Acute Coronary Syndrome ; blood ; Aged ; Cell Membrane ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Thromboplastin ; metabolism

Reverse-genetic engineering of foot and mouth disease virus (FMDV) can improve the productivity, antigen matching, antigen stability, immune response ability, and biological safety of vaccines, so vaccine candidates with anticipated biological characteristics can be promptly achieved. Negative influence in taming of virulent strains can also be decreased or avoided. Reverse genetics not only make up for deficiencies like limitation of viral nature, low success rate, and time and energy consuming, but also realize more active designing of vaccines. Therefore, reverse genetics is significant in improving integral quality and efficiency of vaccines. In this review, we use FMDV vaccines as an example to summarize improvement in biological characteristics of virulent strains and provide a reference for related researches.
Animals ; Antibodies, Viral ; immunology ; Foot-and-Mouth Disease ; immunology ; prevention & control ; virology ; Foot-and-Mouth Disease Virus ; genetics ; immunology ; Reverse Genetics ; Viral Vaccines ; genetics ; immunology

OBJECTIVETo assess suppression effects of vector-based small interfering RNA (siRNA) on vascular endothelial growth factor (VEGF) expression of human tongue squamous carcinoma cell line (Tca8113) in vitro.

METHODSTwo siRNA targeting VEGF constructed in eukaryotic expression vector (Pu-VEGF-siRNA1, Pu-VEGF-siRNA2), eukaryotic expression vector as the experiment control, all of which were transfected into Tca8113 cells with Lipofectamine 2000. Non-transfection cell was used as negative control. VEGF mRNA and protein were detected by reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry and enzyme linked immunosorbent assay (ELISA), respectively.

RESULTSCompared to the experimental and negative controls, the expression of VEGF mRNA and protein were significantly decreased in the Pu-VEGF-siRNA1 group and Pu-VEGF-siRNA2 group. But there were no significant differences between two controls (P > 0.05).

CONCLUSIONVector-based siRNAs targeting VEGF are efficient in down-regulating VEGF expression in Tca8113 cells.

Carcinoma, Squamous Cell ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; RNA, Messenger ; RNA, Small Interfering ; Tongue Neoplasms ; Transfection ; Vascular Endothelial Growth Factor A

OBJECTIVETo examine the antiangiogenic and antitumor effects of vector-based small interfering RNA (siRNA) targeting vascular endothelial growth factor (VEGF) on human tongue squamous cell carcinoma xenografts in vivo.

METHODSTca8113 human tongue cancer nude mice xenograft model was established, and subsequently divided into four groups randomly (5/group). Two siRNA targeting VEGF constructed in eukaryotic expression vector (PU-VEGF-siRNA1, PU-VEGF-siRNA2) were injected intratumor and peritumor with lipofectamine 2000, respectively. No siRNA vector injected and non-injected tumors were used as experimented and negative controlled, respectively. Animals were injected one time every 3 days for a total of 10 times. Three days after the last injection, the weigh and volume of tumors, and intratumor microvessel density (MVD) were measured. The expression of VEGF in xenograft tumors was detected by reverse transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. The cell apoptosis and cell cycle analysis of tumors were detected by Tunel and Flow cytometry, respectively.

RESULTSCompared to the experimental and negative controls, the percentage of cells in the G(1) phase increased (P < 0.05), the expression of VEGF on both mRNA and protein level, the tumor weigh and volume, and MVD decreased in the PU-VEGF-siRNA2 group (P < 0.05), and more apoptosis was induced (P < 0.01). But significant differences were not noted between PU-VEGF-siRNA1 group and two controls (P > 0.05).

CONCLUSIONSVEGF-siRNA can reduce VEGF expression, inhibit tumor growth and angiogenesis, and induce apoptosis in Tca8113 cell carcinoma in vivo. Different VEGF-siRNA may have different effect in vivo.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; blood supply ; genetics ; pathology ; Cell Line, Tumor ; Humans ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; RNA, Small Interfering ; Tongue Neoplasms ; blood supply ; genetics ; pathology ; Vascular Endothelial Growth Factor A ; genetics

BACKGROUNDNeoadjuvant chemoradiation is now considered the standard care for locally advanced rectal carcinoma (T3-4 or/and N1-2 lesions), but the accuracy of staging examinations including endorectal ultrasonography (ERUS) and MRI is far from excellent. In addition, the above staging equipment or professionals who perform the examinations may not be available in some hospitals, while preoperative colonoscopy and biopsy are usually obtainable in most hospitals. The objective of the present study was to investigate the clinical and pathological characteristics of locally advanced rectal carcinoma and identify candidates for neoadjuvant chemoradiation.

