NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

Growth suitability as assessment indicators for medicinal plants cultivation was proposed based on chemical quality determination and ecological factors analysis by Maxent and ArcGIS model. Notopterygium incisum, an endangered Chinese medicinal plant, was analyzed as a case, its potential distribution areas at different suitability grade and regionalization map were formulated based on growth suitability theory. The results showed that the most suitable habitats is Sichuan province, and more than 60% of the most suitable areawas located in the western Sichuan such as Aba and Ganzi prefectures for N. incisum. The results indicated that habitat altitude, average air temperature in September, and vegetation types were the dominant factors contributing to the grade of plant growth, precipitation and slope were the major factors contributing to notopterol accumulation in its underground parts, while isoimperatorin in its underground parts was negatively corelated with precipitation and slope of its habitat. However, slope as a factor influencing chemical components seemed to be a pseudo corelationship. Therefore, there were distinguishing differences between growth suitability and quality suitability for medicinal plants, which was helpful to further research and practice of cultivation regionalization, wild resource monitoring and large-scale cultivation of traditional Chinese medicine plants.
Apiaceae ; growth & development ; China ; Ecosystem ; Environment ; Geographic Information Systems ; Soil ; chemistry ; Temperature

OBJECTIVETo evaluate three-axis otoconia maneuver (TOM) for benign paroxysmal positional vertigo (BPPV).

METHODSThe data from twenty BPPV patients who received three-axis otoconia maneuver treatment and 20 BPPV patients who received canalith repositioning (CRP) maneuver treatment were analyzed retrospectively.

RESULTSThere were 17 patients received 1 TOM session and 3 patients received 2 TOM sessions while 16 patients received 1 CRP session and 4 patients received 2 CRP sessions. The chi-square (X2) test was used in evaluating the association between two independent samples in a contingency table. Both methods had no statistically significant. The significance level for statistical tests was 5% (alpha = 0.05).

CONCLUSIONSThree-axis otoconia maneuver could be effective used in benign paroxysmal positional vertigo with the advantage of repeatedly practicable and instrumental.

Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Semicircular Canals ; Treatment Outcome ; Vertigo ; therapy ; Young Adult

OBJECTIVETo study the effect of chemotherapy with drugs selected according to drug sensitivity test and drug- resistance gene assessment in mice bearing tumors derived from high-passage gastric cancer cells.

METHODSChemosensitivity of the mouse gastric cancer cells to 6 chemotherapeutic drugs was investigated using MTT assay before the 6 drugs were applied in mice with implanted tumors. The implanted tumor volume and tumor inhibition rate were observed, and the expression of the multidrug resistance-1 (MDR1) gene was detected.

RESULTS5-FU, the most sensitive among the 6 drugs, resulted in significantly higher tumor inhibition rates in comparison with the control group, but did not significant affect MDR1 expression. Adriamycin (ADM) had the lowest sensitivity with low tumor inhibition rates and no significant effect on MDR1 expression. Cisplatin (DDP) was identified as the most sensitive drug for mouse gastric cancer with previous 5-FU exposure, and Mitomycin (MMC) was the most insensitive drug for mouse gastric cancer with previous ADM treatment. In mice bearing tumors of the second-passage cells, DDP showed high tumor inhibition rate but scarcely affected MDR1 expression; MMC resulted in low tumor inhibition rate and high MDR1 expression. Application of 5-FU and ADM in these tumor-bearing mice of the second-passage cancer cells resulted in rather low tumor inhibition rates without significant alterations in MDR1 expression as compared with the control group.

CONCLUSIONAppropriate individualized chemotherapy should be implemented in full consideration of the results of drug sensitivity test and drug-resistance gene assessment.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Animals ; Antineoplastic Agents ; pharmacology ; Carboplatin ; pharmacology ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Doxorubicin ; pharmacology ; Drug Screening Assays, Antitumor ; Fluorouracil ; pharmacology ; Immunohistochemistry ; Mice ; Mitomycin ; pharmacology ; Neoplasm Transplantation ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Treatment Outcome ; Tumor Burden ; drug effects

OBJECTIVETo examine the activation and cytotoxicity of human peripheral blood T lymphocyte induced in vitro by human 4-1-BB ligand (4-1-BBL) gene transfected into tumor Tca8113 cells.

