Objective To explore the efficacy and safety of tacrolimus among Chinese Takayasu arteritis (TAK) patients. Methods This was a single center, prospective study of active TAK patients treated with tacrolimus. Clinical manifestations, white blood cell count, hemoglobin level, erythrocyte sedimentation rate (ESR), hypersensitivity C reactive protein (hsCRP), alanine and aspartate aminotransferase and serum creatinine were recorded before and during tacrolimus treatment. Vascular changes were repeated every 6 months during tacrolimus treatment. All data were analyzed by statistical product and service solutions (SPSS) 20.0 statistical software, unpaired t test and Fisher exact probability and Kruskal-Wallis H test were used for statistical analysis. Results A total of 19 consecutive patients with an average age of (26 ±6) years were analyzed in this study. Sixteen of them were women. Pulselessness, fatigue, asymmetric blood pressure and fever were the most common clinical findings. Cervical and subclavian artery were more vulnerable. The most common artery involvement pattern was Numano type Ⅰ, followed by type Ⅱa and type Ⅴ. The median tacrolimus dosage was 2(2, 3) mg. Tacrolimus was effective in 9 out of the 19 patients. Patients who responded to tacrolimus tended to have lower mean ESR [(33±29) mm/1 h vs (42±20) mm/1 h, t=-0.776, P=0.448] and hsCRP [(20 ±31) mg/L vs (54 ±45) mg/L, t=-1.758, P=0.099] levels. However, no statistical significance was observed. During tacrolimus treatment, no drug related side effect was observed. Conclusion Tacrolimus is an alternative and effective therapy for some of the TAK patients.

Objective To observe the expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblast,and to study the role of fibronectin in pathogenesis of chronic fluorosis.Methods Male and female Wistar rats 144 were randomly divided into four groups,which were designated as the control group(normal diets,n =36),fluoride group(normal diets + 100 mg/L fluoride,n =36),lower calcium monophagia group (synthetic diets,n =36) and lower calcium monOphagia with fluoride group(synthetic diets + 100 mg/L fluoride,n =36).Rats were sacrificed 4 and 8 months after beginning of the experiment,respectively,and femur tissue was fixated and paraffin-embedded.The osteoblast isolated from calvaria of neonatal rats was treated with different dose of fluoride(0,1,2,4 mg/L fluoride,respectively) for 48 and 72 h,cell culture supernatant and cells were collected,respectively.The cranial osteoblasts were cultured in vitro and divided into four groups according to different concentration of fluoride added,which were 0(control group),0.01,1.00,and 10.00 mg/L groups.These cells were treated with mineralized induced medium at day 2 and cultured for 3 more weeks whereafter,and then the slides were fixed in alcohol.The expression of fibronectin in rat femur tissue was detected by immunohistochemistry (IHC),and fibronectin mRNA expression was determined by in situ hybridization; the fibronectin levels in supernatant of cultured osteoblast was examined by enzyme-linked immunosorbent assay(ELISA),and the expression of fibronectin mRNA in osteoblasts was detected with RT-PCR; skull mineralized nodule formation of osteoblasts was observed under a light microscopy after stained with 0.1％ red alizarin liquid.Results Little expression of fibronectin (brown granules under light microscope) could be seen in femur tissue of fluorosis rats of control group and lower calcium monophagia group; but abundantly expressed in fluoride group and lower calcium monophagia with fluoride group; the fibronectin was also expressed in osteoblasts,bone cells and bone marrow cells with less red particles in the control group and lower calcium monophagia group,but more in the fluoride group and lower calcium monophagia with fluoride group.The expression of fibronectin protein in supernatant of cultured osteoblasts was significantly increased in the group of 4 mg/L fluoride at 48 h(0.108 ± 0.042,t =0.764,P＜ 0.05) compared with control group(0.081 ± 0.010); the value was also significantly increased in 1,2,4 mg/L groups at 72 h(0.089 ± 0.010,0.087 ± 0.012,0.098 ± 0.023; t =0.765,0.704,0.996; all P ＜ 0.05) compared with control group (0.070 ± 0.014) ; the expression of fibronectin mRNA was much higher in 1,2,4 mg/L groups at 48 h (0.61 ±0.06,0.77 ± 0.07,0.77 ± 0.07) and 72 h(1.61 ± 0.14,2.54 ± 0.20,2.75 ± 0.22) compared with control group [0.48 ± 0.04(t =0.111,0.182,0.182,all P ＜ 0.05),0.97 ± 0.08(t =0.093,0.109,0.108,all P＜ 0.05) ].A lot of mineralized nodules could be seen under light microscope in 1.00 and 10.00 mg/L groups.Conclusions The expression of fibronectin in bone of fluorosis rats and in vitro cultured osteoblasts are increased,and fluoride also promotes the mineralization nodules formation of osteoblasts.These results suggest that fibronectin may regulate the process of bone mineralization,and possibly play a role in the development of skeletal fluorosis.

