Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.

Objective To explore the relationship and internal path between activities of daily living(ADL),sleep quality and mental health of community elderly people in Shanghai.Methods A questionnaire survey was conducted among community residents aged 60 years and older seeing doctors in community health care center of five streets in Shanghai during Sept to Dec,2021 using convenience sampling.Activities of Daily Living(ADL),Pittsburgh Sleep Quality Index(PSQI)and 10-item Kessler Psychological Distress Scale(K10)were adopted in the survey.Single factor analysis,correlation analysis and multiple linear regression were used to analyze the data.The effect relationship between the variables was tested using Bootstrap's mediated effects test.Results A total of 1 864 participants were included in the study.The average score was 15.53±4.47 for ADL,5.60±3.71 for PSQI and 15.50±6.28 for K10.The rate of ADL impairment,poor sleep quality,poor and very poor mental health of the elderly were 23.6%,27.3%,11.9%and 4.9%,respectively.ADL and sleep quality were all positively correlated with mental health(r=0.321,P＜0.001;r=0.466,P＜0.001);ADL was positively correlated with sleep quality(r=0.294,P＜0.001).Multiple linear results of factors influencing mental health showed that ADL(β= 0.457,95%CI:0.341-0.573),sleep quality(β =0.667,95%CI:0.598-0.737)and mental health were positively correlated(P＜0.001).Sleep quality partially mediated the relationship between ADL and mental health(95%CI:0.078-0.124)with an effect size of 33.0%.Conclusion Sleep quality is a mediator between ADL and mental health among community elderly people.Improving ADL and sleep quality may improve mental health in the population.

Objective:To investigate the effect of anoxic cold environment at 4500 m altitude on female men-struation.Methods:From March 1 to March 20,2023,women in a unit at an altitude of 4500 meters were selected for reproductive health questionnaire survey,and were divided into≤6 months group,6 months to 12 months group and≥12 months group according to altitude exposure time.The changes of menstruation in each group were analyzed to explore the relevant influencing factors.Results:The total incidence of abnormal menstruation in working women in hypoxic cold environment was as high as 66.14%,and there was no statistically significant difference between the groups at different high-altitude exposure times(P＞0.05).The highest incidence of dys-menorrhea among the types of menstrual changes was 61.90%,but there was no statistically significant differ-ence between the groups at different high altitude exposure times(P＞0.05).There was a statistically significant difference(P＜0.05)in the proportion of insufficient sleep for at least 3 days per week,nervousness and anxiety,and training during their menstrual period in the women who experienced changes in their menstrual cycle com-pared to those who did not.Conclusions:Hypoxic cold environment can lead to the change of female menstrua-tion,and it is combined with sleep deficiency,tension and anxiety,and menstrual exercise.

Objective To study the expression of miR-205 and miR-367 in cutaneous malignant melanoma(CMM)tissue and their relationships with clinicopathological characteristics and prognosis.Methods The tumor tissues from 85 patients with CMM who were admitted to our hospital from May 2013 to December 2019 were selected as the CMM group,and the nevus tissue samples from 80 patients with benign pigmented nevi who were treated in our hospital during the same period were selected as the control group.The expression level of miR-205 and miR-367 of two groups were detected by qRT-PCR.The clinical and pathological data of patients with CMM were collected,and the relationships between the expression of miR-205 and miR-367 and clinicopathological characteristics were analyzed.Kaplan-Meier was used to draw the survival curves of patients 3 years after surgery,and the survival rate of patients was analyzed by Log-rank test.The influencing factors of prognosis was analyzed by COX proportional risk regression model.Results The expression level of miR-205 of patients in the CMM group was lower than that in the control group,and the expression level of miR-367 was higher than that in the control group(P<0.05).The proportions of patients with ulceration,high clinical stage,tumor invasion,low KPS score,thick primary lesion,high Clark grade,lymph node metastasis in the miR-205 low-expression group were higher than those in the miR-205 high-expression group(P<0.05),the proportions of patients with the above indexes in the miR-367 high-expression group were higher than those in the miR-367 low-expression group(P<0.05).Patietns were followed up for 3 years,with a median time of 28.3 months,and 1 case was lost to followed-up.The Kaplan-Meier survival curve showed that the 3-year survival rate of patients in the miR-205 high-expression group was higher than that in the miR-205 low-expression group(P<0.05),and the 3-year survival rate of patients in the miR-367 low-expression group was higher than that in the miR-367 high-expression group(P<0.05).COX multivariate regression analysis showed that the high clinical stage,low KPS score,high Clark grade,lymph node metastasis,and high expression of miR-367 were the risk factors for prognosis of patients,while high expression of miR-205 was a protective factor(P<0.05).Conclusion Low expression of miR-205 and high expression of miR-367 in CMM tissue are associated with ulceration,high clinical stage,tumor invasion,low KPS score,thick primary lesion,high Clark grade,lymph node metastasis,and poor prognosis.These two indicators may serve as potential biomarkers for prognostic evaluation in CMM patients.

