Objective To study the relationships of Helicobacter pylori (Hp) infection and genetic polymorphisms of glutathione s-transferase P1 (GSTP1) with gastric cancer (GC). Methods The 98 patients with GC and 149 controls with normal finding at endoscopy were enrolled for this study. The rapid urease test (RUT), 13C- urea breath test (13C-UBT) and Giemsa staining of biopsy samples were used to check Hp infection. PCR-based restriction fragment length polymorphisms (PCR-RFLP) was used to detect GSTP1 genotype. Results The rate of Hp infection was higher in GC group than in control group (54.1% vs. 40.9%, x2 =4.11, P＜0. 05). The risk of GC would significantly increase in the GSTP1 homozygous mutant gene (MM) group with Hp infection (OR=5.44, 95%CI 1. 26-26. 79, x2=7.13, P＜0.05). Conclusions Hp infection and GSTP1 genetic polymorphisms are associated with gastric cancer risk in the elderly.

Bevacizumab has been widely used in tumor targeting therapy,while the most common adverse reaction is renal impairment,manifested as proteinuria. The main mechanisms may include interfering podocytes-endothelial vascular endothelial growth factor(VEGF)axis signals,increasing glomerular pressure caused by secondary hypertension,subacute renal thrombotic microangiopathy caused by endothelial damage and so on. Thrombotic microangiopathy is the main renal pathological type,and other rare types include glomer-ular lesions,renal interstitial disease,and benign renal arteriolar nephrosclerosis. Therefore,urine protein excretion and renal function should be closely monitored during bevacizumab treatment period for timely treat-ment,dose reduction or even withdrawal if necessary to ensure renal function.

Objectives:To compare the susceptibility of serum lipoproteins to oxidation and the effects of high-density lipoprotein on oxidative stress in patients with lipid turbulence.　Methods:VLDL, LDL and HDL were isolated using sequential ultracentrifugation from serum of patients with chronic renal failure (CRF) (n=45), myocardial infarction survivor (MIS) (n=33) , cerebral infarction(CI)(n=33) ,type 2 diabetes mellitus(DM)(n=53) and normal individuals (n=44). The degree of lipid peroxidation was estimated using the thiobarbituric acid-reactive substances (TBARS) value and the susceptibilities of lipoproteins to oxidation were assessed by measuring the increased absorbance value at 234 nm due to the conjugated dinene formation. Lipid levels and lipoprotein fractions were measured using standard methods.　Results: VLDL obtained from the patient groups showed significant increase in TBARS values, especially from the patients with MIS (compared with control group, P<0.001). In addition, LDL from MIS and DM groups and HDL from CI and DM groups also showed markedly increase in TBARS content. Significant decrease in lag time was observed in both VLDL and LDL fractions from the four patient groups. However, no change was found in the lag time in HDL fraction from the patient group compared with control group. In addition, HDL from the four patients exhibited significantly decreased inhibitory effects on in vitro oxidation of LDL, with the most significant decrease in HDL from CRF and MIS groups.　 Conclusions:The oxidative modification of lipoproteins in vivo in patients with serum lipid turbulence might be involved in the development of atherosclerosis in these patients.

Objective To assess the value of endoscopic ultrasonography (EUS) and abdominal CT scan on preoperative T and N staging of advanced gastric cancer.Methods A total of 188 patients with advanced gastric cancer received EUS and CT scan to evaluate the T and N staging and lymphatic metastasis before surgical operation.The postoperative pathologic results acted as gold standard for comparison of the two methods.The results consistent with pathologic results were considered as correct otherwise were incorrect.The accuracy of EUS and CT and the each consistency of two methods with pathology were analyzed.Results For T staging, the accuracy of EUS and CT was 87.2%(164/188), 76.6%(144/188), respectively, and Kappa value of EUS and CT was 0.726 and 0.509, respectively.There was a statistical difference between the two methods(χ2=7.181,P=0.007).For lymph node metastasis, the accuracy of EUS and CT was 72.9%(137/188) and 79.8%(150/188), respectively, and Kappa value of EUS and CT was 0.397 and 0.487, respectively.No statistical difference was found between them(χ2=0.963,P=0.326).The consistency test between the two methods for T and N staging and lymph node metastasis revealed that the Kappa value was 0.507 and 0.649, respectively.The accuracy of EUS on T staging was 93.0%, 93.3%, 96.8% and 91.5% in cardiac cancer, gastric fundus cancer, gastric corpus cancer, and gastric antrum cancer, respectively.Corresponding Kappa value in the 4 different positions were 0.843,0.881,0.940 and 0.710, respectively according to consistency tests.The accuracy of gastric antrum cancer with pylorus invasion was 63.2% and the consistency test Kappa value was 0.340.Conclusion EUS is an accurate method for T staging and lymphatic metastasis of advanced gastric cancer, but there is a lower accuracy for T staging of gastric antrum cancer with pylorus invasion.

