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Objective To investigate the growth inhibition effect of evodiamine (Evo) on renal carcinoma 786-0 cells and to explore its molecular mechanism. Methods After treated with Evo, methyl thiazolyl tetrazolium ( MTT) assay was used to detect the vitality of 786-0 cells, flow cytometry was employed to examine the cell cycle distribution in 786-0 cells, and immunoblotting was utilized to determine the expression levels of target proteins related to cell cycle progression. Results Evo remarkably inhibited 786-0 cells vitality in dose-dependent manner. Cell cycle analysis indicated that 786-0 cells were arrested in G2/M phase followed by Evo treatment. Furthermore, the results of immunoblotting showed that Evo up-regulated the protein expression levels of P53, P21 and its downstream target gene CyclinB1 in 786-0 cells. Conclusion Evo treatment can induce 786-0 cell cycle G2/M arrest, and its underlying mechanism might be dependent on the P53/P21 signal pathway.
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Objective To investigate the potential role of Lycium bararum polysaccharide (LBP) with or without interferon -inducible protein 10 ( CXCL10) in inducing dendritic cells ( DC) functional maturation by monitoring the alteration of cytokines for inducing DC maturation in peripheral blood and by detecting the expression of S-100 protein in tumor tissue, thus to reveal its mechanism of inhibiting experimental liver cancer. Methods H22 bearing mice model was established. The mice were randomized into model group, LBP group (50 mg/kg, ig), CXCL10 (right axillary subcutaneous injection of 15 μg/kg), LBP + CXCL10 group (LBP 50 mg/kg, ig, and right axillary subcutaneous injection of CXCL10 15 μg/kg), 5- fluorouracil (5FU) group ( intraperitoneal injection of 12mg/kg) , 12 mice in each group. The mice were administered the corresponding medicine once a day. After treatment for 2 continuous weeks, blood was sampled from infraorbital vein, and the tumor mass, spleen, thymus were extracted for the calculation of anti-tumor rate, thymus index and spleen index separately . The mRNA expression levels of interleukin 12 (IL-12) and tumor necrosis factor-α (TNF-α) in peripheral blood were detected by fluorescence quantitative PCR, the expression of S-100 protein in tumor tissues was detected by immunohistochemical assay. Results Compared with the model group, tumor growth in LBP group and LBP+CXCL10 group was obviously inhibited, and tumor-inhibitory rate was 55.90%, 50.91%, respectively. Meanwhile, the mRNA expression level of IL-12 was 2.94 folds higher in LBP group and 3.39 folds higher in LBP + CXCL10 group, and TNF-α mRNA expression level was 1.55 folds higher in LBP group and 4.74 folds higher in LBP+CXCL10 group than the model group, the differences being statistical significant ( P<0.05 or P<0.01). Results of immunohistochemical assay showed that S-100+DC number in LBP group and LBP+CXCL10 group was larger than that in the model group (P<0.05 ). Conclusion LBP and LBP+CXCL10 exert significant effect on inhibiting experimental liver cancer. The mechanism may be related with inducing the secretion of IL-12 and TNF-α, which plays a key role in inducing DC maturation, and with the increase of the number of DC in tumor microenvironment.
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This study examined the incidence, neuropsychological characteristics and risk factors of cognitive impairment 3 months after stroke in China. Five regions that differed in geography and economy in China were selected. Patients from the hospitals located in the five regions were prescreened at admission, and the demographic data, vascular risk factors and clinical characteristics of stroke were obtained. A battery of cognitive-specific domain tests was performed in the patients who failed to pass cognitive screening 3 months post stroke. Patients were diagnosed as having post-stroke cognitive impairment (PSCI) or no cognitive impairment (NCI) based on the results of the neuropsychological tests. Univariate analysis was performed for suspect risk factors, and significant variables were entered in multivariable logistic regression analysis. Our results showed that a total of 633 patients were recruited 3 months after stroke; complete cognitive tests were performed in 577 of the stroke patients. The incidence of PSCI in these Chinese patients was 30.7%. There were 129 (22.4%) patients with visuospatial impairment, 67 (11.6%) with executive impairment, 60 (10.4%) with memory impairment and 18 (3.1%) with attention impairment. The risk factors associated with PSCI were older age (odds ratio [OR] 1.76, 95% confidence interval [CI] 1.20-2.58), low education level (OR 2.45, 95% CI 1.65-3.64), depressive symptom (OR 1.69, 95% CI 1.09-2.61), obesity (OR 2.57, 95% CI 1.41-4.71), stroke severity 3 months post stroke (OR 1.62, 95%CI 1.10-2.37) and cortex lesion (OR 1.55, 95% CI 1.04-2.31). It was concluded that PSCI occurs commonly 3 months after first-ever stroke in Chinese patients. Visuospatial ability may be the most frequently impaired cognitive domain for the patients with stroke. The critical risk factors of PSCI are older age, low education level, depressive symptom, obesity, stroke severity 3 months post stroke and cortex lesion.
