2.Effect of SOCS3 on progress of liver fibrosis and reversein mice induced by carbon tetrachloride and its mechanism
Jialu XIA ; Xing YAN ; Yaru LIU ; Cheng HUANG ; Jun LI
Chinese Pharmacological Bulletin 2017;33(9):1215-1221
Aim To observe the effects of cytokine signaling inhibition protein-3(SOCS3) on the liver fibrosis progression and reverse.Methods C57BL/6 mouse model was established by subcutaneous injection of carbon tetrachloride(CCl4).After a successful model of fibrosis, one-month normal diet was given to induce the reverse fibrosis model, while normal mice of the same gender and weight were as control group.Mice were sacrificed at 1, 2, 3, 4, 5, 6, 7, 8 weeks, respectively, then the liver tissue was harvested for the observation of its injury by hematoxylin and eosin(HE) staining.Then Masson staining was applied for the detection of changes in collagen, and the immunohistochemistry(IHC) for the observation of type Ⅰ Collagen(Colla-1), alpha smooth muscle actin(α-SMA), transforming growth factor beta 1(TGF-β1) and SOCS3 protein expression.In vitro formation of fibrosis was induced by TGF-β1 stimulating HSC-T6 cell lines, which was then reversed by MDI medium, with co-incubation of HSC-T6 cells with plasmid in the process of the reverse.Western blot was employed to detect SOCS3, Colla-1, α-SMA, TGF-β1 expression.Results The expression of SOCS3 and TGF-β1 increased in mouse model of fibrosis with the worsening fibrosis process and decreased in the reverse process.Over-expression SOCS3 in the reverse process reduced the development of liver fibrosis;meanwhile, the expression of TGF-β1 was also reduced accordingly.Conclusion SOCS3 may influence the development of the liver fibrosis and its reverse via regulating the expression of TGF-β1.
3.Effects of deoxynivalenol on apoptosis of human gastric carcinoma cell line SNU in vitro
Ring LIU ; Xin XING ; Lingxiao XING ; Bingjuan ZHOU ; Xia YAN ; Junling WANG ; Yuehong LI ; Xianghong ZHANG
Cancer Research and Clinic 2009;21(5):295-297
Objective To explore the effects of deoxynivalenol (DON) on apoptosis of human gastric carcinoma cell line SNU in vitro. Methods SNU cells were treated with DON at different concentrations (50, 100, 1000, 2000 μg/L) for 12 hours, and then cells were harvested for cell apoptosis by flow cytometric (FCM) DNA analysis and the expression of Bax, Bcl-2 and Caspase-3 at protein level with FCM and Western blotting. Results FCM results showed that the apoptosis rates of SNU cells in DON treatment groups were all higher than that in control, especially in DON 1000 μg/L and 2000 μg/L groups (P<0.05). In the concentration range from 50 to 2000 μg/L, a significant concentration-depended response correlation could be found between apoptosis rate and DON concentration (r =0.940, P <0.01). FCM and Western blotting showed Bax and Caspase-3 expression in often SNU cells DON treatment for 12 hours were up-regulated while that of Bcl-2 was down-regulated. Conclusion DON can induce apoptosis of SNU cells in vitro in dose-dependent manner, and possible mechanisms of apoptosis induction effects may be up-regulation of the expression of Bax and down-regulation of that of Bcl-2 and activation of the key enzyme of apoptosis Caspase-3.
