Objective To explore the best technology to produce recombinant mouse endostatin by thioredoxin fusion expression system.Methods The recombinant plasmid pThioHis-endo was further transformed into different E.coli.,including BL21,JM109,DH5?.After induction with IPTG of different concentration or for different time period,thioredoxin-endo fusion protein was expressed in E.coli.and the product was identified by SDS-PAGE.The recombinant endostatin expression and extraction,wash,degeneration and refolding of inclusion bodies were carried out by the optimized route,and the product was further purified by affinity chromatography through Ni~(+) column and identified by SDS-PAGE.Results No difference of endostatin expression in BL21,JM109,DH5? was found.The optimal concentration of IPTG was 0.9 mmol/L and optimal inductive phase was 5 h.The soluble recombinant endostatin could be obtained by affinity chromatography through Ni~(+) column.Conclusion Soluble endostatin recombinant fusion protein of high purity and yield could be obtained by the optimized technique route.

This paper is to report the development of a rapid and sensitive method for the determination of s-oleylpropanolamide (OPA) in various tissues of rat (brain, heart, lung, liver, spleen, small intestine, kidney, adipose tissue and muscle), and to assess the applicability of the assay to tissue distribution. OPA was extracted by liquid-liquid extraction method with undecylenoylethanolamide as an internal standard. The concentrations of OPA were determined by LC-MS/MS after a single intragastric dose of 50 mg x kg(-1) at 4 time points (5 rats per group). With multiple reactions monitoring mode (MRM) the limit of quantification (LLOQ) was determined at 1 microg x L(-1). The calibration curve was linear from 1 to 2 x 104 microg x L(-1) (r > or = 0.999 0) for tissue homogenates. Validation parameters such as accuracy, precision and recovery were found to be within the acceptance criteria of the assay validation guidelines. The highest concentration was found in small intestine (the highest time point is 15 min) and heart (the highest time point is 90 min). The assay is rapid, sensitive and applicable to studying tissue distribution of OPA in rats.

Objective Estimating clinical effects of the Therapy of Potassium L-aspartate Injection on Hypokalemia.Methods These hypolalemia patients from the Renji hospital of Shanghai Jiaotong University and Other Two Hospitals,and from March 2007 to May 2008,which were caused from various kinds of reasons.The test group (42 patients) were intravenous dripped by using three rami potassium L-aspartate injection added in 5% Glucose (or saline) 750 mL,the control group (44 patients) were gived two rami chloratum Kalium injection added in 5% Glucose (or saline) 750 mL,the Kalium were 1.10 g/d in both groups,once per day,time of therapy was 7 day.Results The test and control groups effective power after therapy were 86.67% and 91.11%,respectively,there was no significant difference (P=0.3998).The Kalium dose of test group was advanced from (3.18?0.23) mmol/L before testing to (3.73?0.37) mmol/L after therapy,the elevated average was (0.61?0.46)mmol/L,and there was no significant difference (P=0.4722).The accumulation power of Kalium recovery were (51.11?7.45)%,(77.78?6.20)%,(86.67?5.07)%,at the second,the fored and the sevened day,respectively.There was no significant difference between the two groups (P=0.1430).Conclusion The clinical effect was correspond between three rami potassium L-aspartate injection added in 5% Glucose (or saline) 750 ml and the Isodose chloratum Kalium injection,by once per day,and seven days of therapy,which can treat hypokalimeia caused from various kinds of reasons.

Objective To investigate the effect of gender and age on the apparent diffusion coefficient (ADC) of normal adult pancreas.Methods A total of 383 patients with normal pancreas (290 male,93 female,range from 21 to 78 years of age) were enrolled in this study.The subjects were divided into four groups based on different age (≤40 years,41-50 years,51-60 years and ＞60 years) with patient number of 56,108,139 and 80,respectively.Breath-hold single-shot echo-planar DWI (b value =0,500 s/mm2) was performed to determine ADCs on all patients.The average ADCs was calculated by four ADCs measured from the head to tail part of the pancreas in each patient.Patients with different age or gender were analyzed by independent-samples t test.Effect of gender on ADCs was analyzed using the Mann-Whitney U test.Relationship between ADCs and age was analyzed using Spearman rank-order correlation test,and Kruskal-Wallis H test was used to compare the ADCs among 4 age groups.Results The median pancreatic ADC values in female group(n =93) [1.60 × 10-3 mm2/s (1.47 × 10-3-1.77 × 10-3) mm2/s] was higher than that in male group (n =290) [1.57 × 10-3mm2/s(1.41 × 10-3-1.74 × 10-3)mm2/s].Mann-Whitney U test results showed the mean ADCs was similar between the two groups (Z =1.335,P =0.182).The age distribution was similar between the male [(52 ± 10) years of age] and female [(51 ± 11) years of age]groups (t =0.267,P=0.790).The age spectrum showed that there was no correlation between the average ADC values and age (r =0.016,P =0.752).The median ADC values of the four age groups were 1.58 ×10-3,1.54 × 10-3,1.59 × 10-3 and 1.57 × 10-3 mm2/s,respectively.Kruskal-Wallis H test showed no significant difference of mean ADCs among the age groups (x2 =2.15,P =0.542).Conclusion There is no correlation of ADCs between age and gender in normal adult pancreas.