ObjectiveTo clarify the influence of human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein 120 V4 region with mutations at amino acid locuses on its abilities to enter target cells.Methods Based on the facts that ADA strains was a CCR5-tropic strain,only had the ability to infect CCR5 cells; that HXB2 strains was a CXCR4-tropic strain,only had the ability to infect CXCR4 cells,serial glycoprotein 120 mutants with alanine substitution in V4 region of ADA and HXB2 strains,were constructed by overlaping PCR.Eukaryotic expression vectors of mutants and expression vectors of HIV framework gene with luciferase reporter gene were cotransfected into eukaryotic cells to produce pseudoviruse.Concentration of HIV-1 gag P24 in pseudoviruses was detected by enzyme-linked immunosorbent assay(ELISA).U87.CD4.CCR5 and U87.CD4.CXCR4 cells were infected with 20 and 40 ng pseudoviruses,with wild ADA and HXB2 strains as control groups,respectively.The ability to infect cells of pseudovirus of each mutant with HIV-1 V4- region mutated at amine acid locuses 386-417 was measured by detecting the luciferase activity (relative light unit,RLU).ResultsTen mutants with alanine substitution in V4 region of HIV-1 ADA and HXB2 strains were successfully constructed,respectively.Mutants of pseudoviruse with 20 ng and 40 ng at locuses 389-391 and 414-417 with alanine substitution of V4 region in both ADA and HXB2 strains lost completely the abilities to enter CCR5 and CXCR4 expressing cells[ (0 ± 0)％].It was found that introduction of alanine to ADAs 400-403 and ADAs 408-410 increased the ability to infect cells to (124 ± 35)％,(182 ± 29)％ and (127 ± 8)％,( 134 ± 16)％ with pseudoviruse of 20 ng and 40 ng,respectively.Likewise,the ability to infect CXCR4 expressing cells also increased to (144 ± 42 )％ and (121 ± 18 )％ with pseudoviruse of 20 ng and 40 ng,respectively by introduction of alanine to HXB2s 395-397.However,other mutants in V4 region of ADA and HXB2 only maintained partial entry abilities( 15％- 84％).ConclusionsMutants of V4 region of HIV-1 envelope glycoprotein 120 with alanine substitution at locuses 389-391 and 414-417 in both ADA and HXB2 strains have been constructed successfully.They completely lost the ability to enter target cells.

Objective To study the mutations of Env sequence of SIVmac239 after infection of Chinese rhesus monkeys, and compare the differences and characteristics of Gp120 sequences of enterotropic and neurotropic SIV strains. Methods Six strains of simian immunodeficiency virus were analyzed in this study: four separated from peripheral blood mononuclear cells of SIVmac239-infected monkeys and two neurotropic SIVmac251 strains.Isolated and cultured monoclonal virus was obtained by limiting dilution assay.Gp120 sequences were amplified after the RNA extraction and phylogenetic analysis was processed thereafter.So did the Gp120 amino acid sequence and N-glycosylation sites analysis of the enterotropic and neurotropic strains.Results SIVmac239 had different mutations in four rhesus monkeys.The diversity in amino acid sequences of the enterotropic and neurotropic strains concentrated in the V1 and V4 regions of Gp120.The enterotropic strains had an addition of glycosylation site in V4 but the glycosylation site changes of neurotropic strains were located in the conservative regions of C1, C2 and C3.Conclusions The addition of one glycosylation site in V4 region of GP120 and loss of one glycosylation site in C1 region are associated with enhanced enterotropism and neurotropism.The differences between the enterotropic and neurotropic strains are not dipicted in Gp120 V3 region which is closely related with the tropism of strains.

