Background and purpose: Sorafenib, a multiple targeted agent, can inhibit proliferation and induce apoptosis of diverse tumor cell in vitro. It has extensive biological activities, but the pancreatic cancer effect of monotherapy is poor. This may be related to nuclear factor-kappa B (NF-κB) pathway activation in cancer. Therefore, it is necessary to combine with Pyrrolidinedithiocarbamate (PDTC, a NF-κB activation inhibitor) to enhance curative effect. To investigate the influences of cell proliferation, cell cycle and expression of NF-κB via their acting on human pancreatic cancer PANC-1, and explore their possible mechanism. Methods:The experiment groups were divided into sorafenib group with different concentrations (1.5, 3.0, 6.0, 12.0 μmol/L), PDTC group with different concentrations (10.0, 25.0, 50.0, 100.0 μmol/L) and combination group with low concentration (3.0 μmol/L sorafenib+25.0 μmol/L PDTC). The proliferative activity of PANC-1 of each group was measured by MTT assay at different time points of 24

Cancers of the digestive system account for a large portion of malignant tumors in humans,and the trend is on the rise.The formation of neovascularization is the dominant factor in the metastasis of tumors.The vascular endothelial growth factor(VEGF) is well documented as the most potent inducer of angiogenesis.It promotes the formation of new blood vessels in several aspects,such as proliferation of endothelial cells,endothelial cell migration and increased vascular permeability.So VEGF is regarded as an important factor in the development of digestive system tumors.The purpose of this review is to investigate the relationship between VEGF and digestive system tumors.

Objective To analyze the relationship between the change of serum vascular epidermis growth factor(VEGF) level and curative effect of pancreatic cancer treated by gamma rays, and evaluate it clinical significance. Methods The serum VEGF level of thirty patients with pancreatic cancer treated by gamma rays were determined before and one month after treatment. The relationship between curative effect,survival, life quality and the serum VEGF level was analyzed. Results The serum VEGF level of 30 cases before treatment was (624.1 ± 144.3) pg/ml, whereas one month after treatment it was (279.3 ± 83.4) pg/ml(P ＜0.01 ). In the 19 patient whose serum VEGF level decreased by more than 50%, CR + PR was gained in 18 patients, improved quality of life in 7 patients, and in the 11 patient whose serum VEGF level decreased by less than 50%, CR + PR in 7 patients, improved quality of life in 1 patient( P = 0.012, P = 0. 028). In 24patients who completed 2 years follow-up, 11 of 15 patients whose serum VEGF level decreased by more than 50% survived more than 1 year, which was higher than that of the patients whose serum VEGF level decreased by less than 50%. Conclusions The serum VEGF level decreased significantly after gamma rays treatment and the extent of decrease could be used as a surrogate to evaluate the therapeutic efficacy.

Objective To investigate the clinical significance of detecting cardiac troponin Ⅰ (cTnⅠ) and CK -MB in children with hand-foot-and-mouth disease and myocardial injury.Methods 90 children with hand-foot-and-mouth disease (observation group) were detected the level of serum cTnⅠ and CK-MB.At the same time,40 healthy children were chosen as control group.Results Compared with the control group,the levels of CK-MB and cTnl in observation group were significantly higher than those of the control group (t =8.92,5.46,all P ＜ 0.01),which indicated that children with hand,foot and mouth disease was easier to merge myocardial injury.CK-MB and cTnl levels in high-risk group were significantly higher than those in normal children group (t =9.17,6.13,all P ＜ 0.01),and the levels of CK-MB and cTnl were positively correlated with severe degree (r =0.767,0.683,all P ＜ 0.01).For children with hand,foot and mouth disease merged myocarditis,cTnl diagnostic sensitivity (53.8％) was lower than that of CK-MB diagnostic sensitivity (71.8％),but the specificity was better than that of CK-MB,and the diagnosis of cTnl detection window 2 weeks longer than CK-MB,but after 2 weeks both lose their clinical diagnostic significance.Conclusion Children with hand,foot and mouth disease easily merge myocardial damage,dynamic measuring CK-MB and cTnl levels could help early diagnosis of children with hand,foot and mouth disease whether merged myocardial damage,both applications can also complement each other,more timely and accurate reflection of disease progression and recovery,it is worth promoting.

