OBJECTIVETo investigate the effect of T-cell vaccination in murine experimental autoimmune hepatitis (EAH).

METHODSTo induce the EAH model, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally to C57Bl/6 at day 1 and day 7. For T-cell vaccination, splenocytes were removed from animal 2 weeks after induction of EAH and from control animals, and activated in vitro by mitogen stimulation with Concanavalin A (Con A), then inactivated by mitomycin and injected at 5 10(7) cells per animal as T-cell vaccination at 14 and 7 days before first induction of EAH.

RESULTSThe histological grade and serum ALT level of the mice who received T-cell vaccination were decrease significantly, compared with that of model group (1.44+/-0.88 vs. 2.33+/-0.87, t=2.24, P<0.05; 63.0U/L+/-23.4U/L vs. 115.0U/L1+/-39.6U/L, t=2.37, P<0.01, respectively); there was no significant change in mice who received irrelevant T-cell vaccination.

CONCLUSIONT-cell vaccination with T cells from EAH animals, but not with irrelevant T cells, was able to protect animals from EAH.

Animals ; Hepatitis, Autoimmune ; prevention & control ; Male ; Mice ; Mice, Inbred C57BL ; T-Lymphocytes ; immunology ; Vaccination

OBJECTIVETo analyze the maltose clearance in plasma and urine of healthy volunteers with high-performance liquid chromatography.

METHODSMaltose solution was infused to 12 healthy volunteers during a 4-hour period at an infusion rate of 0.2, 0.3, and 0.5 g/(kg x h), Plasma and urine specimens were collected at different time points before and after infusion, and then analyzed with high-performance liquid chromatography.

RESULTSThe coefficients of variation of the precision and accuracy of the analysis method ranged 3.68%-4.58% and 0.44%-4.83% for plasma, respectively, and 2.91%-7.62% and 0.95%-8.27% for urine, respectively. The plasma maltose concentration increased in a dose-dependent manner (r > 0.99). The plasma maltose concentrations returned to the baseline levels 12 hours later. Two hours after injection, the urinary excretion of maltose increased, reached the peak value within 2-4 hours, began to decrease 6 hours later, and became zero 24 hours later.

CONCLUSIONSAn infusion rate of 0.2-0.5 g/(kg x h) of maltose will not remarkably increase the blood glucose level in healthy people. The routine infusion rate should below 0.3 g/(kg x h), unless an emergency exists.

Blood Glucose ; analysis ; Chromatography, High Pressure Liquid ; Humans ; Maltose ; blood ; urine

To study the pathogenic spectrum of hand, foot and mouth disease (HFMD) and the molecular characterizations of human enteroviruses 71 (HEV71) isolated from the clinical specimens of HFMD patients in Inner Mongolia in 2010. A total of 921 clinical specimens including stools and throat swabs were collected from HFMD patients in outpatient service in Inner Mongolia and then viral isolation was performed, the positive viral isolates were identified by using the real-time PCR method (detecting EV, HEV71 and CVA16 in a single tube), and VP4 and VP1 coding region amplification and sequencing was performed with the viral isolates that were identified as non-HEV71, non-CVA16 HEVs. A total of 153 viruses were isolated form 921 clinical specimens, the positive rate was 16.61%, of which 61 (39.87%) were HEV71, 82 (53.59%) were CVA16, 7 (6.53%) were other HEVs(6 were CVB4 and 1 was polio vaccine virus type II) and 3 (1.96%) were adenoviruses. Nine viruses were isolated from severe cases, of which 6 were HEV71 and 3 were CVA16. Thirty two HEV71 isolates were selected from the patients presenting mild symptoms and the patients presenting severe symptoms randomly, and the VP1 coding regions of represented HEV71 isolates were amplified and sequenced. Finally the phylogenetic tree was constructed among the VP1 coding regions of the different genotypes and subgenotypes of HEV71 strains. The nucleotide acid and amino acid of 32 represented HEV71 strains in Inner Mongolia were closed to HEV71 strains isolated from mainland China since 2007, especially from Beijing in 2008, and it showed that all HEV71 strains clustered within the C4a evolution branch of C4 subgenotype. There was slight difference in the nucleotide and the amino acid sequence in VP1 region among the 32 Inner Mongolia HEV71 strains, the identity were 96.4%-100% and 98.14%-100%, respectively, and there was a little difference in the nucleotide acid sequence between the HEV71 strains from Inner Mongolia in 2010 and in 2007, the identity was from 96.95% to 97.87%. Thirty two HEV71 strains were in different lineages in the phylogenetic tree, and it indicated that these strains belonged to many different viral transmission chains. HEV71 and CVA16 were the main pathogens of HFMD in Inner Mongolia in 2010 and most severe cases were caused by HEV71. All the HEV71 strains circulated in Inner Mongolia belonged to C4a evolution branch within C4 subgenotype. Phylogenetic analysis revealed that 2010 Inner Mongolia HEV71 strains were located in different lineages, and had more nucleotide identity with 2008 Beijing HEV71 strains than with 2007 Inner Mongolia HEV71 strains. This indicated that Inner Mongolia HEV71 strains had not evolved independently, but co-evolved with the HEV71 strains in other provinces in mainland China.
Adolescent ; Adult ; Child ; Child, Preschool ; China ; Enterovirus A, Human ; classification ; genetics ; isolation & purification ; pathogenicity ; Female ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Male ; Molecular Sequence Data ; Phylogeny ; Young Adult

