Objective To assess the characteristics of biochcmical parameters of bone metabolism in patients with inflammatory bowel disease (IBD) and the relationship between osteoporosis and the reduction of bone mass so as to identify the risk factors for osteoporosis in IBD. Methods Sixty-nine patients with IBD[49 with Crohn disease (CD) and 20 with ulcerative colitis (UC)] and 20 sex- and age-matched healthy subjects (control group) from the medical examination centre in 2007 were enrolled in the study. The disease activity was estimated according to CD active index and Truelove-Witts score. The biochemical markers of bone metabolism including ostecalcin (OC), 25-bydroxycholecalciferol[-25 (OH)Ds-] and cross-linked N-telopeptides of type I collagen(NTX) were tested using enzyme immunoassay. The body mass index (BMI) and bone mineral density (BMD) were also measured. Results The concentration of 25(OH)D3 was significantly lower in both CD[(44.45±39.38) nmol/L] and UC patients[(34.67±23.79) nmol/L] compared with controls[(98.42±25.84) nmol/L] (P <0.05), while NTX level was significantly higher in both CD [(58.41±15.15) nmol BCE/mmol Cr] and UC[(57. 67±10. 75) nmol BCE/mmol Cr] patients compared with controls[(30. 38±13.35) nmol BCE/mmol Cr] (P<0.05). The levels of OC and 25 (OH)D3 in patients treated with steroids[(17. 3±13. 43) ng/ml and (26. 99 ± 9. 12) nmol/L, respectively] were lower than those in patients untreated with steroids[(27.33 ± 16.86) ng/ml and (45.33±39.03) nmol/L,respectively] . BMD examination revealed that the incidence of osteoporosis in CD or UC groups were 14.3%(7/49) or 3/20,respectively, with no difference between two groups (P>0. 05). The T score of lumbar spine in patients treated with steroids (-1.19±0. 93) was significantly lower than that in patients untreated with steroids (-0.80±1.29) (P<0.05), but there was no difference in T score of femoral between two groups (P>0.05). Stepwise regression analysis revealed that advanced age and reduced BMI were risk factors for osteoporosis in CD patients.Conclusions The lower BMI in patients with IBS contributes to prevalence of osteoporosis, and advanced age and reduced BMI are risk factors for osteoporosis in CD patients. NTX and 25 (OH)D3are important bone metabolic markers to predict osteoporosis in patients with IBD.

Objective To study the cause,clinical characteristics and treatment of HFmrEF,HFrEF and HFpEF patients.Methods Three hundred and eighty-five heart failure (HF) patients aged ≥60 years admitted to our hospital from January 2016 to March 2017 were divided into HFrEF group (n=96),HFmrEF group (n=34) and HFpEF group (n=255) according to their ejection fraction.Their demographic data,HF cause,clinical characteristics,cardiac ultrasonographic data,laboratory testing data and therapies were recorded.Their clinical characteristics were compared.Results The number of males was greater and the cardiac function grade Ⅳ was higher in HFmrEF group than in HFrEF and HFpEF groups.The incidence of hypertension was the highest followed by that of valvular disease.The number of HFmrEF patients who used intravenous nitrates,spironolactone and milrinone was greater than that of HFpEF and HFrEF patients who used intravenous nitrates,spironolactone and milrinone.The serum creatinine level was higher in HFmrEF patients than in HFpEF and HFrEF patients at the time when they were discharged.Conclusion Hypertension and valvular disease are the mian risk factors for HFmrEF.The number of males is greater and the cardiac function grade Ⅳ is higher in HFmrEF patients than in HFrEF and HFpEF patients.The serum creatinine level is higher and the outcome is better in HFmrEF patients than in HFrEF and HFpEF patients at the time when they are discharged.

AIM To investigate weather and how aminophylline induces human trachea smooth muscle cells (TSMCs) to apoptosis. METHODS Human TSMCs were isolated and cultured in DMEM containing 10% fetal bovine serum. Passage 4～6 cell were used in experiment. The cells were cultured with aminophylline or 8 Br cAMP for 24 or 48 h. Light micros copy and electron microscopy were used to observe morphological change. DNA fragmentation was analyzed by agarose gels electrophoresis. SP Immunohistological staing method was performed to detect the changes of expressions of p53,bcl 2 and bax gene. The apoptosis cell percentage were detected by situ end labeling technique(TUNEL) of fragmental DNA. RESULTS ①Aminophylline or 8 Br cAMP decreased the number of viable cells in time and concentration dependent manner; ② Electron microscopic examination showed nuclear contraction, chromatin condensation and apoptotic bodies formation in aminophylline group or 8 Br cAMP group; ③ Agarose gel electrophoresis of fragmented DNA showed a ladder like pattern; ④ The expression of p53 or bax gene in apoptosis group was significantly higher than in control group, but the expression of bcl 2 gene was lower than in control group; ⑤ The positive rate of TUNEL in aminophylline or 8 Br cAMP group was significantly higher than in control group ( P

