BACKGROUND: Studies have confirmed that RhoA/ROCK signaling pathway plays an important role in the process of epithelial-mesenchymal transition. OBJECTIVE: To investigate the effect of Y-27632 (inhibition of rhodopsin kinase of 4-aminopyridines compounds) on the epithelial-mesenchymal transition of rat peritoneal mesothelial cells (RPMCs) induced by transforming growth factor ?1 (TGF-?1). DESIGN,TIME AND SETTING: An animal observational experiment was performed at the Laboratory of Nephrology in the First Affiliated Hospital of Sun Yat-sen University from March 2007 to March 2008. MATERIALS: A total of ten healthy male Sprague Dawley rats were provided by the Animal Experimental Center of Sun Yat-sen University. Y-27632 was the product of Calbiochem Company (German). TGF-?1 was the product of R&D Company (USA). METHODS: Primary RPMCs were cultured in vitro. After synchronization for 24 hours, RPMCs were randomly divided into 4 groups: control group (RPMCs cultured in the DMEM/F12 without serum), TGF-?1 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L TGF-?1), Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632), TGF-?1 +Y-27632 group (RPMCs cultured in the DMEM/F12 without serum, and then added 10 ?g/L Y-27632. 2 hours later, 10 ?g/L TGF-?1 were added). The cells were collected 48 hours after culture. MAIN OUTCOME MEASURES: The mRNA and protein expression of E-cadherin and ?-Smooth muscle actin (?-SMA) were measured by RT-PCR. The protein expression level of E-cadherin, ?-SMA and vimentin was measured by Western blotting. RESULTS: Compared to control group, the expression of E-cadherin significantly decreased in the TGF-?1 group, and the expression of ?-SMA and vimentin significantly increased (P

Objective To explore the effect of transforming growth factor β1 (TGF-β1) on epithelial-mesenchymal transition in rat peritoneal mesothelial cells(RPMCs) and its mechanism.Methods Primary peritoneal mesothelial cells of SP rats were cultured in vitro. After synchronization for 24 h, RPMCs were randomly divided into 2 groups: Group A (control), Group B (TGF-β1, 10 μg/L). RPMCs were stimulated by 10 μg/L TGF-β1 for different time. The mRNA and protein expression levels of E-cadherin, α-smooth muscle actin (α-SMA) and collagenⅠwere measured by RT-PCR and Western blot, respectively. The protein expression level of total RhoA was measured by Western blot. Active RhoA was extracted by Plasma Membrane Protein Extraction Kit, and assessed by Western blot. Results TGF-β1 down-regulated mRNA and protein expression of E-cadherin in RPMCs, and upregulated mRNA and protein expression of α-SMA and CollagenⅠ. TGF-β1 stimulation elicited a robust increase in RhoA activity in a time-dependent manner. RhoA activity peaked at 1 h.Conclusion RPMCs can be transdifferentiated into myofibroblast under the effect of TGF-(β1,)and the mechanism may be related to the activation of RhoA associated signal pathway.

Objective To express and purify recombinant Tp0136 epitope fragment, and study the immunity activity. Methods The Tp0136 selective fragment(Tp0136B) gene was devised by the surface property analysis, solvent-accessible suface calculateions, secondary structure function region analysis, and was inserted between the sites of Nde Ⅰ and Not Ⅰ in pET22b ( + ) . The recombinant plasmid was expressed in E. coli BI21. After nickel ion metal affinity chromatography, the antigenic and immune reactivity of rTp0136B was confirmed. Then indirect ELISA with the rTp0136B as coating antigen was performed to detect the anti-Tp0136 antibody in sera from 100 normal human controls and 131 primary syphilis patients. Results The rTp0136B was soluble expressed with a molecular weight of about 28 000 and was obtained with a purity of ＞98% by chromatography. Western blot proved that the rTp0136B could specifically react with anti-Tp0136 polyclonal antibody. Specific humoral response was elicited by the recombinant protein in Japan negative. The positive detection rate in sera from primary syphilis patients was 85.5%. Conclusion This result suggested that the recombinant Tp0136 epitope fragments have a satisfactory immunocompetence,which may have applications in the serodiagnosis of primary syphilis.

OBJECTIVETo study the mechanisms of tongjingning granule (TJN) on primary dysmenorrhea (PD).

