Intrahepatic cholangiocarcinomas is a type of biliary epithelial cell carcinoma.Hepatolithiasis,viral hepatitis,primary sclerosing cholangitis,Caroli disease,parasitic infections are its risk factors.In recent years,the incidence of intrahepatic cholangiocarcinomas is gradually increased.Ultrasound,CT,MRI can be used to diagnose.The main way to treat intrahepatic cholangiocarcinomas is comprehensive treatment based on liver resection.There is still some controversy in liver transplantation.Adjuvant therapy will play a more and more important role in patients who unable to accept surgery.

Gallstone ileus is a rare complication of cholelithiasis,and is common in older gallstones patients.It lacks the typical clinical presentation,so preoperative diagnosis is very difficult.The best diagnostic modality now is abdominal CT scan.Surgery is the most important and useful treatment.According to patient' s own situation,choose individualized surgical approach.Endoscopic,laparoscopic and other minimally invasive treatment are promising.With the development of related technologies,it maybe increased of the gollstone ileus treatment.

To obtain fusion protein of Mycobacterium bovis with high purity, the recombinant prokaryotic expression vector for Mb2277 gene was constructed and the immunogenicity of its products was initially investigated in the present study.A pair of primer was designed according the gene sequence Mb2277 from the genomic DNA of M.Bovis in GenBank. and was amplified by PCR using DNA of M.Bovis 93006 strain as template. The PCR product and pET-28a(+) was then digested by BamHⅠ and EcoR Ⅰdouble enzyme. To constructed a prokaryotic expression plasmid, the purified Mb2277 was cloned to pET28a(+). Then the recombinant plasmid was transformed into competent cell of E.coli BL21(DE3).The bacteria were induced by IPTG and its lysates were analyzed by SDS-PAGE and Western-blotting. In this way, the prokaryotic expression plasmid for M. bovis Mb 2277 protin was obtained, and a expression band with molecular of 25 ku could be found in SDS-PAGE analysis. As demonstrated by Western blotting this expression product showed excellent reactivity with rabbit immune sera against M. bovis.

Objective To investigate the effects of methylthioadenosine phosphorylase (MTAP) on invasion and migration in breast cancer cells.Methods Human breast cancer cell line MCF-7 cells were treated with MTAPtargeted siRNA to diminish MTAP mRNA.MCF-7 cells proliferation was evaluated by cell counting kit-8,the analysis of cells invasion and migration was performed using Transwell chamber.The expressions of MTAP and matrix metalloproteinase 1 (MMP1) in cell extracts were detected by Western blotting.The experimental divided into blank contrd group,negative control group,MTAP-siRNA experimental group.Results The MCF-7 cells growth was promoted after knockdown the MTAP mRNA.MATP-siRNA experimental group 450 nm absorbance values at 24h,48 hand72 h of the control group were (112.3±11.9)％,(144.4±8.4)％,(169.3±9.4)％ respectively.Cell invasion analysis by Transwell chamber showed 570 nm absorbance values were 0.49 ± 0.06 (control),0.45 ± 0.07 (negative control) and 0.87 ± 0.07 (MTAP-siRNA) respectively.Cell migration analysis by Transwell chamber showed 570 nm absorbance values were 0.46 ± 0.06 (control),0.49 ± 0.08 (negative control)and 0.75 ± 0.07 (MTAP-siRNA) respectively.The expression of MMP1 in MCF-7 cells was upregulated after knockdown the MTAP mRNA.Conclusion The knockdown of MTAP in MCF-7 cell can increase the cells invasion and migration,and this may involve the the MMP1.

This paper reviews the development of appropriate health technology policy from 1992 to 2012 in Zhejiang province.The evolvement of recent twenty years is classified into several stages and each is analysed and evaluated.This study provides reference for the establishment of appropriate health technology policy and the transformation of science and technology policy across the country.

