1.The Ras/Erk pathway in the specially activated human ??T cells by Mycobacterium tuberculosis low molecular peptide antigen
Yanqiang HOU ; Baiqing LI ; Jianguo HU
Chinese Journal of Immunology 2000;0(08):-
Objective:To investigate involvement of the Ras/Erk signal pathway in activation of human ??T cells stimulated by Mycobacterium tuberculosis low molecular peptide antigen(Mtb-Ag).Methods:PBMC were isolated from peripheral blood of healthy subjects and simulated in vitro with Mtb-Ag,CD3mAb or PMA and inomysin(IM).CD69 expression of total T cells and ??T cells were measured by flow cytometry at different hours after stimulation, and the number of expanded ??T cells were counted after 10 days of culture. PD98059,a specific inhibitor for Erk pathway was used to treat PBMC for 30 min before stimulation.Results:The proportions of CD69 expressing cells in total T cells and ??T cells were same as 99% at 6 h after stimulation of PBMC with PMA +IM and similar about 55% at 24 h after stimulation with CD3 mAb,whereas those at 24 h stimulation of PBMC with Mtb-Ag were 75.2% and 16.0%,respectively.CD69 expression and proliferation of ??T cells activated by Mtb-Ag were markedly inhibited by pretreatment of PBMC with PD98059.Conclusion:Mtb-Ag is a specific stimulator for activation of ??T cells, and Ras/Erk signal transduction pathway involves in the activation of ??T cells.
2.Advances in CncRNAs research
Yanzhuo WANG ; Xiaoli LOU ; Yanqiang HOU
Chinese Journal of Laboratory Medicine 2017;40(7):540-543
RNAs are known to regulate diverse biological processes, either as protein-encoding molecules or as non-coding RNAs.However, a new class of bi-functional RNAs, carrying both protein coding and RNA-intrinsic functions, have been identified and termed as 'CncRNAs'.This review discuss three major genomic sources of CncRNAs and describe the dual characteristics and functional mechanisms of them, providing a new perspective to further understand the complexity of the transcriptomics and genomes and the complex gene-regulatory network in organisms.
3.The Roles of Granule Exocytosis and Fas/Fas Ligand Pathway in the Cytotoxicity of Mycobacterium Tuberculosis Antigen Activated Human ??T Cells to Tumor Cell Lines
Jianguo HU ; Yanqiang HOU ; Yong CHEN ; Baiqing LI
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective: To investigate the mechanisms of anti-tumor cytotoxicity of human ??T cells activated by Mycobacterium tuberculosis antigen (Mtb-Ag) . Methods: The peripheral blood mononuclear cells (PBMC) of healthy donors were stimulated by Mtb-Ag in vitro to activate preferentially ??T cells that were expanded in IL-2 contained medium.The high purified ??T cells were isolated by the positive selection on MACS separator. The expression of mRNA of perforin, granzyme B and Fas Ligand in purified ??T cells were measured by RT-PCR technique. Fas Ligand on ??T cells and Fas molecule on tumor cells were detected by flow cytometry. The MTT assay was used to measureanti-tumor cytotoxicity of ??T cells. The inhibitory effects of concanamycin A (CMA) for perforin/granzyme pathway, and brefeldin A (BFA) for Fas/Fas ligand pathway on cytotoxicity of ??T cells were observed. Results: The mRNA of perforin and granzyme B and Fas ligand were highly expressed in purified ??T cells. The cytotoxicity of ??T cells against K562 and PG that expressed low level of Fas molecule (6.5% and 7.8%) was remarkably inhibited by pretreatment of the ??T cells with CMA (200 nmol/L) by 72% and 76%, respectively, while the BFA(30 ?mol/L) showed less inhibitory effects by 32% and 25%, respectively; However, it was showed that the BFA (30 ?mol/L) preferentially inhibited the cytotoxicity of ??T cells against tumor cell lines Raji and A375 that expressed high level of Fas molecule on cell surface (98.5% and 70.6%, respectively).Conclusion: Preferentially, granule exocytosis was a main mechanism by which the Mtb-Ag activated ??T cells showed cytotoxicity aganist tumor cells that expressed low level of Fas molecule, whereas both Fas/Fas ligand pathway and granule exocytosis were involved in the cytotoxicicty of Mtb-Ag activated ??T cells against the tumor cells that expressed high level of Fas molecule.
