Objective To observe a recombinant mutant of HBV core protein for dominant negative gene therapy against HBV encapsidation in vitro. Methods C gene and S gene of HBV were acquired through PCR and subcloned into pGEM T to construct pGEM T C and pGEM T S respectively. After digestion and ligation of these two plasmids, pGEM T CS was constructed. The cloned gene was inserted into pcDNA3.1 + to construct pcDNA3.1 + CS, which was identified by DNA sequencing. The recombinant plasmids were transformed into HepG2 cells, and screened with G418. The resistant HepG2 cell clones were chosen to test the expression of core surface protein by RT PCR, and the expressing HepG2 clones were cultured with 10% HBV DNA positive human serum for 72 hours. The intracellular HBV particles were extracted and the DNA was subjected to dot hybridization. Results The analysis showed that the HepG2 cells expressing mutant C protein had capabilities to resist HBV invasion in varied degrees. The mutant C protein had a dominant negative role in the encapsidation of HBV compared with the naive part of core protein. Conclusions The production of recombinant mutant core protein has a potential value for gene therapy against HBV infection.

This article elucidates the relationship between the human susceptibility to obesity and gene polymorphisms such as peroxisome proliferators-activated receptors(PPARs)and PPAR?coactivator-1,along with milestones in the formation and development of capacity for fat deposition during evolutionary history of human.An biological evolutionary analysis,identifying factors favoring the energy stores,may be helpful to the development of preventive public health strategies.

Objective To evaluate the value of diagnostic negative MDCT cholangiopancreatography in patients with suspected obstructive biliary diseases by receiver operating characteristic(ROC)curve. Methods Dual-phases contrast-enhanced MDCT of the abdomen was performed in 30 patients.Multiplanar reformation,minimum intensity projection and volume rendering were generated using original data in vein phase.By double-blind method,the images were readed by two radiologists.ROC curves were analysed with software SPSS11.5,the different results of reading images between two radiologists was evaluated with Kappa test.Results The Az of the area under the ROC curve of MDCT cholangiopancreatography for the two observers was 0.968 and 0.962 respectively,it was more than 0.9,which showed that the value of diagnosing the suspected obstructive biliary diseases by negative MDCT cholangiopancreatography was good.Kappa index test was satisfactory.Conclusion Negative MDCT cholangiopancreatography may be a routine choice in diagnosing the suspected obstructive biliary diseases.

Objective To prospectively assess the nutritional status in the patients with alimentary tract malignancy,and to elucidate the factors related to malnutrition.Methods The nutritional status of 328 patients with newly diagnosed alimentary tract malignancy was assessed using subjective global assessment(SGA)and serum levels of prealbumin and albumin.And the factors influencing the nutritional status of the patients with alimentary tract malignancy in different locations were analyzed.Results The prevalence of malnutrition was 64.43% in all,75.81% in colon cancer,63.24% in esophageal cancer,62.40% in gastric cancer and 60.27% in rectal cancer.The changes of nutritional status mainly manifested weight loss with the incidence of 67.39%,serum prealbumin level under 200 g/L with the incidence of 24.1% and serum albumin level less than 35 g/L with the incidence of 31.70%.And there was significant difference in weight loss and serum levels of prealbumin and albumin among the patients with different nutritional status(P=0.000).The factors that influence the nutritional status of the patients with alimentary tract malignancy include the location and TNM staging of tumors,and the age,appetite and digestive symptoms of the patients.Conclusion The patients with alimentary tract malignancy are susceptible to malnutrition due to the multiple factors such as the tumor location and metabolic impacts of tumor on host.Nutritional screening,assessment and early intervention should be emphasized in the inpatients with alimentary tract malignancy.

OBJECTIVETo observe the therapeutic efficacy of Dangfei Liganning Tablet (DLT) in the treatment of antipsychotics induced mild hepatic damage.

METHODSTotally 80 mental inpatients with antipsychotics induced mild liver injury were randomly assigned to two groups, the treatment group (40 cases) and the control group (40 cases). Patients in the treatment group took DLT, two tablets each time, three times per day, while those in the control group took Liver-protecting Tablet (LT), four tablets each time, three times per day. The treatment course was 4 weeks for all. Changes of glutamic-pyruvic transaminase (ALT) and glutamic-oxalacetic transaminase (AST) were observed before treatment, week 1, 2, and 4 after treatment. The therapeutic efficacy was compared between the two groups.

RESULTSCompared with the former time point, ALT and AST gradually decreased in the two groups at week 1, 2, and 4 (P <0. 05). The cured rate was 72. 5% and the total effective rate was 97. 5% in the treatment group. They were 62. 5% and 90. 0% respectively in the control group. There was no statistical difference in the two indices between the two group (P >0.05). No obvious adverse reaction occurred in the two groups.

CONCLUSIONDLT could treat antipsychotics induced mild hepatic damage in a safe and effective way.

Alanine Transaminase ; metabolism ; Antipsychotic Agents ; adverse effects ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Humans ; Liver ; metabolism ; Protective Agents ; administration & dosage ; therapeutic use ; Tablets ; therapeutic use

