The recombinant plasmid of mChlL-18 prokaryotic expression was transformed into E.coli BL21(DE3) strain and then induced by IPTG at 37℃.After crushed and washed,the expressing inclusion bodies were thoroughly denatured with 6 mol/L guanidine hydrochloride.Then according to experiment design,the effects of rChlL-18 protein refolding yield at different densities were investigated by the systems of artificial chapercne at different densities.Experiment results indicate that there is a optimal condition on assiting rChIL-18 protein by using the artificial chaperone technique.The optimal condition can improve the refolding yield of rChIL-18 protein,and then the expressed product of fusion chicken IL-18 gene in E.coli has a relativity high bioactivity.

Background Research showed that the morbidity rate of primary angle closure glaucoma (PACG) in Mongolian population is 3.02 times more than Han nationality population.To understand the cause and mechanism of PACG in Mongolia is of an important significance.Objective This study was to investigate the pathogenesis of Mongolian PACG.Methods Thirty-two eyes of 32 PACG patients in Mongolia and 40 eyes of 40 PACG patients of Han peoples were included in Inner Mongolia Autonomous Region People's Hospital according to the diagnosis criteria of glaucoma group of Chinese Medical Ophthalmology Association (version 1987),and 13 eyes of 13 normal Mongolia and 17 eyes of 17 normal Han peoples who suffered with ocular truma were recruited as controls.Intraocular pressure(IOP) was measured before surgery.The trabecular meshwork tissue was obtained from all the eyes during the operation.Annexinv-FITC/PI double staining was performed and the apoptosis rate of trabecula cells was tested with flow cytometry.Written informed consent was obtained initial of the study.Results The IOP value in Mongolia PACG group,Han PACG group,Mengolia normal group and Han normal group was (35.97±7.11)mmHg,(38.70± 6.82) mmHg,(14.69 ± 2.91) mmHg and (13.59 ± 2.91) mmHg,respectively,showing a significant difference among the 4 groups(F=106.144,P=0.000),and the IOP was significantly higher in the Mengolia PACG group and Han PACG group than the normal groups(P＜0.05).The apoptosis rate of the cells was (7.14±0.67)％,(5.40±0.69) ％,(5.86±0.91) ％ and(2.29±0.65) ％ in the Mongolia PACG group,Han PACG group,Mongolia normal group and Han normal group,respectively,with a significant difference among them (F =174.888,P =0.000),and apoptosis rate of the Mongolia PACG group was significantly higher than that of the Han PACG group and the Mongolia normal group (P＜0.05).No significant difference was found between the Mongolia PACG group and the Han PACG group or between the Mongolia normal group and Han normal group (P＞0.05).The cell apoptosis rate was increased with the elevation of IOP (b =0.990,F=10.209,P =0.009) with the regression equition Y =2.788 +0.092X.Conclusions The apoptosis rate of trabecula cells in Mongolian is higher than Han people.If these results are associated with the high incidence of Mengolia PACG is worth of study.

Objective To compare the effect of 4 different cold and hot properties of traditional Chinese medicine (TCM) on body temperature and related factors of yeast induced fever rats,and discuss the thermoregulatory mechanism of cold and hot properties of TCM.Methods 108 male SD rats were randomly divided into a normal group,a yeast-induced group,a R.palmatum treated group,a C.chinensis treated group,a Euodia ruticarpa treated group,and a Alpinia officinarum Hance treated group,with 18 rats in each group.Pyrexia model was induced by injecting yeast suspension subcutaneously on rat.At the 4h,8h and 12h after injection of yeast,the rats were sacrificed,and the blood and hypothalamus were collected.The levels of prostaglandin E2 (PGE2),cyclic adenosine 3',5'-monophosphate (cAMP) and arginine vasopressin (AVP) in hypothalamus and plasma were detected by ELISA assay.Results At the 4h after injection of yeast,the temperature of rats in the model group began to rise,and it reached the peak at 8h,while RheumpalmatumL and Coptis chinensis could significantly reduce the body temperature of yeast-induced rat (P＜ 0.01 or P＜ 0.05).At 8h,the levels of PGE2 and cAMP in hypothalamus increased significantly [respectively (31.55 ± 9.88) pg/mg and (0.17±0.03) pmol/mg] compared with the normal group,while the level of AVP (0.14±0.02) pmol/ml in plasma reduced (P＜0.05).Compared with model group,at 8h RheumpalmatumL and Coptis chinensis could significantly lowered PGE2 [respectively (113.65± 18.60) pg/mg and (127.72 ± 15.75) pg/mg,P＜ 0.01 or P＜0.05],and cAMP [respectively (0.69±0.08) pmol/mg and (0.74±0.10) pmol/mg,P＜0.05] in hypothalamus,and increased AVP levels [respectively (1.08 ± 0.12) pmol/ml and (0.91 ±0.01) pmol/ml,P＜0.05 or P＜0.01] in plasma.Euodia ruticarpa and Alpinia officinarum had no significant effect on both body temperature and the levels of inflammatory factors.Conclusion The two cold property traditional Chinese medicines,R.palmatum and C.chinensis,could significantly reduced the body temperature of yeast-induced rats,which may be related to its effective regulation on levels of PGE2 and cAMP in hypothalamus and AVP in plasma,however,the two hot property traditional Chinese medicine,Euodia ruticarpa and Alpinia officinarum Hance,had no related effects.

