1.Effect of STAT5 Pathway Inhibitor Pimozide on NO and iNOS Expressions in the Inflammation Model of RAW264.7 Cell
Pengjiu YU ; Limei WAN ; Hui XIE
China Pharmacy 2015;(22):3043-3045
OBJECTIVE:To investigate the effect of STAT5 pathway inhibitor pimozide on the expressions of nitric oxide (NO)and nitric oxide synthetase(iNOS)in the model of mouse macrophage RAW264.7 inflammation induced by lipopolysaccha-ride (LPS). METHODS:RAW264.7 cells in logarithmic growth phase were divided into blank control group,drug control group (10μmol/L pimozide),model group(1μg/ml LPS)and the pimozide groups of low,middle and high doses(2.5,5 and 10μmol/L), where the corresponding cells were given pimozide 30 min before the administration of LPS,and then were cultured for 24 h. Griess method was used to determine the content of NO in the supernate of cell culture solutions of all groups,real-time quantita-tive polymerase chain reaction(RT-PCR)to determine iNOS mRNA expression,and Western blot method to determine the protein expression of iNOS and phosphorylated STAT5(p-STAT5). RESULTS:The content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells in the model group were higher than those in the blank control group,with statisti-cally difference (P<0.01). Compared to the model group,the pimozide groups of middle and high doses had lower content of NO,iNOS mRNA and protein expressions and the content of p-STAT5/STAT5 in the cells,with statistically difference(P<0.01 or P<0.05). CONCLUSIONS:STAT5 pathway inhibitor pimozide can inhibit the release of NO by inhibiting iNOS mRNA and pro-tein expressions in cells.
2.Analysis of GDNF(△N39)-R9 Fusion Protein Delivery Across The Cellular Membrane,Blood Brain Barrier and Biological Function
Progress in Biochemistry and Biophysics 2006;0(08):-
In order to study the application of glial cell line-derived neurotrophic factor(GDNF)in clinic,gene mutation,fusion protein expression in E.coli and purification methods have been used to obtain the fragments of GDNF,GDNF(△N39),GDNF(△N39)-R9.Using primary cultured dopaminergic neurons and PC12 cells with transfected with GFR?1 and Ret to observe their biological function and cytotoxicity.Using B-Endo3 cells and Transwell method to analyze their delivery across the cellular membrane and blood brain barrier.The results show that GDNF(△N39)-R9 has the same neurotrophic function with wild GDNF and nearly no cytotoxicity to dopaminergic neurons and PC12-GFR?1-Ret cells and can get through effectually the cellular membrane and simulacrum of blood brain barrier with matrigel and B-Endo3.
3.Inducing differentiation of human amnion-derived mesenchymal stem cells into insulin-secreting cells in vitro
Yujie ZHAO ; Ning FANG ; Daixiong CHEN ; Limei YU ; Huangfei YU ; Weihong WAN ; Chunhua ZHAO
Chinese Journal of Endocrinology and Metabolism 2011;27(12):1013-1016
ObjectiveTo investigate the potential of human amnion-derived mesenchymal stem cells to differentiate into insulin secreting cells in vitro.MethodsThe hAD-MSCs were isolated from human amnion by trypsin-collagenase digestion.The phenotype of the isolated cells was identified by flow cytometry and immunocytochemical staining.The 3rd generation cells were inoculated at density of 2.5 × 106 unit/ml or 5 × 105 unit/ml in 6 well plates or preset coverslip 24 well plates.Induced groups were treated in serum-free HG-DMEM with 10 mmol/L nicotinamide and N2 supplement.The cells in the non-induced groups were incubated in LG-DMEM supplemented with 10% fetal bovine serum.At days 7,14 and 21 after induction,insulin and β2 microglobulin was determined by immunocytochemical stain,the content of insulin in the culture supernatant was assayed by radioimmunoassay,and insulin mRNA and PDX-1 mRNA were detected by reverse transcriptase-polymerase chain reaction.Results( 1 ) The hAD-MSCs highly expressed CD29,CD44,CD73,CD166,and vimentin.( 2 ) At 7,14,and 21 days,the percentages of insulin-positive cells in hAD-MSCs induced groups were 74.67% ± 1.53%,75.00%:1.00%,and 74.33% ±1.53%,respectively.Contents of insulin in the supematant of hAD-MSCs induced groups were ( 331.62 ± 1.76 ),( 330.50 ± 1.22 ),and ( 331.65 ± 0.48 ) μIU/ml,respectively,but non-induced groups were negative.(3) PDX-1 mRNA and PDX-1 protein were expressed before and after the induction of hADMSCs,but insulin mRNA was expressed only in the induced groups.( 4 ) Both hAD-MSCs induced groups and non-induced groups expressed β2 microglobulin ( all P > 0.05 ).ConclusionThe hAD-MSCs have a potential of differentiating into ISCs and thus may become a new cell source of therapy for type 1 diabetes.
