Background and purpose:Brucine is one of the active components from Strychnos nux vomica,with signifi cant analgesic,anti-inflammatory and platelet-aggregating inhibitory properties.Due to its cytotoxic effect,the anti-tumor effect of brucine has increasingly been appreciated.In this study,we investigated the impact of brucine on A549 cells proliferation,apoptosis as well as the underlying mechanisms.Methods:MTT assay was used to examine the cell viability,flow cytometric analysis and fluorescent microscope were applied to examine cell apoptosis,ELISA method was used to examine the effect of brucine on PGE2 release from A549 cells and RT-PCR analysis was used to measure mRNA content,western blotting analysis was used to measure protein expression and luciferase activity was detected to examine the effect of brucine on COX-2 promoter activity.Results:Brucine was able to suppress the proliferation of A549 cells and induce cell apoptosis to time-dependent and dose-dependent manner.To understand the mechanisms,COX-2 was identifi ed to be an important target molecule involved in the apoptosis induced by brucine because brucine could suppress the COX-2 mRNA,protein expressions as well as PGE2 release in A549 cells in a timedependent manner.Furthermore,overexpression of COX-2 abrogated brucine-induced cell apoptosis,in contrast,when A549 cells were transfected with COX-2 siRNA,the apoptotic effect of brucine was dramatically enhanced.Further analysis revealed that brucine was able to suppress COX-2 transcriptional activation.Conclusion:Brucine was able to induce lung cancer apoptosis via downregulation of COX-2.

Objective:To determine the effects of silencing glucose regulated protein ( GRP94 ) on the proliferation and apoptosis of breast carcinoma MCF7 cells. Methods:Chemically synthesized siRNA targeting GRP94 gene and transfected into MCF7 cells used by Liopfectamine RNAIMAX. The mRNA and protein expression levels of GRP94,cyclinD1,Bax and Bcl-2 were detected by Real-time PCR and Western blot. CCK8 assay was used to detect the effect of specific GRP94 siRNA on cell proliferation and the effect on cell cycle and apoptosis were analyzed by flow cytometry and Hoechst 33258 staining. Results:Compared with the siRNA-NC cells, the expression of GRP94 was significantly down-regulated in MCF7 cells. Knockdown of GRP94 in MCF7 cells decreased cell proliferation and promoted cell apoptosis. The expression of cyclinD1and Bcl-2levels were significantly reduced, and Bax level was increased in siRNA-GRP94 MCF7 cells. Conclusion: The siRNA-mediated GRP94 silence significantly inhibits MCF7 cell proliferation,promoted cell apoptosis by down-regulating cyclin D1,Bcl-2 expression and up-regulating the Bax expression in MCF7 cells.

Objective To investigate the value of femoral neck fracture(FNF) typing by digital subtraction angiography(DSA) in prognostic prediction of the fracture healing. Methods From June 2005 to June 2008,45 patients were admitted to our hospital for treatment of single femoral neck fracture.They were 26 males and 19 females,aged from 26 to 60 years(average,45.4 years).Their fractures were all typed according to DSA results.Type Ⅰ ( 15 patients) were DSA pictures which showed 3 to 6 supporting band vascular images crossing the fracture line.Type Ⅱ(14 patients) were DSA pictures which showed one to 2 supporting band vascular images crossing the fracture line.Type Ⅲ(16 patients) were DSA pictures which showed no supporting band vascular images crossing the fracture line.All cases underwent close reduction and internal fixation with cannulated screws under C-arm fluoroscopy.The prognoses of all the patients were evaluated according to their symptoms and imaging examinations in terms of avascular necrosis of femoral head (ANFH) and fracture healing. Results All the patients obtained an average follow-up of 36.7 months (from 24 to 60 months).In patients of DSA type Ⅰ,the ANFH rate and nonunion rate were respectively 0 and 13.3％ (2/15).In patients of DSA type Ⅱ,the ANFH rate and nonunion rate were respectively 7.1％ (1/14) and7.1％(1/14).In patients of DSA type Ⅲ,the ANFH rate and nonunion rate were respectively 100％ (16/16) and 0.The ANFH rate following FNF treatment was negatively related to age and the vascular number showed by DSA.The nonunion ratewas positively related to age but not significantly associated with the vascular number showed by DSA. Conclusion DSA has a significant value in predicting the prognosis for patients with femoral neck fracture.

