Objective To clone and express the immunodominant domain of the chlamydial proteaselike activity factor(CPAF) from Chlamydophila psittaci(Cps) and evaluated the diagnosing value of the recombinant protein in Cps infection.Methods The immunodominant region epitope of CPAF (CPAFm,A196-A450)from Cps was chosen according to bioinformatics analysis and references.The specific primer was designed and the gene was amplified by PCR and then ligated into a pGEX6p-2 vector.Recombinant protein was induced to express by IPTG and analyzed by SDS-PAGE and Western blot.Indirect EL1SA method of serological diagnosis was established with the reorganization protein as coating antigen.One hundred and eighty sera samples from ducks with respiratory tract infection symptoms were detected with the established indirect ELISA and a commercial ELISA-kit to assess the value of the recombinant protein in serodiagnosis.The results were further identified with Western blot.Results Prokaryotic expression vector pGEX6p-2/CPAFm was constructed and a 54x103 fusion protein was attained.The indirect ELISA method was established with reorganization protein for envelope antigen.Using the indirect ELISA to detect Cps lgG positive and negative reference sera,the sensitivity and specificity were both 100％ (20/20).And the recombinant protein has no cross reaction with either Chlamydophila pneumoniae or Chlamydophila trachomatis.The concordance rate between the indirect ELISA and Western blot to 180 ducks sera samples was 100％,while the concordance rate of the commercial ELISA kit was 77.5％-95.0％.Conclusion The prepared recombinant protein of the CPAF immunodominant region epitope gene from Gps can highly benefit on developing new indirect ELISA as methods to detect specific anti-Cps antibodies.

Objective To detect the expression and the biological activity of a recombinant adenovirus expression vector carrying human neurotrophin-3 (NT-3) receptor TrkC gene (Adeno-TrkC) in neural stem cells. Methods The expression of TrkC mRNA in 293 cells infected with Adeno-TrkC was detected by RT-PCR, and the expression of TrkC protein in neural stem cells infected by Adeno-TrkC was detected with immunocytochemistry and Western blotting. The effects of human neurotrophin_3 (NT-3) on the neural stem cells infected by Adeno-TrkC differentiating into neuron-like cells and astrocyte-like cells in vitro were observed. Results The transcription of TrkC mRNA and the expression of TrkC protein was detected in 293 cells and neural stem cells infected by Adeno-TrkC. This kind of TrkC was able to make more neural stem cells differenting into neuron-like cells in vitro with its ligand NT-3 and the percentage of neuron-like cell’s differentiation was 55^2% which was higher than that of other control groups.Conclusion TrkC protein is expressed in neural stem cells transfected with Adeno-TrkC and is able to make neural stem cells differenting into neuron-like cells. The present study may provide the basis on gene therapy of central nervous injury using further NT-3 and TrkC.

Objective To investigate whether nuclear transcription factor κB(NF-κB) through Toll-like receptors 2(TLR2) was activated by lipid-associated membrane proteins(LAMPs) of Mycoplasma genitalium.Methods LAMPs were extractded and THP-1 cells were stimulated.The activation of NF-κBp65 was detected by ELISA and the expression of TLR2 mRNA was detected by RT-PCR.Effects of TLR2 neutralizing antibody on LAMPs induced the activation of NF-κBp65 was analyzed by ELISA.After LAMPs stimulated 293T cells with the co-transfection pFLAG-TLR2,pNF-κB-luc,pRL-TK,the activity of NF-κB firefly luciferase and pRL-TK Renilla luciferase were detected by the dual-luciferase reporter gene,to analyzed the role of TLR2-mediated NF-κB activation by LAMPs in 293T cells.Results The activation of NF-κBp65 was mediated in LAMPs induced THP-1 cells and was significantly increased by LAMPs in a dose dependent manner.when LAMPs was 4.0 μg/ml,the activation of NF-κBp65 was the highest level.TLR2 mRNA expression was up-regulated by LAMPs in THP-1 cells.TLR2 neutralizing antibody could inhibit the activation of NF-κB by 60% in LAMPs stimulated THP-1.NF-κB fluorescence was significantly increased by co-transfection pFLAG-TLR2 in a dose-dependent manner. ConclusionMycoplasma genitalium-derived lipid-associated membrane proteins activate NF-κB via TLR2 and the activation of TLR2-mediated play an important role in pathogenic process of LAMPs.

