Objective Clone and mutagenesis analysis of nhaA gene in Vibrio cholerae O1 and O139. Methods 40 strains of Vibrio cholerae O1 and O139 were collected. A full length nhaA gene fragment was amplified with PCR and cloned into plasmid vector pcDNA3. Homology and mutagenesis of nhaA gene in Vibrio cholerae O1 and O139 were analyzed after sequencing the nhaA gene. Results nhaA gene of Vibrio cholerae O1 and O139 were successfully amplified and cloned. Sequence analysis manifested that nhaA gene of Vibrio cholerae O1 and O139 in China share a high homology with reference sequence of wide-type Vibrio cholerae O1 in GENEBANK(99% and 96% respectively). The amino acid mutagenesis rates of nhaA gene in Vibrio cholerae O1 and O139 were 2% and 11% respectively. The important residues(Asp133,Asp163,Asp164,His225,Leu73 and Gly338 )had no mutation. But in residues 203 and 221 of nhaA gene Vibrio cholerae O1 and O139 had common mutation. Conclusions Mutagenesis of nhaA gene and NhaA protein may be the result of Vibrio cholerae adaptation to survival environment.

Objective To study the clinical effects of human amniotic membrane transplantation loaded with autologons marrow mesenchymal stem cells(MSCs) in treatment of pterygium patient. Methods Totally 10 patient (10 eyes) with primary giant pterygium aged 42 ～60 years(7 males and 3 females) were enrolled. All patients were informed consents. Bone marrow was sterilely collected by bone marrow aspiration. MSCs were harvested, purified and cultured. At the third passage,MSCs were incubated in amniotic membrane. After pterygium resection,amniotic membrane patch with cells surface upward was covered on surface of exposed sclera. Another simple amniotic membrane was sutured to protect transplanted cells. Results 1 ～ 3 days after dissection of corneal epithelial area was repaired,a week around later,migration of conjunctival epithelial gradually grew,about 10 ～ 14 days, absorption and fusion of amniotic membrane began. Follow-up of 6 months to 3 years, there was no recurrence. Conclusion After amniotic membrane loaded with MSCs were transplanted to the ocular surface ,corneal epithelium repaired rapidly ,transplanted amniotic membrane and conjunctival integrated, conjunctival irritation was few, ocular surface scar was light, the pterygium was no recurrence.

Objective To investigate the involvement of VEGFR-3 signaling pathway in lymphatic metastasis of thyroid cancer. Methods The expression of VEGFR-3 mRNA in thyroid carcinoma and normal thyroid tissue was detected by RT-PCR. The expression of VEGFR-3 mRNA and the lymphatic vessel density (LVD) in thyroid carcinoma and normal thyroid tissue was measured by immunohistochemical staining. Results All thyroid cancer samples expressed VEGFR-3 and the expression rate of VEGFR-3 in thyroid carcinoma was significantly higher than that in the normal thyroid group ( P ＜ 0. 05 ). LVD in thyroid cancer was significantly higher than that in normal control. LVD with lymph nodes metastasis ( N + ) was significantly higher than that without lymph node metastasis (NO) (P ＜ 0. 05). Conclusions There is upregulatin of VEGFR-3 expression in thyroid carcinoma. The higher the expression of VEGFR-3, the severer of lymph node metastasis. VEGFR-3 receptor-mediated signal transduction pathway might be an important factor of thyroid cancer lymph node metastasis.

Objective:To study the possibility of dengue virus E gene vaccine.Methods:The recombinant eukaryotic expression plasmid pcDNA3 E was first transfected into NIH3T3 cells by lipofectin SDS PAGE and Western blotting analyzed the expression of E gene Then the recombinant plasmid was intramuscularly injected to BALB/c mice,and the specific humoral and cellular immunity were tested Results:The recombinant plasmid DNA could induce specific immune reactions and the immune response could last a long time Conclusion:The dengue virus E gene vaccine could induce specific immune reaction,which might have provided some material and new experimential data for the further study of dengue vaccines

Through genetic recombination technique, the rat glial cell line-derived neurotrophic factor (rGDNF) cDNA was in-serted into polylinker site of retroviral vector pLXSN, to generate a recombinant plasmid pLXSN-GDNF as transfer vector. Therecombinant plasmid was verified with restriction analysis, PCR, dot blot hybridization and Southern blot hybridization. The re-sults showed that GDNF cDNA was cloned correctly into retroviral vector pLXSN, recombinant retroviral vector was construct-ed. It is concluded that the eukaryotic cell expression vector was constructed successfully for gene therapy of Parkinson's,Alzheimer's and other central nervous system diseases.

AimTo study the effect of interferon gamma( IFN-γ) and tumor necrosis factor alpha(TNF-α) in the immune pathogenesis of dengue virus infection. Method The serum levels of IFN-γ and TNF-α were measured with emzyme linked immunosorbent assay (ELISA) method in 30 cases of the patients with the dengue virus infection in Guangzhou district. The results were treated with t-test of two sample mean. ResultsThe serum levels of IFN-γ and TNF-α in patients with the dengue virus infection were much higher than healthy controls( P ＜ 0.05、 P ＜ 0.01 ). IFN-γ was detectable on the first day of postinfection. Level of IFN-γ reached their peaks on the second day, then declined . The level of TNF-α had an obvious rise from the second day and reached their peaks on the third day, then declined. ConclusionThe data suggest that the IFN-γ and TNF-α may play an important role in the dengue virus pathogenicity and immunity.