4.Analysis on mycoplasma infection status in genital tract and its drug susceptibility
Jinghong PENG ; Jun LIU ; Guoying ZHANG ; Weizhong ZHUANG ; Yanqiang HOU
International Journal of Laboratory Medicine 2014;(13):1738-1739
Objective To understand the status of mycoplasma infection and drug susceptibility to provide reference basis for clinical rational drug use.Methods The reagents kits produced by Shanghai Aopu Biopharmaceutical Co.,Ltd.were adopted to per-form the mycoplasma culture,identification and susceptibility test.Results Among 1 745 specimens,896 cases of mycoplasma in-fection were detected with the total detection rate of 51 .35%,including Ureaplasma urealyticum (Uu)infection in 841 cases (48.19% ),Uu combined Mycoplasma hominis(Mh)infection in 92 cases(5.27% )and Mh infection in 17 cases(0.97%).The de-tection rate was 9.46% in males and 41 .89% in females,the infection rate in females was obviously higher than that of males.The results of drug susceptibility test showed that the sensitive rate to doxycycline,josamycin and minocycline was higher than 95%, while the resistant rate to norfloxacin and lincomycin was higher than 75%.Conclusion Mycoplasma infection in this hospital is dominated by Uu.Doxycycline,josamycin and minocycline can be used as the empirical medication to treat mycoplasma infections in this hospital.But before clinical treatment,the mycoplasma culture and the drug susceptibility test should be performed as possible for rational and standardized medication.
5.A preliminary analysis of plasma microRNA expression profile in patients with primary biliary cirrhosis
Dongyu LIANG ; Deyong GAO ; Xiaoli LOU ; Yanqiang HOU
Chinese Journal of Laboratory Medicine 2014;37(8):608-612
Objective To evaluate the expression and clinical significance of miRNA in plasma of patients with primary biliary cirrhosis.Methods Plasma from 19 PBC patients,10 healthy volunteers and 10 viral hepatitis patients were selected from Shanghai Songjiang Hospital from december 2010 to January 2013.Among them 3 PBC patients' plasma and 3 healthy volunteers' plasma were detected by miRNA microarray for miRNA expression profile examination.Real-time PCR was used to verify the results of microarray,miRNA target gene predictior software was used to predict the target genes of differentially expressed miRNA.ROC was used to determine the clinical value of plasma miRNA.Results According to microarray,a total of 16 miRNAs were found to be differentially expressed.As revealed by qRT-PCR,the expression of miRNA-92a-3p and miRNA-4516 decreased while the expression of miRNA-572 and miRNA-575 were up-regulated in PBC group compared with healthy controls (P < 0.05).In comparison with nonPBC cirrhosis group,only miRNA-92a-3p and miRNA-4516 were down-regulated (P < 0.05).The area under the curve (AUC)of miRNA-92a-3p for the diagnosis and differential diagnosis of PBC were 0.92 and 0.84,respectively.while for The area under the ROC curve of miRNA-4516,the AUC for diagnosis and differential diagnosis PBC were 0.89 and 0.76,respectively.The optimal cut-off values for identifying PBC from healthy controls were defined as 1.26 ng/μl.for miRNA-92a-3p (sensitivity,92% ;specificity,80%)and 1.16ng/ul for miRNA-4516 (sensitivity,85% ;specificity,70%)respectively.The optimal cut-off values for identifying PBC from viral hepatitis were defined as 1.08 ng/μl.for miRNA-92a-3p (sensitivity,89% ; specificity,81%)and 1.06 ng/μl for miRNA-4516 (sensitivity,77% ;specificity,68%).Conclusion The results indicate that plasma from patients with PBC has a unique miRNA exprssion profile and these differentially expressed miRNA can be used as clinical biomarkers of PBC.
