1.Application of Multivariate Statistics Analysis in HPLC Fingerprint of Shenmai Injection
Hao WU ; Yanhua TIAN ; Pingping GUO ;
Chinese Traditional Patent Medicine 1992;0(01):-
Objective: To establish an effective fingerprint analysis. Methods: Cluster analysis and mahalanobis distance analysis were adopted to analyze data of HPLC fingerprint of shenmai injection. Results: Precise classification of 18 samples was done by cluster analysis and mahalanobis distance analysis. Conclusion: Authors believe that multivariate statistics analysis applied to fingerprint can be recommanded in the quality control of shenmai injection.
2.HPLC Determination of Paeoniflorin, Hesperidin and Glycyrrhizic Acid in Yiwei Capsules
Shanfei ZHAN ; Pingping GUO ; Yanhua TIAN ; Hao WU ;
Chinese Traditional Patent Medicine 1992;0(04):-
Objective: To monitor the quality and production procedure of Yiwei Capsules from various aspects. Methods: The method of HPLC gradient elution analysis was adopted to determine paeoniflorin, hesperidin, and glycyrrhizic acid of Yiwei Capsules at the same time. Results: Good results were obtained in recoveries and coefficient of variation of the three substances. Conclusion: In this method the pretreatment is simple and the operation is easy which is helpful in quick and accurate determination.
3.Gab2 effects the invasion and metastasis of breast carcinoma through PI3 K/Akt/ARK5/MMP signal pathway
Hongyan TIAN ; Pingping CHEN ; Xiao LI ; Hongli LI ; Tiantian REN ; Baogang ZHANG ; Chonggao YIN ; Yuqing LIU
Chinese Pharmacological Bulletin 2015;(7):1014-1018
Aim To investigate the molecular mecha-nism of Gab2 in the invasion and metastasis of breast cancer and provide a theoretical basis for clinical pre-vention of breast cancer invasion and metastasis. Methods The Gab2 , MMP-2 and MMP-9 expressions in 80 cases of breast cancer were detected by immuno-histochemistry . Western blot was used to detect the ex-pression of Gab2 protein in MDA-MB-231 cells and MCF-7 cells. The siRNA plasmid was used to transfect MDA-MB-231 cells. Western blot was used to detect the proteins expression of Gab2 , MMP-2 and MMP-9 . Transwell in vitro experiment was used to detect the in-vasion ability of each group transfected MDA-MB-231 cells, Western blot was used to analyze phosphorylation of Akt and ARK5 induced by epithelial growth factor ( EGF ) in transfected cells ( SiGab2/MDA-MB-231 and Scr/MDA-MB-231 ) . Results The expression of Gab2 protein in invasive ductal carcinoma was signifi-cantly higher than in normal breast tissue ( P<0. 01 ) . The expression of Gab2 was dramatically correlated with lymph node metastasis, ER expression, tumor his-tological grade, MMP-2 and MMP-9 (P<0. 05). The expression of Gab2 protein in MDA-MB-231 cells was higher than in MCF-7 cells. The expression of Gab2, MMP-2 and MMP-9 decreased in SiGab2/MDA-MB-231 cells and the invasion ability of SiGab2/MDA-MB-231 cells was significantly decreased ( P<0. 05 ) , and after 5 minutes’ stimulating by EGF, the phosphoryla-tion of Akt and ARK5 was significantly reduced. Con-clusion Gab2 can promote the invasion and metasta-sis of breast cancer by effecting the expression of MMP-2 and MMP-9 through PI3 K/ Akt /ARK5 signal path-way.
4.Mechanism of RING finger protein 34 involved in innate immunity:a preliminary study
Yongjie ZHU ; Pingping ZHANG ; Pengyu ZHOU ; Penghao WANG ; Jiankang CHEN ; Yinyin TIAN ; Xiang HE ; Hui ZHONG
Military Medical Sciences 2017;41(1):43-47,61
Objective To study the mechanism of RING finger protein 34 ( RNF34 ) involved in innate immunity . Methods Recombinant PCR was used and transient expression of the plasmid was achieved in HEK 293T cells.The cells were stimulated with Sendai virus ( SeV) or N-RIG-Ⅰfor the indicated time while luciferase activity was observed using the dual-luciferase reporter assay kit .Results We constructed the plasmid pcDNA 3-Flag-RNF34 and its three mutations .The study found that when stimulated by SeV , RNF34 could inhibit the activity of NF-κB and IFN-βmore significantly than RNF34-ΔFYVE, RNF34-ΔCID and RNF34-ΔRING.We also found that RNF 34 and its three mutants had similar inhibitory effect when the activation of NF-κB and IFN-βwas stimulated by the N-RIG-Ⅰ.Conclusion RNF34 negatively regulates innate immunity by acting on the RIG-Ⅰ-MAVS signaling pathway .