METHODSThis was a retrospective study. Patients who were treated for rectal cancer at Changhai Hospital from January 1999 to July 2008 were identified from our prospectively collected database. Statistical analysis was performed using SPSS Software System (version 15.0). The Mann-Whitney test, chi-square test and multivariate Logistic regression analysis were performed.

RESULTSA total of 1005 cases were included in this research, of which 761 cases were identified as locally advanced rectal carcinoma depending on postoperative TNM staging. The results of multivariate Logistic regression analysis indicated seven independent risk factors that could be used to predict a locally advanced rectal carcinoma independently: a high grade (including poor differentiation and undifferentiation) (OR: 3.856; 95% CI: 2.064 to 7.204; P = 0.000); large tumor size (OR: 2.455; 95% CI: 1.755 to 3.436; P = 0.000); elevated preoperative serum CEA level (OR: 1.823; 95% CI: 1.309 to 2.537; P = 0.000); non-polypoid tumor type (OR: 1.758; 95% CI: 1.273 to 2.427; P = 0.001); the absence of synchronous polyps (OR: 1.602; 95% CI: 1.103 to 2.327; P = 0.013); the absence of blood in stool (OR: 1.659; 95% CI: 1.049 to 2.624; P = 0.030); and a greater circumferential tumor extent (OR: 1.813; 95% CI: 1.055 to 3.113; P = 0.031). Based on these findings, a Logistic equation was established, the accuracy of which was 64% according to the information of the additional 50 cases.

CONCLUSIONSSome independent risk factors related with locally advanced rectal carcinoma were identified, based on which it is possible to establish a Logistic equation as a tool to predict candidates of neoadjuvant chemoradiation. Further research about optimization of the equation is warranted.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Colonoscopy ; methods ; Endosonography ; Female ; Humans ; Logistic Models ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neoplasm Staging ; Rectal Neoplasms ; diagnosis ; pathology ; Retrospective Studies

OBJECTIVEAnalyze Naive and Mermory T cell subsets in HIV/AIDS patients and investigate their relationship with disease development.

METHODST cell subsets from 15 normal control subjects, 79 HIV/ AIDS patients were detected by FCM.

RESULTSWith diesase progression, CD4+ Naive cell counts and ratio was both decreased obviously (P < 0.001); CD4+ Tcm cell counts was significantly decreased (P < 0.001), CD4+ Tcm cell ratio was obviously higher (P = 0.002); CD4 TEM cell ratio was obviously increased( P < 0.001); CD8+ T Naive cell counts and ratio was also decreased obviously (P < 0.05); CD8+ T(CM), T(EM), T(EMRA) are not significantly different.

CONCLUSIONSThe peripheral lymphocyte subsets in HIV/AIDS patients changed obviously. The counts of naive T cell decreased, while the proportion of memory T cell increased significantly. It will help to understand pathogenesis of HIV.

Acquired Immunodeficiency Syndrome ; immunology ; virology ; Adult ; CD4 Lymphocyte Count ; Case-Control Studies ; Female ; HIV ; immunology ; Humans ; Male ; Middle Aged ; T-Lymphocyte Subsets ; immunology

Bacillus are well known antibiotic producers. In this study,dozens of Bacillus strains from different sources were screened. Among them,a strain with strong antifungal activity was found. With 16S rDNA test and Biolog assay,this strain was identified to be Bacillus amyloliquefaciens. The fermentation conditions were optimized in small conical flasks. After ammonium sulfate salting out,dialysis,freezing vacuum dehydration,the crude protein extracts were obtained. The thermal stability,pH stability,protease stability,ion stability and antifungal spectrum of this protein were studied further. Scanning electronic microscope was also used to explore the antifungal mechanism.

BACKGROUNDEstrogen is involved in suppression of colon cancer development and exerts its function via estrogen receptors alpha and beta (ERalpha, ERbeta). The recently identified ERalpha46 resulted from exon 1-deletion from the 66-kDa full length form of ERalpha66 is devoid of the transactivation domain AF-1, whose function remains largely unknown.

METHODSIn this study, we compared the expression of ERalpha46 mRNA in 32 normal colorectal tissues and their matched colorectal cancer tissues by real-time quantitative polymerase chain reaction (PCR). Human colon adenocarcinoma cell HT-29, that has low endogenous expression of ERalpha46, was transfected with ERalpha46-expression vector; methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, DNA fragmentation and TUNEL staining were used to evaluate the proliferation and apoptosis status of the cells in the presence of 17beta-oestradiol.

