Pancreatic adenosquamous carcinoma is a rare exocrine pancreatic cancer,and surgical treatment is the only promising method in survival time extension and life quality enhancement.In July 2013,1 patient with pancreatic adenosquamous carcinoma and sarcomatoid change was admitted to the Shaoxing People's Hospital.A cauda pancreatis cystic nidus was detected by computed tomography 1 year ago,and no any other clinical symptoms were detected.The left upper quadrant of the patient was glutted and unwell,and the levels of CA19-9 and CA125 were 3 630 kU/L and 645 kU/L,respectively.The results of computed tomography and magnetic resonance imaging showed that a huge cystic and solid tumor was located between the cauda pancreatis,the stomach and the spleen;the tumor was mainly cystic and rooted in the tail of the pancreas;the spleen,stomach and surrounding tissues were infiltrated by the tumor.The results of postoperative pathological examination showed that the tumor was pancreatic adenocarcinoma with squamous epithelial differentiation and sarcomatoid change;the invasive squamous carcinoma dominated stomach wall,and the sarcomatoid change dominated spleen and lymph nodes with partial squamous epithelial differentiation and surrounding lymph nodes metastases.Although the surgical resection was complete and the postoperative thymosin injection and traditional Chinese medicine were implemented,tumor recurred at postoperative month 5,and the patient was dead due to cachexia and pernicious anemia at postoperative month 8.The prognosis of patients with pancreatic adenosquamous carcinoma is poor,early detection and surgery are key points for curative effect enhancement.

Objective To investigate the correlation between the frequency of myeloid-derived suppressor cells (MDSC) and the frequency of regulatory T cells (Treg) in the peripheral blood in patients with chronic hepatitis B (CHB) and its clinical significance.Methods A total of 45 CHB patients including 23 mild-to-moderate CHB patients,22 severe CHB patients,and 15 healthy controls were enrolled.The frequencies of MDSC and Treg in the peripheral blood were studied using flow cytometry and its correlation with clinical data was analyzed by Sepearman correlation analysis.Results The median frequency of MDSC in CHB patients was 0.414％,which was significantly higher than that in healthy controls 0.226％ (Z=-2.356,P=0.018 9).The frequency of MDSC in CHB patients was negatively correlated with the level of alanine transaminase (ALT) and aspartate transaminase (AST) (r=-0.480,-0.478; both P＜0.01),but had no relations with hepatitis B virus (HBV) viral load (r=-0.049,P=0.75).An increase frequency of MDSC was observed in CHB patients with an ALT of 5 × upper limits of normal (ULN) or less or AST of 3 × ULN or less.The frequency of MDSC in CHB patients was positively correlated with that of Treg (r =0.345,P =0.02).Conclusions The activation and proliferation of MDSC may facilitate and maintain HBV persistent infection.The change of the frequency of MDSC is in line with that of Treg,indicating that immunosuppressive functions of MDSC may be related with the development of Treg in CHB.

Objective:To develop a sensitive,rapid,simple gold immunochromatography assay for human cardiac troponin I(cTnI).Methods:The cTnI was used to immunize Balb/c mice by route method.The spenocytes were fused with SP2/0 myeloma cell lines.Six hybridoma cell lines,secreting stably McAbs to cTnI antigen have been obtained and screened.The 3F10 was coupled with colloidal golds,the 3A7 biotinylated.Streptavidin was coupled with the nitrocellose strip to prepare an test apparatu.According to red line in nitrocellose,thus can analysis the results.Results:The sensitivity of cTnI measured by this apparatus was 0.4 ng/ml.The 30 patients with acute myocardial infarction(AMI)were detected for cTnI by this apparatus and by the product of PBM Co USA as standard.The coincidence rate of both two assays was 93.6%. Conclusion:This method is a sensitive,specific simple and rapid assay.