4.Role of Sophocarpine in Coxsackievirus B3m Infection in vitro
xing-yu, ZHANG ; shu-xia, CHEN ; jing-xing, LIU ; fu-rong, QIAN
Journal of Shanghai Jiaotong University(Medical Science) 2006;0(08):-
Objective To observe the in vitro anti-Coxsackievirus B3m (CVB3m) effect of sophocarpine(SC) extracted from Sophora flavescens, a traditional Chinese herb. Methods HeLa cells were cultured and the micro-dose cytopathic effect (CPE) assays were applied to detect the toxicity of SC. CPE-inhibitory assays were used to observe the in vitro anti-CVB3m effect of SC. MTT and crystal assays were introduced to examine the anti-CVB3m effect of SC. HeLa cells were infected with CVB3m and added with SC in different concentrations 15 h later.The viability and number of survival of HeLa cells were determined by MTT and crystal violet assays, respectively. Results No toxicity was found on HeLa cells by SC with concentrations 100 ?g/mL, SC could accelerate and aggravate the CPE. SC could protect the CVB3m-infected HeLa cells with concentrations from 1.56 to 25 ?g/mL, and the viability and cell number measured by MTT and crystal violet assay in the SC-handled cells were higher and bigger than those in the virus infected ones. However, the inhibitory effect of virus was exacerbated with higher concentrations (50 and 100 ?g/mL), and the cell number and viability of the SC-handled cells were smaller and lower than those of the infected ones. Conclusion SC with a proper concentration has the in vitro anti-CVB3m effect and may protect HeLa cells from CVB3m infection.
5.Antibiotic resistance phenotypes and genotypes of ESBLs and AmpC ?-lactamases from Klebsiella oxytoca
Wen-Li ZHANG ; Ding-Xia SHEN ; Xing-Ping LI ; Yan-Ping LUO ; Jun LIU ; Xin-Yu WANG ; Xing-Hai SHI ;
Chinese Journal of Laboratory Medicine 2000;0(06):-
Objective To study antibiotic resistance phenotypes and genotypes of extended spectrum ?-lactamases (ESBLs) and AmpC ?-lactamase-producing Klebsiella oxytoca isolated from specimens of respiratory tract in children.Methods Bacterial isolates were identified by API or VITEK32. Agar dilution was used for antibiotic susceptibility test,and ESBLs and AmpC were detected by confirmatory test recommended by CLSI/NCCLS and by 3-aminophenylboronic acid (APB) disk potentiation test, respectively.Microarray was used to determine the genotypes of ESBLs and AmpC ?-lactamases.Genotypes of Klebsiella oxytoca were determined by enterobacterial repetitive intergenic consensus (ERIC)- PCR.Results ESBLs were positive in 129 out of 165 isolates (78.2%).Both ESBLs and AmpC ?- lactamases were positive in 16 out of 165 isolates (9.7%).AmpC ?-lactamase alone producer was not detected in term of phenotype and genotype.CTX-M was the most common type of ESBLs and DHA was the only type of AmpC ?-lactamase in these isolates.Most antibiotic resistant strains of Klebsiella oxytoca possessed the same genotype by ERIC-PCR.Although all strains were susceptible to carbpenem,Klebsiella oxytoca with ?-lactamases were more resistant to other antibiotic agents than those without ?- lactamases.Conclusions There is high prevalence of ESBLs production among Klebsiella oxytoca isolated from children in Urumqi.The main genotypes of ESBLs and AmpC ?-lactamases are CTX-M and DHA.
6.The efficacy and safety of autologous cryopreserved platelet transfusion in management of thrombocytopenia after chemotherapy in hematological malignancy
Guoqiang LIU ; Huifang DING ; Xifing LU ; Min XU ; Jian XING ; Xia ZHAO ; Fang HAN ; Yinghui SHANG
Chinese Journal of Internal Medicine 2012;51(3):188-191
Objective To investigate the efficacy and safety of autologous cryopreserved platelet transfusion in the management of thrombocytopenia after chemotherapy in hematological malignancy.Methods A total of 40 patients diagnosed as hematological malignancy with complete remission were equally assigned into study group and control group.During chemotherapy interval in the study group,when platelet counts exceeded 120 × 109/L,autologous platelets were collected with CS3000 Cell Separator and cryopreserved at-80℃ with 5% dimethylsulfoxide.When platelet counts dropped below 15 × 109/L after chemotherapy,autologous platelets were thawed with 40℃ water bath and transfused back to each patient.In the control group,when platelet counts dropped below 15 × 109/L after chemotherapy,allogeneic fresh platelets were transfused.Median loss during the freeze-thaw-wash procedure in study group was observed,and the 1 h,24 h corrected count increments(CCI)were calculated in the both groups.The hemostatic effects and adverse reactions were also observed.Results In the control group,1hCCI and 24hCCI were (19.3 ±6.1)× 109/L and(12.2 ± 7.0)× 109/L,respectively,with the effective rate of 80% and the transfusion reaction rate of 45%.Totally 20 collection and transfusions were finished in the study group.A total of(3.4-8.5)× 1011 platelet were obtained in each collection.Platelet recovery after freezing and thawing was(73.51 ±9.03)%(62%-83%).1hCCI was(17.4±7.6)× 109/L,24h CCI was(10.5 ±5.8)× 109/L and the effective rate was 85%.There was no significant different between the two groups (P > 0.05).The transfusion reaction rate was 15 %,which was significantly lower than that of the control group(P < 0.05).Meanwhile,adverse reactions were occurred less in the study group.Conclusion This study demonstrates that autologous cryopreserved platelet transfusions can be safely administered for supporting thrombocytopenia in hematological malignancy patients undergoing chemotherapy.