5.Protective Effects of Electro-acupuncture Serum on β-amyloid Protein Induced Damage of Primarily Cultured Rat Hippcampal Neurons
Jing ZHU ; Shuijin SHAO ; Guohong CUI ; Jinxin TIAN ; Pingping LU ; Fangfang MU ; Haidong GUO
Chinese Journal of Information on Traditional Chinese Medicine 2013;(10):38-40,43
Objective To explore the protective effects of electro-acupuncture (EA) serum onβ-amyloid protein (Aβ) induced primary rat hippcampal neurons. Methods The rat models of Alzheimer's disease were established by intracerebral injection of Aβ1-40. After treated them with EA, the serum was harvested. Primary cultured hippocampal neurons were treated with Aβ25-35 to establish neuronal damage model in vitro, and divided into normal group, model group and EA serum group. The proliferation of neurons was detected by MTT test. Neuronal apoptosis was examined by TUNEL staining, and expression of cysteine aspartic acid proteases-3 (Caspase-3) was detected by immunocytochemical staining. Results MTT test showed that the cell viability was significantly decreased after treated with Aβ. While compared with the model group, cell proliferation of EA serum group was significantly enhanced (P<0.01). TUNEL staining showed that the number of apoptotic cells in EA serum group decreased significantly compared with the model group (P<0.01). After 48 h of Aβ treatment, Caspase-3 expression levels were significantly elevated. However, compared with the model group, the number of Caspase-3 positive cells in EA serum group was significantly reduced (P<0.01). Conclusion The EA serum could promote the proliferation of hippocampal neurons, reduce the expression of Caspase-3, counteract the neurotoxicity of β-amyloid protein, and reduce neuronal apoptosis.
6.Expression Changes of Autophagy Associated Protein Beclin-1 and Apoptosis Associated Protein p53 in Rats with Alzheimer's Disease and Regulatory Effects of Electroacupuncture
Qingchun ZHU ; Guohong CUI ; Shuijin SHAO ; Jinxin TIAN ; Xiaojing HAN ; Lisheng ZHANG ; Pingping LU ; Haidong GUO
Chinese Journal of Information on Traditional Chinese Medicine 2014;(9):68-71
Objective To observe the effects of electroacupuncture on the expressions of autophagy related protein Beclin-1 and apoptosis related protein p53 of hippocampus in rats;To explore the mechanism of electroacupuncture on Alzheimer's disease (AD).Methods The rats were randomly divided into the normal group, the sham-operation group, the model group, and the electroacupuncture treatment group. “Baihui” and “Yongquan” points were taken for electroacupuncture treatment and the treatment course was 7 days. The rats were treated once a day for 4 courses. Changes in morphology and number of Nissl positive cells were examined by Nissl staining in hippocampal CA1 regions. Expressions of Beclin-1 and p53 protein were determined by Western blot analysis.Results Number of Nissl positive cells in CA1 region of the model group was significantly less than that of normal group (P<0.01). After electroacupuncture treatment, number of pyramidal cells and expression of Nissl body significantly increased (P<0.05). Expression of Beclin-1 decreased, while expression of p53 increased in the hippocampus of the model group, compared with that in the normal group (P<0.05). However, electroacupuncture treatment could significantly upregulate the expression of Beclin-1 protein (P<0.01), but downregulate the level of p53 (P<0.05).Conclusion Electro-acupuncture treatment could fight against Aβ-induced neuronal apoptosis, and improve the morphological changes of AD’s hippocampus.
7.Regulation of deleted in liver cancer-1 gene domains on the proliferation of human colon cancer HT29 cell
Pingping WU ; Peng WU ; Qiqiang LONG ; Nan LI ; Zhi JIN ; Xiaoqiang TIAN ; Peilin HUANG
Chinese Journal of Digestion 2012;32(11):744-749
Objective To study the role of deleted in liver cancer-1 (DLC-1) gene main domains on the regulation of human colon cancer HT29 cell proliferation.Methods Subcloning recombinant plasmid vectors with Rho GTPase activating protein (RhoGAP),sterile alpha motif (SAM) or steroidogenic acute regulatory-related lipid-transfer (START) domains of DLC-1 gene knockout were constructed and transfected into human colon cancer cell HT29.Wild HT29 cell group (control group),pcDNA3.1-HT29 cell group (vector group) and pcDNA3.1-HT29-DLC-1 cell group (whole DLC-1 gene transfected group) were set as control.The change of cell proliferation was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay and colony formation test.The cell apoptosis was analyzed by flow cytometry.The activity of RhoA protein was detected by pull-down assay.The differences between the groups were analyzed by the analysis of variance.Results At 48 hours after the successful transfection,compared with control group and vector group,cells proliferation and the activity of RhoA protein were significantly suppressed in whole DLC-1 gene transfected group (F=146.36,698.08,both P<0.05) and early cell apoptosis increased (F=294.08,P<0.05).Compared with control group and vector group,there was no significant difference in cell proliferation ability,cell apoptosis and the activity of RhoA protein activity in RhoGAP knockout transfected cells (F=0.99,0.049,5.769,all P>0.05).Compared with whole DLC-1 gene transfected group,the suppression of cell proliferation was more significant in SAM knockout transfected cells (F=31.00,P<0.05),the activity of RhoA protein down regulated (F=92.57,P<0.05) and apoptosis increased (F=130.44,P<0.05).Compared with whole DLC-1 gene transfected group,the ability of cell proliferation increased (F=15.47,P<0.05),apoptosis cell decreased (F=110.23,P<0.05) and the activity of RhoA protein up regulated (F=199.39,P<0.05) in START knockout transfected cells.Conclusions The role of DLC-1 gene in the suppression of cell proliferation in HT29 cells was RhoGAP-dependent.SAM domain may be the self suppression domain for endogenous RhoGAP activity.START domain may take effect through enhancing RhoGAP domain.