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Feminino , Humanos , Masculino , Pessoa de Meia-Idade , China , Epidemiologia , Transtornos Cognitivos , Epidemiologia , Estudos Transversais , Incidência , Fatores de Risco , Acidente Vascular CerebralRESUMO
This study examined the incidence, neuropsychological characteristics and risk factors of cognitive impairment 3 months after stroke in China. Five regions that differed in geography and economy in China were selected. Patients from the hospitals located in the five regions were prescreened at admission, and the demographic data, vascular risk factors and clinical characteristics of stroke were obtained. A battery of cognitive-specific domain tests was performed in the patients who failed to pass cognitive screening 3 months post stroke. Patients were diagnosed as having post-stroke cognitive impairment (PSCI) or no cognitive impairment (NCI) based on the results of the neuropsychological tests. Univariate analysis was performed for suspect risk factors, and significant variables were entered in multivariable logistic regression analysis. Our results showed that a total of 633 patients were recruited 3 months after stroke; complete cognitive tests were performed in 577 of the stroke patients. The incidence of PSCI in these Chinese patients was 30.7%. There were 129 (22.4%) patients with visuospatial impairment, 67 (11.6%) with executive impairment, 60 (10.4%) with memory impairment and 18 (3.1%) with attention impairment. The risk factors associated with PSCI were older age (odds ratio [OR] 1.76, 95% confidence interval [CI] 1.20-2.58), low education level (OR 2.45, 95% CI 1.65-3.64), depressive symptom (OR 1.69, 95% CI 1.09-2.61), obesity (OR 2.57, 95% CI 1.41-4.71), stroke severity 3 months post stroke (OR 1.62, 95%CI 1.10-2.37) and cortex lesion (OR 1.55, 95% CI 1.04-2.31). It was concluded that PSCI occurs commonly 3 months after first-ever stroke in Chinese patients. Visuospatial ability may be the most frequently impaired cognitive domain for the patients with stroke. The critical risk factors of PSCI are older age, low education level, depressive symptom, obesity, stroke severity 3 months post stroke and cortex lesion.
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Objective To determine the serum level of chernokine CCL27 in patients with psoriasis vulgaris,and to analyse its clinical relevance.Methods A total of 61 patients(40 in progressive stage and 21 in stable stage)with psoriasis vulgaris,with an average disease duration of 37.97±14.34 years,were included in this study.Appropriate thempy was given to these patients.Serum samples were collected from the patients before and after therapy,as well as from 45 healthy human controls.ELISA was applied to examine the serum concentration of CCL27.Clinical severity of psoriasis vulgaris was assessed by psoriasis area and severity index(PASI)score.Results Serum level of CCL27 was 670.02±262.15 ng/L in psoriatic patients,compared to 373.10±92.84 ng/L in the controls(t=8.18.P＜0.01).Increased serum level of CCL27 was observed in patients with progressive psoriasis vulgaris compared to those with stable psoriasis (799.94±214.54 ng/L vs 422.57±135.53 ng/L,t=8.39,P＜0.01).After 8 weeks of therapy,a significant decrease was noticed in the serum level of CCL27 in patients who experienced≥70%reduction in PASI score(t=9.95,P＜0.01).but not in those experiencing a PASI reduction of＜70%(t=1.84,P＞0.05).The serum level of CCL27 was positively correlated with PASI score(r=0.58,P＜0.01).Conclusions The serum level of CCL27 is significantly elevated in patients with psoriasis vulgaris,and it is correlated with the disease severity.