4.Analysis on Quality of Life and Its Influencing Factors in Patients with Chronic Gastritis
Ying CHEN ; Limei HE ; Hongying LI ; Qiaoyun HUANG ; Xiaojing YANG ; Chonghua WAN
Journal of Kunming Medical University 2014;(2):8-11,26
Objective To analyze the status of quality of life and influencing factors in patients with chronic gastritis. Methods 124 chronic gastritis patients were measured by the Chronic Gastritis scale of Quality of Life Instruments for Chronic Disease (QLICD-CG) before and after treatments. The scores were compared by paired t-test,and the influencing factors were analyzed by independent sample t-test and One-Way ANOVA. Results There were significant differences in all domains and total scores between the before and after treatments. The quality of life of chronic gastritis patients were influenced by treatments,gender,marital status,occupation and economic conditions ( <0.05) . Conclusion While reasonable treatment is very important to improve quality of life of the chronic gastritis patients, some influencing factors should be addressed, and mental health education and psychological service should be strengthened.
5.Psoralen induces rat mesenchymal stem cell differentiation towards male germ cells in vitro.
Qian WAN ; Hua LU ; Yanli DENG ; Limei LIANG
Journal of Southern Medical University 2013;33(12):1723-1726
OBJECTIVETo test the hypothesis that psoralen induces rat mesenchymal stem cells (RMSCs) to differentiate towards male germ cells.
METHODSRMSCs were induced by psoralen at the final concentration of 3.0 µg/ml, and the morphological changes of the cells were detected microscopically and the cell proliferation changes were measured by MTT assay. The expressions of 7 representative marker genes of male germ cells, namely Oct-4, Stra8, RVLG, SCP3, TNP2, Itgb1, and Itga6, were investigated in the induced cells by real-time quantitative PCR.
RESULTSRMSCs showed no obvious morphological changes after induction with 3.0 µg/ml psoralen for 72 h. Psoralen induction for 5 days did not significantly affect the proliferation of RMSCs; Psoralen induction for 72 h, however, significantly up-regulated the genes Oct-4, Stra8, SCP3, and Itgb1 (P<0.05) without affecting the expressions of the genes RVLG, TNP2, or Itga6 (P>0.05).
CONCLUSIONPsoralen may participate in the differentiation of RMSCs towards male germ cells.