Objective To study the effect of berberine on the damage of renal tubular epithelial cells(RETC)in diabetic in vitro.Methods RTEC were divided into control group,model group(high glucose),low dose experimental group(2 mg·L-1 berberine and high glucose),medium dose experimental group(4 mg·L-1 berberine and high glucose),high dose experimental group(8 mg·L-1 berberine and high glucose),BBM-H+miR-NC(transfected with mimics control group,8 mg·L-1 berberine and high glucose),BBM-H+miR-135b group(transfected with miR-135b mimics,8 mg·L-1 berberine and high glucose).Methyl thiazolyl tetrazolium(MTT)method was used to detect cell proliferation activity,flow cytometry was used to detect cell apoptosis rate,Western blot was used to detect the expression of B-cell lymphoma-2(Bcl-2)and Bel-associated X(Bax)proteins in cells,and reactive oxygen species(ROS)level were detected by chemical fluorescence method,thiobarbituric acid method was used to detect malondialdehyde(MDA)level,and xanthine oxidation method was used to detect superoxide dismutase(SOD)level,the levels of tumor necrosis factor-α(TNF-α),interleukin-8(IL-8)and interleukin-1β(IL-1 β)were detected by enzyme-linked immunosorbent assay(ELISA).Results The proliferative activity(OD value)of RTEC in control group,model group and low,medium,high dose experimental groups were 0.66±0.04,0.36±0.02,0.43±0.03,0.54±0.03,0.63±0.05;the apoptosis rates were(3.62±0.31)％,(29.41±2.33)％,(20.10±1.65)％,(15.02±1.25)％,(9.58±1.43)％;MDA were(1.04±0.12),(5.24±0.29),(3.45±0.22),(2.16±0.13),(1.60±0.11)nmol·mL-1;SOD were(240.22±12.06),(130.56±10.84),(169.62±12.50),(201.97±12.78),(236.74±14.52)U·mL-1;TNF-α were(31.25±2.51),(51.84±4.20),(44.52±2.61),(38.25±1.50),(32.10±1.78)mg·L-1;IL-8 were(10.59±1.14),(19.95±1.74),(16.10±1.03),(13.52±1.25),(11.17±0.92)mg·L-1;IL-1β were(23.01±1.45),(56.92±2.51),(43.20±1.96),(32.05±1.23),(26.37±2.48)mg·L-1.There were statistically significant differences between model group and control group(all P＜0.05).Compared with the model group,the above indexes in low,medium and high dose experimental group were statistically significant(all P＜0.05).The above indexes of BBM-H+miR-NC group were statistically significant compared with those of BBM-H+miR-135b group(all P＜0.05).Conclusion Berberine can reduce diabetic renal tubular epithelial cell damage by down-regulating miR-135b.

Objective To explore the acupoint selection rules of acupuncture and moxibustion for the treatment of female menopausal obesity and to provide reference for clinical acupuncture treatment of female menopausal obesity based on data mining techniques.Methods Clinical literature on acupuncture treatment of female menopausal obesity was searched by computer in CNKI,Wanfang,VIP,SinoMed,and PubMed database,and the time limit of searching was from the establishment of database to the date of 1 August 2023,and the literature was screened and extracted according to the inclusion and exclusion criteria.The association rule analysis was performed using IBM SPSS Moderler 18.0 software,and cluster analysis was performed using IBM SPSS Statistics 27.0 software.Results A total of 50 literature meeting the requirements were screened out,and 83 groups of acupoint prescriptions were extracted.Descriptive analysis showed that the intervention measures of acupuncture and moxibustion in the treatment of female menopausal obesity were mainly acupoint catgut embedding,electroacupuncture and auricular acupuncture.The top 7 acupoints with the highest frequency were Sanyinjiao(SP6),Zusanli(ST36),Shenshu(BL23),Tianshu(ST25),Guanyuan(RN4),Zhongwan(RN12)and Qihai(RN6).Stomach meridian of foot yangming,conception vessel,bladder meridian of foot taiyang and spleen meridian of foot taiyin are the most commonly selected meridians.In terms of specific acupoints,the main acupoints are intersection points,front-mu points and back-shu points.The acupoints are mainly distributed in the chest and abdomen,lower limbs and waist and back.By using data mining technology,the core acupoint pair of acupuncture and moxibustion for menopausal obesity is'Zusanli-Sanyinjiao',and the core acupoint prescription is'Zusanli,Sanyinjiao,Zhongwan,Tianshu,Shenshu,Guanyuan,Qihai';cluster analysis found 7 types of clinical acupoint clustering.Conclusion Acupuncture and moxibustion for the treatment of female menopausal obesity focuses on tonifying the essence of the kidney and restoring the qi mechanism and characteristics of acupoint selection of spleen and stomach qi.