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【Objective】 To explore a method for the construction of antitumor suicide-gene therapeutic system. 【Methods】 The thymidine kinase gene of herpes simplex virus type 1 (HSV1-tk) was orientationally cloned into the retroviral vector plasmid (pLXSN) by DNA recombinant technique, and then the recombinant plasmid of pLXSN-tk was identified by restriction endonuclease cutting and DNA sequencing. Introduced by PolyFect Transfection reagent, the recombinant plasmid was transfected into the packaging cell line PT67. Screened by G418, a cell line, which could produce virus stably, was obtained. The virus was transfected into human gastric carcinomatous cell strain SGC-7901 and the transfected SGC-7901 was applied to evaluate an anti-tumor effect. 【Results】 The identification with restriction endonuclease cutting and DNA sequencing showed that HSV1-tk was successfully inserted into the recombined plasmid of pLXSN-tk. The stable cloned PT67/tk was obtained by screening with G418 and then its amount was enlarged by culture, the titer of the PT67/tk solution being 4?10~4 cfu/mL. The anti-G418 cloned cell line SGC-7901/tk was got by infecting SGC-7901 with the virus. Anti-tumor experiment showed that GCV had an obvious toxic effect on SGC-7901/tk but had no effect on SGC-7901, indicating recombinant retroviral HSV1-tk expressed HSV1-tk gene which had biological activity. 【Conclusion】 Cloning HSV1-tk into the retroviral vector is effective in obtaining the recombinant retroviral HSV1-tk which can express HSV1-tk gene, and also effective in establishing an antitumor suicide-gene therapeutic system. This research naturally lays a foundation for further studying of Chinese medicines having synergistic anti-tumor action with suicide gene.
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AIM To explore the effect of berberine in inhibition of proliferation and induction of apoptosis of gastric cancer cell MGC-803.METHODS The MGC-803 cell morphology and cell counting were studied by Trypan blue, MTT, Methyl green pyronin stain assay, the MGC-803 cell DNA changes were investigated by flow cytometry and agarose gel electrophoresis.RESULTS Berberine could significantly inhibit the growth of MGC-803. After 48 hours of treatment with different concentration berberine (8, 4, 2, 1 mg*L-1), the inhibitive rate of MGC-803 were 86.7%, 65.9%, 20.0%, 0%, respectively. Meanwhile, MGC-803 showed typical apoptosis features: cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. A typical subdiploid peak before G0/G1 phase was observed by flow cytometry. The Trypan blue stain achromatophilia rates of death cells, which with well membranes, were between 60%～82%. Agarose gel electrophoresis analysis, which showed that the chromosomes of MGC-803 were broken by berberine before the structures of membrane were damaged, while DNA broke into long fragments rather than small fragments, supported that the activation effect of berberine on nuclease was mild.CONCLUSION Berberine could significantly inhibit proliferation of gastric cancer cell MGC-803 and induced apoptosis, and the cytotoxicity of berberine on MGC-803 was weak.
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Suicide - gene therapy is now regarded as a new method for tumor with good prospect. Since bystander effect is the main mechanism and is positively correlated with the immune function, stimulating the immune function of tumor carriers and improving the inflammatory microimmune situation are two important keys to good therapeutic effect. As it is known that Chinese herbal medicine is effective in improving human immune function and has less toxic effect, therefore, suicide - gene therapy combined with Chinese herbal medicine may be a new, safe and economic regimen for tumor.
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Objective To compare the pharmacological actions of Liandai Tablet(LT) and its main active components on apoptosis of gastric cancer cells and DNA damage. Methods Effects of Liandai Tablet(LT) and its main active components,berberine and indirubin,on growth and apoptosis of gastric cancer cell strain MGC_803 were explored and their effects on DNA damage were also studied. Results LT serum in high and low dosages and berberine could inhibit the growth of MGC_803 as compared with the control group,and typical morphological features of apoptosis were found in the MGC_803 by methyl green pyronin stain assay.But indirubin at various concentrations showed no obvious inhibitory effects. Agarose gel electrophoresis assay revealed that the MGC_803 cell DNA was split into large fragments when treated with berberine. Conclusion LT serum exerts a similar inhibitory effect on the growth and apoptosis of gastric cancer cells as compared with berberine.The effects of LT at various serum concentrations on MGC_803 DNA was less than that of berberine,and indirubin at the given concentration had no this effect.