6.Study of the serum quality controlling method
Xiaoli LOU ; Wenhua SUN ; Yanqiang HOU ; Hongwei CHEN ; Shanyuan MA ; Liang PENG
International Journal of Laboratory Medicine 2014;(22):3110-3111
Objective To improve the serum quality controlling level ,in order to regularize and standardize the laboratory processes .Methods Gallery of hemolysis ,lipidemia or icterus serum specimens was established by Roche cobas p612 QSI camera system .The trouble serum specimens were screened out through taking photos of interfere serum and comparing with the gallery . The serum indexes of hemolysis ,lipidemia and icterus were detected respectively as reference information of inspection report .Com‐paring the results of 16 710 specimens detected by system screening and visual judgment ,the accuracy of QSI camera system was verified .Results 195 trouble serum pictures were elected in the gallery .The compliance rates of hemolysis ,lipidemia and icterus se‐rum specimen between QSI camera system and visual judgment were 94 .2% ,87 .4% ,and 60 .9% respectively .Conclusion The QSI camera system can replace visual judgment to screen the trouble serum specimens .
7.The efficacy and security of adenosine combined tirofiban on patients with percutaneous coronary intervention
Yanqiang WU ; Fang YUAN ; Qiang FENG ; Hefei LI ; Lin ZHU ; Aijun HOU ; Yuliang SHEN
Clinical Medicine of China 2014;30(12):1271-1274
Objective To compare the egicacy and security of intracoronary administration of tirofiban combined high-dose adenosine during percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (STEMI).Methods Eighty-eight cases with STEMI were randomly divided into observation group(44 cases) who were accepted 2 times intracoronary adenosine(2 mg,10 ml 0.9% NaCl),and control group(44 cases) who were afforded only 10 ml 0.9% NaCl by prospective,double-blind,and random study.The two groups were received10 g/kg tirofiban after aspiration catheter in the culprit lesion distal bolus injection of 3 rain,at the same time,continuous infusion of 0.15 g/(kg · min) for 24 h.The postoperative coronary arteriography and electrocardiogram were evaluated.Meanwhile,the postoperative myocardial blush grade(MBG),thrombolysis in myocardial infarction (TIMI),corrected TIMI frame counts (CTFC),ST-segment elevation resolution (STR) major adverse cardiac events (MACE),and adverse reactions of adenosine were recorded.Results There was no significant difference in terms of postoperative TIMI and STR between two groups (P > 0.05).The CTFC of observation group was (24.4 ± 4.9) frames,significant better than that of control group((21.9 ±3.7) frames;t =2.701,P <0.01).The ratio of MBG in observation group was 24/44,higher than that of control group(14/44 ; x2 =4.632,P < 0.05).There were no significant difference regarding of the ratio of death,MACE,target vessel revascularization,grade of NYHA between observation and control group at followed up for 1 and 12 month (P > 0.05).The ratio of patients with blood pressure decrease ≥ 10 mm Hg,new second degree atrioventricular block in observation group were 15.9% and 20.5%,higher than that in control group (2.3% and 15.9% ; x2 =4.950,7.221 ; P =0.026,0.007).The adverse reaction was transient.Conclusion The intracoronary administration of tirofiban combined high-dose adenosine during PCI in patients with STEMI plays an effective role on improvement of myocardial perfusion.