7.In vitro co-culture of human adipose-derived mesenchymal stem cells and T-lymphocytes from patients with aplastic anemia
Liang WANG ; Min XU ; Muhua ZHANG ; Jian XING ; Xia ZHAO ; Fang HAN ; Guoqiang LIU
Chinese Journal of Tissue Engineering Research 2014;(10):1603-1608
BACKGROUND:Clinical infusion of hematopoietic stem cells and mesenchymal stem cells for treatment of aplastic anemia has been reported. OBJECTIVE:To investigate the effect of human adipose-derived mesenchymal stems cells on the secretion function of T lymphocytes of aplastic anemia patients. METHODS:Human adipose-derived mesenchymal stems cells were extracted from healthy human adipose tissues. T-lymphocytes were harvested from peripheral blood of patients with aplastic anemia by density gradient centrifugation. Human adipose-derived mesenchymal stems cells were co-cultured with T-lymphocytes. The levels of interleukin-2, interleukin-4, interleukin-10 and interferon-γwere detected by enzyme linked immunosorbent assay. T-bet and GATA-3 levels were examined by real-time PCR and western blot. RESULTS AND CONCLUSION:The levels of Th1 type cytokines interferon-γand interleukin-2 in the co-culture group were significantly lower than those in the T-lymphocyte group (P<0.05). But the levels of Th2 type cytokines interleukin-4 and interleukin-10 in the co-culture group were significantly higher than those in the T-lymphocyte group (P<0.05). The T-bet mRNA and protein levels in the co-culture group were significantly lower than those in the T-lymphocyte group, while the GATA-3 mRNA and protein levels were significantly higher in the co-culture group. Human adipose-derived mesenchymal stems cells can mediate an immunoregulation effect on T-lymphocytes of aplastic anemia patients in vitro, which is possibly related with the inhibition of Th1-dominant response due to the disorder of T-bet and GATA-3 gene expression.
8.Effect of combination of bone marrow stromal cells and bcl-2 gene therapy on expression of basic fibroblast growth factor in focal cerebra ischemic rat brain
Yanhui XING ; Zhiyi HE ; Xiaotian ZHANG ; Xia FU ; Lei LI ; Liping LIU
International Journal of Cerebrovascular Diseases 2009;17(2):85-91
Objective To investigate the efficacy of the combination of bone marrow stromal cells (BMSCs) and bcl-2 gene in the treatment cerebral ischemia taxi its effect on the expression of basic fibroblast growth factor (bFGF) in rats.Methods Forty Wistar rats were used to establish middle cerebral artery occlusion model. They were randomly divided into 4 groups: saline control, bcl-2, BMSCs and BMSCs + bcl-2 groups (n = 10 in each group). Every group was redivided into 3- and 14-day after reperfusion subgroups (n = 5 in each subgroup). Neurological scores of the experimental rats were assessed. BMSCs were labeled with bromodeoxyuridine (BrdU). The distribution and numbers of BMSCs, the expressions of Bcl-2 and bFGF were detected by immunohistochemistry, and apoptotic cells were detected with TUNEL staining in rat brain. Results The neurological score at day 3 after reperfusion in the BMSCs + bcl-2 group was significantly lower than that in the saline control group (P < 0. 05), and at day 14, it was significantly lower than that in the other 3 groups (all P <0. 05). A large number of BrdU-positive BMSCs were observed in the infarcted hemisphere in the BMSCs + bcl-2 and BMSCs groups. The numbers of BrdU-positive BMSCs at day 3 and 14 after reperfusion in the BMSCs + bcl-2 group were significantly higher than those in the BMSCs group (all P <0. 05). The expressions of Bcl-2 in the infarcted hemisphere at day 3 and 14 after reperfusion in BMSCs +hel-2 group wre significantly higher than those in the other 3 groups (all P <0. 05). The expressions of Bcl-2 at all time points were increased more significantly than those in the other 3 groups (all P <0. 05). The numbers of apoptosis in brain at all time points in the BMSCs + bcl-2 group were decreased more significantly than those in the other 3 groups (all P <0. 05). Conclusions Both BMSCs and bcl-2 genes have the therapeutic effect on cerebral ischemia. The efficacy of combination of both ,of them is significantly superior to monotherapy. They may significantly improve the neurological function and increase the expression of bFGF in rats. Its mechanism may be that bcl-2 genes have inhibited BMSCs apoptosis at the same time of anti-apoptosis in brain.