Objective To analyze atypical 64-slice spiral CT imaging finings of pancreatic cancer and to improve the ability to identify CT manifestations of pancreatic cancer. Methods A retrospective analysis was performed on the atypical 64-slice spiral CT imaging findings of 12 eases of pancreatic cancer confirmed by pathology after surgery. Results All the twelve cases were pancreatic ductal adenocarcinoma.Among them, 7 cases were moderately differentiated ductal adenocarcinoma, 1 case was well-differentiated ductal adenocarcinoma, 1 case was mucinous adenocarcinoma, 3 cases were adenosquamous carcinoma. Among 8 cases with ductal adenocarcinoma, the lesions were located in the pancreatic head and (or) uncinate process in 7 cases, and in the pancreatic neck of 1 case. Tumors were expressed as isodense or low-density or cysticsolid lesions, the masses showed no enhancement in the enhanced scanning phase. Tumors were clearly exogenous or exogenous tendencies in 5 cases. Five cases had no distal pancreatic duct dilation, 2 patients had common bile duct and intrahepatic biliary dilation, and only 1 patient had atrophy of distal pancreas. There was one case of mucinous carcinoma, plain CT scan showed a cystic lesion in head of pancreas about 5cm in diameter, the solid part below the cystic lesion was slightly enhamced in the enhanced scanning phase and the body and tail pancreatic duct was moderately dilated (7 mm). There was no common bile duct and adjacent blood vessels invasion. Among 3 cases of adenosquamous carcinoma, lesions were located in the pancreatic head of 2 cases and in pancreatic body of 1 case. The maximal diameter of mass ranged 3.0 cm ～ 4.5 cm.Cystic necrotic area was observed within the lesions in 3 cases in enhanced pancreatic parenchymal phase of CT scan. Distal pancreatic duct were mildly dilated (4 ～ 5 mm) in 3 cases. There was no common bile duct and intrahepatic bile duct dilation. Conclusions Pancreatic cancer may show atypical CT imaging findings and great cautions are needed for differential diagnosis.

AIM To investigate the effects of tranfection of IL 2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS Spleen cells were transfected with IL 2R antisense RNA eukaryotic expression plasmids using adhesion assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL 2R mRNA and protein expression level were measured by slot blot hybridization assay and flow cytometry method respectively. RESULTS The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti mIL 2R?? and pciAnti mIL 2R?? transfected group was higher than that of pcAnti mIL 2R? and pcAnti mIL 2R? transfected group; the inhibitory rate of pcAnti mIL 2R? tranfected group was higher than that of pcAnti mIL 2R? tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL 2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION IL 2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL 2R?? chimeric antisense RNA showed higher inhibitory rate than ? or ? antisense RNA. IL 2R? antisense RNA was more effective than ? antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL 2R antisense RNA was exclusively on the growth of cells functionally expressing IL 2R. The inhibitory effect on the spleen cells proliferation was likely due to the blocking of IL 2R expression by antisense RNA.

Objective To explore the values of potential clinical application ofsingle tube/ten colorsflow cytometry for leukocyte differential count in peripheral blood.Methods Utilizing multiple monoclonal antibody combinations and the vavious logical gating strategies,the single tube/12 antibodies with no-wash method for the leukocyte differential count in peripheral blood were determined by using 10 colors flow cytometry.Leukocyte differentials of 142 peripheral blood samples were determined by both Beckman-Coulter LH750 hematology analyzer and 10 colors flow cytometry.The results were then compared to standard microscopic examination as a reference method.The clinical diagnostic efficiency ofsingle tube/10 colorsflow cytometry was calculated.The correlations between standard microscopic cytology,single tube/10 colorsflow cytometry and the hematology analyzer were determined.In addition,the clinical diagnosis efficiency for blast counts ofsingle tube/10 colorswere compared to the results determined by BD FACS Calibur flow cytometer.Results The leukocyte differentials were correlated well between the single tube/10 colorsflow cytometry and standard microscopic cytology(r>0.700,P<0.01) except for basophils.The correlations with neutrophilic granulocytes,lymphocytes,immature granulocytes and blasts were superior(r=0.972,0.951,0.801,0.912,respectively,P<0.01).When 1% was selected as the cut-off point for immature granulocytes determined by standard microscopic cytology,the sensitivity and the specificity ofsingle tube/10 colorsflow cytometry were 92%(57/62) and 79% (63/80),respectively.When 0.5% was selected as the cut-off point for blasts detected by standard microscopic cytology,the sensitivity and the specificity were 99% (67/68) and 92% (68/74).Using the immunophenotyping results from BD FACS Calibur as a standard,the sensitivity for detecting blasts bysingle tube/10 colOrsflow cytometry was 100% (40/40),the specificity was 91% (10/11),the positive predictive value was 98% (40/41),the negative predictive value was 100% (10/10) and the accuracy was 98% (50/51).Conclusions Thesingle tube/10 colorsflow cytometry has a excellent correlation with the standard microscopic cytology when applied on leukocyte differential count in peripheral blood.It may potentially use as a subsequent method for verification of abnormal results of complete blood cell count in the future.

