1.Apoptosis of endometrial adenocarcinoma cell line JEC induced by influenza virus H_3N_2
Huan WANG ; Zhongming WU ; Rui MA ; Yu WANG
Journal of Third Military Medical University 1983;0(03):-
Objective To study the apoptosis-inducing effects and mechanisms of influenza virus A H_ 3 N_ 2 on a self-established endometrial adenocarcinoma cell line JEC. Methods JEC cells were infected with different concentrations of H_3N_2, the apoptosis were detected using the HE staining, DNA agarose gel electrophoresis and flow cytometry. The expressions of Fas, FasL and TGF-? were examined by immunocytochemical staining. Results After infection, the JEC cells showed the morphological apoptosis; DNA agarose electrophoresis demonstrated a ladder-like pattern of DNA fragments; FITC/PI stained FCM showed the apoptotic rate of JEC had been decreased along with the prolonging of infected time and elevated with the increase of the virus concentrations. Immunocytochemical staining showed that enhanced expression of Fas, attenuated expression of TGF-?, and no expression of FasL after H_3N_2 infection. Conclusion Influenza virus A H_3N_2 can induce JEC cells to apoptosis in time- and concentration- dependent manners, which may be related with the expressions of Fas and TGF-?.
2.Changes of liver fibrosis-related miRNAs induced by soluble egg antigen of Schistosoma japonicum
Huan WANG ; Yajing LU ; Yanru GAO ; Shuhong WANG ; Rui ZHOU ; Huifen DONG
Chinese Journal of Schistosomiasis Control 2017;29(2):192-196
Objective To investigate the expression of miRNA associated with hepatic fibrosis induced by Schistosoma ja-ponicum soluble egg antigen stimulation in mouse hepatocytes(AML12),so as to lay the foundation for clarifying the mecha-nism of schistosome infection leading to hepatic fibrosis. Methods The expressions of miR-122,miR-182,miR-23b,miR-27b and KSRP in AML12 cells treated with SEA were measured by q-PCR. KSRP protein in cell lyses was measured by Western blotting. AML12 cells were transfected with miR-27b precursor or anti-miR-27b for 24 h,then q-PCR was adopted to determine KSRP mRNA,and KSRP protein was detected by Western blotting. Results The expressions of miR-182,miR-23b and miR-27b were decreased and miR-122 was increased in AML12 cells following SEA treatment(all P<0.05). An increase of mRNA and protein of KSRP expression was also observed in AML12 cells after SEA stimulation(both P<0.05). In addition,KSRP mRNA expression was not changed significantly in AML12 cells transfected with anti-miR-27b or miR-27b precursor,and miR-27b precursor reduced KSRP protein expression as compared with the control. In contrast,the expression of KSRP protein was increased in the anti-miR-27b group and decreased in the miR-27b precursor group. Conclusions After the stimulation of SEA,the expressions of a variety of liver fibrosis-related miRNAs and KSRP change in murine hepatocytes,including miR-27b. And miR-27b can regulate the expression of KSRP. These findings might lay a foundation for further study on the molecular mechanism of fibrosis induced by schistosome infection.
3.Mechanistic study on the role of microRNA-146a in THP-1 cells-associated inflammatory response induced by Cryptococcus neoformans and Cryptococcus gattii
Yan WANG ; Rui GAO ; Hong CHEN ; Huan CHEN ; Yi JIN ; Ningxin LIAO ; Jianghan CHEN
Chinese Journal of Infection and Chemotherapy 2017;17(4):393-396
Objective To analyze the differential expression of microRNA-146a (miR-146a) in monocyte-macrophage cell line (THP-1 cells) after induction by Cryptococcus neoformans (C.neoformans,reference strain WM148) or Cryptococcus gattii (C.gattii,reference strain R265),and investigate the mechanism of miR-146a in regulating the inflammatory response of cryptococcal meningitis.Methods The cultured THP-1 cells were divided into two groups to be induced by C.neoformans or C.gattii,respectively.THP-1 cells were induced with inactivated WN148 (or R265) strains at multiplicity of infection (MOI) of 5 in all experiments.The supematant and the cell pellet were collected separately after incubation.The expression of miR-146a was measured by real-time quantitative PCR (qRT-PCR) technique.The levels of TNF-u and IL-6 release were assayed by ELISA.Results The expression of miR-146a increased significantly in the C.neoformans induction group compared to 0 h.It reached peak at 3 h (P<0.01),and then declined gradually.The level of TNF-α increased in supematant and reached peak at 12 h.The expression of IL-6 did not change significantly at each time point.The expression of miR-146a and TNF-α increased gradually and reached peak at 12 h in the C.gattii induction group (P <0.01),but the change did not reach statistical significance at 3 h,6 h time points.The expression of IL-6 gradually increased,and reached peak at 12 h time point.Conclusions Following stimulation with C.neoformans or C.gattii,the expression ofmiR-146a in THP-1 cells showed different patterns over time.The expression levels of TNF-α and IL-6 showed different patterns.These findings suggest that there may be different regulatory mechanisms in the THP-1 cells-associated inflammatory response after stimulation by inactivated C.neoformans and C.gattii strains.
