An optimized recombinant HPV16 E6E7 fusion gene(HPV16 ofE6E7)was constructed according to codon usage for mammalian cell expression,and a mutant of HPV16 ofE6E7 fusion gene(HPV16 omfE6E7)was generated by site-directed mutagenesis at L57G,C113R for the E6 protein and C24G,E26G for the E7 protein for HPV16 ofE6E7 [patent pending(CN 101100672)].The HPV16 omfE6E7 gene constructed in this work not only lost the transformation capability to NIH 3T3 cells and tumorigenicity in SCID mice,but also maintained very good stability and antigenicity.These results suggests that the HPV16 omfE6E7 gene should undergo further study for application as a safe antigen-specific therapeutic vaccine for HPV16-associated tumors.

Objective To investigate 4 variants single nucleotide polymorphisms (SNPs) of 5-lipoxygenase-activating protein(ALOX5AP) in lipoxygenase pathway and in cytochrome P450 pathway as susceptibility genes for stroke in a southeastern Chinese population,and evaluate the associations between susceptibility genes and cerebral infarction,to find whether gene-gene interactions increase the risk of cerebral infarction.Methods By case-control study,two hundred and ninety-two patients with cerebral infarction and 259 healthy control subjects were included.Eight variants in 5 candidate genes were examined for stroke risk,including the SG13S32 (rs9551963),SG13S42 (rs4769060),SG13S89 (rs4769874),and SG13Sl14 (rs10507391) variants of the ALOX5AP gene,the G860A (rs751141) variant of the soluble epoxide hydrolase (EPHX2) gene,the A1075C (rs1057910) variant of the CYP2C9 *2 gene,the C430T (rs1799853) variant of the CYP2C9* 3 gene,and the A6986G (rs776746) variant of the CYP3A5 gene.Gene-gene interactions were explored using generalized multifactor dimensionality reduction (GMDR)methods.Results There were no statistically significant differences in the frequencies of the genotypes of the 8 candidate genes.The GMDR analysis showed a significant gene-gene interaction between SG13S114 and A6986G,with scores of 10 for cross-validation consistency and 9 for the sign test (P =0.011).These genegene interactions predicted a significantly higher risk of cerebral infarction (adjusted for age,hypertension,and diabetes mellitus;OR =1.804,95％ CI 1.180-2.759,P =0.006).Conclusions A two-loci gene interaction confers significantly higher risk for cerebral infarction.The combinational analysis used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases.

Objective To explore the prognostic determinants and clinical treatment strategy in 142 patients with severe trauma brain injury(STBI).Methods Retrospective analysis of clinical data of 142 patients with STBI in our department from April 2006 to April 2012.All the patients were divided into good prognosis group(Ⅲ~V grade)and poor prognosis group(I~Ⅱgrade)ac-cording to the GOS classification standard.Age,gender,GCS,encephalocele,morphotogy of the basal cisterns on CT scanning,asso-ciated inj ury,shock,hyoxemia,underlying disease and hyperglycemia were chosen as the observation index.Statistical analysis was performed with Pearson Chi-square Test.Results 52.11% of patients with good prognosis,47.89% of patients with poor progno-sis and 3 1 .6 9% of patients were dead.Age,GCS,encephalocele,morphotogy of the basal cisterns on CT scanning,associated inj ury, shock,underlying disease were the prognostic determinants of STBI(all results P<0.05).Conclusion Age,GCS,encephalocele, morphotogy of the basal cisterns on CT scanning,associated inj ury,shock,underlying disease can determine the prognosis of STBI. Multidisciplinary cooperation treatment depending on the patient′s conditions is the key of improving the outcomes of STBI.

Objective To investigate the interrelations of ALOX5AP SG13S114A/T , COX-2 765G/C , COX-1-50C/T polymorphisms and cerebral infarction. Methods The ALOX5AP SG13S114A/T, COX-2 765G/C and COX-1 50C/T polymorphisms in 411 cases with cerebral infarction and 411 controls were measured by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism method. The generalized multifactor dimensionality reduction (GMDR) method was employed to detect gene-gene interactions. Results Single-gene analysis showed that there were no significant differences in the genotype and allele frequency distributions of ALOX5AP SG13S114A/T, COX-2 765G/C and COX-1 50C/T between two groups. However, in those cases carrying ALOX5AP SG13S114AA as well as COX-2 765CC , the risk of cerebral infarction increased significantly by 2.842 times. Conclusions The combinational analysis among genes used in this study may be helpful in the elucidation of genetic risk factors for common and complex diseases.

