Objective:To discuss the effect of human umbilical cord mesenchymal stem cells culture supernatant(hUCMSCs-Sup)on the proliferation,apoptosis,and endometrium receptivity of the human endometrial stromal cells(hEndoSCs)treated with mifepristone(Ms),and to clarify the possible mechanism.Methods:The hEndoSCs were cultured in vitro and divided into control group and 40,60,80,and 100 μmol·L-1 Ms groups.The survival rates of the cells in various groups were detected by MTT assay.The hEndoSCs were divided into control group,40 μmol·L-1 Ms group,and 60 μmol·L-1 Ms group.The apoptotic rates of the cells in various groups were detected by flow cytometry;the expression levels of apoptosis-related protein B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X protein(Bax)proteins in the cells in various groups were detected by Western blotting method,and the ratio of Bcl-2/Bax was calculated.After treated with hUCMSCs-Sup,the hEndoSCs were divided into control group,Ms group,Ms+hUCMSCs-Sup group,and Ms+hUCMSCs-Sup+3-methyladenine(3-MA)group.The survival rates of the cells in various groups were detected by MTT assay;the apoptotic rates of the cells in various groups were detected by flow cytometry;the expression levels of microtubule-associated protein 1 light chain 3B-Ⅱ(LC3B-Ⅱ)and microtubule-associated protein 1 light chain 3B-I(LC3B-Ⅰ)proteins in the cells in various groups were detected by Western blotting method,and the ratio of LC3B-Ⅱ/LC3B-Ⅰwas calculated;the expression levels of endometrium receptivity marker molecules mRNA in the cells in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)method.Results:Compared with control group,the survival rates of the cells in 40,60,80,and 100 μmol·L-1 Ms groups were significantly decreased(P<0.05)in a time-dependent and dose-dependent manner.Compared with control group,the apoptotic rates of the cells in 40 and 60 μmol·L-1 Ms groups were significantly increased(P<0.05),and the ratios of Bcl-2/Bax were significantly decreased(P<0.05).After treated with hUCMSCs-Sup,compared with control group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms group were significantly decreased(P<0.05),the apoptotic rate was significantly increased(P<0.05),and the expression levels of homeobox A10(HOXA10),leukemia inhibitory factor(LIF),and integrin subunit beta 3(ITGB3)mRNA in the cells were significantly decreased(P<0.05);compared with Ms group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰin the cells in Ms+hUCMSCs-Sup group were significantly increased(P<0.05),the apoptotic rate was significantly decreased(P<0.05),and the expression levels of HOXA10,LIF,and ITGB3 mRNA in the cells were significantly increased(P<0.05);compared with Ms+hUCMSCs-Sup group,the survival rate of the cells and ratio of LC3B-Ⅱ/LC3B-Ⅰ in the cells in Ms+hUCMSCs-Sup+3-MA group were significantly decreased(P<0.05).Conclusion:hUCMSCs-Sup can increase the survival rate and decrease the apoptotic rate of the hEndoSCs after treated with Ms,and increase the endometrium receptivity,and its mechanism may be associated with the activation of autophagy of the hEndoSCs by hUCMSCs-Sup.

ObjectiveTo analyze the epidemic characteristics of measles and rubella in Pudong New Area of Shanghai from 2013 to 2022， and to provide data support for the elimination of measles and rubella. MethodsEnzyme linked immunosorbent assay was used to detect IgM antibodies in serum samples. The sequence of 630 nucleotides at the C-terminal of N gene of measles virus was amplified by reverse transcription-polymerase chain reaction and the phylogenic tree was constructed. ResultsA total of 1 529 suspected cases of measles were detected from 2013 to 2022， among which the positive rate of measles IgM antibody was 33.55% （513/1 529）. The highest positive rate （20.73%） was from March to May ， and the positive rate of rubella IgM antibody was 6.80% （104/1 529）. The positive rate of both IgM was higher in males than that in females （P<0.05）. The IgM against measles was mainly detected in 0‒ years old （63.16%， 96/152） and 20‒ years old （45.61%， 161/353）. The IgM against rubella was mainly detected in 10‒20 years old （27.27%， 18/66）. The IgM antibody could be detected more easily from 4 to 28 days after eruption， and the IgM antibody positive rate of measles/rubella from 2020 to 2022 was significantly lower than previous years （2013‒2019）. There were 2 D8 genotype strains， and the rest were H1a gene subtypes. ConclusionThe positive rate of IgM antibodies against measles/rubella in Pudong New Area of Shanghai decreased significantly. People aged 0‒ years and 20‒ years old are more susceptible to measles， and rubella is concentrated in 10‒ years old. It is necessary to strengthen the vaccination of school-age children， in order to achieve the goal of eliminating measles. The age group with high risk of exposure should be checked for vaccination status to ensure the enhanced immunization， and the surveillance of imported measles cases should be strengthened.