Turkey is a presidential republic country located in the Eurasian continent, which has a universal health coverage since the health reform in 2003. The leading causes of death in this country are ischemic heart disease, stroke, and lung cancer. Besides, lower respiratory infections, chronic kidney disease, and hypertensive heart disease are the diseases which have a fastest growing rate. Chinese acupuncture was officially recognized by Turkey in 1991 with the promulgation of Acupuncture Treatment Legislation. At present, only trained practitioners and dentist could conduct acupuncture treatment, which was stated in Regulation of Tradition and Complementary Medicine Practice. The application of Turkish acupuncture and moxibustion is still applied in a simplified way that lack of TCM theory. Moreover, Chinese herbal medicine is still not officially recognized and still under control of the Ministry of Agriculture. Therefore, it is suggested to introduce TCM theory in the spread of acupuncture, to promote acupuncture research and clinical practice, to clarify the different standards between the two countries, and to cooperate in Chinese medicine researches, especially those related to the local high incidence and refractory disease so as to promote the development of TCM in Turkey and provide medical services for local residents.

Objective To study the relationship between the asthenia TCM syndromes in embryonic development stops and the level of zinc, iron, copper, magnesium, phosphorus, calcium in serum, and provide thoughts for assisting reproduction and preventing miscarriage. Methods Totally 100 patients of embryonic development stops were selected randomly (6-10 weeks gestation) to be the investigated group, with other 100 cases of live fetus as the control group. The contents of trace elements in serum were detected with atomic absorption spectrometry. Results The levels of trace elements in embryonic development stops patients were generally lower than the control group. The serum iron in patients with spleen deficiency syndrome, and serum zinc and iron in patients with kidney deficiency syndrome were significant lower (P<0.05). Conclusion The contents of trace elements in serum have relationships with asthenia TCM syndromes in embryonic development stops. It should be paid attention to supplementing trace elements during the gestation period. For patients with deficiency of kidney and spleen, the supplement of zinc and iron should be given greater prominence.

Objective To evaluate the changes of macrophages and expression of Rac1 in the inflammatory site of Crohn's disease,and to investigate the effects of 6-thioguanine (6-TG) and peptidoglycan on apoptosis of human peripheral blood monocyte-macrophage by regulating Rac1 signaling pathway.Methods Ten patients with Crohn's disease and eight healthy controls diagnosed were enrolled at Department of Gastroenterology and Hepatology,Tianjin Medical University General Hospital from January 2013 to January 2014.The number of macrophages,apoptosis and expression of Rac1 in the inflammation sites and non-inflammation sites of intestinal mucosa were detected in both patients and controls.Peripheral blood mononuclear cells (PBMCs) were sorted by CD14 immunomagnetic beads.The apoptosis of monocytes,expression of Rac1 and related apoptosis signaling molecules were detected in patients treated with peptidoglycan,6-TG and Rac1 inhibitor NSC23766 and another 15 healthy donors.Results The number of macrophages and apoptotic cells significantly increased in the inflammatory group of Crohn's disease patients compared with the non-inflammatory group.The expression of PAK1,downstream molecular of Rac1 signaling pathway of macrophages was also significantly higher in the inflammatory group of Crohn's disease patients than that in healthy controls and non-inflammatory group.Compared with control group,anti-apoptotic signals (NF-κB,Bcl-xL and STAT-3) in PBMCs increased in the peptidoglycan group,while slightly decreased in 6-TG group.6-TG and NSC23766 significantly promoted peptidoglycan-related anti-apoptosis [peptidoglycan group (8.6±3.7)％,peptidoglycan + 6-TG group (42.0±2.7)％,peptidoglycan + NSC23766 group (58.5±6.9)％,P＜0.05].Conclusions Peptidoglycan plays a role in the pathogenesis of Crohn's disease by recruiting macrophages.However,6-TG inhibits peptidoglycan-induced activation of Rac 1 signaling pathway leading to macrophage apoptosis.