Objective To identify the key policies related to the maternal and child population in the 14th Five-Year Plan for Health and Development in each province(region,city)and to understand the policy priorities for different groups of women and chil-dren,providing a reference for the formulation of future maternal and child health policies.Methods Word frequency analysis,coding analysis,and matrix coding were conducted using Nvivo12 qualitative research software to interpret the maternal and child-related policies in 24 health and development plans.Results Health and development plans in all provinces(regions,cities)prioritize maternal and child health and focus on healthcare services(24)and medical services(22)for women and children.However,there is a relatively insufficient emphasis on creating a child-friendly environment,and related safeguard measures need further improvement.Conclusion Provinces(regions,cities),as well as healthcare institutions,should focus on the healthcare of women of childbearing age in future planning,prioritize the construction of a child-friendly environment,and gradually improve the safeguard system.It is important to explore characteristic safeguard measures for maternal and child health.

Objective To investigate and analyze the distribution of serum cytokeratin 19 fragment antigen 21-1(CYFRA21-1)and squamous cell carcinoma-associated antigen(SCCA)levels in healthy pregnant women during pregnancy and to assess their diagnostic value for cervical cancer in pregnancy.Methods A total of 441 healthy pregnant women and 69 patients with cervical cancer in pregnancy who attended the Obstetrics and Gynecology Hospital of Fudan University from Jan 2021 to May 2024 were selected,and 165 healthy women in the Physical Examination Center of the Obstetrics and Gynecology Hospital of Fudan University were included in the same period as the control group.The healthy pregnant women were divided into 143 in early pregnancy(T1 group),147 in middle pregnancy(T2)and 151 in late pregnancy(T3).Serum CYFRA21-1 and SCCA values were detected and analyzed in all groups.One-way ANOVA,independent samples t-test,Mann-Whitney U-test,Kruskal-Wallis H-test,logistic analysis,and ROC curves were used for comparative analysis.Results The CYFRA21-1 and SCCA values were 1.66(1.19-2.17)ng/mL and 0.8(0.6-1.0)ng/mL in the control group,3.07(2.11-4.14)ng/mL and 0.9(0.7-1.3)ng/mL in the healthy pregnant women group,and were 4.33(2.99-7.60)ng/mL and 1.8(0.9-8.5)ng/mL in the patients with cervical cancer in pregnancy group,respectively.There was a statistically significant difference in the two serum values between every two groups(P<0.05).CYFRA21-1 levels were 3.13(2.46-4.05)ng/mL,1.89(1.50-2.53)ng/mL and 4.19(3.48-5.43)ng/mL in the T1,T2,and T3 groups,respectively;and SCCA levels were 0.9(0.7-1.1)ng/mL,0.7(0.6-1.0)ng/mL and 1.2(0.8-1.7)ng/mL,respectively.The results of T1 and T3 groups were higher than those of the control group(P<0.05);however,there was no statistically significant difference between the results of the T2 group and those of the control group(P>0.05).The areas under the ROC curves for the diagnosis of cervical cancer in pregnancy for CYFRA21-1,SCCA,human epididymis protein 4(HE4),anti-carcinoembryonic antigen(CEA)and joint indicators were 0.684,0.724,0.612,0.791 and 0.913,with sensitivities of 36%,48%,38%,57%and 73%,specificities of 96%,97%,89%,86%and 99%,respectively.The cut-off values of each indicator were 6.05 ng/mL,2.60 ng/mL,51.45 pg/mL and 1.75 ng/mL,respectively.Conclusion Serum CYFRA21-1 and SCCA levels were higher in pregnant women during early and late pregnancy compared with non-pregnant individuals,while they were not statistically different from non-pregnant women during mid-trimester.CYFRA21-1 and SCCA have diagnostic value for patients with cervical cancer during pregnancy.