AIM:To clarify if interferon-?(IFN-?), tumor necrosis factor-?(TNF-?)and interleukin-1?(IL-1?)can induce apoptosis of human airway smooth muscle cells (ASMCs) in vitro. METHODS: Human ASMCs were isolated and cultured in DMEM containing 10% fetal bovine serum. Passage 4-6 cell was used in the experiment. IFN-?,TNF-? and IL-1?, were used separately or together in the treatment of human ASMCs. The effects of IFN-?,TNF-? and IL-1? on the growth of the cells was detected by MTT method at the hour 0,24,48 and 72. Light microscopy and electron microscopy were used to examine the morphological change. DNA fragmentation was analyzed by agarose gel electrophoresis. SP immunohistological staing method was performed to detect the change of expressions of p53 , bcl-2 and bax gene. The apoptosis cell percentage were detected by in situ end labeling technique (TUNEL)of fragmental DNA. RESULTS: (1)IFN-? or IFN-? together with TNF-? and IL-1? decreased the number of viable cells in a time dependent manner. (2) Light and electron microscopic examination showed cell shrinkage, membrane blebbing, nuclear contraction, chromatin condensation and nuclear fragmentation in human ASMCs. (3) Agarose gel electrophoresis showed a characteristic"ladder"of DNA bands representing integer multiples of the internucleosomal fragments (about 180-200 bp) in cytokine cotreated human ASMCs. (4)The expression of p53 and bax gene in cytokine cotreated group was significantly higher than in control group, but the expression of bcl-2 gene was lower than in control group. (5)Stimultaneous treatment with IFN-?(4?10 5 U/L),TNF-?(4?10 5 U/L)and /or IL-1? (10?10 4 U/L) induced apoptosis of human ASMCs. Apoptotic index of human ASMCs in cytokine co-treated group was significantly higher than in control group ( P

Objective To investigate the influence of benzopyrene on extracellular matrix(ECM) protein deposition of human airway smooth muscle cells(HASMC) and the related pathway mechanism.Methods HASMC were primarily cultured and the 2-6 generations cells were applied in this experiment.The expression amount of ECM gene and protein was detected by real time PCR and Western Blot the phosphorylation level was analyzed by using the Western Blot method.Results Benzopyrene could to increase the expression of HASMC collagen Ⅰ α1 protein (P＜0.01) and ECM protein (including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA(P＜0.05).Benzopyrene could induce the rapid increase of ERK1/2 phosphorylation level (P＜0.01).Furthermore,the ERK pathway inhibitor PD98059 could significantly inhibit the increase of benzopyrene-induced collagen Ⅰ α1 (P＜0.01)and ECM protein(including collagen Ⅰ α1,versican,fibronectin,laminin α2) mRNA expression(P＜0.01).Conclusion Benzopyrene induces the ECM protein deposition of HASMC by activating the ERK1/2 pathway,blocking the ERK1/2 signal pathway can inhibit the benzopyrene-induced airway remodeling.

Objective To explore the mechanism of notch signaling pathway in the treatment of cervical cancer with notch receptor blocker(DAPT).Methods After treatment of Me180 cells with different concentrations of DAPT combined with curcumin-photodynamic therapy (PDT),3-(4,5 Dimethylthiazlo-yl)-2,5 Dimethylthiazol-2-yl)-2,5 dipheny tetrazo lium bromide(MTT)assay was used to detect the effect on the proliferation of Me180 cells,annexin V-FITC/PI combined with flow cytometry was used to detect the effect on apoptosis,and real-time fluorescence quantitative PCR and Western blot were used to detect the mRNA and protein expressions of Notch1 gene respectively.Results Both DAPT(1.0 μmol/L)and curcumin-PDT groups could inhibit the proliferation and induce apoptosis of cervical cancer cell line Me180,and the synergistic effect of the two groups was significant (P＜0.05 or＜0.01).DAPT and curcumin-PDT group reduced the mRNA expression level of Notch1 in cervical cancer Me180 cells with inhibition ratio of 40.0％ and 32.3％ respectively.DAPT and curcumin-PDT group inhibited the protein expressions of Notch1,nuclear factor-kappa B(NF-kB),vascular endothelial growth factor (VEGF),and the synergistic effect of the two groups was statistically significant.Conclusions Both DAPT and curcumin-PDT may effectively block the conduction of Notch signaling pathway,which is associated with down-regulation of the expression of Notch1 and NF-kappa B.Notch signaling pathway may be one of the targets of curcumin photodynamic therapy.

