Objective: To investigate the inhibitory effect of eukaryotic expression vector (attenuated salmonella typhimurium) carrying tumor necrosis factor-related apoptosis inducing ligand (TRAIL) and Chicken anemia virus VP3 gene on gastric cancer cells in vitro and in vivo. Methods: The cloning vectors pBud-TRAIL, pBud-VP3, and pBud-TRAIL-VP3 were transformed into attenuated Salmonella typhimurium by electric transformation technique. The S. typhimurium-based carriers were then transfected into gastric cancer cells, line SGC-7901 after stability assay. The expression of fusion green fluorescent protein was examined using fluorescent microscopy after 24 h. MTT assay was used to examine the inhibition of cell growth. Flow cytometry was used to detect cycle distribution and apoptosis rates of cells. The expression of caspase-3 and caspase-9 was assayed by immunohistochemistry method. Salmonella typhimurium carrying recombinant plasmid was administrated orally in sarcoma-bearing mice; 8 weeks later RT-PCR was used to detect the expression of cloning vectors in tumor tissue. Meanwhile, the sizes of tumors were also determined. Results: The recombinant plasmids were stably transformed into attenuated Salmonella typhimurium, and the plasmids was satisfactorily expressed in gastric cancer cells via attenuated Salmonella typhimurium. TRAIL and VP3 inhibited the proliferation of gastric cancer cells after 48 h. Flow cytometry analysis showed that the pBud-TRAIL-VP3 obviously enhanced apoptosis rates of gastric cancer cells. TRAIL and VP3 jointly increased the expression of caspase-3 and caspase-9. In vivo study showed that TRAIL and VP3 genes were expressed in tumor tissue and could inhibit the tumor growth(P<0.05). Conclusion: Attenuated Salmonella typhimurium-mediated TRAIL and VP3 transfection of gastric cancer cells can inhibit cell growth in vitro and in vivo. The joint effect of TRAIL and VP3 is correlated with the increase of caspase-3 and caspase-9 expression.

OBJECTIVE To investigate the relationship between HBV DNA and serum IL-18 level in patients with hepatitis B virus. METHODS They were divided by MEIA method into 3 groups. group A:HBsAg+, HBeAg+ and HBcAb +(A); group B:HBsAg+, HBeAb+ and HBcAb+(B); and group C:HBsAb+(as normal controls ). Serum IL-18 level was determined by ELISA method in hepatitis patients and normal controls and HBV DNA was determined continuously . RESULTS Concentration of serum IL-18 in patients with different type of viral hepatitis was significantly higher than those in healthy volunteers(P

OBJECTIVETo investigate the effect of curcumin on the injury in hippocampal neurons and the expression of regulated upon activation nonnal T-cell expressed and secreted (RANTES) in hippocamp during cerebral ischemia/reperfusion (I/R) in rats with spontaneous hypertension (SH).

METHODSMale Wistar-Kyoto (WKY) rats and spontaneous hypertension rats (SHR) were randomly divided into five groups (n = 6): sham group (W-Sham and S-Sham group), ischemia/reperfusion group (W-/R and S/R group), curcumin group (S-Cur group) . Each group was splitted into 5 subgroups of 3 h,12 h, 1 d, 3 d and 7 d according to the time interval before reperfusion. Global brain ischemia/reperfusion model was established by 4-VO method. Hematoxylin-eosin staining (HE staining) was used to observe the vertebral cell morphology in hippocampal CA1 region. Nissl staining was applied to detect the average density of cone cells in hippocampal CA1 region. The expression of RANTES in hippocamp was determined by ELISA. The behavior of the rats was evaluated at 7 days after reperfusion. Results: Compared with the sham group rats, the ability of learning and memory was significantly decreased in ischemia/reperfusion group rats, the number of injured neurons were greatly elevated , the protein expression levels of RANTES was significantly increased (P < 0.05). Compared with W-I/R group rats, the ability of learning and memory in S-I/R group rats was greatly reduced, the number of injured neurons increased extremely, the protein expression level of RANTES was significantly enhanced( P <0.05). The number of injured neurons declined significantly in S-Cur group rats, the ability to learn and remember of these rats was improved and the RANTES protein content decreased significantly (P < 0.05).

CONCLUSIONSHR are more susceptible to ischemia/reperfusion induced hippocampal neuronal injury which may be improved by curcu min. Its underlying mechanism is possibly associated with the inhibition of RANTES protein expression level.