OBJECTIVETo evaluate the role of nutritional status on serum immunoglobulins, body weight and postoperative infectious-related complications in patients with Crohn's disease receiving perioperative parenteral nutrition (PN).

METHODS32 patients with Crohn's disease receiving perioperative parenteral nutrition in our department between 1984 and 1994 were enrolled in this survey. 16 patients with loss of body weight in the range of 15%-30% were assigned to the malnutrition group, the other 16 patients with normal weight or loss of body weight less than 15% to the control group. Serum IgM, IgG and IgA levels were measured before and after PN by enzyme-linked immunosorbent assays. Liver function, body weight changes and postoperative complications were also analyzed.

RESULTSIgM levels were elevated before PN in both groups [control group: (133 +/- 16) mg/dl, malnutrition group: (139 +/- 41) mg/dl; normal value: (110 +/- 35) mg/dl; P = 0.04], decreased to normal value [(105 +/- 29) mg/dl, P = 0.02] in the malnutrition group while having no obvious changes in the control group [(129 +/- 13) mg/dl, P = 0.34]. No significant changes in concentrations of IgG and IgA were found (P in the range of 0.20-0.57). The average weight gain was 1.862 kg in malnutrition group [before PN: (45.8 +/- 8.9) kg, after PN: (48.0 +/- 8.8) kg; P = 0.005] and no significant changes in the control group [before PN: (55.6 +/- 6.1) kg, after PN: (56.3 +/- 6.0) kg; P = 0.46]. There was an increase in infectious complications in the control group (control group: 4 cases, 25%, malnourished group: 2 cases, 12.5%; P = 0.13).

CONCLUSIONSPerioperative parenteral nutrition ameliorated the humoral immunity, increased the body weight in patients with obvious malnutrition, whereas it had little value for those without or with mild malnutrition.

Adult ; Aged ; Body Weight ; Crohn Disease ; immunology ; surgery ; therapy ; Female ; Humans ; Immunoglobulins ; blood ; Male ; Malnutrition ; etiology ; Middle Aged ; Nutritional Status ; Parenteral Nutrition ; Pneumonia ; etiology ; Postoperative Complications ; etiology

OBJECTIVETo compare the postoperative analgesic efficacy and safety of the non-addictive propacetamol hydrochloride (Pro-Bufferin) injection and dolantin in a prospective, randomized, double blind and controlled clinical trial.