Aim To investigate ALEX1 gene expres-sion in cervical cancer tissues and adjacent non-can-cerous tissues, and to explore the ALEX1 genetic influ-ence on cell proliferation,cycle and apoptosis of human cervical cancer cell line HeLa. Methods ALEX1 protein expression in cervical cancers and in non-can-cerous cervical tissues was evaluated using immunohis-tochemical method. A small interference RNA targeting ALEX1 gene was transfected into HeLa cells′, and the effect of ALEX1 interference on HeLa cells′ cycle and apoptosis was analysed by flow cytometry. The effect of ALEX1 interference on HeLa cells′ proliferation and sensitivity to resveratrol was analysed by CCK-8 assay. Results ALEX1 protein expression was significantly increased in cervical cancer tissues compared with non-cancerous tissues. HeLa cells′proliferation was inhibi-ted compared with control group and blank group. He-La cells′ sensitivity to resveratrol was enhanced com-pared with control group blank group. Conclution SiRNA silencing of ALEX1 gene could significantly in-hibit HeLa cells′ proliferation and enhance resveratrol ability of inhibiting HeLa cells′proliferation.

Objective:To investigate the effects of ALEX1 overexpression on cell proliferation and apoptosis of human breast cancer cell line MCF-7.Methods: MCF-7 cells were infected recombinant lentivirus LV5-ALEX1 and the negative control lentivirus LV5-NC,respectively.After 72 h, the expression of ALEX1 was detected by Real-Time PCR and Western blot.CCK8 assay were performed to observe the proliferation ability after 24 h, 48 h, 72 h, 96 h.The effect of overexpression ALEX1 on apoptosis was determined by flow cytometry.The level of Bax,BCL-2 and active caspase3 was detected by Western blot.Results:The mRNA level of ALEX1 markedly increased after 72 h(165.81±12.14 vs 52.29±2.32,P<0.01).In CCK8 assay,the results revealed that the cell pro-liferation was inhibited compared with control group in 48 h,72 h,96 h( P<0.05).The results revealed that overexpression of ALEX1 enhanced MCF-7 apoptosis(20.55%±2.50 % vs 3.60%±1.614%,P<0.05).The results by Western blot showed that the protein levels of Bax and active caspase were increased in LV5-ALEX1 group compared with LV5-NC group.However,the protein levels of BCL-2 was decreased in LV5-ALEX1 group compared with LV5-NC group.Conclusion:Overexpression of ALEX1 significantly reduced MCF-7 cancer cells proliferation and induced MCF-7 cells apoptosis.

AIM:Increasing evidence indicates that inflammation contributes to the initiation and perpetuation of atrial fibrillation ( AF) .Al-though tumor necrosis factor ( TNF)-αlevels are increased in patients with AF , the role of TNF-αin the pathogenesis of AF remains unclear.Recent research has revealed that T-type Ca2+currents ( ICa,T ) play an important role in the pathogenesis of AF .METH-ODS:In this study , we used the whole-cell voltage-clamp technique and biochemical assays to explore the role of TNF-αin the regula-tion of ICa,T in atrial myocytes.RESULTS:We found that compared with sinus rhythm (SR) controls, T-type calcium channel (TCC) subunit mRNA levels were decreased , while TNF-αexpression levels were increased , in human atrial tissue from patients with AF .In murine atrial myocyte HL-1 cells, after cultured for 24 h, 12.5, 25 and 50 μg/L TNF-αsignificantly reduced the protein expression levels of the TCC α1G subunit in a concentration-dependent manner .The peak current was reduced by the application of 12.5 or 25μg/L TNF-αin a concentration-dependent manner [from ( -15.08 ±1.11) pA/pF in controls to ( -11.89 ±0.83) pA/pF and (-8.54 ±1.55) pA/pF in 12.5 and 25 μg/L TNF-αgroups, respectively].TNF-αapplication also inhibited voltage-dependent inactivation of ICa,T shifted the inactivation curve to the left .CONCLUSION:These results suggest that TNF-αis involved in the path-ogenesis of AF, probably via decreasing ICa,T function in atrium-derived myocytes through impaired channel function and down -regula-tion of channel protein expression .This pathway thus represents a potential pathogenic mechanism in AF .

Objective To analyze the clinical characteristics and follow-up data of catheter ablation of recurrent atrial tachycardias (ATs) after Mini-Maze surgery,and to explore prognostic factors for recurrence.Methods 59 patients in Guangdong General Hospital with ATs post Mini-Maze and concomitant open-heart surgery from April.2010 to June.2015 were included.According to high density precise mapping,activation mapping,voltage mapping and entrainment mapping,they underwent electrophysiological study and ablation which was guided by three-dimensional mapping system.All patients were followed up regularly.We explored the prognostic factors for recurrence by the Cox regression analysis.Results There were 88 types of ATs being mappedwith mean (1.49 ± 0.75) types of ATs identified per case.Most ATs were macro-reentry ATs(67/88,76.1％)and focal ATs (20/88,22.7％),respectively.56 patients (94.9％) achieved immediate ablation success.In a mean follow-up of (30.8 ± 17.7) months,recurrences were observed in 12 patients after the first time catheter ablation.Recurrent time was 3.5 (1.3,12.0) months and the overall ablation success rate was 74.6％ (44/59).6 patients received second ablation and the achievement of freedom from arrhythmias reached 79.7％ (47/59).Multivariate analysis showed that the LA diameter was the independent predictor for recurrence (HR 1.108,95％ CI 1.002 to 1.226,P =0.045).Conclusion Catheter ablation of ATs post Mini-Maze with concomitant surgery is save and feasible.LA diameter is the independent predictor for recurrence.