Objective To investigate the risk factors of hepatitis B virus(HBV) re-infection after orthotopic liver transplantation(OLT)and to evaluate the therapeutic efficacy of hepatitis B immunoglobulin(HBIG)combined with nucleos(t)ide analogues. Methods The study included 160 patients with HBVrelated liver diseases who underwent OLT in the Third Affiliated Hospital of Sun Yat-sen University from October 2003 to Augest 2007, 117 of whom were treated with nucleos(t)ide analogues before OLT;and all patients were received HBIG i. m and nucleos(t)ide analogues treatment after OLT. Preoperative data of the patients were retrospectively reviewed, and HBV re-infection was assessed prospectively. Independent t test was used to compare normally distributed data and Fisher's exact test was used for the comparison of rates among groups. Results HBV re-infection Was observed in 19 patients after OLT with a rate of 11. 88%(19/160), which was not correlated with HBV DNA loads, HBeAg and the duration of antiviral therapy before OLT(r=0.108, 0.127 and 0.033, P>0.05). Of 19 patients with HBV re-infection, 17 were treated with lamivudine after OLT, and HBV YMDD mutants were detected in 8. The YMDD positive group had a higher HBV DNA level than YMDD negative group(7.0 ± 2.0 log copies/mL vs 3.2 ± 2.5 log copies/mL, t = 3.531, P=0.003). Among above 17 patients, 12 received adefovir add-on treatment, and3 received entecavir instead of lamivadine; all achieved satisfactory responses. Conclusions Low dose of HBIG combined with long-term use of nucleos(t)ide analogues can effectively prevent HBV re-infection after OLT. HBV YMDD mutation may be the primary reason for HBV re-infection in the patients treated with lamivudine after OLT.

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.

Objective To investigate the feasibility and technique in laparoscopic cholecystectomy and exploration of common bile duct (CBD) in patients who have previous subtotal gastrectomy.Methods From January 2001 to October 2010,22 patients who had a history of subtotal gastrectomy received laparoscopic cholecystectomy and CBD exploration in our hospital.In addition,these patients received cholangioscopic lithotomy,electrohydraulic lithotripsy,T-tube drainage,or primary suturing of the CBD wound.Results The procedures were completed successfully in 18 patients.In 4 patients,conversion to laparotomy was necessary because of multiple stones in a stenosed CBD (n=1),severely congested CBD wall with edema (n=1),CBD carcinoma associated with impacted stones (n=1) or Mirzzi syndrome (n =1).T tube drainage was employed in 16 patients,and primary suturing in 2 patients at the end of the operation.There was no hemorrhage,biliary leakage or abdominal infection.At a mean follow-up of 13 (range 3 to 60) months there was no residual stone,biliary stenosis or cholangitis.In 3 patients Stone recurred,and another 3 patients were lost to follow-up.Conclusions A history of subtotal gastrectomy is not a contradiction to laparoscopic cholecystectomy and CBD exploration.With choledocoscopy,surgery can be safe and feasible even for patients with previous gastectomy.

Aim To explore the role of endoplasmic reticulum stress(ERS) in Astragaloside Ⅳ-induced cardioprotection in H9c2 cardiac cells, and to explore the potential mitochondrial mechanism.Methods Conventional culture was performed of rat heart tissue-derived H9c2 cells.Experiment was randomly divided into the control group, the ERS inducer 2-Deoxy-D-Glucose(2-DG) group, Astragaloside Ⅳ and 2-DG combination group, Astragaloside Ⅳ group.Confocal microscopy was used to observe the changes of TMRE fluorescence intensity so as to confirm the influence of ERS on the mitochondrial potential, and further speculate on the opening of the mitochondrial permeability transition pore(mPTP).Western blot analysis was applied to detect the expressions of ERS proteins GRP 78, GRP 94 and IRE1.Transmission electron microscopy was used to observe the ultrastructure of the cells.Results Different doses of 2-DG could mimic the mPTP opener atractyloside to induce the mPTP opening with the peak at 100 μmol·L-1;Astragaloside Ⅳ significantly reduced 2-DG-induced mPTP opening, the expression of GPR 78, GRP 94 and IRE1 and reduced injury of mitochondria and endoplasmic reticulum.Conclusions Endoplasmic reticulum stress could be induced by 2-DG.Astragaloside ⅳ-induced mitochondrial cardioprotection involves inhibition of the ERS through GRP 78, GRP 94 and IRE1 by prevention of the mPTP opening.

Objective To test the effect of Undaria pinnatifida soluble dietary fiber on endothelial function in hyperlipidemic rats. Method Forty rats were divided into 4 groups(n=10) :control group,hyperlipidemic model group,low dose dietary fiber-treated group(5%) ,high dose dietary fiber-treated group(10%) . After treatment for 8 w. Endothelium-dependent vasorelaxation in isolated aortic rings,the content of plasma malondialdohyde(MDA) ,nitric oxide(NO) and endothelin-1(ET-1) were determined. The protein expression of endothelial NO synthase(eNOS) was measured by Western blotting. Results Undaria pinnatifida soluble dietary fiber treatment significantly decreased MDA and ET-1 level. It also significantly improved endothelial function and plasma NO level concomitantly with unregulation of the expression of eNOS protein. Conclusion In hyperlipidemic rats Undaria pinnatifida soluble dietary fiber could improve vascular endotheliual function,which might be explained by its action to decreasing plasma ET-1 level and increased NO production.

