Objective To study on the expression of the eukaryotic recombinant vector carrying HlyX gene of Leptospira serovar Lai in mammalian cell and explore the humoral immune response in BALB/c mice immunized with the recombinant plasmid. Methods The HlyX gene was amplified from Leptospira serovar Lai genomic DNA by PCR and inserted into pcDNA3.1 vector. After transformed into E. coli DH5α,the recombinant plasmid was assayed for identification by PCR analysis,restriction nuclease enzyme digestion and sequencing. The recombinant plasmid was transfected into COS-7 cells,then RT-PCR and Western blot were performed to test the expression of the target gene. The recombinant plasmid was injected intramuscularly into BALB/c mice for three times at intervals of two weeks,and the antibody titer was measured by ELISA. Results PCR showed the full length HlyX gene was about 1100 bp. PCR analysis,restriction nuclease enzyme digestion and sequencing indicated the recombinant vector was constructed successfully. After the plasmid Was transfected into COS-7 cells,a fragment about 1100 bp was found by RT-PCR and a specific band relative molecular mass(Mr)about 40×103,which was consistent with the expected size of the target proteins was showed by Western blot. ELISA showed the antibody titer in BALB/c mice immunized by the HlyX gene of Leptospira serovar Lai can elicit high-titer antibody in BALB/c mice,which has laid the foundation for the application of the DNA vaccine.

Objective To explore the effects of different doses of topiramate (TPM) on the expression of nerve cell adhesion molecule (NCAM) and growth-associated protein 43 (GAP- 43) mRNA in hippocampus of rats with epilepsy. MethodsForty-eight rats were randomly divided into normal control group, kainic acid (KA) group, 10 mg/kg TPM group, 40 mg/kg TPM group, 100 mg/kg TPM group and 400 mg/kg TPM group (n=8). The models of rats with epilepsy treated by different doses of TPM were established. The behavior of rats was observed, and the expression of NCAM and GAP- 43 mRNA in hippocampus of rats was determined by Real-time PCR. Results The expression of NCAM and GAP- 43 mRNA in KA group was significantly higher than that in normal control group (P<0.01), while there was no significant difference between 10 and 40 mg/kg TPM groups and KA group, that in 100 and 400 mg/kg TPM groups was significantly lower than that in KA group (P<0.01), and that in 400 mg/kg TPM group was significantly lower than that in 100 mg/kg TPM group (P<0.01). Conclusion KA can up-regulate the expression of NCAM and GAP- 43 mRNA in hippocampus of rats with epilepsy. Higher dose of TPM can inhibit the expression of NCAM and GAP- 43 mRNA, and the inhibitory effect is related with the dose of TPM.

To improve ethanol production in Saccharomyces cerevisiae,an integration plasmid pUPGKAT with PGK promoter(phosphoglycerate kinase promoter),adh1 gene(the coding sequences of alcohol dehydrogenaseⅠ) and CYC1 terminator(Cytochrome c transcription terminator) was constructed.Firstly,a fusion fragment composed of PGK promoter and adh1 gene was generated by over lap extension PCR and ligated into pUG6 resulting in plasmid pUPGKA.Subsequently,CYC1 termi nator was amplified from pSH65 by PCR and ligated to the SpeⅠand SacⅡrestriction site of pUPGKA.To integrate PGK-adh1-CYC1 into S.cerevisiae genome,pUPGKAT was digested by TthⅢⅠand the lin-earized plasmid was used to transform S.cerevisiae YS2-△adh2(adh2 disrupted strain) by lithium acetate method.The yeast mutant YS2-△adh2-adh1 which had the adh1 gene placed under the PGK promoter and harbored the adh2 deletion was constructed.Anaerobic fermentation showed overexpression of adh1 by PGK promoter resulted in a 8.84% higher ethanol production compared to YS2-△adh2.

Acyl carrier protein is an essential component involved in the biosynthesis of DHA(Docosahexaenoic Acid) via PKS(Polyketide synthase) pathway,which takes the growing acyl chain from one enzyme to another.One cDNA clone,with high homology of ACP,was isolated from Schizochytrium sp.FJU-512 cDNA library.The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4'-phosphopantetheine prosthetic(4'-PP) binding site.The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp.The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG.SDS-PAGE analysis demonstrated that ACP was effectively expressed.

0.05); the levels of LL, LI, RRS in CBA group and CBA+IBT group were significantly lower than those in control group(P

Background and purpose:Head and neck cancer is common worldwide. This article aimed to describe the patterns of incidence of head and neck cancer and their changes in Shanghai from 2003 to 2012, in order to provide reference for prevention programs, research and control strategies on cancer.Methods:The data of lip, oral cavity and pharynx cancer cases were collected by the Shanghai Cancer Registry. The distributions of Shanghai lip, oral cavity and pharynx cancer incidences from 2003 to 2012 were described. The patterns were described according to gender, age, basis of diagnosis, histological type, diagnostic stage in detail. We compared incidences of recent 5 years (from 2008 to 2012) with those of previous 5 years (from 2003 to 2007).Results:On average from 2003 to 2012, 1105 new cases of head and neck cancer were diagnosed in Shanghai each year, with 2.08% among the total cancer cases. The crude rate was 8.01 per 100000 and the age-standardized rate was 4.45 per 100000. Nasopharyngeal cancer was the major subtype of the head and neck cancer, with 50.81% among the total head and neck cases. The crude and age-standardized rates among males were higher than those among females. The histologically verified percentage was 85.77%. The squamous carcinoma was the major histological type, with 57.58% among the total cases. The age-stan-dardized rate of nasopharyngeal cancer was in decline.Conclusion:The incidence of head and neck cancer was low in Shanghai during the past 10 years. Male cases were more than female cases. The major histological type was squamous carcinoma. Half of new cases were nasopharyngeal cancer which appeared to affect patients at a relatively young age. Patients with nasopharyngeal cancer were diagnosed at relatively advanced stages.