Animals ; Brain Ischemia ; metabolism ; pathology ; physiopathology ; Chemokine CCL5 ; metabolism ; Cognition ; drug effects ; Curcumin ; pharmacology ; Hippocampus ; cytology ; metabolism ; pathology ; Hypertension ; metabolism ; pathology ; physiopathology ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Reperfusion Injury ; metabolism

Structural modifications were performed with natural product of oleanolic acid to search for novel anticancer drugs. Ten oleanolic acid derivatives were designed and obtained by the reaction of oxidation, acylation or hydrolyzation, etc. The cytotoxic activity of derivatives was evaluated against HeLa, HepG2 and BGC-823 cells in vitro by MTT assay, gefitinib and etoposide used as a positive control. The results showed that compound 5a was particularly active to inhibit HepG2 cells growth, and anti-tumor activity of compound 7 on HeLa cells was significantly stronger than oleanolic acid. They are worthy to be studied further.

Objective To establish an HPLC method for determination of low-molecular-weight carbonyl compounds in the oil-containing herbs. Methods After carbonyl compounds in the samples were extracted with water, the solution reacted with 2, 4-dinitrophenylhydrazine (DNPH) in an acidic medium to form 2, 4-dinitrophenylhydrazone derivatives, which were separated on Kromasil KR100-5 C18 column (250 mm× 4.6 mm, 5 μm). The mobile phase A was water-acetonitrile-tetrahydrofuran-isopropanol (59∶30∶10∶1), mobile phase B was water-acetonitrile (35∶65), gradient elution. The flow rate was 0.8 mL/min, detection wavelength was 365 nm, and column temperature was 30 ℃. Results Good linearities were obtained in corresponding concentration ranges, with correlation coefficient over 0.999. The limits of detection of the eight DNPH derivatives were 0.002-0.008 μg/mL, and the average recoveries were 88.49%-93.65%. Conclusion The method is simple, accurate and reliable, with good reproducibility.

Objective To find out the status of HIV infection,sexual behaviors with both men and women as well as condom use among currently married men who have sex with men (MSM) so as to document for HIV intervention targeting this sub-population,in Beijing,Chengdu,Harbin and Zhengzhou cities.Methods “Snowballing” sampling method was used to recruit subjects in four cities.Participants were investigated by anonymous questionnaire containing demographics,sexual orientation,both homosexual and heterosexual behaviors and condom use during the past 6 months.Blood samples from participants were used to test HIV antibodies.Results Of the 858 participants for four cities,the average age was 38.3 (SD=9.1) years.36.7％ of them had completed the junior high school or under.The percentage of participants who identified themselves as homosexual,bisexual,and heterosexual or undecided were 40.9％,54.3％ and 4.8％,respectively.The overall HIV prevalence was 8.0％,and among four cities the HIV prevalence in Chengdu was the highest ( 13.3％ ).Results from the Multiple logistic regression analysis showed that never used condoms when having anal sex with men during the past 6 months among participants who did not have a city residential card,identifying himself as having homosexual orientation,living in Harbin,having first anal sex experience before 18 years of age the range of OR value was 1.5-2.1.In addition,the faetors associated with never using condoms in vaginal sex with women during the past 6 months were as follows:being over 46 years old,not having a city residential card,living in Beijing,identifying himself as heterosexual orientated or with unknown sexual orientation,never using condoms in anal sex with men during the past 6 months,the range of OR value was 1.7-5.9.Conclusion The prevalence of HIV infection seemed to be quite high among those currently married MSM.High rates of unprotected homosexual and heterosexual behaviors might accelerate the spreading of HIV from currently married MSM to their wives.

OBJECTIVETo investigate the therapeutic efficacy of interferon (IFN) therapy and risk of long-term administration for chronic hepatitis C (CHC) patients who cannot tolerate the standard treatment.

METHODSForty-six CHC patients who had proven intolerant to standard treatments were treated with low-dose IFN (non-pegylated IFN: 60 to 300MIU QOD, or pegylated IFN: 50 to 90 mug/w) plus ribavirin (RBV; 0.6g to 0.9 g/d) for 72 weeks.

RESULTSForty-three (93.5%) of the patients were able to tolerate the long-term treatment with low-dose IFN plus RBV. Only three patients experienced severe side effects (low white blood cell and platelet counts) that required treatment withdrawal. The virology response rates over treatment time were: rapid virologic response (RVR): 10.9%; early virus response (EVR): 30.4%; 24 week virologic response: 45.7%; and, 48 week virologic response: 47.8%. B-sonographic imaging revealed that three patients experienced improved liver morphology through the treatment course. The patients who achieved RVR, EVR, or 24 weeks virologic response also attained higher 48 week virologic response. The 24 week virologic response had the strongest predictive value of good prognosis.