9.Effect on cellular immune functions in peripheral blood of immunotherapy of dendritic cell and cytokine-induced killer cell combined with chemotherapy for multiple myeloma
Xia ZHAO ; Qing XU ; Huifang DING ; Min XU ; Jian XING ; Guang LU ; Caifeng SUN ; Guoqiang LIU
Chinese Journal of Immunology 2015;(4):490-496
Objective:To valuate the treatment value and analyse the effect on the cellular immune functions by studying the differences of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood after adoptive immunotherapy ( dendritic cells and cytokine-induced killer cells,DC-CIK) combined with chemotherapy on MM.Methods:50 patients with MM were randomly divided into two groups.24 patients in chemotherapy group were treated by chemotherapy only,26 patients in joint group were treated by adoptive immunotherapy( DC-CIK) combined with chemotherapy,and the clinical outcomes and the levels of T-lymphocyte subsets and CD4+CD25+Treg cells in peripheral blood between two groups were compared.Moreover,the differences of cellular immune indicators (Th1/Th2,the ratio of AgNOR,and TGF-β)between two groups were also compared.Results: After treatment,quality of life,clinical index and survival in joint group were better than in chemotherapy group( P<0.05);the proportion of CD3+CD8+,the ratios of CD4+CD25+,CD4+CD25+/CD4+and the level of TGF-βof joint group wes clearly lower than chemotherapy group(P<0.05),and the ratios of CD3+CD4+/CD3+CD8+, Th1/Th2 and AgNOR of joint group wes clearly higher than chemotherapy group .Conclusion: DC-CIK combined with chemotherapy could be an effective and promising treatment to patients with MM,and it maybe strengthen the anti-tumor action of bodies by regulating the balance between Th1 and Th2 reaction.
10.The application of improved CHQS for mass epidemiology study on hearing impairment
Cheng LIU ; Guangqian XING ; Xia XU ; Zhibin CHEN ; Han ZHOU ; Dengyuan WANG ; Huiqin TIAN ; Xingkuan BU
Journal of Clinical Otorhinolaryngology Head and Neck Surgery 2010;(1):19-20,24
Objective:To develop and evaluate the improved Chinese hearing questionnaire for school children(CHQS)for mass epidemiology study on hearing impairment in China.Method:Using the probability proportion to size(PPS) method, 8 412 residents were investigated in 40 clusters in Jiangsu province with the WHO ear diseases and hearing disorders survey protocol.87.9% of the residents aged 7 years and over answered the questionnaire and accepted the pure tone audiometry.Result:The prevalence of hearing impairment was 12.9% by the questionnaire. Compared with golden standard(pure tone audiometry), Sen=58.5%, Spe=96.7%, PV+=78.9%, PV-=91.7%, overall accuracy=90.0% . The sensitivity for women was higher than men.Conclusion:The questionnaire produced high efficiency and specificity values.It could be used in mass hearing screening, particularly in remote and rural area, although the sensitivity was as low as most questionnaires.