OBJECTIVE: This study aimd to evaluate the application and clinical effect of improved submental island flap in hypopharyngeal cancer reserved laryngeal function surgery. METHOD: A retrospective review of clinical data was performed on 38 patients of hypopharyngeal cancer reserved laryngeal function using sumental island flaps, by the way of improving in design of vascular pedicle, reconstructive mode of laryngeal and hypopharyngeal function and closing of wound of neck following hypopharyngeal cancer resection. Meanwhile, the effect and prognosis was comprehensively assessed on patients with hypopharyngeal cancer reserved laryngeal function using improved submental island flaps. RESULT: The submental flaps kept alive in all 38 cases. During the follow-up period, 18 cases were dead, and of them, 7 cases died of the second primary carcinoma, included 4 cases of esophagus cancer, 1 case of cancer of soft palate, 2 cases of nasopharyngeal carcinoma; and 5 cases died of cervical or parapharyngeal lymph nodes recurrence; 2 cases died of hepatic metastasis; and 4 cases died of pulmonary metastasis. The overall 5-years survival rate was 52.6%. CONCLUSION Improved submental island flap repairing postoperative defect of hypopharyngeal cancer reserved laryngeal function has many advantages including higher success rate, more security, easy and simple to operate as well as good clinical effects, and is worth to widespread using.
Humans ; Hypopharyngeal Neoplasms ; surgery ; Hypopharynx ; surgery ; Larynx ; Neck ; surgery ; Neoplasm Recurrence, Local ; Prognosis ; Reconstructive Surgical Procedures ; Retrospective Studies ; Surgical Flaps ; Survival Rate

ObjectiveTo explore the clinical characteristics of pathological fracture in extremities caused by bone tumors or tumor-like lesions. MethodsFrom August 2002 to December 2010, 139 patients with pathological fractures were entered in the study, including 79 males and 60 females with an average age of 31.1 years. Fractures included tumor-like lesion in 55 cases, benign tumor in 13, giant cell tumor (GCT)in 7, primary malignant tumors in 28, and metastatic tumors in 36. Forces induced to fractures were classified into four grades: spontaneous fracture, functional activity, minor injury, severe injury. Age, fracture location, histological results, fractures forces, prodromes, and misdiagnosis were all observed. Chi-square test were use to compare forces and prodromes within different tumors. ResultsThe highest morbidity rate is 32.4％(45/139) which lies in 11-20 years old. The cites of fractures including femurs in 71 cases, humerus in 36, tibia in 15, fingers in 7, radiuses in 4, fibula in 3, ulnas in 2, and metatarsus in 1. Fracture forces include spontaneous fractures in 29 cases, functional activity in 42, minor injuries in 65, and traumatic injuries in 3. Sixty-seven patients(48.2％) had local prodromes. The prodromes of both malignant tumors and metastatic tumors were more than benign tumors. Twenty cases experienced misdiagnosis with average delay time of 12 weeks. ConclusionMinor injury forces and local prodromes are clinical key features of pathological fractures. Both of them are key points of avoiding misdiagnosis.

OBJECTIVETo research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.

METHOD48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).

RESULTAll of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.

CONCLUSIONThe higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; PTEN Phosphohydrolase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Xenograft Model Antitumor Assays

The paper reports the establishment of a method for simultaneous determination of peimisine and sipeimine contents in Fritillaria walujewii Regel and Fritillaria pallidiflora Schrenk. The analyses were performed on an ultra-performance liquid chromatography with evaporative light scattering detection (UPLC-ELSD), equipped with a binary solvent manager, a sampler manager and a column compartment, and connected to Waters Empower 2 software. An Acquity UPLC BEH C18 column (100 mm x 2.1 mm, 1.7 microm) was used for all analysis. The investigated compounds were separated with gradient mobile phase consisting of acetonitrile-0.02% triethylamine-water. The temperature of sample manager was set at 25 degrees C. Drift tube temperature was 40 degrees C, and spray parameter was 40% with injection volume of 1 microL. The investigated compounds including peimisine and sipeimine had good linearity (r > or = 0.9991) over the tested ranges. The average recovery was 94.5% and 98.1% with RSD < or = 2.36%. The UPLC-ELSD method is simple, sensitive and accurate with good repeatability, which is available for quality control of F. walujewii Regel and F. pallidiflora Schrenk.
Alkaloids ; analysis ; Cevanes ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Fritillaria ; chemistry ; Light ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Scattering, Radiation