Objective To investigate the therapeutic effects of recombinant yeast hepatitis B virus(HBV)vaccine combined with interferon(IFN)α-1b and determine the rational dosage of HBV vaccine for the further clinical study with larger sample.Methods Two hundreds and sixteen patients with chronic hepatitis B(CHB)were enrolled in this randomized,multi-center,double-blinded and placebo-controlled clinical trial.All the subjects were not treated with antiviral drugs within 6 months and randomly divided into 90μg,60μg and placebo groups with a ratio of 1:1:1.All the patients were intramuscularly administrated with 90μg or 60μg recombinant HBV vaccine or placebo at week 0,2,6,10,14,18,22,respectively.Meanwhile,they were also treated with IFNα-1b 50μg,3 times a wcek for 24 weeks.All patients were followed up for 24 weeks after withdrawal of anti-HBV therapy.Serum HBV DNA level,HBeAg titer and liver function were monitored frequently.Interferon-γ secreting lymphoeytes were detected by Enzyme-linked immunospot(ELISPOT)in part of the patients.Results The serum HBV DNA levels were(6.03±1.79),(5.52±1.82)and(6.29±1.70)log10 copy/mL at week 24 of treatment in high dose,low dose and placebo groups,respectively (P=0.458).And the serum HBV DNA levels were(5.92±1.98),(5.49±1.99)and(6.16±1.76)log10 copy/mL at weck 24 after withdrawal of treatment,respectively(P=0.720).The rates of patients whose HBV DNA＜1×105 copy/mL in these three groups were 30.4%,39.4% and 20.8% at week 24 of treatment,respectively and there was significant difference between high dose group and low dose group(P=0.015).The rate of patients whose HBV DNA＜1×105 copy/mL at week 24 after withdrawal was highest in low dose group,but no significant differences before and after treatment in these three groups(P=0.257).The HBV DNA negative rates were 17.4%,25.4% and 6.9% in these three groups,respectively,which were significantly different(P=0.012).At week 24 of treatment and week 24 after withdrawal of treatment,the alanine aminotransferase normalization rate,HBeAg seroconversion rate were highest in low dose group,but no significant differences in these three groups.ELISPOT positive rates at week 24 of treatment and week 24 after withdrawal of treatment in high close and low dose groups were higher than that in placebo group(P＜0.05).The incidence of adverse events was similar and there was no drug related severe adverse events in each group.Conclusion Recombinant HBV vaccine maybe contribute to anti-HBV therapy and 60μg of dosage seems to be rational.

We screened differential expression bone-related microRNAs (miRNAs) in serum of patients with osteogenesis imperfect (OI). First, we selected the reference gene (s) fit for quantitative detection of serum miRNAs by using geNorm and several other programmes. Then real-time fluorescent quntitative PCR was used to detect the expression level of bone-related miRNAs gained by means of miRanda, Targetscan and Pictar softwares caculation and reading literature. Then, the results were analyzed with the matched t test. All 6 candidate reference genes had a stable expression level in serum of healthy controls and patients with different characters, and the optimal number of reference genes is 4 (miR-16, let-7a, snRNAU6, miR-92a) after Pairwise Variations analysis (V4/5 = 0.133 < 0.15). For validating the universality of expression stability, we detected the relative expression value of miR-16, let-7a, snRNAU6 and miR-92a in another 8 healthy controls and 16 patients with OI and the result revealed that the expression of 4 genes remained stable (M < 1.5). After measuring serum levels of more than 100 bone-related miRNAs in patients with real-time qPCR, 11 miRNAs showed differential expression, and bioinformatic analysis suggested these altered expressional mioRNAs had possibilities to participate in the process of OI. So the experiment indicated that there existed many differential expression bone-related miRNAs in serum of patients with OI, and these miRNAs had potentials to be promising biomarkers for serologic tests and diagnosis of OI.
Biomarkers ; blood ; Case-Control Studies ; Child ; Female ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Male ; MicroRNAs ; blood ; genetics ; Osteogenesis Imperfecta ; blood ; genetics