Objective To study the clinical value of treatment for malignant bone tumors by interventional chemoembolization combined radiotherapy.Methods Forty-four patients with malignant bone tumors were divided into two groups.In the study group(group A),29 cases were treated by infusion of lipiodol with chemotherapeutic drugs,and the tumor feeding arteries were embolized with PVA partials and Gelatin sponge or steel coils,then regional radiotherapy were administered regularly.In the control group(group B),15 cases received radiotherapy and systemic chemotherapy.All the cases were followed up for 6 to 36 months.Results Marked improvement of the symptoms and tumors decreased in size and volume in patients of group A was noticed.Six of them were operated successfully.The 6-,12-,36-month local control rates in group A were 3.5%,13.8% and 51.7%.and the(6-),12-,36-month survival rates were 100.0%,93.1% and 48.3%.The difference of local control rates and the difference of survival rates between the two groups were significant(P

To evaluate the efficacy and safety of an oxaliplatin, fluorouracil (5 FU), and folinic acid (FA) combination in treatment of patients with metastatic or advanced gastric cancer,we used oxaliplatin 130 mg/m 2 (2 hour intravenous infusion,1 d)and FA 200 mg/m 2 (2 hour intravenous infusion,1 to 5 d) followed by 5 FU 500 mg/m 2 (4 hour continuous infusion,1 to 5 d) every 3 weeks. Efficacy of treatment was evaluated after 3 cycles and the responders were confirmed 4 weeks later. In 26 case evaluated, patial release was achieved in 11 cases, stablized in 9 cases, progressed in 6 cases. The overall response rate was 42.3%. The main adverse effects were gastrointestinal tract toxicity, neurosensory toxicity and bone marrow suppression. This trail suggested that the short term curative effect of this regimen was similar to those of others and showed good efficacy and an acceptable safety profile. It is a new option for the treatment of metastatic and advanced gastric cancer.

Objective:To investigate the association of plasma epoxyeicosatrienoic acids (EETs) with early neurological deterioration (END), and whether EETs are mediated by EPHX2 gene variants in patients with minor ischemic stroke (MIS).Methods:This is a prospective, multi-center observational study in patients with acute MIS in the Chinese population. Acute MIS patients with the first onset and onset within 24 hours who were admitted to Deyang People′s Hospital, the Second Affiliated Hospital of Wenzhou Medical University and the Third Affiliated Hospital of Wenzhou Medical University from March 2013 to June 2015 were recruited. Plasma EETs levels were measured on admission. Single nucleotide polymorphisms of EPHX2 gene rs751141 were genotyped using mass spectrometry. The primary outcome was END within 10 days after admission. END was defined as an increase in National Institutes of Health Stroke Scale score of 2 or more points.Results:A total of 322 patients were enrolled, of which 85 (26.4%) patients experienced END. EETs levels were significantly lower in patients with END [(60.3±7.3) nmol/L] compared to patients without END [(68.4±8.1) nmol/L , t=8.464, P<0.001]. Frequency of EPHX2 gene rs751141 GG was higher in patients with END [66/85(77.6%)] than in patients without END [123/237(51.9%),χ2=17.130, P<0.001], and patients with EPHX2 gene rs751141 GG genotype showed lower EETs levels [GG: (59.6±7.8) nmol/L, AG:(67.9±8.2) nmol/L, AA:(68.8±3.2) nmol/L, F=9.285, P<0.001]. Low level (≤64.3 nmol/L) of EETs was an independent predictor of END (31.5-51.3 nmol/L group: OR=2.96,95% CI 1.18-8.77, P=0.02; 51.4-64.3 nmol/L group: OR=2.46,95% CI 1.06-6.89, P=0.03) in multivariate analyses. END was associated with a higher risk of poor outcome (modified Rankin Scale scores 3-6) at 3 months ( OR=1.82,95% CI 1.46-2.35, P=0.02). Conclusion:END is fairly common and associated with poor outcomes in acute MIS. EPHX2 gene variants may mediate EETs levels, and low levels of EETs are related to END in acute MIS.

