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.

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.

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

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.

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.

【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.

Objective To investigate the value of EBUS-TBNA (endobronchial ultrasound-guided trans-bronchial needle aspiration) in diagnosis of lung cancer and analyze its complications. Methods A retrospective study was carried out in our hospitalfrom March 2013 to March 2015. A total of 171patients received C-TBNA (Con-ventional transbronchial needle aspiration) first and got negative results. Therefore, they received EBUS-TBNA next. Weanalyzedthe sensitivity, specificity, accuracy of malignancy with EBUS-TBNA and positive (malignant) lymph node sites biopsied with EBUS-TBNAand the incidence of its complications. Results Lung cancer was proven in 107 patients who had received EBUS-TBNA, while 16 were benign cases. Thirty-onenegative patients received CT guided needle biopsy or surgeryfor finally pathology. Based on above statistical data , we calculated EBUS-TBNA′s sensitivity, specificity and accuracy in 88.42%, 100.00%and 94.21%respectively. A totalof 818 lymph nodes were punctured by EBUS-TBNA, 408 were malignancy. The rate of diagnosis for malignancy was 49.88%. The complica-tioncontained 1 in hemorrhage, 1 in infectionand no life-threatening conditionandno death. Conclusion The value ofdiagnosis in lung cancerisrelatively high with EBUS-TBNAin safety and efficient.

OBJECTIVE:To establish the HPLC fingerprints for Dangua yangmu cream. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.026% phosphoric(gradient elution)at flow rate of 0.8 ml/min,the detection wavelength was 270 nm,the column temperature was 25 ℃ and the injection volume was 20 μl. The chromatographic peak of aurantio-obtusin was used as reference peak to determine the 10 batches of Dangua yangmu cream,and Similarity Evalua-tion System for Chromatographic Fingerprint of TCM(version 2.0)was conducted to identify common peaks and evaluate similari-ty. RESULTS:There were totally 25 common peaks in the 10 batches of Dangua yangmu cream,and the similarity was not lower than 0.921. The validation results showed the fingerprints of 10 batches of Dangua yangmu cream had good consistency with the ref-erence fingerprints. COMCLUSIONS:The established method is specific and reliable,and can provide basis for the quality evalua-tion and control of Dangua yangmu cream.

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.

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.