Objective To investigate the effect of long?chain non?coding RNA Fez family zinc finger protein 1 antisense RNA1 ( lncRNA FEZF1?AS1) on the biological function of hepatocellular carcinoma (HCC). Methods SMMC771 and BEL?7402 cells were transfected with sh?FEZF1?AS1 and OE?FEZF1?AS1, respectively. The expression of lncRNA FEZF1?AS1 was detected by real?time quantitative PCR. Cell proliferation was detected by Cell Counting Kit?8 ( CCK?8), and apoptosis was detected by flow cytometry. The effects of lncRNA FEZF1?AS1 on invasion and migration were detected by Transwell and wound healing assays. The expression levels of adhesion molecules were detected by Western blot. The effect of lncRNA FEZF1?AS1 on the in vivo growth was verified by nude mice xenograft experiments. Results The silencing or ectopic expression of lncRNA FEZF1?AS1 inhibited or promoted the proliferation of hepatocellular carcinoma cells. CCK?8 assay showed that the proliferation abilities of SMMC7721 and BEL?7402 cells in sh?FEZF1?AS1 transfection group significantly decreased, achieving (35.43± 4.06)% and ( 34.68± 3.97)%, respectively, on the fifth day. There were significant differences between sh?FEZF1?AS1 group and sh?NC group [52.21 ± 8.46)% and (53.76 ± 7.64)%] ( all P＜0.05). In contrast, the proliferation ability of SMMC7721 and BEL?7402 cells transfected with OE?FEZF1?AS1 was significantly increased, achieving (83.49±6.92)% and (80.31 ± 3.13)%, respectively, on the fifth day. There were significant differences between OE?FEZF1?AS1 and OE?NC group [53.03 ± 8.84)% and ( 55.11 ± 7.09)%] ( all P＜0.05). The subsequent flow cytometry results showed that cell apoptotic rates of SMMC7721 and BEL?7402 cells transfected with sh?FEZF1?AS1 were ( 13.02 ± 1.38)% and ( 11.88 ± 1.29)%, respectively, which were significantly higher than those in sh?NC groups [(5.57±1.46)% and (8.06±1.42)%, respectively, all P＜0.05]. In contrast, the apoptotic rates of SMMC7721 and BEL?7402 cells transfected with OE?FEZF1?AS1 were (3.01 ± 0.39)% and ( 3.22 ± 0.43)%, which were significantly lower than those in OE?NC groups [(6.68±0.96)% and (6.63±0.45)%, all P＜0.05]. In addition, knockdown or overexpression of lncRNA FEZF1?AS1 expression inhibited or enhanced the migration and invasion abilities as well as the levels of adhesion molecules in hepatocellular carcinoma cells.After 30 days of feeding under the same conditions, the tumor volumes of sh?FEZF1?AS1 and sh?NC SMMC7721 cells xenograft mice models were (0.26±0.03) cm3 and (0.63±0.06) cm3, respectively, showing significant difference (P＜0.05). The tumor volumes of sh?FEZF1?AS1 and sh?NC BEL?7402 cells were (0.31±0.02) cm3 and (0.72±0.08) cm3, and the difference was statistically significant ( P＜0.05). Conclusion lncRNA FEZF1?AS1 may strengthen the growth, migration and invasion of hepatocellular carcinoma cells.