5.Recombinant human interferon alpha 2b broad-spectrum anti-respiratory viruses pharmacodynamics study in vitro.
Hui-Qiang WANG ; Lin-Lin MA ; Jian-Dong JIANG ; Rui PANG ; Yu-Jun CHEN ; Yu-Huan LI
Acta Pharmaceutica Sinica 2014;49(11):1547-1553
This study is to investigate the effect of recombinant human interferon alpha 2b against broad-spectrum respiratory viruses in vitro. At the cellular level, the effect of the recombinant human interferon alpha 2b on influenza A virus was detected using real-time fluorescence quantitative RT-PCR. The effects of the recombinant human interferon alpha 2b on influenza B virus, parainfluenza virus, respiratory syncytial virus (RSV) and coronavirus were detected using cytopathic effect (CPE) method. In this study, the therapeutic index of recombinant human interferon alpha 2b anti-HPIV was 1476.63, the therapeutic index of recombinant human interferon alpha 2b anti-RSV was 141.37, the therapeutic index of recombinant human interferon alpha 2b anti-coronavirus was more than 2820.76, and the antiviral effect of recombinant human interferon alpha 2b was better than ribavirin (RBV). Recombinant human interferon alpha 2b has a stronger inhibitory effect on different influenza A virus RNA than drug control. The therapeutic index of recombinant human interferon alpha 2b anti-influenza B virus was 2.74, with modest effect. Recombinant human interferon alpha 2b in vitro has broad spectrum antiviral activities, low toxicity and high therapeutic index. Recombinant human interferon alpha 2b is expected to become the efficient medicine in clinical against respiratory viruses, as well as provide better services for prevention and treatment of respiratory viruses' infections.
Antiviral Agents
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pharmacology
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Humans
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Influenza A virus
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drug effects
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Influenza B virus
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drug effects
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Interferon-alpha
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pharmacology
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Parainfluenza Virus 1, Human
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drug effects
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Recombinant Proteins
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pharmacology
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Ribavirin
6.Determinations of mifepristone and its metabolites and their pharmacokinetics in healthy female Chinese subjects.
Yan-ni TENG ; Rui-qian DONG ; Ben-jie WANG ; Huan-jun LIU ; Zhi-mei JIANG ; Chun-min WEI ; Rui ZHANG ; Gui-yan YUAN ; Xiao-yan LIU ; Rui-chen GUO
Acta Pharmaceutica Sinica 2011;46(10):1241-1245
The aim of this study is to establish an HPLC method for simultaneous determinations of mifepristone and its metabolites, mono-demethylated mifepristone, di-demethylated mifepristone and C-hydroxylated mifepristone in plasma and to evaluate the pharmacokinetic characteristics of mifepristone tablet. Twenty healthy female Chinese subjects were recruited and a series of blood samples were collected before and after 0.25, 0.5, 1.0, 1.5, 2.0, 4.0, 8.0, 12.0, 24.0, 48.0, 72.0 and 96.0 hours administration by a single oral dose of 75 mg mifepristone tablet. Mifepristone and its three metabolites were extracted from plasma using ethyl acetate and determined by high performance liquid chromatography. The main pharmacokinetic parameters of mifepristone and its metabolites, including Cmax, tmax, MRT, t(1/2), V, CL, AUC(0-96 h) and AUC(0-infinity), were calculated by Drug and Statistical Software Version 2.0. The simple, accurate and stable method allows the sensitive determinations of mifepristone and its metabolites in human plasma up to 4 days after oral administration of 75 mg mifepristone tablet and the clinical applications of their pharmacokinetic studies.