Objective To investigate hypolipemic treatment of hyperlipidemic panereatiti(HLP)with integrated traditional Chinese and western medicine.Methods Cinical data of 20 patients of HLP were analyzed retrospectively.Eight patients in the control group were treated with conventional therapies,while 12 patients in the treatment group were treated as follows:①Enema with 180ml solution(50% magnesium sulfate 30 ml.Glycerin 60ml,water 90ml).②Rhubarb gastrogavage with 9 g tid.③Intravenous drip with 24 g salvia miltiorrhiza qd.Results The treatment group had significant difference comparing with the control group in terms of the serum TG in 48 hours(P<0.01),time of autonomous bowel movement recover(P<0.01),days of abdominal pain disappear(P<0.05),days of hospitalization(P<0.01).Conclusion The treatment of Enema with 180 ml solution.Rhubarb gastrogavage with 9g tid,and Intravenous drip with 24 g salvia miltiorrhiza qd can relieve the symptoms of HLP and decrease blood-fat greatly.

Objective To discuss the application of combined probiotics pretreatment and late triple therapy with helicobacter pylori infection in children.Methods 300 children with helicobacter pylori infection were randomly divided into three groups according to the way of random table.Group A was treated with standard triple therapy, omeprazole,amoxicillin and clarithromycin treatment,and 10 d was 1 treatment course.B group was given probiotic pretreatment,prior to the triple therapy using compound lactobacillus acidophilus piece 1 piece /time,cold water to take after meals,taking 2 weeks,follow -up treatment with triple therapy for 10d.In group C,triple therapy before using compound lactobacillus acidophilus piece 1 piece /time,cold water to take after meals,taking 2 weeks,follow -up treatment with triple therapy for 10d.To take one week after the completion of compound lactobacillus acidophilus piece 1 piece /time,cold water after meals.The clinical therapeutic effects were recorded.Results The helicobacter pylori clearance rate of group A was 55.00%,that of group B was 86.00%,that in group C was 89.00%,the helico-bacter pylori clearance rate of group B and group C was significantly higher than that in group A,the difference was statistically significant (χ2 =23.103 7,28.670 6,all P <0.05).The treatment effect between group B and group C had no statistically significant difference (χ2 =0.411 4,P >0.05 ).In A group,nausea and vomiting occurred in 5 cases,2 cases of diarrhea,abdominal distension abdominal pain in 4 cases,skin rash in 5 cases.In group B,nausea and vomiting occurred in 2 cases,0 cases of diarrhea,abdominal distension abdominal pain in 1 case,skin rash in 1 case.In group C,nausea and vomiting occurred in 1 case,0 cases of diarrhea,abdominal distension abdominal pain 0 cases,skin rashes in 1 case.The incidence rate of adverse reactions in group B and group C was lower than that in group A,the difference was statistically significant (χ2 =11.965 8,8.000 0,all P <0.05).The incidence rate of ad-verse reactions between group B and group C had no statistically significant difference (χ2 =0.687 3,P >0.05). Conclusion Joint probiotics pretreatment and late triple therapy application in helicobacter pylori in children can promote helicobacter pylori clearance,reduce the triple therapy drug adverse reactions,it is worthy of popularization and application in clinic.

AIM: To investigate the effect of phospholamban antisense RNA (asPLB) on the activity of sarco-endoplasmic reticulum (SR) Ca 2+-ATPase, and the change of intracellular free Ca 2+ concentration ([Ca 2+]i) in rat cardiomyocytes by adeno-associated virus(AAV) vector. METHODS: rAAV-asPLB and rAAV-LacZ were constructed by AAV Helper-Free System. RT-PCR and Western blotting were used to determine the mRNA and protein expression of PLB. The activity of SR Ca 2+-ATPase and the [Ca 2+]i were measured. RESULTS: Compared to controls, the PLB mRNA and protein expression reduced in rat cardiomyocytes transfected with rAAV-asPLB. The activity of Ca 2+-ATPase was increased. In rest state, the level of [Ca 2+]i in rAAV-asPLB transfected group was decreased. The level of [Ca 2+]i was increased when induced by isoproterenol. CONCLUSION: rAAV-asPLB vector disrupts the expression of PLB, enhances the activity of Ca 2+-ATPase, reduces the resting [Ca 2+]i and enhances the isoproterenol-induced [Ca 2+]i.