Objective To investigate the effect of body mass index(BMI)on the clinicopathological characteristics of patients with idiopathic membranous nephropathy(IMN).Methods A total of 261 patients with IMN were divided into the normal group(66 cases),the overweight group(105 cases)and the obese group(90 cases)according to BMI.Clinical and renal pathological data of patients were compared between the three groups.The correlation between BMI and clinicopathological indexes was analyzed by Pearson or Spearman's correlation.The influencing factors of estimated glomerular filtration rate(eGFR)were analyzed by multiple linear regression,and the influencing factors of interstitial fibrosis(IF),tubular atrophy(TA),glomerulosclerosis(GS)and mesangial cell proliferation(MCP)were analyzed by binary Logistic regression.Results Compared with the normal group,the prevalence of diabetes mellitus,triglycerides(TG)and low-density lipoprotein cholesterol(LDL-C)were elevated in the overweight group.The prevalence of hypertension,hemoglobin(HGB),uric acid(UA),LDL-C,TG,24-h urinary protein(UTP)and serum complement 3(C3)were elevated,and high-density lipoprotein cholesterol(HDL-C)was decreased in the obese group(P＜0.05).The prevalence of hypertension,UA,TG and serum C3 were elevated in the obese group compared to the overweight group(P＜0.05).The glomerular basement membrane(GBM)thickness was higher in the obese group and the overweight group than that in the normal group,and the proportion of GS and IF was higher in the obese group than that in the normal group(P＜0.05).BMI was positively correlated with hypertension,TG,LDL-C,serum C3,UTP,GS,IF,MCP and deposition in the mesangial region of C3,and negatively correlated with HDL-C(P＜0.05).Multiple linear regression analysis showed that age,blood urea nitrogen(BUN),anti-phospholipase A2 receptor antibody(anti-PLA2R),UTP and TA were independent risk factors of eGFR.Binary Logistic regression analysis showed that elevated BMI,age,UTP and serum creatinine(Scr)were independent risk factors for IF.Age,Scr and elevated UA were independent risk factors for TA.Elevated BMI and decreased eGFR were independent risk factors for GS.Elevated BMI was an independent risk factor for MCP.There was no significant difference in the treatment protocol of IMN patients between the three groups.Conclusion Obesity can exacerbate multiple clinical and pathological outcomes in IMN patients.

Purpose: This study aims to investigate the current status of affiliate stigma among parents of autistic children, analyze the influencing factors, explore the relationship among mindfulness, coping styles, and affiliate stigma, and verify the mediating role of coping styles between mindfulness and affiliate stigma in parents of children with autism in China.MethodBetween February and April 2023, the Child Development Behaviour Centre of a public hospital in China recruited 345 parents of children with autism. These parents completed the general information questionnaire, the Mindful Attention Awareness Scale, the Affiliate Stigma Scale, and the Simple Coping Style Questionnaire. We then adapted the Hayes Process Macro and Bootstrap methods to examine the mediating effects of coping styles between mindfulness and affiliate stigma. Results: (1) The total affiliate stigma score of parents of children with autism was 48.53 (standard deviation:: 10.74). Parents' age, monthly family income, duration of care, mindfulness, and coping styles were the influencing factors of parental affiliate stigma. (2) Mindfulness was positively correlated with positive coping style (r = 0.33, p < .01) and negatively correlated with negative coping style, affiliate stigma (r = −0.38, −0.39, p < .01), whereas affiliate stigma was negatively correlated with positive coping style (r = −0.34, p < .01) and positively correlated with negative coping style (r = 0.41, p < .01). (3) Positive coping style and negative coping style play a parallel mediating role between mindfulness and affiliate stigma of parents of autistic children. Conclusions Parents of children with autism experience significant levels of affiliate stigma. Mindfulness has a direct impact on associated stigma in parents of children with autism and also indirectly predicts associated stigma through the intermediary influence of positive and negative coping styles. Healthcare professionals could perform mindfulness interventions from an optimistic psychology viewpoint to boost parents' mindfulness and coping abilities, thereby accomplishing the objective of mitigating affiliate stigma.