METHODSOne hundred and twenty PD patients were treated with TJN, and 40 patients treated with aspirin as controls. The levels of serum estrogen (E2) and progestin (P) content in the luteal metaphase and anaphase (MI/AI) as well as endothelin (ET) and calcitonin gene related peptide (CGRP) in the metaphase and menstrual stage before and after treatment in partial patients were determined.

RESULTSThe therapeutic effect , and the curative markedly-effective rate and improvement rate of main accompanied symptoms of severe and moderate PD patients in the treated group were superior than those in the control group (P < 0.01). The level of E2 and ET were significantly decreased, and the content of P and CGRP were significantly increased in the treated group after treatment respectively (P < 0.01).

CONCLUSIONJTN not only can regulate ovarian hormone, ET and CGRP, but shows advantages in adjusting spirits and emotions, improving homeostasis and consolidating the therapeutic effect.

Adolescent ; Adult ; Calcitonin Gene-Related Peptide ; blood ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Dysmenorrhea ; blood ; drug therapy ; Endothelin-1 ; blood ; Estradiol ; blood ; Female ; Humans ; Phytotherapy ; Progesterone ; blood

Objective To compare the bio-medical parameters in SIV infected Chinese rhesus monkeys with diverse disease progression,by which the pathogenesis of simian AIDS were to be investigated.Methods Sixteen Chinese rhesus monkeys were inoculated intravenously with SIVmac239 and followed-up for 18 months.Based on their progression patterns and plasma viral loads,animals were divided into 3 groups,including 1 rapid progressor( RP),13 normal progressors(NP),and 2 elite controllor(EC).Their parameters of haematology,virology,immunology and pathology were examined and compared. Results Compared with other animals,RM449(RP) showed higher viral load,unresponsive humoral immunity,and higher level of auto-antibodies against lymph node,thymus,and spleen.Additionally,its effector memory CD4 count was lower,with the transformation progress being blocked-like from naive/central memory subsets to effector memory subset,as the flow-cytometry assay showed.Notable decrease in its peripheral B cell was also observed,especially to the sub-population of tissue-like memory B cells and activated memory B cells.Pathological examination showed the depletion of lymphoid tissue,atrophy of spleen and loss of thymus.Moreover,most of these parameters of RM450 and RM453 (EC) changed opposite to that of RP.Conclusion The hallmarks of RM449 were higher viraemia and lower SIV specific IgG level,which may due to the disturbance of T cells and B cells development and differentiation.Moreover,destructions of organs of the immune system may contribute to the disturbance.Our study suggest that the change of micro-environments of thymus induced by SIV infection,which is necessary in T cell and B cell development and differentiation,may contribute at least partially to the AIDS pathogenesis.

Objective To evaluate the efficacy of Ganoderma lucidum preparation on the behaviors,biochemistry,and autoimmunity parameters of mouse models of APP/PS-1 double transgenic Alzheimer's disease(AD).Methods A total of 44 4-month-old APP/PS-1 double transgenic AD mice were randomly divided into AD model group,Aricept group,Ganoderma lucidum middle-dose(LZ-M)group,and Ganoderma lucidum high-dose(LZ-H)group,with 11 mice in each group.In addition,10 4-month-old C57BL/6 mice were used as the control group.Water maze test was conducted to observe the behavior changes,and the protein expressions in brain tissues were detected by Western blot analysis.The autoimmune indicators were detected by indirect immunofluorescence method.Results In the navigation experiment,the time of finding the platform was gradually shortened since the 2day in the control,LZ-H,and LZ-M groups,and the time of searching the platform in the AD model group gradually increased.On the 5day,the time of finding platform was significantly shorter in control group (t=5.607,P=0.000) and LZ-H group(t=2.750,P=0.010)than AD model group.In the space exploration experiment,the number of crossing the target platform(t=2.452,P=0.025)and the residence time in the target quadrant(t=2.530,P=0.020)in AD model group mice was significantly smaller/shorter than those in control group;in addition,the number of crossing the target platform in the AD model group was significantly smaller than that in LZ-H group(t=2.317,P=0.030)and LZ-M group(t=2.443,P=0.030),while the residence time in target quadrant decreased significantly(t=2.770,P=0.020)compared with LZ-H group;the number of crossing through the target platform quadrant(t=2.493,P=0.022)and residence time in the target quadrant(t=2.683,P=0.015)in LZ-H group were significantly higher than in Aricept group.Western blot analysis showed that the expression of ApoA1 in the brain tissues of mice in LZ-H and LZ-M groups were significantly higher than those in AD model group(P<0.01,P<0.05);Aβ-40 expression in LZ-H group was significantly lower than that in AD model group(P<0.05);the expressions of Syt1,ApoE,and ABCA1 in brain tissues of mice in LZ-H group were significantly higher than those in model group(P<0.01,P<0.05).The plasma IgG level in Aricept group(t=30.945,P=0.000),LZ-M group(t=25.639,P=0.000)and LZ-H group(t=4.689,P=0.001)were significantly higher than that in the control group.Conclusion Ganoderma lucidum preparation can improve behavior disorders of AD model mice,promote the expressions of ApoA1,ApoE and Syt1,inhibit the expression of Aβ-40 protein,and improve the autoimmune function.