Objective To discuss the influences of fasting on the embolism adverse reactions, sense of hunger, blood amylase, blood glucose and anxiety of patients with hemoptysis of pulmonary tuberculosis after embolism operation. Methods Selected 92 cases of patients with hemoptysis of pulmonary tuberculosis who have taken bronchial artery embolism in Pulmonary hospital of Changsha central hospital from June 2014 to December 2015,randomly divided the patients into three groups according to operating date, namely, the non-fasting group, 6h fasting group and 24h fasting group. The non-fasting group have routine diet after the operation; the 6h fasting group and 24h fasting group respectively fasted for 6 hours and 24 hours. Then compared the embolism adverse reactions, sense of hunger, blood amylase, blood glucose and anxiety of the patients in the three groups so as to check whether there is difference. Results Before operation, there was not significant difference in adverse reaction of embolism, sense of hunger, anxiety, blood glucose and amylase between the three groups (P > 0.05). After operation, there was significant difference in hunger sense of difference between the three groups(F=13.308,P<0.05), the hunger sense of 24 h fasting group was more serious than 6h fasting group than non-fasting group;the anxiety score of 24h fasting group is higher than the other two(F=6.509,P<0.05), and the value of the next morning fasting blood glucose in fasting group 24h is higher than other two(F=6.642,P<0.050). comparing the score of anxiety in 6h fasting group and non-fasting group, there was not significant difference, and comparing the post-embolism adverse reaction, 72 h blood amylase of all three groups and these groups one to one, there was also not significant difference ( P>0.05). Conclusion Fasting will cause and intensify the sense of hunger, anxiety and nutrition disorder of patients with hemoptysis of pulmonary tuberculosis after embolism.

Objective To investigate the chemical constituents of the Radix Campylotropidis Hirtellae ( RCH ) . Methods 60%ethanol extract of the RCH was separated by column chromatography on silica gel, ODS and Sephadex LH-20 to obtain aimed compounds.Structures of the compounds were investigated by modern spectroscopy.Results Nine compounds were separated from Campylotropis hirtella ( Franch.) Schindl, and the structure of the new compound 1 was determined by spectral analysis and named hedysarimcoumestanⅠ( 1 ) , while the others were identified as the known isotrifoliol(2), hedysarimcoumestan B(3), umbelliferone(4), orobol(5), quercetin(6),(+) catechin(7), kaempferol (8) and dihydrokaempferol(9) .Conclusion Compound 1 is a new compound and 3-5 are separated from C.hirtella for the first time.The screening results of inhibition of H5N1 virus activity show that compound 5 has some inhibitory activity at a level of 50.0 μg /ml.

Objective To explore the role of type Ⅶ collagen (COL7A1) gene in the pathogenesis of pretibial dominant dystrophic epidermolysis bullosa (DDEB-Pt).Methods Peripheral blood samples were obtained from a sporadic Chinese patient of Han nationality with DDEB-Pt,his parents and 100 healthy human controls.A modified salting-out method was used to extract genomic DNA from the blood samples,and PCR was performed to amplify 118 exons of the COL7A1 gene followed by DNA sequencing.Results A G→A mutation was identified at position 6109 (G6109A) in exon 78 of the COL7A1 gene in this patient,which caused a change from GCT to ACT at codon 2037 in the triple helix region,and resulted in the substitution of glycine (Gly) by arginine (Arg) (p.Gly2037Arg).Conclusion A novel glycine substitution mutation was identified in the COL7A1 gene in the patient with DDEB-Pt,which may be a pathogenic mutation.