Administration, Oral
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Area Under Curve
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Asian Continental Ancestry Group
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Biological Availability
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Chromatography, High Pressure Liquid
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methods
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Female
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Humans
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Mifepristone
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administration & dosage
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metabolism
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pharmacokinetics
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Tablets
7.Comparison of the early control process of H7N9 avian influenza in Shanghai and COVID-19 in Wuhan
Li LUO ; Ge BAI ; Rui-ming DAI ; Qian WANG ; Huan-huan ZHU ; Xin LIU ; Ping WU ; Ying WANG ; Tian-tian ZHANG
Shanghai Journal of Preventive Medicine 2020;32(6):478-
Objective To analyze and estimate the possibility of early control in Shanghai if COVID-19 had begun in Shanghai. Methods Comparison was made in the processes of early control between H7N9 avian influenza in Shanghai in 2013 and COVID-19 in Wuhan in 2019.The early incidence data of Korean COVID-19 was simulated and analyzed to predict whether the medical resources needed in Shanghai were available. Results If it had occurred in Shanghai, it would have taken 22 days from the first case to the government′s emergency response.It was estimated that there would have been 602-763 patients with cumulative onset and onset after incubation period.At least 500 beds of infectious diseases could have been allocated in Shanghai in case of emergency.Through adding beds and resources reallocation in the whole city, patients could have been fully admitted and treated. Conclusion If COVID-19 epidemic had occurred in Shanghai, it′s early control would have been possible though there might have difficulties.
8.Primary development of visual uroflow scale.
Wei Yu ZHANG ; Huan Rui WANG ; Xian Hui LIU ; Tao WANG ; Jing Wen CHEN ; Yi Ran SUN ; Xiao Peng ZHANG ; Hao HU ; Ke Xin XU
Journal of Peking University(Health Sciences) 2020;52(4):684-687
OBJECTIVE:
To develop the visual uroflow scale (VUS), analyze the relationship of VUS score and index of free uroflowmetry, assess urination function preliminarily and improve the work efficiency in the clinic.
METHODS:
Male lower urinary tract symptoms (LUTS) patients, who attended the Department of Urology in Peking University People's Hospital from March 2016 to March 2017, were assessed for their urination function according to the Visual Uroflow Scale without help from clinicians before undertaking a free uroflowmetry test. And afterwards, a free uroflowmetry was undertaken, and variables including maximal flow rate (Qmax), the average flow rate (Qave) and voiding volume (VV) was obtained. During the study, 124 cases were collected and 53 cases met the inclusion and exclusion criteria and were included in the study cohort. The Spearman correlation analysis was used for analyzing the correlation of VUS scores with free uroflowmetry variables and age. The validity of VUS was evaluated.
RESULTS:
Most of the patients could choose the very figure matched with self-condition by first instinct without any help from the clinician. The data were analyzed by Spearman correlation analysis. In the present study, voiding time was positively correlated with the VUS score (correlation coefficient, 0.62, P < 0.05). In the present cohort, the patients chose the third and fourth figures to take longer time to urinate, implying worse LUTS situation. Flow time and VUS scores were positively correlated (correlation coefficient, 0.61, P < 0.05). The patients with higher VUS scores would spend more time on urinate, no matter how long urinary hesitation was. Both Qmax and Qave were negatively correlated with the VUS score (correlation coefficient -0.54, -0.62, P < 0.05). The study illustrated that the VUS score suggested that the Qmax basically and further reflected the urination function. And its relationship to age revealed the decreased urination function of aging male, which had reached a consensus.
CONCLUSION
Development of VUS has helped the clinician assess the urination function preliminarily at the first time. Patients are assessed for a VUS score before getting surgery or receiving the drug for treatment, and can be re-assessed after. The VUS score can provide an objective quantitative basis to evaluate the treatment efficacy. In addition, considering that it is convenient, timesaving and easy to understand, the VUS is available for follow-up.
Cohort Studies
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Humans
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Lower Urinary Tract Symptoms
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Male
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Urination
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Urodynamics
9.Changes in Ca(2+)concentration and caspase-3 expression and their relationship in Raji cells exposed to electromagnetic radiation.
