2.Role of solasodine hydrochloride in AS2O3 induced HeLa cells apoptosis as well as its effect on cell telomerase activity in vitro
Jin-Xia, AI ; Liang, LIU ; Ping, WANG
Chinese Journal of Endemiology 2011;30(3):279-283
Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.
3.Cultural anthropology of traditional Chinese medicine
Xia WAN ; Jianping LIU ; Yanke AI ; Liuji LI
Journal of Integrative Medicine 2008;6(7):674-7
Biological, psychological and sociological model of medicine substantializes the old model lacking the social humane attributes. The new medical model makes people take medical anthropology into research and highly evaluate traditional medical system. Cultural anthropology of traditional Chinese medicine (TCM) is part of medical anthropology with three major characteristics: wide research scope, specificity, and integration. It has developed its own research methods, such as field investigation, comprehensive inspection and comparison study. Cultural anthropology provides an efficient research method for TCM, and its application would further develop TCM theory and form comprehensive evaluation on TCM effects.
4.Clinical outcome assessment and the effect measure
Hongwei ZHANG ; Jianping LIU ; Xia WAN ; Yanke AI
Journal of Integrative Medicine 2007;5(5):497-501
Effect of clinical intervention on human body is multistage and multifaceted, involving physiology, psychology, social function and the surrounding resources, etc. Therefore, the range of clinical outcome assessment includes patient, his or her family and care giver. The evaluation of clinical intervention mainly focuses on its effectiveness, safety and health economics. Effects of clinical interventions are measured by comparisons of the outcome of intervention and control groups. Different effect measures come from different comparisons.
5.Methods of selecting outcome variables for clinical intervention
Xia WAN ; Jianping LIU ; Hongwei ZHANG ; Yanke AI
Journal of Integrative Medicine 2007;5(1):11-4
Abstract: There are two stages for selecting the clinical intervention outcome variables. One is crude collection and the other is precise collection. The selection methods include brainstorming method, Delphi method, Gordon method, systematic review, systematic analysis method, mathematic model method and so on. Each method has some advantages or disadvantages. When selecting the clinical intervention outcome variables, researchers should make sure the research aim, the representative and the sensitivity of the variables, and select all kinds of recommended methods comprehensively, then decide which method should be used.
6.Stroke-induced acroparalysis treated with xunjingcuiqi needling technique: a randomized controlled trial.
Yu-Hua CHAI ; Rui-Xia ZHANG ; Cheng-Ai XUE ; Yong-Qin LIU ; Xiang-Xiang WANG
Chinese Acupuncture & Moxibustion 2014;34(6):534-538
OBJECTIVETo observe the impacts on the muscle strength in the patients of stroke-induced acroparalysis treated with Xunjingcuiqi needling technique.
METHODSOne hundred patients were randomized into a Xunjingcuiqi group and a routine acupuncture group, 50 cases in each group. In the routine acupuncture group, the routine acupuncture technique was adopted at the main acupoints, such as Shangxing (GV 23), Baihui (GV 20), Dicang (ST 4), Quchi (LI 11), Huantiao (GB 30) and Zusanli (ST 36), etc. In Xunjingcuiqi group, on the basis of the routine acupuncture technique, Xunjingcuiqi needling technique (pricking technique was quickly applied with the filiform needle along the running course of meridian to promote the conduction of meridian qi) was added. For the patients being hard to feel the needling sensation and with the muscle strength of 0 to 1 degree, Dongzhencuiqi technique was supplemented at shu-stream points of yang meridians (after qi arrival, the needling manipulation with limb movement was given to promote the conduction of meridian qi). The treatment was given once every day in the two groups. Ten treatments made one session. Three sessions of treatment were required. At the end of each session treatment, the muscle strength and clinical efficacy were assessed.
RESULTSIn the 1st, 2nd and 3rd sessions of treatment, 20, 24 and 36 cases achieved the 3 to 5 degrees muscle strength in Xunjingcuiqi group, respectively; and 6, 10 and 15 cases achieved the 3 to 5 degrees muscle strength in the routine acupuncture group. The differences were significant statistically in comparison of the two groups (P < 0.01, P < 0.05). The markably effective rates were 60.0% (30/50), 64.0% (32/50) and 70.0% (35/50) after the 1st, 2nd and 3rd sessions of treatment in Xunjingcuiqi group, respectively; and those were 38.0% (19/50), 44.0% (22/50) and 46.0% (23/50) in the routine acupuncture group, respectively. The differences were significant in the 1st and 3rd sessions of treatment between the two groups (both P < 0.05).