Objective To investigate clinical significance of diagnosis of the anti saccharomyces cerevisiae antibody (ASCA), pancreatic acini antibody (PAB)resistance,resistance to small goblet cell antibody (GAB),anti neutrophil cytoplasm anti-bodies (ANCA)for inflammatory bowel disease (IBD)and the differential diagnosis of ulcerative colitis (UC)and crohn's disease (CD).Methods Collected 200 cases of test group sets of inflammatory bowel disease,serum by indirect immunofluo-rescence (IF).Results In the serum of 200 patients,106 cases with positive or weakly positive.Among them,the positive ASCA/weak positive 24 cases,14 cases of PAB,GAB 63 cases,28 cases ANCA,and included in ASCA group respectively and PAB group,GAB group,ANCA group.Positive rate of ANCA and GAB in the diagnosis of UC were 34% and 58%. Positive rate of ASCA and PAB in the diagnosis of CD were 28.6% and 38.1%.ANCA associated with GAB detection in the diagnosis of UC specific degree was 60%,ASCA associated with PAB detection in the diagnosis of CD specific degree was 75%.Conclusion Serum inflammatory bowel disease antibody spectrum in ASCA,ANCA,GBA and PAB four antibod-ies of joint detection has important guiding value to the diagnosis of IBD,also can be used as one of UC and CD in the differ-ential diagnosis methods.

Objective To establish a human HepG2 cell growth model under the low oxygen environment induced by cobalt chloride in order to observe the impacts of human HepG2 cell proliferation,cellular cycles and apoptosis,namely cellular growth conditions,under the low oxygen environment induced by cobalt chloride with different concentrations and to study the HepG2 cell growth mechanism under the low oxygen environment induced by cobalt chloride.Methods The human HepG2 cells in the logarithmic phase were randomized grouping as control group and CoCl2 experimental group with different concentrations (50 μm/L,100 μm/L,150 μm/L and 200 μm/L).① HepG2 cell proliferation was tested by MTT assay to calculate cell's suppression rate and draw HepG2 cell growth inhibition curves.② The move ability of HepG2 cells was observed by scratch test.③ The cellular apoptosis and periodic changes were detected using the flow cytometer Annexin-V FITC/PI double-staining and PI single staining methods.④HepG2 cell's Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and Bcl-2 protein expression were detected by Western Blot.Results ① The test results obtained via MTT assay showed that CoCl2 suppressed the human HepG2 cell proliferation within a certain amount of time and concentration and presented a time-dose dependent relation.② Scratch damage trial suggested that the cobalt chloride suppressed the HepG2 cell migration and wound repair capacity and presented a concentration dependent relation.③ Flow cytometer' s test results revealed that the apoptosis rates (％) in control group and experimental group with different concentrations (50 μm/L,100 μm/L,150 μm/L and 200 μm/L) were 3.42,7.74,13.07,20.56,28.53 and 44.45 (P ＜0.05),respectively.The apoptosis rate of the experimental group was significantly increased compared with the control group,as well as showing a concentration dependency.The results of cellular cycles revealed that the cobalt chloride significantly suppressed the HepG2 cell's periodic changes along with increases of concentration,as well as blocked the cell cycle staying in phase G1,thereby suppressing cell proliferation.④Western Blot test:Compared with the control group,the Bcl-2 protein expression was significantly decreased in the experimental group after treatment of cobalt chloride with different concentrations.Conclusion Within a certain range,CoC12-indiuced low oxygen environment can suppress the human HepG2 cell proliferation and healing migration capacity,induce apoptosis and present a time-dose dependent relation.The mechanism is likely associated with decreases of Bcl-2 protein expression.