8.Thyroid autoantibody and risk of preterm birth: a meta-analysis of prospective cohort studies
Zengfang WANG ; Jiying HU ; Pingping WANG ; Ying TAN ; Wenjing LIAN ; Huaxiang TIAN ; Bin WANG
Chinese Journal of Perinatal Medicine 2013;(5):257-261
Objective To evaluate the relationship between positive thyroid autoantibody and risk of preterm birth.Methods Literature search was done in PubMed,Embase,China Academic Journal Network Publishing Database,Wanfang Medical Database and China Biology Medicine disc databases from January 1st,1989 to January 26th,2012.Criteria for inclusion included:(1) Prospective cohort study; (2) The exposure was positive thyroid autoantibody and outcome was preterm birth; (3) The enrolled subjects were pregnant women without cardiovascular or rheumatic disease; (4) Relative risk (RR) and its 95% confidence interval (95%CI) of preterm birth were provided in the study.Meta-analysis was performed by Stata 12.0.The relationship between positive thyroid autoantibody and risk of preterm birth was evaluated by RR and 95% CI.Results Ten cohort studies were enrolled.One thousand six hundred and fifty seven cases of preterm birth occurred among 25 081 pregnant women.Heterogeneity among the 10 studies was found in meta-analysis (I2 =79.2%,P<0.01).The risk of preterm birth in pregnant women with positive thyroid autoantibody was higher than those in control group by random effects analysis (RR=1.61,95%CI:1.18-2.20,P<0.05).Subgroup analysis was further performed.In five studies,the cases of control group were pregnant women with normal thyroid function; heterogeneity was not found in these five studies (I2=39.1%,P=0.160); and RR of the risk of preterm birth was 2.55 in pregnant women with positive thyroid autoantibody (95 % CI:2.04 3.19,P<0.01).In the other five studies,the cases of controlgroup were pregnant women who had not been ruled out the possibility of thyroid dysfunction;heterogeneity was not found in these five studies either (I2 =0.0%,P =0.970); and RR was 1.18(95 % CI:1.01-1.37,P<0.05).After excluding two low-quality studies,RR of the risk of preterm birth was 1.72 in pregnant women with positive thyroid autoantibody (95%CI:1.18 2.53,P<0.05).The funnel plots presented symmetrical graphics,indicating that there was no publication bias.Conclusions Positive thyroid autoantibody in pregnant women is a risk factor of preterm birth.
10.WT1 downregulation during K562 cell differentiation and apoptosis induced by bufalin.
Yunpeng LIU ; Xiujuan QU ; Pingping WANG ; Xin TIAN ; Ying LUO ; Shizhou LIU ; Xianglan LU
Chinese Journal of Hematology 2002;23(7):356-359
OBJECTIVETo study the change of WT1 gene expression during human leukemic K562 cell differentiation and apoptosis induced by bufalin.
METHODSCell viability was determined by trypan blue exclusion, cell differentiation and apoptosis by nitro blue tetrazolium (NBT) reduction test, morphology and flow cytometry, expression of WT1 protein by Western blot analysis, and expression of WT1 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).
RESULTS(1) The cell proliferation was inhibited by bufalin and the IC(50) at 24, 48, 72 h were 0.026, 0.032 and 0.006 micro mol/L, respectively. (2) Bufalin induced K562 cell differentiation towards macrophage/monocyte within concentration from 0.01 to 0.05 micro mol/L and apoptosis at higher than 0.026 micro mol/L. (3) The expression of WT1 protein and mRNA were downregulated by bufalin in the initial stage of differentiation and apoptosis induced by bufalin.
CONCLUSIONK562 cell differentiation and apoptosis induced by bufalin might relate to the downregulation of WT1 expression.
Apoptosis ; Cell Differentiation ; Cell Proliferation ; Down-Regulation ; Humans ; WT1 Proteins ; genetics