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 investigate the regulatory effect of vitamin D on the HMGB1/RAGE pathway and adipokine levels in a mouse model of obesity and asthma.Methods:This study was conducted at the Experimental Center of the Second Affiliated Hospital of Jiaxing University and the laboratory of Jiaxing University from February to September 2023. Thirty mice were marked with digital ear numbers and were randomly divided into five groups, with six mice in each group: Group Ⅰ (normal control group), Group Ⅱ (asthma group), Group Ⅲ (obesity and asthma group), Group Ⅳ (asthma + vitamin D group), and Group V (obesity and asthma + vitamin D group). An obesity mouse model was induced using a high-fat diet, while an asthma mouse model was induced through sensitization via intraperitoneal injection of ovalbumin and aerosol inhalation. The vitamin D intervention consisted of continuous intragastric administration of vitamin D (1 mL/d) for 2 weeks. Blood levels of interleukin-4, interleukin-6, interleukin-10, adiponectin, and leptin were determined using the enzyme-linked immunosorbent assay method. The expression of genes encoding high mobility group protein B1 (HMGB1) and the receptor for advanced glycation end products (RAGE) was detected using a quantitative reverse transcription polymerase chain reaction. All the obtained results were statistically analyzed using SPSS software.Results:The white blood cell count in bronchoalveolar lavage fluid (BALF) of Group Ⅱ was (1.34 ± 0.48) × 10 5/L, which was significantly lower than (4.07 ± 0.14) × 10 5/L in Group Ⅳ ( t = -18.28, P < 0.001). The white blood cell count in BALF in Group Ⅲ was (9.61 ± 0.91) × 10 5/L, which was significantly higher than (4.89 ± 0.38) × 10 5/L in Group V ( t = 11.75, P < 0.001). The percentage of eosinophils in BALF of Group II was (28.75 ± 1.94)%, which was significantly higher than (11.51 ± 1.99)% in Group Ⅳ ( t = 15.20, P < 0.001). The percentage of eosinophils in BALF of Group V was (12.50 ± 1.42)%, which was significantly lower than (29.80 ± 1.96)% in Group Ⅲ ( t = 17.74, P < 0.001). The ELISA results demonstrated that serum levels of interleukin-4, interleukin-6, interleukin-17, interleukin-1β, immunoglobulin E, and tumor necrosis factor-α in Group V were significantly lower than those in Group Ⅲ ( t = 15.24, 9.65, 2.26, 5.83, 10.86, 2.50, all P < 0.001). The serum level of interleukin-10 in Group Ⅲ was (4.97 ± 0.25) pg/mL, which was significantly lower than (8.84 ± 0.64) pg/mL in Group V ( t = -13.89, P < 0.001). The serum level of adiponectin in Group V was (1.95 ± 0.85) mg/L, which was significantly higher than (1.15 ± 0.13) mg/L in Group Ⅲ ( t = -12.67, P < 0.001). The HMGB1 expression level in lung tissue of Group Ⅳ was 1.42 ± 0.09, which was significantly lower than 1.91 ± 0.16 in Group Ⅱ ( t = 6.55, P < 0.001). The expression level of RAGE mRNA in lung tissue in Group Ⅳ was 1.35 ± 0.11, which was significantly lower than 1.55 ± 0.152 in Group Ⅱ ( t = 4.19, P < 0.05). The expression level of HMGB1 in lung tissue in Group V was 1.51 ± 0.10, which was significantly lower than 2.44 ± 0.10 in Group Ⅲ ( t = 1.02, P < 0.001). Conclusion:Vitamin D may alleviate lung injury by up-regulating the expression of HMGB1 and RAGE in a mouse model of obesity and asthma. This provides a new concept and method for the treatment and prevention of obesity and asthma.