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Objective To investigate the possibility of establish a combined regimen of Chinese medicine and suicide gene therapy,we observed the killing action of medicated serum of Liuwei Dihuang Bolus (LDB)combined with HSV-tk/GCV suicide gene therapy system on rats hepatoma cell line CBRH7919.Methods The HSV-tk/GCV suicide gene therapy system was constructed and the working solution of ganciclovir (GCV)at 39.2 ?mol/L was detected firstly.Then medicated or non-medicated serum was prepared from SD rats treated with or without LDB by gavage.Meanwhile,the effects of medicated and non-medicated serum on cell proliferation were detected.The control groups without serum added were blank control group,GCV group and suicide gene therapy (SGT)group.Blank serum groups,blank serum + GCV groups,blank serum + SGT groups,medicated serum groups,medicated serum+GCV groups and medicated serum+SGT groups all had two serum concentrations of 5% and 7.5%,respectively.Six double holes were set for each group.CBRH7919/ tk+ and CBRH7919/ tk-were mixed together,and the tk+ cell percentage was adjusted to 0%,5% and 10%,respectively.Then the mixture of CBRH7919/ tk+ and CBRH7919/ tk-was implanted into the 96-hole plates at a density of 3?103 cells/hole,cultured with complete medium for 24 hours,and then treated with medicated or non-medicated serum for 12 hours and sequentially with GCV at 39.2?mol/L for another 60 hours.The number of surrival cells was determined by MTT assay.Q value (the ratio of measured and theoretical pharmacological action)was used to analyze the synergism of Chinese medicine and suicide gene therapy:additive action when 0.85≤Q1.15.Results No cytotoxicity was found when blank serum or medicated serum was below the concentration of 20% (volume fraction).When the concentration of serum was at 5% and 7.5%,the killing action of SGT + medicated serum group was stronger and the cell survival rate was higher those that of medicated serum group and SGT + blank serum group (P1.15).Conclusion HSV-tk/GCV suicide gene therapy system combined with LDB has a synergistic killing action on rats hepatoma cells.
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AIM To explore the effect of berberine in inhibition of proliferation and induction of apoptosis of gastric cancer cell MGC 803.METHODS The MGC 803 cell morphology and cell counting were studied by Trypan blue, MTT, Methyl green pyronin stain assay, the MGC 803 cell DNA changes were investigated by flow cytometry and agarose gel electrophoresis.RESULTS Berberine could significantly inhibit the growth of MGC 803. After 48 hours of treatment with different concentration berberine (8, 4, 2, 1 mg?L -1 ), the inhibitive rate of MGC 803 were 86 7%, 65 9%, 20 0%, 0%, respectively. Meanwhile, MGC 803 showed typical apoptosis features: cell shrinkage, nuclear condensation, DNA fragmentation and formation of apoptotic bodies. A typical subdiploid peak before G 0/G 1 phase was observed by flow cytometry. The Trypan blue stain achromatophilia rates of death cells, which with well membranes, were between 60%～82%. Agarose gel electrophoresis analysis, which showed that the chromosomes of MGC 803 were broken by berberine before the structures of membrane were damaged, while DNA broke into long fragments rather than small fragments, supported that the activation effect of berberine on nuclease was mild.CONCLUSION Berberine could significantly inhibit proliferation of gastric cancer cell MGC 803 and induced apoptosis, and the cytotoxicity of berberine on MGC 803 was weak.
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Aim To explore the proliferation-inhibited,apoptosis-induced and cell cycle-regulated effect of evodiamine on human hepatoma cell line HepG2.Methods MTT,Dapi assay,flow cytometry analysis,comet assay were used.Results Evodiamine could significantly inhibit the growth of human hepatoma cell line HepG2.After 72 hours of treatment with evodiamine at different concentrations(64,16,4,1,0.25 ?mol?L-1),the inhibitory rate of HepG2 was 74.0%,69.0%,60.5%,44.0% and 16.4%,respectively.Meanwhile,HepG2 showed typical apoptosis.After 24 and 36 hours' treatment with evodiamine(1 ?mol?L-1),a typical subdiploid peak before G0/G1 phase was observed by flow cytometry and cell cycle was arrested in the G2/M phase,while the rate of apoptosis was 4.4%,18.0% and 30.3% of treatment with evodiamine for 12,24 and 36 hours respectively.After 24 and 36 hours' treatment with evodiamine(1 ?mol?L-1),the average optical density was lower than that of the control and the length of tail increased compared with the control.Meanwhile the changes were related with time.Conclusion Evodiamine inhibits the proliferation and induces apoptosis of HepG2.
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Aim To investigate the curative effect of Danshen injection combining with HSV-tk/GCV system on rats′ hepatocarcinoma cells and murine transplanted hepatocarcinoma.Methods ① Rats′ hepatocarcinoma cell line CBRH7919(tk~-),CBRH7919/tk(tk~+) and the 5% tk~+ mixed cells were treated with diverse concentrations of Danshen injection,GCV separately,and Danshen injection plus GCV(n=3).The survival rate of each groups was examined using MTT Assay and was analyzed using paired comparison.Q-value analysis method was used to estimate the synergistic effect of Chinese herbal on the suicide gene system.Q-value is a ratio of the actural effect of combination treatment to its theoretical effect.It is thought to be an additive effect when 0.85≤Q