Wei WANG ; Huan-xin LIU ; De-wen WANG ; Hong-yan ZUO ; Rui-yun PENG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2013;31(2):104-107
OBJECTIVETo study the effects of electromagnetic pulse (EMP), S-band high power microwave (S-HPM), and X-band high power microwave (X-HPM) on the Ca(2+) concentration and caspase-3 expression in Raji cells and the relationship between Ca(2+) concentration and caspase-3 expression, and to investigate the regulatory mechanism of electromagnetic radiation damage.
METHODSRaji cells were cultured conventionally. Some cells were irradiated by EMP, S-HPM, and X-HPM in the logarithmic growth phase for 6 hours and then collected; others received sham irradiation as a control. The Ca(2+) concentration in the cells was measured by laser scanning confocal microscope; the caspase-3 expression in the cells was evaluated by Western blot.
RESULTSCompared with the control group (Ca(2+) fluorescence intensity = 43.08 ± 2.08; caspase-3 expression level = 0.444 ± 0.13), the EMP,S-HPM, and X-HPM groups had significantly increased Ca(2+) concentrations, with Ca(2+) fluorescence intensities of 69.56 ± 1.71, 50.06 ± 1.89, and 70.68 ± 1.59, respectively (P < 0.01), and had upregulated caspase-3 expression, with expression levels of 0.964 ± 0.12, 0.586 ± 0.16, and 0.970 ± 0.07, respectively (P < 0.01). Each of the EMP and X-HPM groups had significantly higher Ca(2+) fluorescence intensity and caspase-3 expression level than the S-HPM group (P < 0.01), but there were no significant differences between the EMP and X-HPM groups. The linear regression analysis showed that the caspase-3 expression was upregulated as the Ca(2+) concentration increased, with a positive correlation between them (P < 0.01).
CONCLUSIONEMP, S-HPM, and X-HPM cause damage probably by increasing the Ca(2+) concentration in cells and in turn inducing caspase-3 overexpression.
Calcium ; metabolism ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Electromagnetic Radiation ; Humans
10.Prophylaxis of neonatal respiratory distress syndrome by intra-amniotic administration of pulmonary surfactant.
Jian-ping ZHANG ; Ying-lan WANG ; Yun-hui WANG ; Rui ZHANG ; Huan CHEN ; Hao-bin SU
Chinese Medical Journal 2004;117(1):120-124
BACKGROUNDNeonatal respiratory distress syndrome (NRDS) is caused by a deficiency in pulmonary surfactant (PS) and is one of the main reasons of neonatal mortality. This study was conducted to evaluate the efficacy and safety of intra-amniotic administration of pulmonary surfactant for prophylaxis of NRDS.
METHODSForty-five pregnant women who were due for preterm delivery and whose fetuses' lungs proved immature were divided into two groups. Fifteen women (study group) were administered one dose of pulmonary surfactant injected into the amniotic cavity and delivered within several hours. Nothing was injected into the amniotic cavity of 30 women of the control group. The proportion of neonatal asphyxia, NRDS, mortality and the time in hospital were analyzed to determine if there was any difference between the two groups.
RESULTSThere was no significant difference between the two groups for neonatal asphyxia. Foam tests showed that higher proportion of neonates in the study group than in the control group (56.3% vs 13.3%, P < 0.05) had lung maturity. A greater number of control neonates (11/30, 32.3%) had NRDS, compared with the neonates given PS via the amniotic cavity before delivery (1/16, 6.3%, P < 0.05). The neonates in the study group spent nearly 10 days less in hospital than the control group [(32.4 +/- 7.6) days vs (42.0 +/- 15.7) days, P < 0.05], but the difference in mortality between the two groups was not statistically significant.
CONCLUSIONSIntra-amniotic administration of pulmonary surfactant can significantly reduce the proportion of NRDS and the time in hospital of preterm neonates. Whether this method can reduce the mortality of preterm neonates needs to be evaluated further. Intra-amniotic administration of pulmonary surfactant provides an additional effectual means for NRDS prophylaxis.
Amnion ; Female ; Humans ; Infant, Newborn ; Pulmonary Surfactants ; administration & dosage ; Respiratory Distress Syndrome, Newborn ; prevention & control ; Safety ; Treatment Outcome