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.

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.

ObjectiveTo systematically evaluate the effect of action observation therapy (AOT) on upper limb function in children with cerebral palsy. MethodsRelevant literatures about the effect of AOT on upper limb function in children with cerebral palsy were retrieved from the databases of PubMed, Embase, Cochrane Library, Web of Science and CNKI, from the establishment to July 9, 2022. ResultsEleven articles involving 497 patients were included, which were mainly published in the past ten years. The studies included hospital-based studies with therapist supervision and home-based studies without therapist supervision, mainly related to the improvement of upper limb function of AOT in children with cerebral palsy. Experimental group performed actions related to activities of daily living, while control group mainly watched video clips excluding actions, 15 to 120 minutes a time, three to five times a week, with most of the intervention periods of three to four weeks. AOT improved the upper limb function of children with cerebral palsy in terms of body structure and function, and activity and participation, specifically grip strength, muscle tension, and hand dexterity and function. ConclusionHospital-based AOT with therapist supervision can improve upper limb function in children with cerebral palsy, while the effect of home-based AOT without therapist supervision and the long-term effect of AOT need to be further studied.

Objective: To understand the epidemiological and clinical features of patients with hemorrhagic fever and renal syndrome (HFRS) in Shenzhen, and to accumulate experience in the diagnosis and treatment of HFRS in this area. Methods: A retrospective analysis was conducted by collecting the clinical data from 46 patients who were confirmed with HFRS and admitted to the Department of Infectious Diseases of Shenzhen People's Hospital from January 2015 to December 2018. The demographic characteristics, epidemiological, clinical manifestations, examinations, treatments and prognosis, and other characteristics were analyzed. Results: All the 46 patients with HFRS were residens in Shenzhen, with a male-to-female ratio of 6.67∶1.00(40∶6), aged (40.18 ± 15.63) years old, and 38 patients (82.61%) aged 23-45 years old. There were 41 patients (89.13%) with a history of HFRS epidemiology, and there were mice in their houses or workplaces. The houses of 39 patients (84.78%) were rented, and 34 patients(87.18%) rented their houses in urban villages. There were morbidity throughout the year, and 33 patients (71.74%) were ill from January to June. In clinical classification, 44 cases (95.65%) were mild, 2 cases (4.35%) were medium, and there were no severe or critical cases. The clinical manifestations were that all patients were hospitalized due to fever mainly with hyperthermia. Thirty-nine patients (84.78%) were presented with systemic aches, headaches, low back pain and eyelid pain, and 28 patients (60.87%) had skin and mucous membrane hyperemia flushing. Clinical stages showed that all patients had pyretogenesis stage and polyuria stage, including pyretogenesis stage [(7.34 ± 6.82) d], polyuria stage [(9.94 ± 5.77) d], only 4.35% (2/46) patients with hypotension shock stage, all patients did not have oliguric stage. On the next day of admission, the number of white blood cells in 46 patients was (8.17 ± 3.19) × 10⁹/L, and 38 cases (82.61%) in the normal range; platelet was (61.92 ± 32.53) × 10⁹/L, and 42 cases (91.30%) were decreased; the procalcitonin was (1.62 ± 0.38) ng/ml, and 41 cases (89.13%) were increased; C-reactive protein was (74.33 ± 30.48) mg/L, and 46 patients (100.00%) were elevated; creatinine was (176.25 ± 55.15) μmol/L, and 19 cases (41.30%) were increased. Abnormal liver function was manifested by increased enzymology, alanine aminotransferase was (137.58 ± 46.76) U/L, and aspartate aminotransferase was (129.82 ± 40.29) U/L. All patients were positive for Hantavirus IgM. B-ultrasound results showed that in 58.70% (27/46) patients, both kidneys were plump and the parenchymal echo was enhanced. Liver injury occurred in 39 patients (84.78%) and sinus bradycardia in 2 patients (4.35%). All the 46 patients were clinically cured. Conclusion The epidemiological characteristics of patients with HFRS in Shenzhen are typical, the clinical manifestations are mild, and the curative effect and prognosis are good.

Objective To analyze the clinical features of patients with acute stage brucellosis in Shenzhen,and provide a scientific basis for prevention and control of brucellosis in immigrant city.Methods A retrospective analysis was conducted to collect clinical data of patients with brucellosis admitted to the Department of Infectious Diseases,Shenzhen People's Hospital from May 2013 to May 2018.The patient's epidemiology manifestations,pathogen and laboratory examination results,diagnosis and treatment outcomes and prognosis were analyzed.Results Among the 39 patients with brucellosis,males were predominant,with a male to female ratio of 1.4 ∶ 1.0 (23 ∶ 16),an age of (44.91 ± 17.18) years and 24 cases were non-Guangdong natives.There were 23 cases with epidemiological history,including 14 cases with mutton,sheep viscera and goat milk history;the disease occurred throughout the year,mainly from February to July,a total of 26 cases.The clinical manifestations of the patients were mainly fever,sweating,fatigue,joint and muscle pain,weight loss,and liver or spleen or lymph nodes swelling.The blood culture was identified as 38 cases of Brucella melitensis and 1 case of Brucella suis.All strains were sensitive to common antibiotics in vitro.All cases were diagnosed as acute stage of brucellosis,2 cases with orchitis,1 case with brucellosis meningoencephalitis,3 cases with spondylitis,and 3 cases with misdiagnosis.Thirty-nine patients were cured according to the "Brucellosis Diagnosis and Treatment Guidelines (Trial)" and were followed up for 1 year.Conclusions Patients with brucellosis in Shenzhen are mainly infected with Brucella melitensis;fever,sweating,joint and muscle pain are the main clinical symptoms;the patient's efficacy and prognosis are better after treatment;for the occurrence of occasional misdiagnosis,it is recommended that in immigrant cities,medical staff should strengthen their understanding of brucellosis.