METHODSAfter the pain intensity was assessed when the patients were undergone thoracic and abdominal selective surgery became fully conscious, 40 consecutive patients with moderate to severe postoperative pain (equivalent to Pain Grade I and II of American Anesthesia Association classification) were randomized into the study against the control groups. The two groups were similar for age, sex, height/weight, disease categories, operation categories, anesthesia methods and duration, vital signs, hepatorenal function, and blood cell count (P = 0.06-0.93). In the study group, 2 g propacetamol in 100 ml normal saline (NS) intravenously with 1.0 ml NS intramuscularly as the placebo control to dolantin were administered. In the control group, 1.6 g mannitose in 100 ml NS intravenously as the placebo control to propacetamol with 50 mg dolantin (1.0 ml) intramuscularly as the positive control to propacetamol were administered. The intensity change of postoperative pain was then evaluated 10 times with visual analog scale and verbal describing scale during 6 h from the beginning of propacetamol infusion. Vital signs and adverse reactions were also documented. After all data were put into the computer, the blinding codes were decoded and the statistic analysis was then made.

RESULTSThere was no significant difference (P = 0.93) about the area under the curve of "Pain Relieve Score vs. Time". The "starting to effect" time (15-30 min), analgesic duration (6 h) and the percentage of excellent or good analgesic effect (90%) in the two groups were the same. Adverse reactions didn't reached the statistic different level (P = 0.35).

CONCLUSIONSPropacetamol HCL injection 2 g intravenously could be an alternative to dolantin 50 mg intramuscularly for moderate to severe postoperative pain with its advantage of being non-addictive.

Acetaminophen ; analogs & derivatives ; therapeutic use ; Adult ; Aged ; Analgesics ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Male ; Meperidine ; therapeutic use ; Middle Aged ; Pain, Postoperative ; drug therapy

Our previous study demonstrates that hypoxia promotes human bone marrow-derived mesenchymal stem cell (hMSC) proliferation. The aim of the present study was to investigate the gene profile involved in this process by using cDNA microarray. Cultured hMSCs were treated with hypoxia (3% O(2)) for 4 h, 12 h, 24 h, 36 h, 48 h and 72 h, respectively. Then these cells were collected to prepare total RNA. Hypoxia-induced gene expression profile was examined and analyzed by GenePix Pro 4.0 software. Some of cDNA microarray results were confirmed by RT-PCR. Microarray analysis identified that 282 genes expressed differentially, of which most were involved in metabolism. The number of differentially expressed genes at different hypoxia time points was different, and most genes were regulated after 24-hour hypoxia. Among the 282 differentially expressed genes, 4 hypoxia-inducible factor 1 (HIF-1) targeted genes and 10 genes that changed at 3 continuous time points were found. The results obtained indicated that 4 HIF-1 targeted genes, i.e., transforming growth factor beta3 (TGFbeta3), phospho-glycerate kinase 1 (PGK1), insulin-like growth factor binding protein 3 (IGFBP3) and BCL2/adenovirus E1B 19 kDa interacting protein 3 (BNIP3), displayed up-regulated pattern at 36 h under hypoxia. BNIP3 displayed a dynamically up-regulated pattern at 12, 36 and 72 h under hypoxia. However, TGFbeta3 and PGK1 were down-regulated at 72 h. In addition, the gene expressions of adenylate kinase 3-like 1 (HAC), neurofilament light polypeptide 68 kDa (NEFL), N-myc downstream regultated gene 1 (NDRG1), discoidin domain receptor family member 1 (DDR1), tribbles homolog 3 (TRIB3), nucleoprotein (AHNAK) and eukaryotic elongation factor selenocyteine-tRNA-specific (EESTS) were up-regulated. Moreover, the gene expressions of EESTS, NEFL were up-regulated at 5 different time points under hypoxia. Furthermore, it was found that the gene expressions of histone cluster 1 (HIS1) and transferring receptor (TFRC) were down-regulated. These results suggest that the proliferation of hMSCs induced by hypoxia is a complex process in which a number of genes may be involved.
Bone Marrow Cells ; cytology ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Oligonucleotide Array Sequence Analysis ; Oxygen ; metabolism ; Transcriptome

BACKGROUNDChemotherapy is the most frequently adopted adjuvant therapy of pancreatic ductal adenocarcinoma (PDAC), but the development of drug resistance reduces its effectiveness. Clarification of the mechanism of multidrug resistance (MDR) development in PDAC is needed to improve the therapeutic effect of chemotherapy. This study was aimed to investigate the molecular mechanism of MDR of PDAC and to identify genes associated with MDR development.