Objective To investigate the mechanism of brain derived neurotrophic factor (BDNF) regulated by differ-ent isoforms of tyrosine kinase receptor B (TrkB) in epileptic hippocampal neurons. Methods Primary hippocampal neu-rons were cultured in vitro for 7 days, and divided into two groups, ALLN (calcineurin inhibitor) group and Anisomycin (trans-lation inhibitor) group. ALLN group included control group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+ALLN group, epilepsy+ALLN group and epilepsy+ALLN+BDNF group. Anisomycin group was sub-divided into con-trol group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+Anisomycin group, epilepsy+Anisomycin group and epilepsy+Anisomycin+BDNF group. The immunofluorescent technique was used to identificate the hippocampal neurons. Epileptiform discharges were detected by electrophysiological techniques. Western blot assay was used to deter-mine the protein expression of TrkB and phosphorylated TrkB (p-TrkB) in all cell groups. Results (1) In ALLN group, the gray value of p-TrkB/TrkB was higher in control+BDNF group compared with that of control group, the value was higher in epilepsy+BDNF group than that of epilepsy group but was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was lower in epilepsy+ALLN+BDNF group than that of epilepsy+BDNF group, but no significant difference compared with that of epilepsy+ALLN group. (2) In Anisomycin group:the gray value of p-TrkB/TrkB was higher in control+BDNF group than that of control group. The gray value of p-TrkB/TrkB was higher in epilepsy+BDNF group than that of epilepsy group, but which was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was higher in epilepsy+Aniso-mycin+BDNF group than that of epilepsy+BDNF group and epilepsy+Anisomycin group. Conclusion The decreased ex-pression of TrkB.T can improve the inhibition of BDNF/TrkB signaling, and BDNF can activate BDNF/TrkB signal pathway in epileptic hippocampal neurons. The increased TrkB.FL protein level by ALLN can’t improve the inhibition of BDNF/TrkB signal pathway.

Matrix isopotential synchronous fluorescence spectrometric (MISF) method combined with derivative technique has been proposed to determine two spectrally-badly-overlapped anthracene derivatives, 1-chloroanthracene and 9-bromoanthracene,simultaneously. An optimal scanning route of MISF spectra was chosen. By using the novel method, 1-chloroanthracene and 9-bromoanthracene have been determined directly and simultaneously without tedious pre-separation in its mixture.The recoveries of 1-chloroanthracene and 9-bromoanthracene for the analysis are 83.5％～94% and 90.0%～94.0% respectively and their limits of detection are 0.69μg/L and 10μg/L respectively.

Objective To investigate the expression and significance of Insulin like growth factor-ⅡmRNA binding protein3 (IMP3) in gastrointestinal stromal tumors (GIST). Methods One hundred and fifteen cases of GIST were subjected to immunohistochemical staining for IMP3 (SP method) and ki67 (iVisionTM method ). The relationship between IMP3 expression with clinicopathologic parameters and Ki67 proliferation indexes were analyzed. In situ hybridization detection of IMP3 mRNA was performed in 20 cases of GIST positive. Results The expression rate of IMP3 was 56.2%(65/115) in GIST positive. A significant correlation can be found between IMP3 expression and tumor of infiltration , coagulative necrosis , metastasis , nuclear atypia , diameter , mitotic count , risk grade of NIH (P < 0.05), while there was no significant correlation between IMP3 and gender, age, primary location, categories of cell type (P>0.05). There was positive correlation of IMP3 expression and Ki67 labeling indexes by Spearman analysis. Positive expression with IMP3 can be observed in 3 cases of recurrence.It may play important role in invasive and development and may be an important factor of prognosis in GIST.

[Objective]To forecast the sponge mechanism mediated by LOC389023 in patients with intractable temporal lobe epilepsy(TLE),through investigating the expression of microRNA interacted with dipeptidyl peptidase 10(DPP10)and LOC389023.[Methods]The expression of DPP10 and Kv4.3 were detected in 15 temporal neocortex from patients with brain trauma (control group)and in 26 temporal neocortex from patients with intractable TLE(epilepsy group)by western blot(WB)and immunohisto?chemical(IHC)staining. The location of DPP10 and voltage dependent potassium channel 4.3(Kv4.3)was detected by immunofluo?rescent(IF)staining. The interaction between DPP10 and Kv4.3 was testified by co-immunoprecipitation(Co-IP). The expression of microRNA obtained by softwares(miRanda,Pita,TargetScan and miRDB)was detected by qPCR.[Results]IHC and WB showed an increased expression of DPP10(P<0.05)and a decreased expression of Kv4.3(P<0.05)in the epilepsy group. IF showed that the DPP10 and the Kv4.3 co-expressed in the membrane and the cytoplasm of neurons. Co-IP showed obvious interaction between the DPP10 and the Kv4.3.Five microRNA(miR-32-5p,miR-140-5p,miR-367-3p,miR-25-3p,miR-4325)were obtained by soft?wares. No significant differences in the expression of miR-32-5p and miR-4325 were found between epilepsy group and control group by qPCR(P>0.05). But decreased expression of LOC389023 and miR-140-5p and increased expression of miR-25-3p and miR-367-3p were found in epilepsygroup compared to control group (P < 0.05).[Conclusion]miR-25-3p and miR-367-3p may be regulated by LOC389023 through the sponge mechanism followed by altered expression of DPP10 in intractable temporal lobe epilepsy.

Objective: To investigate the efficacy and mechanism of EGDT against NPC cell lines. Methods: MTT assay was used to assess cell proliferation inhibition of EGDT. The apoptotic induction and cell cycle arrest were detected by flow cytometry. Western blot was adopted to detect the protein levels. Quantitative Real-time PCR was used to determine the mRNA expressions. The NPC xenografts were established to evaluate the tumor growth inhibition of EGDT. Immunohistochemistry was applied to analyze the EGFR expression in the tumor tissues. Results: EGDT showed proliferation inhibition on the NPC cell, induced G0/G1 phase arrest and cell apoptosis in vitro. EGDT decreased the protein and mRNA levels of EGFR and its downstream RAF/MEK/ERK and PI3K/AKT pathways in time- and dose-dependent manner. Furthermore, EGDT also showed a sound antitumor activity in NPC xenograft in vivo. Conclusion: The treatment of EGDT displays EGFR and its mediated downstream signaling pathway blockade through decreasing the protein and mRNA levels, suggesting a promising strategy in treating human NPC.