Objective:To investigate the effect of withdrawal time on postpartum liver function in pregnant women receiving tenofovir disoproxil fumarate (TDF) therapy for blocking mother-to-child transmission of HBV.Methods:A prospective study was conducted in Hangzhou First People’s Hospital from June 2016 to August 2018. A total of 84 pregnant women with HBsAg and HBeAg positive were enrolled and divided into two groups according to simple randomized grouping method with 42 cases in each group. In group A TDF was withdrawn immediately after delivery and in group B TDF was withdrawn 12 weeks after delivery. Finally, 66 patients completed the follow-up for 24 weeks postpartum, 35 cases in group A and 31 cases in group B. All patients were administered TDF from week 24-28 of pregnancy. HBV DNA loads and ALT levels were regularly measured and compared. Multivariate logistic regression was used to explore the risk factors of postpartum ALT flare. SPSS 26.0 statistical software was used for statistical processing.Results:Compared with the baseline levels, the HBV DNA loads at 16 weeks postpartum had no significant changes in both groups( Z=-0.742 and -1.891, both P>0.05). Postpartum ALT flare was observed in 21 of the 66 patients, 9 cases (25.71%, 9/35) in group A, and 12 cases (38.71%, 12/31) in group B ( χ2=1.280, P>0.05); and there was no significant difference in the severity of postpartum ALT flare between the two groups ( χ2=0.527, P>0.05). Binary logistic regression analysis showed that increased ALT level during pregnancy was an independent risk factor of postpartum ALT flare ( OR=13.75, 95% CI 1.49-126.85, P<0.05). Conclusions:When TDF was used for preventing mother-to-child HBV transmission, withdrawal at different times after delivery had no effect on postpartum liver function. ALT flare during pregnancy is a risk factor for postpartum ALT flare, so TDF should be discontinued carefully and liver function should be closely monitored postpartum for such patients.

Objective:To investigate the status of frailty and activities of daily living (ADL) in maintenance hemodialysis (MHD) patients, and to explore the effect of frailty phenotype on ADL.Methods:The patients who underwent MHD in Kidney Disease Center of the First Affiliated Hospital from March 2019 to March 2020 were enrolled in this study. The demographic and laboratorial data were collected by cross-sectional survey method. Fried frailty phenotype scale and ADL scale were used to evaluate the frailty and ADL, respectively. The differences of basic data and different frailty phenotypes between the normal function group and the function decline group were compared in terms of ADL, physical self-maintenance ability and instrumental ADL ability. Pearson correlation analysis was used to analyze the correlation between frailty and ADL, and binary logistic regression analysis was used to analyze the influencing factors of ADL.Results:A total of 676 MHD patients were included in this study, including 434 males (64.2%) and 242 females (35.8%). The age was (59.2±19.4) years old, and the median dialysis age was 59.0 (25.3, 110.0) months. There were 159 frailty patients (23.5%), 230 pre-frailty patients (34.0%), and 287 non-frailty patients (42.5%). The ADL was decreased in 163 patients (24.1%), including 131 patients (19.4%) with decreased physical self-maintenance ability and 161 patients (23.8%) with decreased instrumental ADL ability. Pearson correlation analysis showed that the frailty score was positively correlated with total ADL score ( r=0.728, P<0.001), physical self-maintenance ability score ( r=0.669, P<0.001) and instrumental ADL ability score ( r=0.729, P<0.001). Binary logistic regression analysis results showed that older age and lower physical activity, fatigue, slowed steps and lower grip strength in the frailty phenotypes were the independent influencing factors of ADL, physical self-maintenance ability and instrumental ADL ability (all P<0.05). Conclusions:The prevalence of frailty is 23.5% in MHD patients, and 24.1% of MHD patients have decreased ADL. Elder age and lower physical activity, fatigue, reduced step counts, and lower grip strength in frailty phenotypes are the independent influencing factors for poor ADL, poor physical self-maintenance ability and poor instrumental ADL ability.