Objective To compare the results of anti-cardiolipin (ACA)measuring by two commercial available kits.Methods ACA in total of 66 serum samples were both determined by kits from Euroimmun and YHLO simultaneously,then the re-sults were analyzed comparatively and correlatively.The Euroimmun kit was applied to determination the level of ACA-IgA/G/M,and the YHLO kit determined ACA-IgG/M and anti-β2-glycoprotein I antibody (β2GPI IgG).Results The positive rate by Euroimmun kit was 37.88% (25/66),while 31.82% (21/66)was positive (one positive among ACA-IgG,IgM andβ2GPI IgG)when determined by YHLO kit,and there was no significant difference between the two kits.The accordance rate of the two kits was 87.88% (58/66).The ACA-IgA/G/M value by Euroimmun kit and the summation of ACA-IgG, IgM andβ2GPI IgG by YHLO kit showed well linear correlation (r 2 =0.892,P <0.01).Conclusion Results from the two kits were consistent and correlated well,and they are suitable for the clinical application;these two kits have their own char-acteristics,which could be used by individual or combination accordingly.

BACKGROUND:Procolagen type 1 N-terminal propeptide (P1NP) and β-colagen special sequence(β-CrossLaps) are two bone metabolic markers that are closely related to osteoporosis. Combined detection of bone metabolic markers and bone mineral density is of clinical significance for the diagnosis of osteoporosis. Bone metabolic markers are ideal indicators to predict fractures, which can compensate for the lack of bone density test. OBJECTIVE:To introduce the application of bone metabolic markers in the monitoring of drug efficacy on the treatment of osteoporosis as wel as in the prediction of fracture risks in recent 20 years and to explore the clinical values of P1NP and β-CrossLaps to assess the therapeutic efficacy on osteoporosis and risks for osteoporotic fractures. METHODS:A computer-based search of CNKI and SCI databases were performed for relevant articles published from 2000 to 2014 using the keywords of “serum bone metabolic markers; osteoporosis; bone mineral density” in Chinese and English, respectively. Finaly, 44 articles meeting the inclusive criteria were reviewed. RESULTS AND CONCLUSION:This paper analyzes the source and detection mechanisms of P1NPand β-CrossLaps and then compares their advantages in the therapeutic effect assessment of osteoporosis. Serum bone metabolic markers cannot only reflect the dynamic changes of bone metabolism, but also have earlier changes than the bone mineral density. Both P1NPand β-CrossLaps are very important for assessing the early diagnosis of osteoporosis as wel as anti-osteoporosis drug efficacy.

Objective:To explore the effect of bismuth containing triple therapy on serum gastrin (Gas), transforming growth factor-α (TGF-α), and high-sensitivity C-reactive protein (hs-CRP) levels in children with Helicobacter pylori (Hp) positive peptic ulcers.Methods:A total of 96 children with Hp positive peptic ulcers admitted to the Second Hospital of Jiaxing from January 2020 to December 2021 were selected as the study subjects. They were randomly divided into two groups using the remainder of a random number table. The control group (48 cases) received treatment with omeprazole, clarithromycin, and amoxicillin, while the observation group (48 cases) received treatment with bismuth containing triple therapy (amoxicillin+ metronidazole+ bismuth potassium citrate). After 10 days of treatment, the clinical efficacy of the two groups was evaluated The improvement time of clinical symptoms, Hp conversion rate, serum indicators (Gas, TGF-α, hs-CRP) before and after treatment, and incidence of adverse reactions were observed.Results:The total effective rate and Hp conversion rate of the observation group were significantly higher than those of the control group [95.83%(46/48) vs 81.25%(39/48), 97.92%(47/48) vs 83.33%(40/48), P<0.05]. The improvement time of upper abdominal pain, heartburn, and acid reflux symptoms was significantly shorter than that of the control group (all P<0.05). After 10 days of treatment, the serum Gas and hs-CRP levels in both groups significantly decreased compared to before treatment (all P<0.05), and the observation group was lower than the control group after treatment ( P<0.05); The levels of TGF-α in both groups increased compared to before treatment (all P<0.05), and the observation group was higher than the control group after treatment ( P<0.05). There was no statistically significant difference in the incidence of adverse reactions between the two groups [4.17%(2/48) vs 2.08%(1/48), P>0.05]. Conclusions:The triple therapy with bismuth containing agents has a better therapeutic effect on children with Hp positive peptic ulcers, and can promote ulcer mucosal repair by improving inflammatory response, with good safety.