Objective To detect serum levels of thyroid hormones in patients with thyroid nodules (TN), and investi-gate their relationship with the nature of TN. Methods A total of 245 patients with TN were recruited in the study. Accord-ing to levels of thyroid antibodies and postoperative pathological results, all patients were divided into nodular goiter (NG) group, thyroid adenoma (TA) group and thyroid cancer (TC) group. TC group was further classified as the TC with increased level of thyroid antibodies (TC-AB+group) and the TC with normal level of thyroid antibodies (TC-AB-group). The serum levels of free T3 (FT3), free T4 (FT4) and thyroid stimulating hormone (TSH) were detected for all patients before operation, and differences of thyroid hormones were analyzed between different groups. Results The serum level of TSH was signifi-cantly higher in TC group than that of NG group and TA group (P<0.05). The serum level of FT3 was significantly lower in TC-AB+group than that of NG group and TC-AB-group. The serum level of FT4 was significantly lower in TC-AB+group than that of NG group, and the serum TSH level was significantly higher than that of other groups (P<0.05). Conclusion The increased serum levels of TSH were found in some patients with TC, which may partly attribute to their coexistence with autoimmune thyroiditis and subsequent hypothyroidism. The increased serum TSH level may not be the inherent characteris-tics of TC.

Objective To reveal the research trends and hotspots of China's liver neoplasms literature from 2005 to 2010. Methods A bibliometric analysis was made based on the results of retrieving from the Chinese Medical Citation Index (CMCI). Results The number of China' s liver neoplasms literature increased regularly in the last 5 years. The fund articles accounted for 30% of total articles. This paper reports the highly cited articles, authors, institutions and journals. The top 5 provinces of publishing articles are Guangdong, Shanghai, Jiangsu, Shandong and Beijing. There are some major aspects of liver neoplasms research, such as treatment modalities, analysis of causes of disease, evaluation after radiation therapy, and experimental research. 3-dimensional conformal radiotherapy, radiofrequency ablation and chemoembolization have been becoming research hotspots. Conclusions The researches on liver neoplasms have been widely and deeply developing in China. A lot of research progresses were made. As the higher mortality of liver neoplasms, the research of comprehensive treatment is to be the further trend.

In order to investigate the expression and functional role of HERG1 K+ channels in leukemic cells and leukemic stem cells (LSCs), RT-PCR was used to detect the HERG1 K+ channels expression in leukemic cells and LSCs. The functional role of HERG1 K+ channels in leukemic cell proliferation was measured by MTT assay, and cell cycle and apoptosis were analyzed by flow cytometry. The results showed that herg mRNA was expressed in CD34+/CD38-, CD123+ LSCs but not in circulating CD34+ cells. Herg mRNA was also up-regulated in leukemia cell lines K562 and HL60 as well as almost all the primary leukemic cells while not in normal peripheral blood mononuclear cells (PBMNCs) and the expression of herg mRNA was not associated with the clinical and cytogenetic features of leukemia. In addition, leukemic cell proliferation was dramatically inhibited by HERG K+ channel special inhibitor E-4031. Moreover, E-4031 suppressed the cell growth by inducing a specific block at the G1/S transition phase of the cell cycle but had no effect on apoptosis in leukemic cells. The results suggested that HERG1 K+ channels could regulate leukemic cells proliferation and were necessary for leukemic cells to proceed with the cell cycle. HERG1 K+ channels may also have oncogenic potential and may be a biomarker for diagnosis of leukemia and a novel potential pharmacological target for leukemia therapy.

Objective To assess the differences in carotid artery stiffness properties and endothelium-independent dilation (EID)between elderly and young patients,and evaluate the echotracking (ET)system for vascular stiffness at different ages.Methods A total of 79 outpatients with multiple cardiovascular risks were recruited.Clinical data including medical history,height,weight,blood pressure,fasting blood glucose and blood lipid were collected.We evaluated the arterial stiffness parameters of carotid artery and EID using an ultrasonic ET system in 46 elderly subjects,compared with 33 sex-matched non-elderly subjects.The impaired EID function was defined as brachial artery nitroglycerin mediated dilation (NMD)below 4％.Results All stiffness parameters including pressure-strain elasticity modulus stiffness index β (Ep),pulse wave velocity β (PWVβ)and augmentation index (AI)were significantly increased in elderly group compared with the non-elderly group [(138.9±64.7)kPa vs.(100.6±30.8)kPa,(10.9±4.7)vs.(8.2±2.3),and (6.9±1.4)m/s vs.(6.1±0.9)m/s,P＜0.05 respectively],while the exception of arterial compliance (AC)was reduced (0.9±0.3)mm2/kPa vs.(1.0±0.5)mm2/kPa(P＜0.05).The incidence of impaired EID in elderly group was higher than in non-elderly group [56.5％ (26 cases) vs.33.3％ (11 cases),P＜0.05].ET parameters including Ep,stiffness index β,PWVβ,AC and AI were related to age (r=-0.44,-0.45,-0.40,-0.40,0.34,all P＜0.01)； Ep,stiffness index β,PWVβ and AC were also related to impaired EDI (r=-0.38,-0.40,-0.34,-0.29,all P＜0.01).Conclusions Arterial stiffness properties and EID measured by ET system was more serious in elderly with multiple cardiovascular risks than in non-elderly subjects.As a convenient and accurate assessment of stiffness parameters,ET system is optimal option for measuring arterial stiffness and EID in elderly people.