Hyperlipidemia plays a vital role in cardiovascular disease, and threatens our lives. The aim of this paper is to study the effects of Shuanghua granules on blood lipid in normal mice and different hyperlipidemia models. Acute and endogenous hyperlipidemia was induced in mice with yolk and Triton WR-1339 respectively. The model of hyperlipidemia in rats was set up by feeding high cholesterol diet. Then preventive effects of Shuanghua granules was observed compared with lovastatin and Zhibituo. We found that Shuanghua granules 5.6, 11.3, 22.5 g x kg (-1) could significantly reduce the serum TG level in normal mice (P < 0.01, P < 0.05). No significant difference was found in liver index, serum TG and HDL-C levels. When the mice were treated with either yolk or Triton WR-1339 in the presence of Shuanghua granules, the plasma lipoprotein levels (TC and LDL-C) were significantly reduced (P < 0.01, P < 0.05). Shuanghua granules could reduce the serum TC, TG, LDL-C, MDA, NEFA and liver TC, TG, LDL-C levels, simultaneously raise serum and liver HDL-C, serum SOD, LPL, HL, LA levels of hyperlipidemia rats (P < 0.01, P < 0.05). Shuanghua granules also significantly reduced whole blood viscosity, RV, etaP, IER and IEA (P < 0.01, P < 0.05), and lowered fatty degeneration of liver tissue. Compared with hyperlipidemia model, there was no significant increase in faeces lipoids concentrations. The results confirmed the mechanism of blood lipid regulating effects of Shuanghua granules is probably related with its antioxidation, regulating hemorheology and improving LPA, HL, LA enzymatic activity.
Animals ; Blood Viscosity ; drug effects ; Cholesterol ; blood ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Hyperlipidemias ; blood ; drug therapy ; prevention & control ; Lipid Peroxidation ; drug effects ; Lipids ; blood ; Lipoprotein Lipase ; blood ; drug effects ; Liver ; drug effects ; pathology ; Lonicera ; Male ; Mice ; Phytotherapy ; Plant Preparations ; Rats ; Triglycerides ; blood

Objective:Based on the cost-effectiveness,it aimed to assess the health benefits amd economic value of screening,di-agnosis,treatment,and optimal delicacy management of liver disease in hepatocellular carcinoma(HCC)patients.Methods:A Deci-sion tree-Markov model was developed to compare the cost-effectiveness of HCC screening and long-term surveillance versus no screening in population at risk from the health care system perspective.Results:It is found that HCC screening was a cost-effective approach compared to no screening(Incremental Cost-Effectiveness Ratio[ICER]:17 790 yuan/QALY).Scenario analyses suggested that initiating HCC screening at the age of 40,as recommended by clinical guidelines,and implementing long-term surveillance based on risk stratification were more cost-effective.Conclusions:For the implementation of HCC screening programs,attention should be paid to improving participation and compliance among the population at risk,incorporating advanced screening methods,improving management efficiency with digital tools,and introducing innovative payment methods to reduce economic burden.

This paper summarized clinical experience of treating recurrent subacute thyroiditis based on the theory of internal toxin. It is believed that the core pathogenesis of recurrent subacute thyroiditis is the accumulation of internal toxin. The dysfunction of the spleen in transportation leads to the internal retention of turbidity， which serves as the basis for the formation of toxins. The combination of fire， dampness， and toxins is the key to the disease mechanism. Prolonged accumulation of dampness and turbidity leads to the formation of phlegm， fire， and stasis， and prolonged accumulation of pathogenic factors transforms into secondary toxins. For patients with severe fire， dampness， and toxins， the treatment focuses on dispersing stagnated liver qi， invigorating spleen， and differentiation treatment of fire and water， using a self-designed Chaihu Jiedu Decoction （柴胡解毒饮）. For patients with spleen deficiency and toxin accumulation， the treatment focuses on invigorating the spleen， eliminating turbidity， and cutting off the source of toxins， using a self-designed Shiwei Baizhu Powder （十味白术散）. For patients with yang deficiency and toxin stagnation， detoxification is combined with warming yang， nourishing blood， and supporting toxin expulsion， using a self-designed Heyang Baidu powder （和阳败毒散）.