RESULTSHigher ERalpha46 mRNA levels were observed in normal colorectal tissues than in the corresponding cancer tissues. ERalpha46-transfected cells showed a significantly decreased growth rate than control cells and an accumulation of cells in the G(0/1) phase and a reduced proportion of cells in G(2)/M phase after exposed to 10(-8) mol/L 17beta-oestradiol. There were also more positive TUNEL stained cells in ERalpha46-transfected cells than the control cells in the presence of 17beta-oestradiol (P < 0.05).

CONCLUSIONSThese data suggest that ERalpha46 may be involved in the development and/or progression of colorectal cancer via mediating growth inhibition and apoptosis of cancer cells in the presence of 17beta-oestradiol.

Adult ; Aged ; Apoptosis ; Colorectal Neoplasms ; genetics ; pathology ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; genetics ; Female ; G1 Phase ; HT29 Cells ; Humans ; Male ; Middle Aged ; Mutation

OBJECTIVETo investigate whether the WNK lysine deficient protein kinase 4 (WNK4) gene C1155547T polymorphism is associated with essential hypertension (EH) in Xinjiang Kazakhs and to assess the effect of the interaction between this polymorphism and environment factors on EH.

METHODSThe study covered 556 hypertension patients and 341 normotensive controls. The C1155547T was determined by Taqman probe real-time PCR method. Some biochemical index such as glucose, triglyceride and total cholesterol were also measured. All of these results were analyzed with Logistic regression analysis. Additive model was applied to assess the effect of interaction between the WNK4 gene C1155547T polymorphism and environment factors on hypertension.

RESULTSThe C1155547T polymorphism was consistent with Hardy-Weinberg equilibrium in both the case and control groups. There was significant difference in the genotype frequencies (P=0.003). The T allele frequency was significantly higher in the patient group (P=0.002). Logistic regression analysis revealed that the age, body mass index (BMI), total cholesterol as well as the CT+TT genotype frequency conferred increased risks for EH. Positive interaction between the C1155547T polymorphism and gender, BMI, glucose was observed. The ORs were 3.85 (95%CI:1.23-12.04), 5.91 (95%CI:1.99-17.57) and 8.77 (95%CI:1.04-73.93), respectively.

CONCLUSIONThe result suggested that the exon 7 C1155547T polymorphism in WNK4 gene might be associated with EH in Xinjiang Kazakhs, the T allele might be the risk factor of essential hypertension. There were interactive effects between the WNK4 gene C1155547T polymorphism and gender, BMI and glucose.

Adult ; Asian Continental Ancestry Group ; ethnology ; genetics ; China ; Female ; Humans ; Hypertension ; ethnology ; genetics ; Male ; Middle Aged ; Point Mutation ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics

OBJECTIVETo determine the effect of tyrosine kinase A (TrkA) and vascular endothelial growth factor receptor 2 (VEGFR2) in the invasion and metastasis of salivary adenoid cystic carcinoma (SACC).

METHODSThe expression of TrkA and VEGFR2 were detected by immunohistochemical staining in 47 cases of SACC of salivary glands. Clinical data were reviewed by multivariate prognostic analysis.

RESULTSThe positive rate of TrkA and VEGFR2 in SACC was 87.23% (41/47) and 85.11% (40/47) respectively. Express of TrkA and VEGFR2 in perineural invasion and recurrence group were higher than non-perineural invasion and non-recurrence group. Significant difference was found in microvessel density (MVD) and VEGFR2 expression within different groups (P < 0.05). MVD in perineural invasion group (25.14 +/- 2.83) was significantly higher than that in none perineural invasion group (18.81 +/- 1.33) (P < 0.05). MVD in recurrence or metastasis group (26.58 +/- 2.38) was significantly higher than that (19.06 +/- 1.39) in none recurrence nor metastasis group (P < 0.05).

CONCLUSIONPositive correlation between expression of TrkA, VEGFR2 and nerve invasion and vessel metastasis of SACC indicate that TrkA and VEGFR2 play important roles in the invasion and metastasis of SACC. It is possible that TrkA and VEGFR2 could be an aid for evaluating the prognosis of SACC patients.

Carcinoma, Adenoid Cystic ; metabolism ; pathology ; Humans ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Receptor, trkA ; metabolism ; Salivary Gland Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor Receptor-2