METHODSThe eukaryotic expression vector pEGFP-h4-1-BBL was transfected into human oral carcinoma cell line Tca8113 by Lipofectamine 2000. The transfected cells were then selected in medium containing G-418, cloned by limited dilution and named as Tca8113-4-1-BBL. Human 4-1-BBL mRNA and protein expression of transfected cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting respectively. The tumor cell vaccines (TCV) were obtained by treatment with mitomycin (MMC). Human peripheral blood mononuclear cells (PBMC) were prepared from lymphoprep, and then stimulated with anti-CD-3 mAb and incubated with non-transfected or transfected TCV-Tca8113 cells, respectively. The proliferation of T cells was evaluated by trypan blue exclusion; the CCK-8 was used to detect the cytotoxic effect of T lymphocytes. Meanwhile, the secretion of interferon-gamma (IFN-gamma) and interleukin (IL)-2 in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSThe Tca8113 cells transfected by pEGFP-h4-1-BBL could express human 4-1-BBL efficiently. As compared with wild type Tca8113 cells, the transfected Tca8113 cells could markedly promote proliferation, IL-2 and IFN-gamma production and cytotoxic activity of lymphocytes.

CONCLUSIONSThe transfection of human 4-1-BBL gene in Tca8113 cells is effective in enhancing its immunogenicity and inducing antitumor immune response in vitro.

4-1BB Ligand ; genetics ; Carcinoma, Squamous Cell ; genetics ; immunology ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Interferon-gamma ; immunology ; Interleukin-2 ; immunology ; Lymphocyte Activation ; Mouth Neoplasms ; genetics ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Transfection

The in situ gel systems can form gel in situ after administration to achieve sustained release, thus provides a promising strategy for drug delivery systems. The aim of this study was to design and prepare in situ gel systems for the oral delivery of ibuprofen (IBU-ISG) and study its pharmacokinetics in Beagle dogs. The characteristics of the basic material of gellan gum (Kelcogel, Kel) and sodium alginate (Manugel, M) were studied through investigating the complex viscosity of the Kel or M solution with or without different concentrations of calcium ion or sodium citrate to ascertain the amount range of the excipients. The measurement of complex viscosity of the solution (0. 5% Kel and 1% M) with different concentrations of sodium citrate and calcium ion was carried out to select the suitable proportion of calcium ion and sodium citrate. The formulation of binary IBU-ISG was optimized by monitoring the complex viscosity before gelling in vitro release property. The optimized formulation contains 1.0% sodium alginate, 0.5% gellan gum, 0. 21% sodium citrate and 0.056% calcium chloride. A single oral dose of IBU-ISG and reference formulation (IBU suspension) were given to each of the 6 healthy Beagle dogs, ibuprofen in plasma at different sampling times was determined by RP-HPLC. The pharmacokinetics parameters in 6 Beagle dogs were calculated. The Tmax of IBU-ISG and reference formulation were (1.8 +/- 0.6) and (0.4 +/- 0. 1) h. The Cmax values were (29.2 +/- 7.6) and (37.8 +/- 2.2) microg x mL(-1). The T(1/2) were (2.3 +/- 0.5) and (2.0 +/- 0.9) h, and the AUC(0-t) were (131.0 +/- 38.6) and (117.3 +/- 23.1) microg x mL(-1) x h, respectively. The binary IBU-ISG was successfully prepared.
Administration, Oral ; Alginates ; chemistry ; Analgesics, Non-Narcotic ; administration & dosage ; blood ; pharmacokinetics ; Animals ; Area Under Curve ; Calcium Chloride ; chemistry ; Citrates ; chemistry ; Delayed-Action Preparations ; Dogs ; Drug Compounding ; methods ; Drug Delivery Systems ; Excipients ; Female ; Glucuronic Acid ; chemistry ; Hexuronic Acids ; chemistry ; Ibuprofen ; administration & dosage ; blood ; pharmacokinetics ; Male ; Polysaccharides, Bacterial ; chemistry ; Viscosity