METHODSThe gene expression profiles of cell line SW1990 and three drug-selected pancreatic chemoresistant sub-lines, SW1990/5-Fu, SW1990/ADM and SW1990/GEM, were obtained using an oligonucleotide microarray (Affymetrix HG U133 2.0 plus) that contained approximately 38,000 human genes. The microarray results were validated by real-time quantitative polymerase chain reaction and Western blot analysis.

RESULTSThere were 165 genes and expressed sequence tags, some of which have never been linked to drug resistance, that were up- or down-regulated at least 2-fold in all resistant sub-lines when compared with SW1990. According to Gene Ontology annotation, differentially expressed genes related to MDR in pancreatic cancer belong to many functional families and with diverse biological processes. Genes related to antioxidant activity, apoptosis, the cell cycle, signal transduction and intracellular adhesion may undergo epigenetic changes preceding MDR development. A hierarchical clustering was conducted and several interesting clusters were discovered that may be primarily related to cell cycle and developmental regulation. A prediction rule was built from the expression profiles of 117 genes after support vector machine (SVM) analysis, and the prediction result was examined by cytotoxic testing. As a result, a differential gene expression pattern was constructed in multidrug resistant pancreatic cancer cells.

CONCLUSIONSThe findings of this study prove that construction of a chemoresistance prediction rule, based on gene expression patterns, is practical. These data provide new insights into the molecular mechanism of pancreatic cancer MDR development and may be useful for the detection and treatment of MDR in pancreatic cancer patients.

Cell Cycle Proteins ; genetics ; Cell Line, Tumor ; Computational Biology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Gene Expression Profiling ; Glutathione Peroxidase ; genetics ; Glutathione Transferase ; genetics ; Humans ; Microtubule-Associated Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; methods ; Pancreatic Neoplasms ; drug therapy ; genetics ; Tankyrases ; genetics

OBJECTIVETo develop a diagnostic model comprising clinical and serum markers for assessing HBV-related liver fibrosis.

METHODS270 chronic hepatitis B patients were randomly allocated to either an estimation group (195 cases) or a validation group (75 cases). Liver biopsies were done and staging of fibrosis was assessed. Twenty-six common clinical and serum markers were analyzed initially in the estimation group to derive a predictive model to discriminate the stages of fibrosis. The model created was then assessed with ROC analysis. It was also applied to the validation group to test its accuracy.

RESULTSAmong 13 variables associated with liver fibrosis selected by univariate analysis, age, gamma glutamyltranspeptidase (GGT), hyaluronic acid (HA), and platelet count (PLT) were identified by multivariate logistic regression analysis as independent factors of fibrosis. A fibrosis index constructed from the above four markers was established. In ROC analysis, the AUC was 0.889 for the estimation group and 0.850 for the validation group for discriminating > or =S3 from < or=S2. Using the optimal cutoff score 3.0, the sensitivity of the index was 90.2%, the specificity 76.1%, and the accuracy was 82%. There was a positive linear relationship between the index scores and the fibrosis stages (r = 0.731, P<0.001). The AUC for identifying > or=S2 was 0.873 with sensitivity/specificity of 79%/82%, cutoff score 2.2; The AUC for identifying S4 was 0.872 with sensitivity/specificity of 83%/75%, cutoff score 5.4. There were no significant differences in diagnostic efficacy in the model between the estimation and the validation group (P>0.05).

CONCLUSIONA model for assessment of liver fibrosis was established with easily accessible markers. It appears to be sensitive, accurate and reproducible, suggesting it could be used to assist or replace liver biopsy to detect dynamic changes of HBV-related liver fibrosis.