Objective To explore the relationship between substance P(SP),somatostatin(SS) expression and change of morphology structure in jejunum of arsenism rats.Methods Acoording to sex and body mass,forty five clean grade SD rats were divided into control(0.0 mg.kg-1.d-1),low-dose arsenic(0.4 mg.kg-1.d-1) and high-dose arsenic(10.0 mg.kg-1.d-1) groups,n =15.The rats in low-and high-dose groups were treated with As2O3(2,50 mg/L) through drinking water for 4 months,respectively.Morphology changes of jejunum were observed by histological technique-HE staining and SABC immunohistochemistry.SP and SS positive cells in the jejunum were observed and counted,and its average gray value was analyzed with image analysis software (Biomias).Results Some jejunal villi were irregular in arsenism rats; with some brush border loss and irregular; goblet cells increased; infiltration of inflammatory cells in the lamina propria; and vacuoles in some intestinal gland cells.The differences of SP and SS positive cells between groups were statistically significant (F =608.54,227.59,all P ＜0.05).Compared with the control group (0.94 + 0.21,1.14 + 0.14),SP and SS positive cells in low-and highdose arsenic groups(1.85 + 0.25,1.83 + 0.24 and 4.24 + 0.33,3.31 ± 0.41) were significantly higher(all P ＜0.05),and high-dose arsenic group was significantly higher than the low-dose arsenic group(all P ＜ 0.05).The differences of average gray values of SP and SS positive cells between groups were statistically significant(F =68.43,26.57,all P ＜ 0.05).Compared with the control group(133.76 ± 3.61,137.57 ± 5.49),SP and SS positive cells in low-and high-dose arsenic groups(125.13 + 2.35,131.28 ± 5.66 and 118.30 ± 4.58,124.03 ± 3.94) were significantly lower(all P ＜ 0.05),and high-dose arsenic group was significantly lower than the low-dose arsenic group (all P ＜ 0.05).Conclusions Up-regulation of SP,SS may be related to jejunal mucosal injury and morphology structure in arsenic poisoning rats.

Objective To study the molecular mechanism of renal injury of chronic arsenic poisoning rats induced by the expression of cysteine caspase-8 and P53 in renal proximal tubular epithelial cells.Methods Sixty healthy SD rats were divided into three groups,high-,low-dose group,and control group,n =20 in each group.The rats in high and low dose groups were treated with As203 through drinking water,10.0 and 0.4 mg/kg,respectively.The control rats were given distilled water.Four months later,serum and urinary arsenic level was determined,and kidney specimens were taken.The expression of cysteine caspase-8 and P53 in renal proximal tubular epithelial cells was detected by histological technique-HE staining and SABC immunohistochemistry.In addition,cell number counting and image analyses were used in the study.Results The number of caspase-8 positive cells of renal proximal tubule in control group,low-and high-dose group was 3.33±1.32,31.14±8.02 and 46.50±7.20 cell number/visual fields,respectively,which was increased with dose increasing(all P ＜0.05);the average gray value was 151.34±6.40,133.58±4.63 and 128.34±16.28,respectively,decreased with dose increasing(all P ＜0.05).The number of P53 positive cells was 3.17±1.59,26.29±4.23 and 47.00±6.22 cell number/visual fields,respectively,increased with dose increasing (all P ＜ 0.05) ; the average gray value was 142.54±8.06,121.48±5.68 and 101.89±6.35,respectively,decreased with dose increasing (all P ＜ 0.05).Conclusion The increase of caspase-8 and P53 positive cells is one of the molecular mechanisms of renal injury induced by arsenic poisoning.

OBJECTIVE:To demonstrate challenges of pharmacy automation reconstruction so as to set solutions. METHODS:Based on literature review,analysis of pharmacy automation setting and features,this paper gave the suggestions and solutions on construction cost,management model change,equipment maintenance and emergency response,etc. according to the practice of the hospital. RESULTS&CONCLUSIONS:Pharmacy automation construction should be stick to the requirements of new health re-form to lower the cost by using the out resources and interior optimal allocation,to improve efficiency by unified planning and proper design,and to ensure the system running efficiently by sufficient maintenance and contingency plan.