Objective To explore the associations of body mass index (BMI), waist circumference (WC), waist height ratio (WHtR) and the prevalence of hypertension in elderly residents over 60 years in Baodi district, Tianjin. Methods Residents over 60 who underwent medical examinations in the Koudong Health Center, Baodi district, Tianjin, were all invited to participate in the study from April to May, 2018. Participants were asked to fill out structured questionnaires and undergo physical examinations. Stratified analysis and logistic regression analysis were applied to examine joint effects and interactions of BMI and WC (or WHtR) on the risk of hypertension. Results A total of 1 417 residents (83.75%) out of 1 692 residents participated in the study. The prevalence of hypertension in the participants was 46.36%. 66.50% of the participants were BMI overweight or obese. Participants with central obesity accounted for 74.66% (measured by the WC) and 75.38% (by the WHtR). Compared to the normal weight measured by the BMI or the WC, BMI overweight (OR=1.65, 95%CI: 1.19-2.30) or obesity (OR=3.41, 95%CI: 2.23-5.20) and WC central obesity (OR: 1.49, 95%CI: 1.00-2.23) were associated with increased risk of hypertension. The joint effects of BMI and WC (OR=2.49, 95%CI: 1.78-3.46), or BMI and WHtR (WHtR overweight: OR=2.05, 95%CI: 1.41-2.99; WHtR obesity: OR=2.37, 95%CI: 1.50-3.76) were greater than the single effect of the latter (WC overweight/obesity: OR=1.39, 95%CI: 0.90-2.15; WHtR overweight: OR=1.02, 95%CI: 0.62-1.66; WHtR obesity: OR=1.44, 95%CI：0.55-3.81). Conclusions Of the three indices, BMI is strongly correlated with the risk of hypertension. BMI overweight or obesity has enhanced the association of WC or WHtR and the risk of hypertension, suggesting that weight control in the normal range, especially measured by the BMI index, may prevent and control hypertension.

OBJECTIVETo investigate the antireflux effects of a modified Nissen fundoplication following esophagectomy for cancer.

METHODSFrom March 2006 to March 2007, 70 patients with esophageal cancer were divided into two groups randomly. Esophagogastrostomy with a stapler only was perform in 35 patients as controls (group C), and a modified Nissen fundoplication was added after esophagogastrostomy with stapler in the other 35 patients as observed group (group O). There were 48 male and 22 female, ranging in age from 47 to 77 years (mean 60.1 years). The operative morbidity and mortality were recorded. Fourty-nine patients were followed at 3 months after surgery, and the questionnaire of life quality (EORTC QLQ C-30) was conducted in 24 patients in group C and 25 patients in group O. Thirty patients were examined with esophageal manometry, 24 h pH monitoring and gastroscopy. There were 16 patients in group C and 14 patients in group O.

RESULTSThere was no significant difference in postoperative morbidity between the two groups (P > 0.05). However, the scores of heart burn and regurgitation in the group O were less than in group C (P = 0.041 and 0.034 respectively), but there was no difference in scores of dysphagia between the two groups (P = 0.677). The pressure at the anastomotic site was higher than that in the stomach in group O (P = 0.032), but not in group C (P = 0.448). DeMeester score in group O was 53 ± 46, compared to 140 ± 103 in group C (P = 0.043). The score of esophagitis was 0.9 ± 0.8 in group O, which was lower than 1.6 ± 1.0 in group C (P = 0.041).

CONCLUSIONSAddition of modified Nissen fundoplication after esophagectomy and esophagogastrostomy for cancer significantly increases the pressure at the anastomotic site, thus reduces the extent of gastroesophageal reflux, which leads to the reduction of the extent of reflux esophagitis and the improvement of the quality of life.

Aged ; Anastomosis, Surgical ; methods ; Esophageal Neoplasms ; surgery ; Esophagectomy ; Esophagus ; surgery ; Female ; Follow-Up Studies ; Gastroesophageal Reflux ; etiology ; prevention & control ; Humans ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; Stomach ; surgery