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 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.

Porto-sinusoidal vascular disease (PSVD) is an entity characterized with portal hypertension (PH) in the absence of cirrhosis, the related risk factors, and imaging evidence of obstructed portal vein, hepatic vein and inferior vena cava. Its prevalence varies significantly between East and West countries. Until now, the etiologies have been classified as autoimmune, hematologic, and prethrombotic conditions, infections, toxins or drugs, and genetic or metabolic disorders. However, the definite cause remains unknown. Diagnosis is based on three histopathological features: obliterative portal venopathy, nodular regenerative hyperplasia, and incomplete septal fibrosis. The clinical manifestations of early PSVD are nonspecific, whereas those at a late stage are similar to cirrhosis. The imaging detection mainly reveals the PH signs and complications, but the liver stiffness is normal or slightly increased, necessitating a liver biopsy for PVSD diagnosis. PSVD treatment is similar to liver cirrhosis; however, the prognosis is better. In order to gain a thorough understanding of PSVD, the epidemiology, pathogenesis, clinical diagnosis, and treatment are discussed in this article.

Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) at Neiguan on dexmedetomidine-induced bradycardia in patients.Methods:Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients, aged 20-50 yr, weighing 48-60 kg, scheduled for elective gynecological surgery under general anesthesia, were divided into 2 groups ( n=30 each) using a random number table method: control group (group C) and TEAS group (group T). Dexmedetomidine 1 μg/kg was infused intravenously over 10 min followed by intravenous infusion 0.5 μg·kg -1·min -1 in two groups, and the patients in group T simultaneously received TEAS (frequency 2/100 Hz, disperse-dense wave, intensity 5-10 mA according to the current that could be tolerated) at bilateral Neiguan acupoints.The stimulator was only connected, and no current was given in group C. Before the infusion of dexmedetomidine (T 0) and at 10 min of dexmedetomidine infusion (T 1), mean arterial pressure (MAP) and heart rate (HR) was recorded, and electrocardiogram (ECG) was collected to calculate the PR interval, QT interval, QT interval, Tp-e interval and index of cardiac electrophysiological balance (iCEB). The development of arrhythmia was recorded. Results:Compared with the baseline value at T 0, HR was significantly decreased, and QT interval and PR interval were prolonged at T 1 in two groups, and iCEB was increased, and Tp-e interval was prolonged at T 1 in group C ( P<0.05). Compared with group C, HR was significantly increased, PR interval and Tp-e interval were shortened at T 1, and the incidence of bradycardia and atrioventricular block was increased in group T ( P<0.05). Conclusion:TEAS at Neiguan can decrease the risk of bradycardia induced by dexmedetomidine, and the mechanism may be related to shortening atrioventricular conduction time and reducing heterogeneity of ventricular repolarization in patients.