ObjectiveTo understand the experience， knowledge and attitudes of e-cigarette users in Shanghai， and to provide evidence for the development of intervention programs for e-cigarette use. MethodsThe Respondent-Driven Sampling （RDS） method was used to recruit 40 e-cigarette users for qualitative interviews from November 2020 to February 2021. NVivo （11.0） software was used to code interview records， and the thematic pattern and relationship analysis were conducted. ResultsIn terms of the reasons and user experience of e-cigarette use， most respondents used e-cigarettes for the first time after recommendation of friends； The reasons for the current use of e-cigarettes focus on the subjective feelings generated by the use of e-cigarettes （easy to use， replacing traditional tobacco， and having many flavors）. Compared with traditional tobacco use， respondents said e-cigarettes tasted good and had no physical impact （they could be used all the time without making them uncomfortable）. In terms of the awareness of e-cigarette use related knowledge， the respondents' opinions on whether e-cigarettes caused health risks were mostly based on their own experience of using them， and they said it was difficult to identify real information online. As for whether they were interested in relevant knowledge， most respondents said they wanted to learn about the professional knowledge related to e-cigarettes. Regarding the attitude related to the use of e-cigarettes， most respondents believed that e-cigarettes are addictive， e-cigarettes are not a safe alternative to traditional tobacco， ande-cigarettes couldn't help them quit smoking. Three out of ten respondents said they supported including e-cigarettes in the Regulation on Smoking Control in Public. ConclusionThere are certain characteristics of e-cigarette users' feelings， related knowledge and attitude towards e-cigarette use that should be targeted to carry out publicity， education and intervention.

Objective:To study the current status of longitudinal extrauterine growth restriction (EUGR) in extremely preterm infants (EPIs) and to develop a prediction model based on clinical data from multiple NICUs.Methods:From January 2017 to December 2018, EPIs admitted to 32 NICUs in North China were retrospectively studied. Their general conditions, nutritional support, complications during hospitalization and weight changes were reviewed. Weight loss between birth and discharge > 1SD was defined as longitudinal EUGR. The EPIs were assigned into longitudinal EUGR group and non-EUGR group and their nutritional support and weight changes were compared. The EPIs were randomly assigned into the training dataset and the validation dataset with a ratio of 7∶3. Univariate Cox regression analysis and multiple regression analysis were used in the training dataset to select the independent predictive factors. The best-fitting Nomogram model predicting longitudinal EUGR was established based on Akaike Information Criterion. The model was evaluated for discrimination efficacy, calibration and clinical decision curve analysis.Results:A total of 436 EPIs were included in this study, with a mean gestational age of (26.9±0.9) weeks and a birth weight of (989±171) g. The incidence of longitudinal EUGR was 82.3%(359/436). Seven variables (birth weight Z-score, weight loss, weight growth velocity, the proportion of breast milk ≥75% within 3 d before discharge, invasive mechanical ventilation ≥7 d, maternal antenatal corticosteroids use and bronchopulmonary dysplasia) were selected to establish the prediction model. The area under the receiver operating characteristic curve of the training dataset and the validation dataset were 0.870 (95% CI 0.820-0.920) and 0.879 (95% CI 0.815-0.942), suggesting good discrimination efficacy. The calibration curve indicated a good fit of the model ( P>0.05). The decision curve analysis showed positive net benefits at all thresholds. Conclusions:Currently, EPIs have a high incidence of longitudinal EUGR. The prediction model is helpful for early identification and intervention for EPIs with higher risks of longitudinal EUGR. It is necessary to expand the sample size and conduct prospective studies to optimize and validate the prediction model in the future.