Objective: To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages. Methods: LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay. Results: Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels. Conclusion: In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.

objective To investigate the epidemiological and molecular typing features of the pathogenic Haemophilus influenzae(H.influenzae)by biotyping,serotyping and pulsed-field gel electrophoresis(PFGE).Methods A total of 273 invasive isolates of H influenzae were collected from the pediatric patients with pneumonia at Chengdu Children Hospital of Sichuan province from 1988 and 2004 to 2007.The idenbfication of H.influenzae strains were done according to the laboratory standard methodology described by Manual of Clinical Microbiology(American).All strains were biotyped according to Kilian's classification with the API[R]NH system.And serotyped by a slide agglutination assay with type a to f specific antlaerum as described by Pittman.PCR method for identification of H.influenzae were performed as described by Falla.One hundred of 273 strains were analyzed by PFGE as described by Saito with some modifications.The resuIts of PFGE were analyzed by Bionumerics soft(Version 4.0,Applied Maths BVBA,Belium).Restilts 78.2%of 273 cases occurred under 1 years old.Eight biotypes were found among the 273 H.influenzae isolates.17.6%(48/273)of all isolates belonged to biotype Ⅰ,43.6%(119/273)were biotype Ⅱ,22.7%(62/273)were biotype Ⅲ,7.3%(20/273)were biotype Ⅳ,5.9%(16/273)were biotype Ⅴ,0.4%(1/273)were biotype Ⅵ,1.8%(5/273)were biotype Ⅶ and 0.7%(2/273)were biotype Ⅷ.respeetively.99.6% of all 273 isolates were nontypeable.There was only one isolate was serotvpe f Ninty-six PFGE genotypes were obtained in this study.One hundred strains demonstrated a variety of genomic Datterns by PFGE.The most isolates of the flame PFGE genotype(type 35)was 3 isolates.Each of93 PFGE genotypes was represented by only a single isolate.The genotypes distribution didn't correlate with the time distribution of the strains were isolated.Conclusion Nontypeable H.influenzae primarily caused acute Dneumoma in children under 1 years old.They mostly belonged to biotype Ⅰ,Ⅱ and Ⅲ biotypes.The nontypeable H.influenzae strains appeared to more heterogeneous patterns by PFGE genotyping.Genotyping may helP understand the molecular characteristics of outbreak and endemicity according to the results of PFGE.PFGE genotyping proved to have a much stronger discriminatory power than either serotyping or biotyping.Our findings suggest that PFGE analysis is useful for the epidemiologieal study of H.influenzae infections.

Objective To establish cell strain expressing the genes of HIV vpr and mutant HIV vpr-FS, and to explore cell apoptosis ability by HIV Vpr and Vpr-FS. Methods The recombinant plasmids were constructed by cloning HIV vpr and HIV vpr-FS genes into the eukaryotic expression vector pcDNA3.1respectively. To determine the primary structures of HIV vpr and HIV vpr-FS, plasmids were cleaved by restriction enzymes. After the plasmids were transfected into HeLa cells by liposome, the HeLa cells were selected with G418 selective medium, mRNA expression of HIV vpr or HIV vpr-FS of transfected cells was detected by RT-PCR, and Vpr and Vpr-FS protein expression were detected by Western blot assay respectively. The DNA content and the percentage of apoptosis in HeLa HIV vpr cell, HeLa HIV vpr-FS cell and HeLa pcDNA3.1 cell were monitored by flow cytometry and the DNA fragmentation was analyzed by agarose gel electrophoresis. Results BamH Ⅰ and Hind Ⅲ cleavaged products of pcDNA3.1-vpr and pcDNA3.1-vpr-Fincluded 342 bp length fragments suggesting that the length of DNA sequence containing HIV vpr (HIV vpr-FS) within pcDNA3.1 was the same as theoretical length. The HeLa cells transfected by pcDNA3.1-vpr or pcDNA3, l-vpr-FS and selected with G418 could express HIV vpr or HIV vpr-FS by RT-PCR, and express HIV Vpr or HIV Vpr-FS protein by Western blot. The results of flow cytometry and DNA fragmentation showed that there was significant different in the number of apoptotic cells between HeLa HIV vpr cell and HeLa HIV vpr-FS cell, but the difference between HeLa HIV vpr-FS cell and control group was not obvious. Conclusion Recombinant plasmids pcDNA3.1-vpr and pcDNA3. 1-vpr-FS were constructed successfully, and the cell strain expressing HIV Vpr and HIV Vpr-FS proteins was established. The HIV Vpr could induce host cell apoptosis, while the mutant of Vpr did not or weakened this ability. This study provides foundation for further study on HIV vpr gene.