AIM: To clone the extracellular domain of N-cadherin cDNA, and to observe the antigenicity of the expressed protein. METHODS: Total RNA was extracted from CD34+ cells separated from fetal liver and bone marrow cells. The extracellular domain of N-cad cDNA was amplified with RT-PCR and inserted into a vector pOPE101-215. The recombinant pOPE-N-cad was expressed with IPTG induction. Then, mice were immunized with the protein. RESULTS: The extracellular domain of N-cad cDNA from CD34+ cells was identified by DNA sequencing. The recombinant pOPE-N-cad in host XL1-blue expressed a 70 kD protein after induced with IPTG, and anti-N-cad antibody was detected in serum of the immunized mice after 5 times injection of the recombinant N-cad protein. CONCLUSION: CD34+ cells bore N-cad gene and the recombinant protein of the extracellular domain of N-cad cDNA shows good immunogenicity.

To construct a scFv antibody mini-library by phage display technique from the spleen cells of BALB/c mice immunized with extracellular domain of N-cadherin (N-cad), CD34 and AC133, the extracellular domain genes of N-cad, CD34 and AC133 were cloned into a phagemid pSEX81 respectively, and were then displayed on the phagemid in the form of fusion protein with p III protein. After being effectively immunized with the fusion protein, the spleen of the mice were harvested, and total RNA were extracted from the spleen. The cDNA of VH and VL genes were amplified by RT-PCR, and a scFv-phage display antibody library was constructed with the amplified V-genes. The content, multiplicity and expressing potential of the library were examined. As a result, we had produced a scFv library containing 1.4 x 10(6) individual clones which showed different patterns after being digested with restriction endoneuclease Mua I. The surface display expression of the library was also verified. It indicated that the capacity and diversity of the library was sufficient for screening specific scFv antibody against N-cad, CD34 and AC133. The library will be useful for isolating corresponding specific scFv antibodies.
AC133 Antigen ; Animals ; Antibodies ; genetics ; Antigens, CD ; immunology ; Antigens, CD34 ; immunology ; Base Sequence ; Cadherins ; immunology ; Female ; Glycoproteins ; immunology ; Immunoglobulin Fragments ; biosynthesis ; genetics ; Immunoglobulin Variable Region ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Molecular Sequence Data ; Peptide Library ; Peptides ; immunology

The metastasis-associated lung adenocarcinoma transcription 1 (MALAT1) is a highly conserved long non-coding RNA (lncRNA) gene. However, little is known about the pathological role of lncRNA MALAT1 in glioma. In the present study, we explored the expression level of lncRNA MALAT1 in primary glioma tissues as well as in U87 and U251 glioma cell lines. Using qRT-PCR, we found that the expression of lncRNA MALAT1 was significantly increased in glioma tissues compared with that of paracancerous tissues. Meanwhile, the expression of MALAT1 was highly expressed in U98 and U251 cells. In order to explore the function of MALAT1, the expression of MALAT1 was greatly reduced in U87 and U251 cells transfected with siRNA specifically targeting MALAT1. Consequently, cell viability of U87 and U251 cells were drastically decreased after the knockdown of MALAT1. Concomitantly, the apoptosis rate of the two cell lines was dramatically increased. Furthermore, the expression levels of some tumor markers were reduced after the knockdown of MALAT1, such as CCND1 and MYC. In summary, the current study indicated a promoting role of MALAT1 in the development of glioma cell.
*Apoptosis ; Biomarkers, Tumor/genetics/metabolism ; Blotting, Western ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Cyclin D1/genetics/metabolism ; Down-Regulation ; Flow Cytometry ; Glioma/metabolism/pathology ; Humans ; Proto-Oncogene Proteins c-myc/genetics/metabolism ; *RNA Interference ; RNA, Long Noncoding/antagonists & inhibitors/genetics/*metabolism ; RNA, Small Interfering/metabolism ; Real-Time Polymerase Chain Reaction

OBJECTIVE:To study the protective effects and the mechanism of Rabdosia serra water extract(RWE)on hepatic fibrosis(HF)induced by carbon tetrachloride(CCl4)in rats. METHODS:Sixty male SD rats were randomly divided into normal group,model group,colchicine group(0.12 mg/kg),and RWE low-dose,medium-dose and high-dose groups(4,8,16 g/kg,by crude drug),with 10 rats in each group. Except for intraperitoneal injection of olive oil for normal group,other groups were given 40% CCl4olive oil solution intraperitoneally to induce HF model. Since the first day of modeling,each treatment group was given relevant medicine (10 mL/kg) intragastrically, while normal group and model group were given constant volume of water intragastrically,once a day,for consecutive 6 weeks. After medication,biochemical process or ELISA were used to determine the contents of ALT,AST,HA,LN,PCⅢ and Ⅳ-C in serum,the activities or contents of SOD,GSH-Px,MDA,TNF-α,IL-6 and IL-1β in liver tissue. Pathological changes of liver tissue in rats were observed by HE staining. The expression of α-SMA and TGF-β1 in liver tissue were detected by Western blot. RESULTS:Compared with normal group,the contents of ALT,AST,LN,HA,PCⅢ and Ⅳ-C in serum,the contents of MDA,TNF-α,IL-6 and IL-1β in liver tissue were all increased significantly in model group (P＜0.01);the activities of SOD and GSH-Px in liver tissue were decreased significantly(P＜0.01). Liver fibrosis was obvious, and the relative expression of α-SMA and TGF-β 1were increased significantly (P＜0.01). Compared with model group,the contents of ALT,AST,HA,LN,PCⅢ and Ⅳ-C in serum as well as the contents of MDA,TNF-α and IL-6 in liver tissue in colchicines group and RWE groups,the contents of IL-1 β in liver tissue of rats in colchicines group,RWE medium-dose and high-dose groups were all decreased significantly (P＜0.05 or P＜0.01). The activities of SOD and GSH-Px in liver tissue of rats were increased significantly in colchicines group and RWE groups(P＜0.05 or P＜0.01). The fibrosis degree of liver tissue was significantly reduced, while the relative expression of α-SMA and TGF-β 1decreased significantly (P＜0.01). CONCLUSIONS:RWE can protect CCl4-induced HF model rats,the mechanism of which may be associated with regulating lipid metabolism,relieving liver lipid peroxidation injury and anti-oxidative stress response,inhibiting the release of inflammatory factors and the expression of TGF-β1.