OBJECTIVETo conduct genetic diagnosis for a family affected with hamophilia A.

METHODSPotential mutations of the F8 gene were analyzed with PCR and Sanger sequencing. Carriers of the mutation were identified through linkage analysis using short tandem repeat (STR) markers. Suspected mutations were verified among 100 healthy controls to rule out genetic polymorphism. Prenatal diagnosis was provided based on the above results.

RESULTSSequencing analysis has identified two mutations, c.1 A>T and c.4 C>T, which have replaced the start codon (ATG) with leucine (TTG) and glutamine (GAA) with the stop codon (TAA), respectively. The same mutations were not found among the 100 healthy controls. The patient's mother and sister were heterozygous for the same mutations. Upon prenatal diagnosis, the fetus was determined as a male and did not harbor the above mutations. Linkage analysis also confirmed that the fetus has inherited the non-risk X chromosome from his maternal grandfather.

CONCLUSIONDetection of pathogenic mutations can enable prenatal diagnosis for the disease.

Adult ; Factor VIII ; genetics ; Female ; Genetic Linkage ; genetics ; Hemophilia A ; genetics ; Humans ; Male ; Mutation ; genetics ; Prenatal Diagnosis ; methods ; Young Adult

OBJECTIVETo use combined G-banding and array-comparative genomic hybridization (aCGH) for the prenatal diagnosis of a fetus with 5q35 deletion syndrome.

METHODSChromosomal karotypes of the fetus and parents were analyzed with G-banding analysis. aCGH was performed to detect minor chromosomal structural abnormalities.

RESULTSThe karyotype of the fetus was ascertained as 46, XY, t(5;10)(q35;p13), and the karyotypes of the parents were normal. aCGH has identified a de novo 1.68 Mb deletion at 5q35.2q35.3 and a 1.44 Mb duplication at 10p14p13.

CONCLUSIONaCGH has a higher resolution and greater accuracy for mapping chromosomal aberrations and is a useful supplement for G banding karyptyping analysis.

Adult ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 5 ; genetics ; Comparative Genomic Hybridization ; Cri-du-Chat Syndrome ; diagnosis ; embryology ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Karyotyping ; Male ; Prenatal Diagnosis ; Trisomy ; diagnosis ; genetics

OBJECTIVETo explore the effects of uterine adenomyosis on the clinical outcomes of infertility patients treated with in vitro fertilization/intracytoplasmic sperm injection-embryo transfer (IVF/ICSI-ET).

METHODSA retrospective study was conducted of 61 IVF/ICSI-ET cycles as the study group, diagnosed with uterine adenomyosis by transvaginal ultrasound, and 164 IVF/ICSI-ET cycles of patients with tubal infertility as the control group. The baseline characteristics, ovary response and clinical outcomes were compared between the two groups.

RESULTSThe implantation rate, clinical pregnancy rate and live birth rate decreased significantly in the study group (P<0.05), and early abortion rate increased significantly (P<0.05). For patients with adenomyosis, GnRH-antagonist cycles tended to decrease clinical pregnancy rate and increase abortion rate (25.0% vs 45.0%, P=0.184; 66.7% vs 27.8%, P=0.247), and significantly decrease live birth rate (0% vs 30.8%, P=0.025), compared with GnRHa agonist cycles.

CONCLUSIONUterine adenomyosis decreases implantation rate, clinical pregnancy rate and birth rate, and increases abortion rate significantly in patients with IVF/ICSI-ET. GnRH-antagonist cycles have adverse effects on the outcomes of adenomyosis; GnRH agonist long protocol cycles may increase clinical pregnancy rate and decrease abortion rate.

Adenomyosis ; complications ; Embryo Implantation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Gonadotropin-Releasing Hormone ; agonists ; antagonists & inhibitors ; Hormone Antagonists ; Humans ; Infertility, Female ; Pregnancy ; Pregnancy Complications ; physiopathology ; Pregnancy Rate ; Retrospective Studies ; Sperm Injections, Intracytoplasmic

Objective To improve chronic obstructive pulmonary disease (COPD﹚ related to the early diagnosis and prevention of osteoporosis. Methods Report 2 cases of COPD related to osteoporosis. Results Two cases of COPD pa-tients have significant osteoporosis, but repeatedly due to lack of consciousness of medical personnel related to the misdiagnosis. Conclusion Early diagnosis and intervention on COPD combined osteoporosis, for its ability to maintain and improve the quality of our life, and even delay the death is of great significance.