Animals ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Ficusin ; pharmacology ; Germ Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Rats ; Up-Regulation
6.Study on the status quo and correlation between body image and stigma of drug-resistant tuberculosis patients treated with Clofazimine
Fanghui XIE ; Limei LEI ; Rong YAO ; Xia ZHAO ; Bin WAN ; Meng MENG
Chinese Journal of Practical Nursing 2023;39(7):486-491
Objective:To investigate the current situation of body image and stigma of drug-resistant tuberculosis patients treated with Clofazimine, and analyze the correlation between them.Methods:A cross-sectional study was conducted using convenience sampling method to investigate 150 patients with drug-resistant tuberculosis treated with Clofazimine in tuberculosis ward of Chengdu Public Health Clinical Medical Center from October 2020 to October 2021. The general questionnaire, Body Image Scale (BIS) and Tuberculosis Related Stigma Scale were used to conduct a questionnaire survey.Results:A total of 130 questionnaires were effectively collected. The body image score of 130 patients with drug-resistant tuberculosis treated with Clofazimine was (20.51 ± 6.80) points; the score of stigma was (17.78 ± 6.92) points. There was a positive correlation between the total score of disease shame and the total score of body image ( r=0.544, P<0.05). Conclusions:Patients with drug-resistant tuberculosis treated with Clofazimine have body image disorder and stigma, and the two are positively correlated. Caregivers should carry out psychological assessment and intervention at an early stage to improve the patient′s mental health level.
7.Three-month Follow-up Study of Survivors of Coronavirus Disease 2019after Discharge
Limei LIANG ; Bohan YANG ; Nanchuan JIANG ; Wei FU ; Xinliang HE ; Yaya ZHOU ; Wan-Li MA ; Xiaorong WANG
Journal of Korean Medical Science 2020;35(47):e418-
Background:
Most patients including health care workers (HCWs) survived the coronavirus disease 2019 (COVID-19), however, knowledge about the sequelae of COVID-19 after discharge remains limited.
Methods:
A prospectively observational 3-month follow-up study evaluated symptoms, dynamic changes of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) IgG and IgM, lung function, and high resolution computed tomography (HRCT) of survivors of COVID-19 after discharge at Wuhan Union Hospital, China.
Results:
Seventy-six survivors (55 females) with a mean age of 41.3 ± 13.8 years were enrolled, and 65 (86%) were HCWs. A total of 69 (91%) patients had returned to their original work at 3-months after discharge. Most of the survivors had symptoms including fever, sputum production, fatigue, diarrhea, dyspnea, cough, chest tightness on exertion and palpitations in the three months after discharge. The serum troponin-I levels during the acute illness showed high correlation with the symptom of fatigue after hospital discharge (r = 0.782; P = 0.008) and lymphopenia was correlated with the symptoms of chest tightness and palpitations on exertion of patients after hospital discharge (r = −0.285, P = 0.027; r = −0.363, P = 0.004, respectively). The mean values of forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC), FEV1/FVC, total lung capacity and diffusion capacity were all normal (> 80% predicted) and lung HRCTs returned to normal in most of the patients (82%), however, 42% of survivors had mild pulmonary function abnormalities at 3-months after discharge. SARS-CoV-2 IgG turned negative in 11% (6 of 57 patients), 8% (4 of 52 patients) and 13% (7 of 55 patients), and SARS-CoV-2 IgM turned negative in 72% (41 of 57 patients), 85% (44 of 52 patients) and 87% (48 of 55 patients) at 1-month, 2-months and 3-months after discharge, respectively.
Conclusion
Infection by SARS-CoV-2 caused some mild impairments of survivors within the first three months of their discharge and the duration of SARS-CoV-2 antibody was limited, which indicates the necessity of long-term follow-up of survivors of COVID-19.
8.Psoralen induces rat mesenchymal stem cell differentiation towards male germ cells in vitro
Qian WAN ; Hua LU ; Yanli DENG ; Limei LIANG
Journal of Southern Medical University 2013;(12):1723-1726
Objective To test the hypothesis that psoralen induces rat mesenchymal stem cells (RMSCs) to differentiate towards male germ cells. Methods RMSCs were induced by psoralen at the final concentration of 3.0 μg/ml, and the morphological changes of the cells were detected microscopically and the cell proliferation changes were measured by MTT assay. The expressions of 7 representative marker genes of male germ cells, namely Oct-4, Stra8, RVLG, SCP3, TNP2, Itgb1, and Itga6, were investigated in the induced cells by real-time quantitative PCR. Results RMSCs showed no obvious morphological changes after induction with 3.0μg/ml psoralen for 72 h. Psoralen induction for 5 days did not significantly affect the proliferation of RMSCs; Psoralen induction for 72 h, however, significantly up-regulated the genes Oct-4, Stra8, SCP3, and Itgb1 (P<0.05) without affecting the expressions of the genes RVLG, TNP2, or Itga6 (P>0.05). Conclusion Psoralen may participate in the differentiation of RMSCs towards male germ cells.