We developed vectors expressing two antigen of H5N1 influenza virus. Based on the human H5N1 avian influenza virus strain A/Anhui/1/2005 isolated in China, we amplified the matrix protein 2 (M2) and Hemagglutinin (HA) genes by PCR and subcloned them into pStar vector to construct two genes co-expressing recombinant DNA vaccine pStar-M2/HA. After transfection of 293 cells with the plasmid, we confirmed with indirect immunofluorescence assay (IFA) that M2 and HA genes cloned on plasmid pStar co-expressed successfully. Using Ad-Easy adenovirus vector system, by homologous recombination in bacteria and packaging in 293 cells, we constructed two recombinant adenoviruses, namely Ad-M2 and Ad-HA. After infection of 293 cells with the recombinant adenoviruses, we confirmed with IFA that M2 and HA genes cloned into adenoviruses expressed successfully. We then combined the recombinant DNA vaccine and adenoviral vector vaccines in immunization of BALB/c mice with a prime-boost regime. On day 0 and day 28, we immunized the mice with DNA vaccine and on day 14 and day 42, with recombinant adenovirus vaccines. We took blood samples before each injection and 14 days after the final injection. On day 56, we collected splenocytes from the mice. ELISA and hemagglutination inhibition (HI) assay showed that the vaccines successfully induced specific IgG antibodies against HA protein in serum of the immunized mice. ELISPOT confirmed that the vaccines successfully induced the special cellular immune response to M2 and HA protein of H5N1 influenza virus. The study on combined immunization with M2 and HA genes provided basis for development of novel influenza vaccine.
Adenoviridae ; genetics ; metabolism ; Animals ; Female ; Genetic Vectors ; genetics ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; immunology ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Vaccination ; Vaccines, DNA ; immunology ; Viral Matrix Proteins ; biosynthesis ; genetics

Objective To observe the effects of percutaneous endoscopic gastrostomy (PEG)on mortality and complications in patients with persistent dysphagia after stroke using a points scoring system for selecting PEG indication.Methods A total of 75 patients were divided into low score group without PEG,high score group without PEG and low score group with PEG (n=25 each).The follow-up period was 18 months,and the differences in complications,mortalities and survival periods among groups were compared.Results The number of times of aspiration pneumonia was (1.36± 1.44) in low score group,(1.96±2.28) in high score group,(0.36±0.64) in low score group with PEG,with statistically significant differences among three groups (H=7.148,P=0.028).No difference in the morbidity of aspiration pneumonia was found between low score group and high score group (P=0.189).The number of times of aspiration pneumonia was decreased in low score groups after PEG versus in low score group without PEG (P=0.030) and in high score group (P＜0.01).The numberof times of gastrointestinal hemorrhage was (0.48± 0.77)in low score group,(0.64± 0.91) in high score group,(0.12±0.33) in low score group with PEG,with statistically significant differences among three groups (H=5.532,P =0.063).No statistically significant difference in gastrointestinal hemorrhage was found between low score groups and low score group after PEG (P=0.430),as well as between low score group and low score group with PEG (P=0.079).The morbidity of gastrointestinal hemorrhage was lower in low score group than in high score group (P=0.012).The survival rate at the observation end was 88.0％ (22/25),52.0％ (13/25) and 92.0％ (23/25) in low score group,high score group and low score group with PEG,respectively,with statistically significant difference among the three groups (x2 =7.906,P =0.001).Kaplan-Meier survival curve showed that the survival period were longer in the low score group with or without PEG than in high score group (P＜0.01),but no statistically significant difference was found between low score groups with or without PEG (P=0.626).Conclusions The reasonable evaluation using a points-scoring system before PEG might predict the prognosis of such patients:the higher score would indicate higher mortality.PEG operation for low score group with better condition could decrease the aspiration pneumonia and decrease gastrointestinal hemorrhage significantly,but could not prolong general survival time and decrease general mortality.