CONCLUSIONSOur study demonstrated that long-term treatment with low-dose interferon plus ribavirin is effective for patients who are otherwise intolerant to standard treatment. In these patients, low-dose IFN plus RBV can obtain a high virologic response rate at 48 week. Furthermore, the 24 week virologic response is sufficiently predictive of treatment success. As with any treatment regimen, it is important for healthcare workers to monitor the disease status and potential side effects throughout the course of therapy.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Hepacivirus ; Hepatitis C, Chronic ; drug therapy ; virology ; Humans ; Interferons ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo investigate the effects of BmkTXK(beta), a newly purified 'long chain' peptide inhibitor of K(+) channels from the Chinese scorpion Buthus martensi Karsch (BmK), on the electrophysiological properties of isolated rabbit atrial myocytes.

METHODSThe standard whole-cell patch-clamp technique was used to study the effects of multiple concentrations of BmkTXK(beta) on potassium currents and action potentials.

RESULTSBmkTXK(beta) produced concentration-dependent prolongation of action potential duration at 20%, 50%, and 90% repolarization (APD(20,50,90)) without any use-dependence. Meanwhile, it had no significant effect on RMP, APA, or V(max) (n = 9). At a dose of 1 micro mol/L, BmkTXK(beta) decreased I(to) by 41.4% (n = 10, P < 0.01) at a membrane potential of +50 mV [from (13.63 +/- 0.87) pA/pF to (7.98 +/- 0.78) pA/pF]. I(to) was reduced significantly with an IC(50) value of 1.82 micromol/L (95% confidence interval: 1.47 - 2.17 micro mol/L), in a clear concentration-dependent manner. BmkTXK(beta) blocked I(Ks) and I(Ks),tail with an IC(50) of 20.15 micromol/L and a 95% confidence interval of 16.93 - 23.37 micromol/L. At a concentration of 10 micromol/L, BmkTXK(beta) blocked both I(Ks) (mean reduction 37.3% +/- 4.2%, P < 0.01, n = 7) and I(Ks), tail (mean reduction 35.8% +/- 4.1%, P < 0.01, n = 7). At 0 mV, 10 micromol/L BmkTXK(beta) inhibited both I(Kr) (mean reduction 40.5% +/- 2.6%, P < 0.01, n = 6) and I(Kr), tail (mean reduction 42.3% +/- 2.9%, P < 0.01, n = 6). Blocking of I(Kr) by BmkTXK(beta) occurred in a concentration-dependent manner, with an IC(50) of 17.21 micromol/L (95% confidence interval: 14.76-19.66 micromol/L). An absence of effects on I(K1) was observed for BmkTXK(beta), with no change in reversal-potential (n = 6, P > 0.05).

CONCLUSIONSBmkTXK(beta) exerts direct blocking effects on several potassium channels involved in cardiac repolarization, and has a strong effect on prolonging the repolarization of rabbit cardiomyocytes without reverse frequency dependence. This finding suggests that BmkTXK(beta) could be a promising class III drug for anti-arrhythmic therapy without the risk of proarrhythmia.

Action Potentials ; drug effects ; Animals ; Cells, Cultured ; Heart Atria ; Myocardium ; cytology ; Patch-Clamp Techniques ; Rabbits ; Scorpion Venoms ; pharmacology

To investigate the urination-reducing effect and mechanism of Zhuangyao Jianshen Wan (ZYJCW). In this study, SI rats were subcutaneously injected with 150 mg · kg(-1) dose of D-galactose to prepare the sub-acute aging model and randomly divided into the model group, the Suoquan Wan group (1.17 g · kg(-1) · d(-1)), and ZYJCW high, medium and low dose groups (2.39, 1.20, 0.60 g · kg(-1) · d(-1)) , with normal rats in the blank group. They were continuously administered with drugs for eight weeks. The metabolic cage method was adopted to measure the 24 h urine volume and 5 h water load urine volume in rats. The automatic biochemistry analyzer was adopted to detect urine concentrations of Na+, Cl-, K+. The ELISA method was used to determine serum aldosterone (ALD) and antidiuretic hormone (ADH). The changes in P2X1 and P2X3 mRNA expressions in bladder tissues of rats were detected by RT-PCR. According to the results, both ZYJCW high and medium dose groups showed significant down-regulations in 24 h urine volume and 5 h water load urine volume in (P <0.05, P <0.01), declines in Na+ and Cl- concentrations in urine (P <0.01), notable rises in plasma ALD and ADH contents (P <0.05, P <0.01) and remarkable down-regulations in the P2X1 and P2X3 mRNA expressions in bladder tissues (P <0.01). The ZYJCW low dose group revealed obvious reductions in Na+ and Cl- concentrations in urine (P <0.01). The results indicated that ZYJCW may show the urination-reducing effect by down-regulating the P2X1 and P2X3 mRNA expressions in bladder tissues of rats with diuresis caused by kidney deficiency.
Aging ; physiology ; Animals ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression Regulation ; Kidney Diseases ; drug therapy ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Receptors, Purinergic P2X1 ; genetics ; Receptors, Purinergic P2X3 ; genetics ; Urinary Bladder ; metabolism