Objective To observe effects of different extracts of Jianpi Huogu Formula (JPHGF) on proliferation and differentiation of bone marrow mesenchymal stem cell (BMSCs). Methods Whole bone marrow adherent was used to screen, culture, and isolate BMSCs. Extracts from different parts (water, chloroform, ethyl acetate and n-butanol parts) of JPHGF were administrated for a certain time. MTS was used to detent cell proliferation;ALP staining was used to detect ALP activity;ARS staining was used to detect the formation of calcium nodules;oil red O staining was used to detect fat cell formation. Results Extracts from different parts of JPHGF could promote cell proliferation of BMSCs in different levels, followed by its strength in water, chloroform, ethyl acetate, and n-butanol parts;ALP staining results showed that the intensity of ALP expression of the order is water, acetic acid ethyl, chloroform and n-butanol parts;in promoting the formation of calcium nodules, ARS staining results showed that its intensity were water, chloroform, ethyl acetate, and n-butanol parts;oil red O staining results showed that inhibition intensity of fat cells interaction strength was formed from ethyl acetate, water, chloroform to n-butanol parts. Conclusion Extracts from different parts of JPHGF have different effects on BMSCs proliferation and differentiation. Water extraction has the strongest osteogenic differentiation and proliferation, and ethyl acetate has the best effect on the inhibition of cell formation.

Objective To evaluate the clinical application of the whole blood interferon γ(IFN-γ) release assay of QuantiFERON TB Gold in tube (QFT-GIT) in diagnosis of tuberculosis. Methods From October 2014 to October 2015, 109 patients with tuberculosis (45 cases of confirmed patients and 64 cases of clinically diagnosed patients) and 70 patients with non-tuberculosis were enrolled in Tianjin Medical University General Hospital. In order to evaluate diagnosis value between two kinds of tests, and to compare the differences between two groups, QFT-GIT test and colloidal gold anti tuberculosis antibody (TB-Ab) were employed to detect in two groups of patients. The ROC curve of IFN-γrelease quantity was analyzed in two groups. Results The sensitivity and specificity of QFT-GIT were 93.58% and 85.71% respectively. The positive rate was significantly higher in QFT-GIT than that of TB-Ab (χ2=43.68,P<0.01). The sensitivity of combined detection of the two methods decreased to 52.3% (57/109), but the specificity increased to 90.0% (63/70). The release quantity of IFN-γwas significantly higher in tuberculosis group than that in the non-tuberculosis group (U=330,P<0.05). The area under the ROC curve of IFN-γrelease quantity was 0.913 (95%CI:0.864-0.963). Conclusion The whole blood IFN-γrelease assay of QFT-GIT is a sensitive and specific assay for detecting tuberculosis infection. The combination QFT-GIT with TB-Ab can improve the specificity further, which could be a useful tool for the diagnosis of tuberculosis .

Objective To evaluate the changes in mRNA expressions of Toll-like receptor 4 (TLR4) and its down-stream cytokines IL-1β,IL-6 and TNF-a in incisional tissues from a rat with postoperative pain.Methods Incisional pain was induced in 74 male adult SD rats weighing 200-250 g.Paw mechanical withdrawal threshold (PMWT) around the wound on the operated and nonoperated sides was measured at 1 day before operation and at 0.5,1,2,6 and 12 hours as well as 1,2,3,5 and 7 days after operation.Skin incisional tissues were removed for determination of mRNA expressions of TLR4,IL-1β,IL-6 and TNF-a using real-time quantitative PCR at 1 day before operation and at 2 and 8 hours as well as 1,2,3,5 and 7 days after operation.Results Compared with the baseline value before operation,PMWT on the operated side was significantly decreased at 0.5 hours-5 days after operation,mRNA expression of TLR4 around the wounds on the operated side was down-regulated at 2 hours after operation followed by a gradual increase,mRNA expressions of IL-1β,IL-6 and TNF-α on the operative side were up-regulated at 2 and 8 hours as well as 1,2,3 and 5 days after operation (P ＜ 0.05),but no significant changes were found in PMWT and mRNA expressions of TLR4,IL-1β,IL-6 and TNF-α on the non-operated side(P ＞ 0.05).PMWT on the operated side was lowest at 6 hours after operation followed by the gradual increase,mRNA expression of TLR4 on the operated side peaked at 2 days after operation,and mRNA expressions of IL-1β,IL-6 and TNF-a respectively peaked at 2 hours,1 day and 3 days after operation (P ＜ 0.05).mRNA expressions of TLR4,IL-1β,IL-6 and TNF-a were negatively correlated with PMWT on the operative side (r =-0.501,-0.743,-0.893,-0.657,P ＜ 0.05),and mRNA expressions of IL-1 β,IL-6 and TNF-a were positively correlated with the level of TLR4 mRNA(r=0.764,0.283,0.667,P＜0.05).Conclusion mRNA expressions of TLR4 and its down-stream cytokines IL-1 β,IL-6 and TNF-a in skin incisional tissues are up-regulated,which may be involved in the development and maintenance of postoperative pain.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.