Objective To explore the effect of Jianbu-Huqian ointment combined with proximal femoral nail antirotation for the elderly patients with intertrochanteric fracture of femur with osteoporosis. Methods A total of 85 elderly patients with intertrochanteric fracture of femur with osteoporosis were randomly divided into the control group (40 patients) and the observation group (45 patients. Both groups of patients were treated with proximal femoral nail antirotation, and the observation group was added with Jianbu-Huqian ointment. The pain, fracture healing, joint function, bone mineral density and complications were compared between the two groups. Results The VAS pain score at 1st week (6.2 ± 1.6 vs. 7.9 ± 1.6; t=3.683, P<0.05), 3rd week (3.1 ± 1.1 vs. 5.4 ± 1.4; t=4.239, P<0.05) and 7th week (2.3 ± 0.9 vs. 4.0 ± 1.3; t=3.571, P<0.05) after operation of the observation group were significantly lower than those of the control group. The occurrence time (42.4 ± 5.9 d vs. 49.2 ± 6.2 d; t=4.139, P<0.05), walking time (6.9 ± 1.1 weeks vs. 8.7 ± 1.2 weeks;t=3.695, P<0.05), fracture healing time (13.9 ± 2.2 weeks vs. 16.2 ± 3.1 weeks, t=3.473, P<0.05) of the observation group were significantly lower than those of the control group. Harris hip score (89.8 ± 7.21 vs. 81.4 ± 5.93; t=3.728, P<0.05) in the observation group was significantly higher than that in the control group. The bone density of the femur (0.782 ± 0.142 g/cm2 vs. 0.691 ± 0.135 g/cm2; t=3.759, P<0.05) and the area of ward (0.628 ± 0.071 g/cm2 vs. 0.513 ± 0.059 g/cm2; t=4.386, P<0.05) of the observation group were significantly lower than those of the control group. The incidence of postoperative complications in the observation group was 15.0%(6/40), and the control group was 37.8% (17/45). The difference was statistically significant (χ2=5.567, P<0.05). Conclusions The Jianbu-Huqian ointment combined with proximal femoral nail antirotation could reduce the pain, promote fracture healing, and prevent the occurrence of postoperative complications for the elderly patients with intertrochanteric fracture of femur with osteoporosis.

Objective To investigate the clinical and genetic characteristics of a progressive symmetric erythrokeratoderma (PSEK).Methods The clinical and genetic characteristics of a PSEK were compared and analyzed with clinical collection and review of 17 PSEK families reported in China since 1980.Results PSEK was consistent with autosomal dominant inheritance.PSEK had the feature with incomplete penetrance and variable expression.The clinical features included hyperkeratotic plaques with distinct border and strikingly symmetric distribution pattern on the extremities.Part of patients was extended to other areas of the body.The onset of the disease commonly started in infancy or childhood.The general health condition was not affected.PSEK might be associated with other clinical symptoms.The diseased potential increased in the family of consanguineous marriage.Conclusions PSEK has genetic heterogeneity.Its causative genes have not been determined.Further studies are needed.

Objective To explore the value of three antibodies in the differential diagnosis of papillary thyroid carcinoma ,by de‐tecting the expression of HBME‐1 ,CK19 and CD117 in papillary thyroid cacinoma ,thyroid follicular adenoma and Hashimoto′s thy‐roiditis tissues .Methods Totally 85 cases were collected from January 2013 to December 2015 ,including papillary thyroid cacino‐ma ,thyroid follicular adenoma and Hashimoto′s thyroiditis .They were immunohistochemical stained by HBME‐1 ,CK19 and CD117 .SPSS16 .0 software was used to analyze the relationship between the staining results with different pathological changes . Results The positive rates of HBME‐1 ,CK19 and CD117 were 87 .3% ,98 .2% ,and 7 .3% ,respectively .The positive expression of them in benign and malignant groups had significant difference (P< 0 .05) and their consistency checking Kappa were 0 .582 , 0 .551 ,and 0 .874 ,respectively .Conclusion In the differential diagnosis of papillary thyroid carcinoma and benign lesions ,CD117 is better than HBM E‐1 and CK19 .It′s possible to use a combination of them in practice .

Aim To study the effect of different doses of l-Tetrahydropalmatine( l-THP) on morphine-induced conditioned place preference(CPP) and observe whether l-THP itself induces CPP.Methods ①♂SD rats were administered with morphine (5.00 mg?kg -1 ,sc) or saline and trained for 8 days;on d 9,the rats were tested CPP with no treatment or 40 min after they were given different doses of l-THP(1.25～5.00 mg?kg -1,ip) to observe the effect of l-THP on morphine-induced CPP;② With daily injection of l-THP (ip) at different doses,effect on the extinguishment of morphine-induced CPP was tested weekly; ③ Normal saline (NS) or l-THP (1.25～5.00 mg?kg -1 , ip) was used as a training drug to test whether l-THP could induce CPP in the rats. Results 5.00 mg?kg -1 morphine (sc) induced CPP; l-THP of 2.50 mg?kg -1 and 5.00 mg?kg -1 administered prior to the testing reduced the expression of morphine-induced CPP significantly (P