CONCLUSIONXunjingcuiqi needling technique combined with routine acupuncture achieves the apparent superior efficacy on acroparalysis induced by ischemic stroke as compared with the simple routine acupuncture. Xunjingcuiqi needling technique obviously improves muscle strength and shortens the duration of sickness.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Aged ; Female ; Humans ; Male ; Middle Aged ; Paralysis ; etiology ; therapy ; Qi ; Stroke ; complications ; Treatment Outcome
7.Correlation of TLR2 and TLR4 gene polymorphisms with the susceptibility and recurrence of condyloma acuminatum.
Ji-feng LIU ; Bin QU ; Xiang-dong WANG ; Qi WANG ; Xiao-xia ZHAO ; Ai-e XU
National Journal of Andrology 2015;21(8):708-712
OBJECTIVETo explore the correlation of the gene polymorphisms of Toll-like receptor 2 ( TLR2) and TLR4 with the susceptibility and recurrence of condyloma acuminatum (CA).
METHODSUsing Snapshot, we detected the gene polymorphisms of TLR2 597(T/C), 1350(T/C), 15607(A/G), and 2258(G/A) and TLR4 896(A/G) and 1196(C/T) in the peripheral blood of 140 CA patients and 105 HPV-negative controls. We made comparisons between the CA patients and controls as well as between the cases of recurrent CA and those of non-recurrence at 6 months after treatment.
RESULTSThere were 72, 48, and 20 cases of genotype TT, TC, and CC of TLR2 597 (T/C), respectively, in the CA patients, as compared with 71, 31, and 3 cases in the controls. The gene frequency of mutant C was 31. 43% in the patients, significantly higher than 17.62% in the controls (χ2 = 12.04, P < 0.01), and it was 38.68% in the recurrent cases, remarkably higher than 27.01% in the non-recurrent cases (χ2 = 4.16, P < 0.05). There were 74, 49, and 17 cases of genotype TT, TC, and CC of TLR2 1350( T/C), respectively, in the CA patients, as compared with 73, 29, and 3 cases in the controls. The gene frequency of mutant C was 29. 64% in the patients, significantly higher than 16. 67% in the controls (χ2 =11.05, P < 0.01), and it was 36.79% in the recurrent cases, markedly higher than 25. 29% in the non-recurrent cases (χ2 = 4.18, P < 0.05). There were 44, 66, and 30 cases of genotype AA, AG, and GG of TLR2 15607(A/G), respectively, in the CA patients, as compared with 26, 58, and 21 cases in the controls. There was no significant difference in the gene frequencies of mutant G between the two groups (χ2 = 0.33, P > 0.05). No mutant genes of TLR2 2508 (G/A) or TLR4 896(A/G) and 1196(C/ T) were detected in either the CA patients or the controls. Linkage disequilibrium analysis showed a tight linkage between TLR2 597 (T/C) and 1350(T/C) (D' = 1, r2 = 0.93).
CONCLUSIONTLR2 597(T/C) is tightly linked to 1350(T/C), which is correlated with both the susceptibility and the recurrence of condyloma acuminatum.
Aged ; Case-Control Studies ; Condylomata Acuminata ; genetics ; Gene Frequency ; Genetic Linkage ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Genetic ; Recurrence ; Toll-Like Receptor 2 ; genetics ; Toll-Like Receptor 4 ; genetics
8.Effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction on myocardial apoptosis and relevant protein expressions in miniature swine phlegm and blood stasis type coronary heart disease model.
Ai-guo YAN ; Jian-xun LIU ; Xin-zhi LI ; Xiao-xia DONG ; Hong-kun LI
China Journal of Chinese Materia Medica 2015;40(11):2174-2179
There have been very few studies on the effect of Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction in inhibiting apoptosis in myocardial ischemial injury caused by coronary heart disease. In this experiment, Gualou Xiebai Banxia decoction combined with-Xuefu Zhuyu decoction were used to intervene the miniature swine phlegm and blood stasis type coronary heart disease model, in order to observe the effect of the combined prescription on the myocardial apoptosis and the expressions of Bcl-2, Bax, Caspase-3, Caspase-9 in the model. Totally 15 Chinese experimental miniature swine were adopted and randomly divided into the control group, the model group and the phlegm and stasis-treating group. The model group and the stasis-treating group were fed with high fat diets for two weeks, intervened with the coronary artery injury and then given drugs and high fat diets for eight weeks. The control group was fed with ordinary diets for 10 weeks, without the coronary artery injury. After the experiment, myocardia at the juncture of infracted areas were collected and made into formalin-fixed paraffin sections. The TDT-mediate dUTP nick end labeling (TUNEL) assay was used to detect the myocardial apoptosis. The immunohistochemistry (IHC) technique was applied to detect Bcl-2, Bax, Caspase-3, Caspase-9 levels in myocardial tissues. According to the findings, the apoptosis indexes (AI) for the control group, the model group and the phlegm and stasis-treating group were 0.92%, 27.68%, 17.28%, respectively. The AI of the phlegm and stasis-treating group was significantly lower than that of the model group (P < 0.01). Compared with the model group, the phlegm and stasis-treating group showed significantly higher Bcl-2 protein expression (P < 0.01) and lower Bax, Caspase-3 and Caspase-9 protein expressions (P < 0.01). In conclusion, Gualou Xiebai Banxia decoction combined with Xuefu Zhuyu decoction have a significant protective effect against the myocardial apoptosis in miniature swine phlegm and blood stasis type coronary heart disease model.