OBJECTIVETo investigate the effects of Gastric bypass surgery on the apoptosis of islet β-cells in type 2 nonobese diabetic (NOD) rats and its mechanisms.

METHODSSeventy-two 8-week-old GK rats were randomly divided into four groups:operation group (group O, n = 18), sham operation group (group S, n = 18), diet control group (group F, n = 18) and control group (group C, n = 18). The levels of fasting, postprandial blood glucose, insulin and glucagon-like peptide-1 (GLP-1) were measured and compared among the 4 groups before the operation and at 1, 2, 4 and 8 weeks following the operation. The blood samples were collected at 2, 4 and 8 weeks after the operation for the measurement of postprandial blood glucose, and then the rats in batches (6 rats in each group) were decapitated to retrieve the pancreas. The apoptosis of the islet β-cells was detected by using TUNEL assay, and the expression of apoptosis-related proteins Bcl-2, Bax was measured with immunohistochemistry.

RESULTSAs for group O, the fasting blood glucose level decreased from (16.2 ± 0.8) mmol/L before the operation to respectively (9.2 ± 0.6) mmol/L and (9.7 ± 0.7) mmol/L at 4 and 8 weeks after the operation; postprandial blood glucose decreased from (31.1 ± 1.1) mmol/L before the operation to respectively (13.1 ± 0.7) mmol/L and (12.3 ± 0.7) mmol/L at 4 and 8 weeks after the operation. Fasting insulin level increased from (28.0 ± 1.2) mU/L before the operation to respectively (62.8 ± 1.9) mU/L and (61.7 ± 1.4) mU/L at 4 and 8 weeks after the operation; and at 4 and 8 weeks after the operation postprandial insulin level was (77.4 ± 1.1) mU/L and (77.1 ± 1.0) mU/L. At 2 weeks from the operation, the fasting GLP-1 in group O increased from (10.7 ± 1.0) pmol/L to (13.5 ± 0.8) pmol/L, and respectively to (26.1 ± 0.9) pmol/L and (25.3 ± 1.2) pmol/L at 4 and 8 weeks after the operation. The differences in the above-mentioned items before and after the operation were all significant in group O (P < 0.05), and the differences in the items among group O and the other three groups (P < 0.05) were all significant as well. In group O, the apoptosis rate of pancreatic islet cell decreased to (5.9 ± 0.7)% at 4 weeks from the operation, and (6.3 ± 1.1)% at 8 weeks from the operation (P < 0.05). The expression of Bcl-2 protein in group O was 31.3 ± 1.5, 35.7 ± 1.0 and 35.8 ± 0.8 at 2, 4 and 8 weeks post operation, which was significantly higher in statistics than those of the same time point in the other three groups (P < 0.05). The expression of Bax protein in group O was 13.3 ± 0.9, 10.8 ± 0.9 and 10.9 ± 1.1 at 2, 4 and 8 weeks from the operation, which was significantly lower in statistics than those of the same time point in the other three groups (P < 0.05).

CONCLUSIONSGastric bypass surgery can significantly reduce the blood glucose level and promote the secretion of GLP-1, and therefore inhibit the apoptosis of the islet β cells in diabetic rats through the Bcl-2 pathway.

Animals ; Apoptosis ; Blood Glucose ; Diabetes Mellitus, Type 2 ; pathology ; surgery ; Disease Models, Animal ; Gastric Bypass ; Glucagon-Like Peptide 1 ; blood ; Insulin ; blood ; Islets of Langerhans ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; bcl-2-Associated X Protein ; metabolism