Objective To construct a novel recombinant human IFN-ε,and to analyze its physi-cal,chemical properties and biological characteristics.Methods Human genomic DNA was used as the template to synthesize IFN-ε gene by PCR.The sequence was cloned into plasmid vector pET-32a(+),and the recombinant plasmid pET-32a(+)/IFN-ε was transformed into E.coli BL2l(DE3).The form of ex-pression product was inclusion bodies.After purification and renaturation.high purity active protein IFN-εwas achieved.The final product was tested for its physical.chemical properties and biological characteristics including anti-viral and anti-proliferative acfivities.Results IFN-ε was expressed in inclusion body in E.coli.After the protein renaturation and purification,the purity was more than 95%.The rhIFN-ε protein had a specific anti-viral activity of about 1.2×103 IU/mg.Its anti-poliferative activity is obvious and can in-duce cells to produce anti-viral protein MxA.Conclusion Human IFN-ε protein was expressed successful-ly.and this protein has anti-virus and anti-prolireration activity.

Objective To investigate the histogenesis and clinicopathologic features of carcinosarcoma of the urinary bladder. Methods Two cases of carcinosarcoma (one man and one woman)were included.They were admitted to hospital with discontinuous,painless,macroscopic hematuria.Clinicopathalogic features of all findings of the 2 cases were reviewed. Results Cystoscopy of both cases showed that the tumors displayed polypous or cauliflower-like shape,and grew invassively.CT examination of the bladder wall confirmed the presence of parenchymatous tumor.Both of the 2 cases underwent partial cystectomy.Intraoperative examination showed the tumors were similar to carcinoma of bladder.Histopathology showed biphasic neoplasms with distinct high grade epithelial and mesenchymal components.Morphologic characteristic of the tumor was abnormally proliferative and the mitotic figures could be seen.One of the patients died within 11 months and the other,16 months following the operation. Conclusions Carcinosarcoma of the urinary bladder is a kind of rare malignant tumor that is most often in an advanced stage at presentation and has an unfavorable prognosis. It should be identified promptly and treated appropriately.Those who have discontinuous,painless,macroscopic hematuria should be warned of the risk of the disease.

Objective To prospectively study the clinical significance of the MELD scoring system in surgical treatment of obstructive jaundice.Methods 112 patients with obstructive jaundice who were admitted into our hospital from January 2009 to December 2013 were divided into two groups:group A (Stage Ⅰ PTCD and stage Ⅱ open operation,n =53) and group B (1 stage open surgery,n =59).The amount of intraoperative bleeding blood loss,operation time,postoperative complications,duration of hospitalization,mortality rate,and the changes in liver function after surgery were compared between the two groups.Results The differences in the liver function index of the two groups on the same postoperative date were significantly different (P ＜ 0.05).The liver function of group A recovered faster than group B.Patients in group A with a MELD ＜ 10 points stayed in hospital significantly longer when compared with patients in group B.For patients in group A with MELD ＞ 10 points,the operation time,bleeding volume,postoperative complications and hospitalization were significantly less than the patients in group B (P ＜ 0.05).There were 3 patients (group B) who died with MELD ≥ 20 points after operation.Conclusions In patients with obstructive jaundice with a MELD score greater than 10 points,especially those with a score equal to or greater than 20,PTCD should be performed first to relieve biliary tract obstruction,followed by a stage Ⅱ open surgical operation after the liver function had improved.MELD had important clinical significance in the evaluation of operation risk in patients with obstructive jaundice.

Objective To investigate the distribution of CYP2C19 metabolism phenotype in patients with clopidogrel treatment in Huangshi.Methods The peripheral blood samples were obtained from 76 patients with clopidogrel treatment in Cardiovascular Department.CYP2C19 genotypes were determined by the gene chip,CYP2C19 metabolism phenotype were investigated and com-pared with the data of healthy Han Chinese from the published papers.Results There were three kinds of metabolizers about the metabolism phenotypes of CYP2C19,which were extensive metabolizers,intermediate metabolizers and poor metabolizers.In the 76 patients,the ratios of these metabolism phenotypes were 39.47%,44.74% and 15.79%,respectively.The result coincided with the healthy Han Chinese.Conclusion The clinical individualized medication of healthy Han Chinese could be refered to the clinical indi-vidualized medication of patients with clopidogrel treatment in Huangshi.

Objective: To investigate the feasibility and accuracy of sentinel lymph node biopsy(SLN) following neoadjuvant chemotherapy.Methods:Sentinel lymph node biopsy was evaluated following two or three cycles of neoadjuvant chemotherapy with pharmorubicin /cyclophosphamide/5-FU(CEF,48 patients) or pharmorubicin /cyclophosphamide / taxotere(CET,12 patients).Two ml 1% methylene blue was subcutaneously injected at 4 sites of the skin over the tumor before radical operation.Pathohistological results were assessed for all the specimen including blue lymph node and other axillary lymph node(ALND). Results:Among the 66 patients SLN was identified in 60 of them(90%).Residual metastatic disease was identified in 23 cases on SLN,including 15 positive and 8 negative node on ALND,37 cases were completely negative on SLN including 35 negative and 2 positive node on ALND,and SLN was falsely negative in 2 cases(8%).Conclusion:The success rate of sentinel node identification and the predict ability of the SLN for the pathologic status of the adjacent non-SLNs do not seem to be altered after neoadjuvant therapy.