8.Intrapulmonary rotational power-driven thrombectomy therapy for acute massive pulmonary thromboembolism
Yuliang SHEN ; Lingling LIU ; Yufeng GUO ; Yanqiang WU ; Fang YUAN ; Aijun HOU
Clinical Medicine of China 2013;29(9):978-980
Objective To investigate the effect and safety of rotational power-driven thrombectomy therapy through intrapulmonary for acute massive pulmonary thromboembolism.Methods Sixteen patients of acute massive pulmonary thromboembolism diagnosed by CT and pulmonary angiography were treated with Straub Rotarex system.The successful rate,release of clinical manifestations and the blood hemodynamic changes were observed and analyzed.Results The clinical manifestations were improved remarkably in all the 16 patients,arterial partial pressure of oxygen,saturation of arterial blood oxygen,shock index,Miller score and mPAP were (56.7± 13.4) mm Hg,84.1 ± 10.4)%,(1.27 ±-0.39),(22.7±11.4) and (36.3 ±9.4) mm Hg respectively before treatment,and (92.2 ± 8.6) mm Hg,(96.6 ± 12.7) %,(0.57 ± 0.42),(12.1 ± 7.8)points and (21.9 ± 7.3) mm Hg respectively after treatment,which were all improved significantly (t =-2.794,2.601,-2.592,-2.638,-2.617,P < 0.01).Conclusion Rotational power-driven thrombectomy therapy through intrapulmonary is an effective and safe technique for the treatment of acute massive pulmonary embolism.
9.Effect of Persistent Atrial Fibrillation on Circadian Rhythm of Blood Pressure in Patients With Essential Hypertension
Jianjun DENG ; Guangyan MEI ; Ping XIE ; Xiaohuan WANG ; Tingting HOU ; Yanqiang AN ; Zhongwei LI ; Wenjing ZHANG
Chinese Circulation Journal 2015;(10):981-983
Objective: To explore the effect of persistent atrial ifbrillation (AF) on circadian rhythm of blood pressure (BP) in patients with essential hypertension (EH).
Methods: A total of 173 EH patients treated in Gansu Provincial Hospital from 2013-02 to 2014-01were studied. The patients were divided into 2 groups: EH group,n=88 and Persistent AF combining EH group,n=85. The baseline information was studied and the risk factors of persistent AF combining EH were investigated by multivariate logistic regression analysis.
Results: Compared with EH group, the Persistent AF combining EH group showed decreased average daytime DBP, minimum daytime SBP, minimum daytime DBP and the average 24-hour DBP, while increased maximum nighttime SBP and the percentage of reverse dipper in DBP, allP<0.05. There were no significant differences for the average of daytime SBP, maximum daytime SBP, maximum daytime DBP, the average 24-hour SBP, average nighttime SBP, average nighttime DBP, maximum nighttime DBP, minimum nighttime SBP, minimum nighttime DBP and the percentage of reverse dipper in SBP between 2 groups, allP>0.05. Multivariate logistic regression analysis indicated that the maximum nighttime SBP was obviously related to persistent AF combining EH (OR=1.038, 95 CI 1.014-1.062,P=0.001).
Conclusion: Persistent AF may incur daytime BP dropping, such change was not obviously observed for nighttime BP in EH patients.
10. Hepatitis B virus x protein promote human hepatocellular carcinoma cells proliferation by activating p38 and JNK signalling pathway
Chinese Journal of Experimental and Clinical Virology 2019;33(4):394-399
Objective:
To investigate the effects and mechanism of hepatitis B virus x protein (HBx) on human hepatocellular carcinoma cells proliferation.
Methods:
Eukaryotic expression vector HBx-pEGFP-C1 was constructed. HepG2 cells were transfected transiently using Lipofectamine 2000. HBx expression in transfected cells were measured by RT-PCR and Western blot. Cells proliferation and apoptosis were detected by using growth curves and TUNEL staining. The protein levels of caspase-3, p-p38, p-Akt and p-JNK were measured by Western blot.
Results:
HBx was successfully expressed in HepG2 cells. Growth curve result showed that HBx promoted cell proliferation (