9.Estimation of the number of workers with occupational noise-induced hearing loss from manufacturing industry in China, 2020
Limei AI ; Xinxin LI ; Anqi LIU ; Dan WANG ; Jingxin WANG ; Weijiang HU ; Xia WAN
China Occupational Medicine 2023;50(5):512-517
{L-End}Objective To estimate the number of workers with occupational noise-induced hearing loss (ONHL) in the manufacturing industry of China in 2020. {L-End}Methods Multiple data sources were integrated to collect information on workers in the manufacturing industry from the Occupational Diseases and Hazards Monitoring Information System under China Disease Prevention and Control Information System. The total number of workers was split based on age from the 2018 Fourth National Economic Census and the 2020 National Survey of Occupational Disease Hazards. Additionally, data on the prevalence of hearing loss in the general population was adjusted based on the age composition of Jilin Province, and noise prevalence was standardized based on the age composition of the employed population in the Seventh National Population Censu. Attribution fractions (AF) was estimated. The prevalence ratio (PR) was calculated by the prevalence of hearing loss in the occupational noise-exposed workers and general population. The proportion and attributable cases (AC) of ONHL cases in all hearing loss cases were estimated for occupational noise-exposed workers. The number of ONHL was estimated based on AF and the total number of workers with hearing loss. {L-End}Results In 2020, a total of 30 059 525 workers were exposed to occupational noise in the manufacturing industry in China, with noise-exposed prevalence and standardized noise-exposed prevalence of 28.94% and 28.52%, respectively. The prevalence of hearing loss among occupational noise-exposed workers increased with age, and a total of 8 054 789 workers suffered from hearing loss. Most of the cases were at the age between 45-<55 years old. The total PR and 95% uncertainty interval (UI) was 2.83 (2.58-3.06) and the total AF and 95%UI was 64.63% (61.22%-67.30%), and both peaks were in the age of 30-<45 years. The AC and 95%UI was 5 205 980 (4 930 620-5 420 345) persons, and most of the cases were at the age between 40-<55 years. {L-End}Conclusion Occupational noise poses a serious threat to the hearing health of workers in the manufacturing industry of China, especially among middle-aged and young adults.
10.Psoralen induces rat mesenchymal stem cell differentiation towards male germ cells in vitro
Qian WAN ; Hua LU ; Yanli DENG ; Limei LIANG
Journal of Southern Medical University 2013;(12):1723-1726
Objective To test the hypothesis that psoralen induces rat mesenchymal stem cells (RMSCs) to differentiate towards male germ cells. Methods RMSCs were induced by psoralen at the final concentration of 3.0 μg/ml, and the morphological changes of the cells were detected microscopically and the cell proliferation changes were measured by MTT assay. The expressions of 7 representative marker genes of male germ cells, namely Oct-4, Stra8, RVLG, SCP3, TNP2, Itgb1, and Itga6, were investigated in the induced cells by real-time quantitative PCR. Results RMSCs showed no obvious morphological changes after induction with 3.0μg/ml psoralen for 72 h. Psoralen induction for 5 days did not significantly affect the proliferation of RMSCs; Psoralen induction for 72 h, however, significantly up-regulated the genes Oct-4, Stra8, SCP3, and Itgb1 (P<0.05) without affecting the expressions of the genes RVLG, TNP2, or Itga6 (P>0.05). Conclusion Psoralen may participate in the differentiation of RMSCs towards male germ cells.