Objective:To evaluate the effect of electroacupuncture postconditioning on the expression of microRNA-1 (miRNA-1) in rats with arrhythmias induced by myocardial ischemia-reperfusion (I/R).Methods:Twenty-four clean-grade healthy adult male Sprague-Dawley rats, aged 2-3 months, weighing 280-320 g, were divided into 3 groups ( n=8 each) using a random number table method: sham operation group (S group), I/R group and electroacupuncture postconditioning group (EP group). The model of myocardial reperfusion arrhythmia was established by ligating the left anterior descending branch of coronary artery for 30 min followed by 30-min reperfusion in anesthetized rats.The artery was only isolated without ligation after thoracotomy in group S. Bilateral Neiguan acupoints were stimulated for 30 min starting from the time point immediately after reperfusion in EP group.The occurrence of arrhythmia during reperfusion was recorded.The rats were sacrificed at 30 min of reperfusion, and hearts were removed for examination of the pathological changes of the myocardium and for determination of miRNA-1 expression (by quantitative real-time polymerase chain reaction) and expression of Kir2.1 and connexin 43 (Cx43) (by Western blot). Results:Compared with S group, the incidence of arrhythmia was significantly increased, the expression of miRNA-1 was up-regulated, and the expression of Kir2.1 and Cx43 was down-regulated in I/R and EP groups ( P<0.05). Compared with I/R group, the incidence of arrhythmia was significantly decreased, the expression of miRNA-1 was down-regulated, and the expression of Kir2.1 and Cx43 was up-regulated in EP group ( P<0.05). The pathological changes of myocardium were accentuated in I/R and EP groups when compared with S group, and the pathological changes were significantly attenuated in EP group when compared with I/R group. Conclusion:The mechanism by which electroacupuncture postconditioning reduces the occurrence of reperfusion arrhythmia is related to up-regulation of Kir2.1 and Cx43 expression after down-regulation of miRNA-1 expression in rats.

Objective To evaluate the effect of sevoflurane on the electrophysiological stability of i-solated rat hearts subjected to hypothermic perfusion. Methods Clean-grade healthy adult male Sprague-Dawley rats, weighing 280-360 g, were heparinized and anesthetized with pentobarbital sodium. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95％ O2-5％CO2 at 37℃. Twenty-four Langendorff-perfused hearts were divided into 4 groups ( n=6 each) using a ran-dom number table method: control group ( C group ) , sevoflurane group ( S group ) , 32 ℃ hypothermia group ( H group) and 32℃ hypothermia plus sevoflurane group ( HS group) . After 15 min of equilibration, the isolated hearts were continuously perfused for 30 min with K-H solution at 37℃, with K-H solution con-taining 2. 3％ sevoflurane at 37 ℃, with K-H solution at 32 ℃, and with K-H solution containing 2. 3％sevoflurane at 32℃ in C, S, H and HS groups, respectively. Heart rate and monophasic action potential in three layers of the left ventricular anterior wall were recorded at 15 min of equilibration ( T0 ) and 30 of con-tinuous perfusion ( T2 ) , the transmural dispersion of repolarization ( TDR) were calculated, and the occur-rence of arrhythmia was observed. Results Compared with C and S groups, the heart rate was significantly decreased and TDR was enlarged at T1 , and the incidence of arrhythmia was increased in H and HS groups ( P<0. 05) . Compared with H group, TDR was significantly reduced at T1 , and the incidence of arrhythmia was decreased in HS group ( P<0. 05) . Conclusion Sevoflurane can improve the electrophysiological in-stability of isolated rat hearts subjected to hypothermic perfusion, and thus decrease the development of ar-rhythmia.

Objective To evaluate the effects of different concentrations of desflurane on the electrophysiological stability of hearts in female patients.Methods Forty female patients,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 20-50 yr,weighing 45-77 kg,scheduled for elective surgery with general anesthesia,were divided into 2 groups (n =20 each) using a random number table method:0.6 MAC group (group D1) and 1.3 MAC group (group D2).The intravenous access was opened after admission to the operating room,midazolam 0.1 mg/kg,vecuronium 0.1 mg/kg,fentanyl 3 μg/kg and etomidate 0.3 mg/kg were injected intravenously,and mechanical ventilation was performed after tracheal intubation.The concentrations of desflurane reached 0.6 and 1.3 MAC in D1 and D2 groups,respectively.Twelve-lead electrocardiograms were collected before anesthesia induction (T1),at 5 min after intubation (T2) and at 20 min after reaching the set concentration of desflurane (T3).The QT interval and Tpe interval were measured,the QTc interval,Tp-e/QT ratio and index of cardiac electrophysiological balance were calculated,and the development of arrhythmia was also observed.Results The QT interval and QTc interval were significantly longer,and the Tp-e/QT ratio was lower at T3 than at T1,2 in two groups (P＜ 0.05).There was no significant difference in QT interval,Tp-e/QT ratio or index of cardiac electrophysiological balance at each time point between two groups (P＞0.05).No arrhythmia was found in neither group.Conclusion The clinical concentration of desflurane affects the electrophysiological stability of hearts to some extent in female patients without causing arrhythmia.