Objective To investigate the effect of long?chain non?coding RNA Fez family zinc finger protein 1 antisense RNA1 ( lncRNA FEZF1?AS1) on the biological function of hepatocellular carcinoma (HCC). Methods SMMC771 and BEL?7402 cells were transfected with sh?FEZF1?AS1 and OE?FEZF1?AS1, respectively. The expression of lncRNA FEZF1?AS1 was detected by real?time quantitative PCR. Cell proliferation was detected by Cell Counting Kit?8 ( CCK?8), and apoptosis was detected by flow cytometry. The effects of lncRNA FEZF1?AS1 on invasion and migration were detected by Transwell and wound healing assays. The expression levels of adhesion molecules were detected by Western blot. The effect of lncRNA FEZF1?AS1 on the in vivo growth was verified by nude mice xenograft experiments. Results The silencing or ectopic expression of lncRNA FEZF1?AS1 inhibited or promoted the proliferation of hepatocellular carcinoma cells. CCK?8 assay showed that the proliferation abilities of SMMC7721 and BEL?7402 cells in sh?FEZF1?AS1 transfection group significantly decreased, achieving (35.43± 4.06)% and ( 34.68± 3.97)%, respectively, on the fifth day. There were significant differences between sh?FEZF1?AS1 group and sh?NC group [52.21 ± 8.46)% and (53.76 ± 7.64)%] ( all P＜0.05). In contrast, the proliferation ability of SMMC7721 and BEL?7402 cells transfected with OE?FEZF1?AS1 was significantly increased, achieving (83.49±6.92)% and (80.31 ± 3.13)%, respectively, on the fifth day. There were significant differences between OE?FEZF1?AS1 and OE?NC group [53.03 ± 8.84)% and ( 55.11 ± 7.09)%] ( all P＜0.05). The subsequent flow cytometry results showed that cell apoptotic rates of SMMC7721 and BEL?7402 cells transfected with sh?FEZF1?AS1 were ( 13.02 ± 1.38)% and ( 11.88 ± 1.29)%, respectively, which were significantly higher than those in sh?NC groups [(5.57±1.46)% and (8.06±1.42)%, respectively, all P＜0.05]. In contrast, the apoptotic rates of SMMC7721 and BEL?7402 cells transfected with OE?FEZF1?AS1 were (3.01 ± 0.39)% and ( 3.22 ± 0.43)%, which were significantly lower than those in OE?NC groups [(6.68±0.96)% and (6.63±0.45)%, all P＜0.05]. In addition, knockdown or overexpression of lncRNA FEZF1?AS1 expression inhibited or enhanced the migration and invasion abilities as well as the levels of adhesion molecules in hepatocellular carcinoma cells.After 30 days of feeding under the same conditions, the tumor volumes of sh?FEZF1?AS1 and sh?NC SMMC7721 cells xenograft mice models were (0.26±0.03) cm3 and (0.63±0.06) cm3, respectively, showing significant difference (P＜0.05). The tumor volumes of sh?FEZF1?AS1 and sh?NC BEL?7402 cells were (0.31±0.02) cm3 and (0.72±0.08) cm3, and the difference was statistically significant ( P＜0.05). Conclusion lncRNA FEZF1?AS1 may strengthen the growth, migration and invasion of hepatocellular carcinoma cells.

Objective:To explore guidance effect of CYP2C19 gene polymorphism on anti-platelet therapy after per-cutaneous coronary intervention (PCI).Methods:A total of 105 patients with coronary heart disease,who received clopidogrel after PCI,were selected.According to results of CYP2C19 gene polymorphism detection,they were di-vided into normal metabolism group (n=51,fast metabolism type)and poor metabolism group (n=54,medium and slow metabolism type).Platelet inhibition rate induced by adenyl diphosphoric acid (ADP)measured by throm-boelastogram (TEG)and clopidogrel resistance (CR)rate were analyzed and compared between two groups.Re-sults:Normal metabolism group and poor metabolism group occupied 48.57% and 51.43% respectively among the 105 patients (medium metabolism type and slow metabolism type occupied 43.81% and 7.62% respectively in poor metabolism group).Compared with normal metabolism group,there was significant reduction in platelet inhibition rate induced by ADP [(78.14 ± 17.86)% vs.(41.67 ± 12.05 )%] and significant rise in incidence rate of CR (11.76% vs.59.26%)in poor metabolism group,P =0.001 all.Conclusion:CYP2C19 gene polymorphism is an important influencing factor for clopidogrel resistance.Therapeutic effect of clopidogrel after PCI in normal metab-olism group is significantly better than that of poor metabolism group,therefore,CYP2C19 gene polymorphism de-tection is help to guide anti-platelet therapy after PCI.