Animals
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Apoptosis
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drug effects
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Caspases
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metabolism
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Coronary Disease
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drug therapy
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Disease Models, Animal
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Drug Therapy, Combination
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Drugs, Chinese Herbal
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administration & dosage
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Female
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Male
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Medicine, Chinese Traditional
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Myocardium
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pathology
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Phytotherapy
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Proto-Oncogene Proteins c-bcl-2
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analysis
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Swine
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Swine, Miniature
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bcl-2-Associated X Protein
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analysis
9.Relationship between serum homocysteine levels and contrast induced nephropathy after coronary angiography
Min AI ; Guangxue LIU ; Changfu YAN ; Fuchun XIA ; Shuanglu ZHOU ; Jian HE
Chinese Journal of Interventional Cardiology 2017;25(3):144-148
Objective To explore the relationship between serum homocysteine (Hcy) level before coronary angiography(CAG) and contrast induced nephropathy (CIN) after CAG.Methods We included 2264 cases of suspected coronary heart disease from May 2013 to May 2016 and all patients received CAG examination.According to whether CIN has developed or not after CAG, the patients were divided into the non-CIN group (n=2162) and the CIN group (n=102).We analyzed and compared the clinical baseline data, serum Hcy and creatinine (Cr) levels and the estimated glomerular filtration rate between the 2 groups eGFR.Results Patients in the non-CIN group were younger and with less comorbidities of diabetes and chronic kidney disease (all P<0.05).The volume of contrast media consumed in the non-CIN group was less than the CIN group [(122±21)ml vs.(147±24)ml, P=0.012).Hcy level in the non-CIN group (12.81±6.71) μmol/L was lower than that in the CIN group (21.74±11.9)μmol/L before CAG (P<0.05).No significant differences in serum Cr level and eGFR before CAG (P>0.05).At 72 hours after CAG, Cr level of the non-CIN group (69.34±19.54 μmol/L) was lower than that of the CIN group (87.34±21.38) μmol/L (P<0.05).eGFR was higher in the non-CIN group (79.34±19.54)ml/min than that in the CIN group (67.34±21.38)ml/min (P<0.05).Linear regression analysis showed that Hcy level before CAG were positively correlated with Cr level after CAG (r=0.547,P<0.01) and negatively correlated with eGFR after CAG (r=-0.271,P<0.01).Conclusions Hcy level before CAG can be used as one of an effective parameter to predict CIN.
10.Effect of reduced glutathione on the proliferation,oxidative stress and transforming growth factor?1 expression of human hepatic stellate cells
Mei LIU ; Lun-Gen LU ; Ai-Xia DOU ; Weihua CHEN ; Ruidan ZHENG ; Minde ZENG ; Jingyuan FANG ;
Chinese Journal of Digestion 2001;0(10):-
Objective To investigate the impact of reduced glutathione(GSH) on the prolifera- tion,oxidative stress and transforming growth factor?1(TGF-?1) expression of human hepatocytes and hepatic stellate cells(HSCs)(LX-2 cell line).Methods Human hepatocytes and HSCs were incubated with various concentrations of GSH(0.5—50 mmol/L or 0.5—10 mmol/L).The effects of GSH on the proliferation of hepatocytes and HSCs were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphennyhera- zolium bromide colorimetric assay.Human hepatocytes and HSCs were co-cultured with GSH and ferric nitrilotriacetic acid,superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents were detected.HSCs were incubated with high(5.0 mmol/L),media(2.5 mmol/L) and low (0.5 mmol/L) concentrations of GSH,the expressions of TGF-?1 mRNA and protein were detected by ELISA and real- time PCR.Results In concentration ranged from 2.5 to 10 mmol/L,the GSH could promote the pro- liferation of hepatocytes but no HSCs,significantly increased the activity of SOD and decrease the con- tents of MDA in hepatocytes and HSCs,and inhibited the expression of TGF-?1 in HSCs.Conclusions GSH can not only promote the proliferation of hepatocytes,but also protect hepatocytes and HSCs from oxidative stress,and inhibit the secretion of TGF-?1 in HSCs.GSH may play a role in hepatocellular protection,antioxidation and anti-fibrosis.