U251/TR cells, inhibit cell proliferation, invasion and migration and increase the radiosensitivity of U251/TR cells. miR-205-5p mimics transfection combined with with E2F1 down-regulation exerted anti-tumor effect and decreased cell tolerance by suppressing the Wnt/β-catenin signaling pathway activity.Conclusions:The glioma radiation-resistant cell line U251/TR, U87/TR can be established by X-ray gradual ascending and intermittent induction method. The miR-205-5p/E2F1 signal axis exerts tumor-suppressing effect through the classical Wnt/β-catenin signaling pathway, which can be used as an therapeutic target to increase the radiosensitivity of glioma.

Objective:To explore the preventive effects of hyperbaric oxygen combined with upper limb functional exercise on venous thrombosis in patients with peripherally inserted central catheter (PICC).Methods:A total of 144 patients with PICC admitted to the First Department of General Surgery of Hengshui People’s Hospital from December 2019 to June 2020 were retrospectively selected as the research subjects and divided into control group ( n=70) and observation group ( n=74) according to treatment methods. The control group was given upper limb functional exercise on the basis of conventional treatment, while the observation group was given hyperbaric oxygen treatment on the basis of the treatment of the control group. The incidences of venous thrombosis after treatment were compared between the two groups. The changes of coagulation and fibrinolytic functions and of hemorheology indicators of patients were detected before and after treatment. The predictive value of each indicator for the occurrence of venous thrombosis was evaluated by receiver operating characteristic (ROC) curve. Results:Compared with those before treatment, the prothrombin time (PT), the activated partial thromboplastin time (APTT), the thrombin time (TT), and the levels of tissue plasminogen activator (t-PA) in the two groups after treatment were significantly prolonged or increased, and the levels of fibrinogen (FIB), D-dimer, fibrinogen degradation product (FDP), and plasminogen activator inhibitor (PAI) in the two groups after treatment were significantly decreased ( P<0.05); compared with those in the control group after treatment, PT, APTT, TT, and the level of t-PA in the observation group after treatment were significantly prolonged or increased, and the levels of FIB, D-dimer, FDP, and PAI in the observation group after treatment were significantly lower; and all the differences were statistically significant ( P<0.05). Compared with those before treatment, the values of peak systolic velocity (PSV) and resistance index (RI) in the two groups after treatment were significantly lower, and the values of mean flow velocity (Vm) were significantly higher; compared with those in the control group after treatment, the values of PSV and RI in the observation group after treatment were significantly lower, and the value of Vm was significantly higher; and all the differences were statistically significant ( P<0.05). ROC curve analysis showed that the areas of FIB, D-dimer, FDP, and t-PA under the ROC curve predicting the occurrence of venous thrombosis were 0.823, 0.794, 0.684, and 0.787, respectively; the sensitivities were 75.0%, 66.7%, 61.2%, and 64.4%, respectively; the specificities were 81.5%, 83.3%, 74.6%, and 78.3%, respectively; and the accuracies were 74.3%, 72.6%, 68.4%, and 70.2%, respectively. The accuracy of FIB for the diagnosis of venous thrombosis was better than those of D-dimer, FDP, and t-PA. Conclusion:Hyperbaric oxygen combined with upper limb functional exercise can improve the coagulation function, fibrinolysis, and hemorheology of patients. Clinically, it can help predict the occurrence of venous thrombosis in upper limb by detecting the levels of FIB, D-dimer, FDP, and t-PA.

Objective:To explore the preventive effects of hyperbaric oxygen combined with upper limb functional exercise on venous thrombosis in patients with peripherally inserted central catheter (PICC).Methods:A total of 144 patients with PICC admitted to the First Department of General Surgery of Hengshui People’s Hospital from December 2019 to June 2020 were retrospectively selected as the research subjects and divided into control group ( n=70) and observation group ( n=74) according to treatment methods. The control group was given upper limb functional exercise on the basis of conventional treatment, while the observation group was given hyperbaric oxygen treatment on the basis of the treatment of the control group. The incidences of venous thrombosis after treatment were compared between the two groups. The changes of coagulation and fibrinolytic functions and of hemorheology indicators of patients were detected before and after treatment. The predictive value of each indicator for the occurrence of venous thrombosis was evaluated by receiver operating characteristic (ROC) curve. Results:Compared with those before treatment, the prothrombin time (PT), the activated partial thromboplastin time (APTT), the thrombin time (TT), and the levels of tissue plasminogen activator (t-PA) in the two groups after treatment were significantly prolonged or increased, and the levels of fibrinogen (FIB), D-dimer, fibrinogen degradation product (FDP), and plasminogen activator inhibitor (PAI) in the two groups after treatment were significantly decreased ( P<0.05); compared with those in the control group after treatment, PT, APTT, TT, and the level of t-PA in the observation group after treatment were significantly prolonged or increased, and the levels of FIB, D-dimer, FDP, and PAI in the observation group after treatment were significantly lower; and all the differences were statistically significant ( P<0.05). Compared with those before treatment, the values of peak systolic velocity (PSV) and resistance index (RI) in the two groups after treatment were significantly lower, and the values of mean flow velocity (Vm) were significantly higher; compared with those in the control group after treatment, the values of PSV and RI in the observation group after treatment were significantly lower, and the value of Vm was significantly higher; and all the differences were statistically significant ( P<0.05). ROC curve analysis showed that the areas of FIB, D-dimer, FDP, and t-PA under the ROC curve predicting the occurrence of venous thrombosis were 0.823, 0.794, 0.684, and 0.787, respectively; the sensitivities were 75.0%, 66.7%, 61.2%, and 64.4%, respectively; the specificities were 81.5%, 83.3%, 74.6%, and 78.3%, respectively; and the accuracies were 74.3%, 72.6%, 68.4%, and 70.2%, respectively. The accuracy of FIB for the diagnosis of venous thrombosis was better than those of D-dimer, FDP, and t-PA. Conclusion:Hyperbaric oxygen combined with upper limb functional exercise can improve the coagulation function, fibrinolysis, and hemorheology of patients. Clinically, it can help predict the occurrence of venous thrombosis in upper limb by detecting the levels of FIB, D-dimer, FDP, and t-PA.

Objective:To investigate the effect of pulsed dye laser combined with triamcinolone acetonide on hypertrophic scar after thyroid surgery.Methods:From August 2016 to October 2017, 76 patients (22 males and 54 females, aged between 18 and 48 years, with an average of 31 years) with hypertrophic scar after thyroid surgery in the Department of Dermatology and Cosmetology, Weihai City Hospital, Shandong Province, were divided into two groups. In the observation group, 38 patients were subjected to 595 nm pulsed dye laser combined with triamcinolone acetonide injection. In the control group, 38 patients were only locally injected with triamcinolone acetonide. Once every 4 weeks, the patients who did not recover or had a recurrence continued treatment, and the efficacy and adverse reactions were judged for 5 times in total. The patients were followed up for 1 year.Results:After treatment, 21 cases (55.26%) were cured in the observation group. 32 cases were effective, accounting for 84.21%. 4 patients in the control group were cured, accounting for 10.53%, and 16 patients were effective, accounting for 42.11%. The cure rate and effective rate in the observation group were higher than those in the control group ( χ2=5.371, 4.989, P<0.01). Conclusions:595 nm pulsed dye laser combined with local triamcinolone acetonide injection is effective in the treatment of postoperative hypertrophic scar.