Objective To evaluate clinical and radiographic outcomes associated with total hip arthroplasty (THA) using metal-on-metal prosthesis with large diameter femoral head.Methods From October 2007 to December 2007,41 patients (49 hips) underwent large diameter femoral head metal-on-metal THA in our hospital were involved in this study.Clinical outcomes measures were Harris score,hip range of motion and incidence of complications.Abduction angle and anteversion angle of cup were measured on radiological films.The radiolucent line and osteolysis around the prosthesis were also recorded.Results Thirty-nine patients (47 hips) were followed up at least 2 years.The average Harris hip score had improved from (43.8±13.1) points preoperatively to (92.0±5.4) points at final follow-up.All the patients had attained satisfactory results.No late complication happened.For the rang of motion at final follow-up:flexion of the hip had improved from 79.8° to 113.2°,abduction had improved from 20.9° to 40.2°,external rotation had improved from 12.0° to 30.8°.Radiological measurement showed the mean abduction angle of cup was 39.5°±4.9°,the mean anteversion angle of cup was 14.5°±2.1°.No radiolucent line and osteolysis were found after THA.Conclusion The short-term effects of THA using metal-on-metal prosthesis with large diameter femoral head is encouraging,especially for young patients.

OBJECTIVETo investigate the clinical effect and safety of chiropractic in treating cervicogenic sudden hearing loss.

METHODSFrom January 2011 to October 2013, 90 patients with cervicogenic sudden hearing loss were randomly divided into treatment group and control group according to the random number table produced by SPSS 19.0 software. In the treatment group, there were 17 males and 28 females, aged from 31 to 62 years old with an average of (47.57±9.43) years; course of disease was from 1 to 3 days with an average of (1.43±0.68) days; pure-tone audiometry score was from 46.5 to 77.8 dB with the mean of (61.20±9.83) dB; Northwick Park Neck Pain Questionnaire (NPQ) score was from 17 to 31 scores with an average of (23.46±7.18) scores. In the control group, there were 15 males and 30 females, aged from 28 to 64 years old with an average of (45.77±6.99) years; course of disease was from 1 to 3 days with an average of (1.50±0.73) days; pure-tone audiometry score was from 48.1 to 75.0 dB with the mean of (63.91±8.05) dB; Northwick Park Neck Pain Questionnaire (NPQ) score was from 20 to 29 scores with an average of (25.61±10.43) scores. The patients of control group were treated with dexamethasone intravenous drip of 10 mg, 3 days later, decreased to 5 mg, 3 days again. And with the methycobal intravenous drip of 500 μg, treatment continued for 10 days. The patients of treatment group were treated with chiropractic additionally except for the therapeutic methods of control group. Chiropractic included local muscle loosening, attacking point, bilateral pulling atlanto-axial joint, and continuous treatment for 10 days. The pure-tone audiometry score and NPQ score were compared between two groups after treatment.

RESULTSAfter the treatment, pure-tone audiometry score and NPQ score in treatment group improved to (40.23± 8.14) dB and (12.70±8.29) scores respectively, which were obviously better than that of control group's (37.70±10.61) dB and (21.24±11.13) scores (P<0.05).

CONCLUSIONCompared with routine method for cervicogenic sudden hearing loss, additional chiropractic can improve hearing and relieve neck pain effectively.

Cervical Vertebrae ; Female ; Hearing Loss, Sudden ; therapy ; Humans ; Male ; Manipulation, Chiropractic ; methods ; Medicine, Chinese Traditional ; Middle Aged

Aurantii Fructus is the dried and immature fruit of Citrus aurantium and its cultivars. To investigate the chemical constituents of Aurantii Fructus, the separation and purification of constituents were performed by column chromatography on silica gel LH-20, HW-40, ODS, PHPLC and PTLC. Fourteen flavonoids, including four flavone glycosides and ten polymethoxyflavones (PMFs) were isolated from the EtOAc fraction and Petroleum ether fraction of Aurantii Fructus and their structures were identified by physicochemical properties and spectral data (NMR and MS) as (2R) -and (2S)-6"-O-acetylprunin (1,2), naringenin-7-O-β-D-glucopyranside (3), 5,7,4'-trihydroxy-8,3'-dimethoxyflavone-3-O-6"-(3-hydroxyl-3-methylglutaroyl)-β-D-glucopyranoside(4), 4'-hydroxy-5,6, 7-trimethoxyflavone (5), natsudaidain (6), nobiletin (7), sinensetin (8), 5,6,7,4'-tetramethoxyflavone (9), 5,7,8,4'-tetramethoxyflavone (10), 3,5,6,7,8,3',4'-heptamethoxyflavone (11), tangeretin (12), 5-demethyl nobiletin (13), and 5-hydroxy-6,7,3', 4'-tetramethoxyflavone (14). Compound 3-5 s were isolated from this plant for the first time and compound 1 was a new one.
Citrus ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Fruit ; chemistry ; Mass Spectrometry ; Molecular Structure