Objective We propose a method using total variation (TV) regularization in deconvolution for partial volume correction in PET imaging. In the degraded image model, we used TV regularization procedure in Van Cittert (VC) and Richardson-Lucy (RL) deconvolution algorithms. These methods were tested in simulated NCAT images and images of NEMA NU4-2008 IQ phantom and tumor-bearing mouse scanned by Simens Invoen microPET. The simulated experiment and tumor-bearing mouse experiment showed that the algorithms using TV regularization provided superior qualitative and quantitative appearance compared with traditional VC and RL algorithms. When the mean intensity of the tumor increased by (10 ± 1.8)%, the SD increase percentage was decreased from 49.98%to 14.26%and from 42.76%to 4.70%, suggesting the efficiency of the proposed algorithms for reducing PVEs in PET.

Objective We propose a method using total variation (TV) regularization in deconvolution for partial volume correction in PET imaging. In the degraded image model, we used TV regularization procedure in Van Cittert (VC) and Richardson-Lucy (RL) deconvolution algorithms. These methods were tested in simulated NCAT images and images of NEMA NU4-2008 IQ phantom and tumor-bearing mouse scanned by Simens Invoen microPET. The simulated experiment and tumor-bearing mouse experiment showed that the algorithms using TV regularization provided superior qualitative and quantitative appearance compared with traditional VC and RL algorithms. When the mean intensity of the tumor increased by (10 ± 1.8)%, the SD increase percentage was decreased from 49.98%to 14.26%and from 42.76%to 4.70%, suggesting the efficiency of the proposed algorithms for reducing PVEs in PET.

OBJECTIVETo investigate the epidemiological status of cholestasis in first-hospitalized patients with chronic liver disease in Shanghai, and to provide a scientific basis for developing prevention and treatment measures.

METHODSFrom April 2005 to September 2014, 5,146 first-hospitalized patients in Shanghai with a diagnosis of chronic liver disease were enrolled in this study. Clinical data of the 4,660 patients who fit the study criteria for participation were collected for retrospective analysis.Diagnosis of cholestasis was made according to serum alkaline phosphatase (ALP) levels higher than 1.5 times the upper limit normal (ULN) and gamma-glutamyltransferase (GGT) levels higher than 3 times the ULN. The incidence rate of cholestasis was assessed for relation to age, sex, etiology, and type of liver disease, and statistically compared to the general clinical data and specific biochemical indicators with potential sex-related differences. T-test and chi-square test were performed for the statistical analyses.

RESULTSOf the 4,660 study participants, 10.26% had cholestasis; the prevalence of cholestasis increased with increasing age in male patients. The distribution of the cholestasis incidence according to the type of chronic liver disease was: 75.00%, primary sclerosing cholangitis; 42.86%, primary biliary cirrhosis; 35.97%, hepatic tumor; 30.77%, autoimmune hepatitis; 28.31%, drug-induced liver disease; 16.46%, alcoholic hepatitis; 13.98%, cryptogenic cirrhosis; 12.99%, schistosomal cirrhosis; 7.53%, alcoholic cirrhosis; 7.32%, mixed cirrhosis; 5.94%, viral liver cirrhosis; 2.70%, nonalcoholic fatty liver disease. There was no significant difference in the prevalence of cholestasis between the two sexes. In the patients with cholestasis, the levels of GGT and total bilirubin were significantly different between the two sexes.

CONCLUSIONThe incidence rate of cholestasis in first-hospitalized patients with chronic liver disease was 10.26%, and the rate increased with increased age. Patients with primary sclerosing cholangitis or primary biliary cirrhosis had higher incidence rates of cholestasis. Incidence rates of cholestasis of the various chronic liver diseases were not related to sex.

Bilirubin ; China ; Cholestasis ; Chronic Disease ; Humans ; Incidence ; Liver Diseases ; Male ; Prevalence ; Retrospective Studies ; gamma-Glutamyltransferase