Tanshinones are the bioactive components of the Chinese medicinal herb Salvia miltiorrhiza, while its biosynthetic pathway remains to be characterized. Rapid identification and characterization of the genes correlated to tanshinones biosynthesis is very important. As one of the intermediates of tanshinones biosynthesis, the ferruginol content is relative low in both root and engineered bacteria. It is urgent to construct an efficient system for conversion of miltiradiene to ferruginol to obtain large amount of ferruginol as the substrates for further identifying other downstream genes involved in tanshinones biosynthesis. In this study, we constructed the whole-cell yeast biocatalysts co-expressing miltiradiene oxidase CYP76AH1 and cytochrome P450 reductases (SmCPR1) from Salvia miltiorrhiza, and then characterized it with RT-PCR. After permeabilization, the yeast whole-cell could catalyze turnover of miltiradiene to ferruginol efficiently through single-step biotransformation with a conversion efficiency up to 69.9%. The yeast whole-cell biocatalyst described here not only provide an efficient platform for producing ferruginol in recombinant yeast but also an alternative strategy for identifying other CYP genes involved in tanshinones biosynthesis.

Objective To observe the clinical effects of external application of Chinese medicine ion introduction through acupiont in the prevention and treatment of hyperamylasemia and pancreatitis following endoscopic retrograde cholangio pancreatography (ERCP). Methods Three hundred subjects were divided into treatment group and control group randomly, and treated with routine therapy after surgery, moreover, treatment group was treated with external application of Chinese medicine ion introduction through acupiont. The incidence of hyperamylasemia and pancreatitis, the time of serum amylase returned to normal in patients with hyperamylasemia and pancreatitis was observed, and the abdominal pain after ERCP was scored. Results Excluding 17 cases according to the exclusion criteria, the treatment group included 142 cases and the control group included 141 cases. The hyperamylasemia and pancreatitis happened less frequently in the treatment group than in control group, but there was no significant difference (P>0.05). The time of serum amylase returned to normal in patients with pancreatitis was (4.25±0.95)d in treatment group and (5.28±1.11)d in control group, with no significant difference (P>0.05). The time of serum amylase returned to normal in patients with hyperamylasemia was (2.88 ± 0.78)d in treatment group and (3.81±1.62)d in control group, showed a significant difference (P<0.05). The score of abdominal pain was 0.95±1.04 in treatment group and 1.21±1.12 in control group, showed a significant difference (P<0.05). Conclusion External application of Chinese medicine ion introduction through acupoint can promote the recovery of the patients postoperative ERCP combined with hyperamylasemia or pancreatitis, and can better alleviate abdominal pain.

Objective To explore the protection of early autophagy activation on podocyte injury induced by aldosterone.Methods In vitro cultured mouse podocyte clones (MPC5) were treated with aldosterone for 6,12,24,48 h respectively.Apoptosis of podocytes was detected by Annexin V combined with flow cytometry.After 24 h treatment with aldosterone,the existence of apoptotic body and autophagosome was observed by electron microscopy.The protein expressions of LC3,caspase-3 and nephrin were examined by Western blotting.The mRNA expression of Beclin-l was detected by real-time PCR.Results The induction of apoptosis and autophagy by aldosterone in podocytes was in timedependent mannner.After 24 h treatment with aldosterone,the apoptosis was increased by 26.5％ (P ＜ 0.05)and the expression of nephrin was decreased by 28.0％ (P ＜ 0.05) compared to control group.Aldosterone remarkably induced the expression of Beclin-1 at 6 h and promoted the transformation of LC3-Ⅰ to LC3-Ⅱat 12 h (P ＜ 0.05).Compared to simple aldosterone treatment,the apoptosis rate of podocyte was increased by 39.0％ (P ＜ 0.05) and the expression of nephrin was declined by 19.5％ (P ＜ 0.05) after 3-methyladenine (3-MA) pre-treatment.Conclusions Aldosterone can induce autophagy and apoptosis in podocytes.Autophagy occurs earlier (12 h) than apoptosis (24 h).The occurrence of autophagy can inhibit the apoptosis,so the autophagy pathway may be a new research topic of glomerular disease treatment.

Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.