Objective To investigate the clinical efficacy and safety of six-hole moxibustion box therapy for stable chronic obstructive pulmonary diseases(COPD).Methods Sixty patients with thoracic facet joint disorder were randomly allocated to treatment and control groups, 30 cases each. The control group inhaled tiotropium bromide inhalation powder spray and the treatment group received six-hole moxibustion box therapy in addition. The COPD Assessment Test (CAT) score and the dyspnea score were recorded and peripheral blood oxygen saturation was measured in the two groups before and after treatment. The adverse reactions were monitored in the two groups. Results There were statistically significant pre-/post-treatment differences in the CAT score and the dyspnea score in the two groups (P<0.05). There were statistically significant post-treatment differences in the CAT score and the dyspnea score between the treatment and control groups (P<0.05). There were statistically significant differences in pre-/post- treatment CAT score difference value and dyspnea score difference value between the two groups (P<0.05). There was a statistically significant pre-/post-treatment difference in peripheral blood oxygen saturation in the treatment group (P<0.05). There was a statistically significant post-treatment difference in peripheral blood oxygen saturation between the two groups (P<0.05).Conclusion Six-hole moxibustion box therapy plus tiotropium bromide inhalation powder spray is safe and effective in treating stable COPD.

The glycoproteins in the biological sample are low abundance and are susceptible to be inhibited and interfered by other non-glycoproteins.An enrichment step is usually required before the glycoprotein analysis, but the operation steps of conventional solid-phase-based glycoprotein enrichment methods are difficult to be compatible with the most classical enzyme-linked immune sorbent assay (ELISA) technique.In this study, a novel water-soluble dendrimer based boronic acid capture (DBC) material was developed using PAMAM 4.0 as the carrier and boronic acid as the affinity group.The method was applied to the detection of glycoproteins in human liver microsomes using ELISA.In this study, the DBC enrichment conditions were optimized by model glycoprotein, and then its sensitivity and anti-interference ability were investigated.This method was applied to the enrichment of glycoproteinsin human liver microsomal.The results showed that the enrichment selectivity of DBC for glycoprotein could be up to 100000 folds, and the enrichment signal of glycoprotein could be increased by 100 times.Therefore, the ELISA method using DBC as a novel enrichment material for glycoprotein had high sensitivity and selectivity in detection of biological samples with only one simple incubation step, which was useful for glycoproteins researches.

Objective To clone and express recombinant outer membrane protein P6, determine its optimal expression conditions and to investigate the immunoprotective effects of the P6 protein on mice. Methods The P6 gene of nontypeable Haemophilus influertzae(NTHi) was amplified by PCR from the NTHi genome and cloned into expression vector pET-32a (+) to generate the pET-32a-P6 recombinants. They were confirmed by nuclease digestion and sequence analysis. The verified recombinant was transformed into E. coli BL21 (DE3). Its optimal expression conditions were determined such as engineering strains, the concentration of IPTG, inducing temperature, inducing time, different medium etc. The recombinant protein was purified by Q Sepharose~(TM) XL ion exchange and gel filtration chromatography. The protein was analyzed by SDS-PAGE, Western blot and sequencing. BALB/c mice were immunized with recombinant protein be-fore challenged by NTHi through intraperitoneal injection. Then the mortality rate of different group was com-pared. Results The recombinant P6 of NTHi was successfully constructed and expressed in E. coli at a rel-atively high level. The purity was up to 95% after purification. The relative molecular mass of the protein is 14 145. 848. The recombinant protein was confirmed to show specific reaction on the antiserum through Western blot. The animal experiments showed the mortality rates of immunization groups were significantly lower than that of the control group (P < 0.05). Conclusion The successful expression of the recombinant P6 will be very helpful for the further study on development of vaccine, its purified, immunological activity and antibody preparation.

Objective To assess the prediction value of nonbronchial systemic arterial supply in hemoptysis patients with non-enhanced CT. Methods Fifty-six consecutive patients with hemoptysis underwent non-enhanced CT. Thickness of pleural adjacent to parenchymal lesion larger than 3 mm was regarded as index of nonbronchial systemic arterial supply. Conventional angiography was used as the standard of reference. CT findings were compared with those of conventional angiography. The sensitivity, specificity, and accuracy of CT for predicting nonbronchial systemic arterial supply were assessed. Results The sensitivity, specificity and accuracy of CT for predicting nonbronchial systemic arterial supply were 72.73%, 95.00% and 91.11%, respectively. Sensitivity was higher when nonbronchial systemic arterial supply located in superolateral and posterolateral lung, and lower in anteromedial and inferior lung. Specificity and accuracy were high for predicting nonbronchial systemic arterial supply in every locations. Conclusion Non-enhanced CT can predict nonbronchial systemic arterial supply in patients with hemoptysis, which is helpful for selecting angiography and embolization.

Objective To explore the selection of palliate operation styles for unresectable malignant obstructive jaundice diseases. Methods The clinical data of 112 cases of unresectable malignant obstructive jaundice diseases in the last 5 years were analyzed retrospectively. Results The rate of operation mortality was 7.1％ and the incidence rate of post-operative cholangitis was 17.3 ％.The survival time in the Roux-en-Y choledochojejunostomy group was (9.4±1.6) months, and there were no significant differences among laparotomy stent internal drainage group[(9.8±12.5)months]and the PTCD stent internal drainage group [(9.0± 3.1)months]. But survival time in the laparotomy bridge internal drainage group [(6.8±1.7)months]was significantly lower (P＜0.05). The survival times in the ERCP stent drainage group [(3.5±2.2)months]and exploratory laparotomy group [(2.8±2.7)months]were even more significantly lower (P＜0.01).Conclusion As a palliative operation for unresectable malignant obstructive jaundice diseases, Roux-en-Y choledochojejunostomy applies to the middle and distal obstruction,laparotomy stent internal drainage applies to hilar obstruction,ERCP stent drainage only applies to distal obstruction,and PTCD stent internal drainage applies to any part obstruction of the bile duct. The Roux-en-Y choledochojejunostomy, laparotomy stent internal drainage, and PTCD stent internal drainage would improve the life time and life quality of these patients.