Adolescent ; Adult ; Aged ; Female ; Forecasting ; Hepatitis B, Chronic ; complications ; diagnosis ; Humans ; Liver Cirrhosis ; diagnosis ; etiology ; Logistic Models ; Male ; Middle Aged

To study on the molecular evolution of Coxsackie virus A16 (CVA16)isolated from clinical speci-mens of Hand, foot and mouth Disease( HFMD) patients in Inner Mongolia in 2010. A total of 921 clinical specimens including stools, throat swabs and vesicle fluids were collected from 888 HFMD patients in out-patient service in Inner Mongolia and viral isolation was then performed, the positive viral isolates were identified by using the real-time PCR method detecting CVA16. A total of 50 CVA16 isolates were selected from the patients presenting mild symptoms, severe symptoms and the death patients randomly, and the VP1 coding regions of representative CVA16 isolates were amplified and sequenced. Finally the phylogenetic tree was constructed among the VP1 coding regions of the different genotypes and subgenotypes of CVA16 strains. Eighty two viruses were isolated form 921 clinical specimens, the positive rate was 8. 90%, of which 3 viruses were isolated from severe cases and 1 viruses was from death cases. The nucleotide acid of 50 representative CVA16 strains in Inner Mongolia were closed to CVA16 strains isolated from mainland China since 1998, especially from Beijing in 2009 and from Henan in 2010, the identity were 96. 18% approximately 98. 88% and 94. 94a approximately 98. 76%, respectively. There was a little difference in the nucleotide acid between the CVA16 strains from Inner Mongolia in 2010 and in 2007, the identity were 91. 68% approximately 96. 52% The phylogenetic tree showed that all CVA16 strains clustered within Bla and B1b evolution branch of B1 genotype. There was slight difference in the nucleotide and the amino acid in VP1 region among the 50 Inner Mongolia CVA16 strains, the identity were 89. 99% approximately 100% and 98. 31% approximately 100%, respectively, indicating that these strains belonged to many different viral transmission chains. The CVA16 strains circulated in Inner Mongolia in 2010 were all belong to B1a and B1b evolution branch of B1 genotype, and the two evolutionary branchs of Coxsackie virus A16 were co-evolved and co-prevailed in Inner Mongolia Autonomous Region.
Adolescent ; Adult ; Animals ; Capsid Proteins ; genetics ; Cell Line, Tumor ; Cercopithecus aethiops ; Child ; Child, Preschool ; China ; epidemiology ; Coxsackievirus Infections ; epidemiology ; mortality ; virology ; Enterovirus ; classification ; genetics ; isolation & purification ; Evolution, Molecular ; Feces ; virology ; Female ; Genotype ; Hand, Foot and Mouth Disease ; epidemiology ; mortality ; virology ; Humans ; Infant ; Male ; Phylogeny ; RNA, Viral ; genetics ; Sequence Analysis, DNA ; Vero Cells ; Young Adult

OBJECTIVETo investigate the malignant tendency of liver basophilic cells in rats by examining the liver dynamic pathological changes during the hepatocarcinogenesis induced by diethylnitrosamine (DEN).

METHODSOne hundred forty male Sprague-Dawley rats, about 200 g each, were randomly divided into a normal group and a model group. The model group rats were administered 1% DEN intragastrically once a week for 14 weeks. The normal control group rats were given saline instead of DEN. Seven to ten rats of the model group were sacrificed at 2, 3, 5, 8, 10, 12, 14, 18 weeks. The remaining rats were followed to the end of the experiment at 26 weeks. Histopathological changes of the livers were analyzed, and the localization of GST-P and PCNA in the livers were detected in situ by immunohistochemistry.

RESULTSAccording to the characteristics of the lesions in the model group, histological liver change patterns were categorized into three phases: (1) liver injury phase (2 to 5 weeks) with centrilobular necrosis, a small amount of collagen deposition in the necrotic regions with fibrous septa development and cell proliferation; (2) the cirrhosis phase (8 to 12 weeks) with significant hepatocellular regeneration and collagen deposition. As the regenerative nodules and fibrous septa formed, cirrhosis with uniform sized nodules developed in all the rats at 12 weeks. In the regenerative nodules, significant hepatocellular metamorphosis was seen; (3) In the carcinomatous transformation and nodular remodeling phase (after 14 weeks), two types of cancer, namely hepatocellular carcinoma and cholangiocarcinoma were found. Incidence of the cancer was 62.5% at 18 weeks. Basophilic cell lesions appeared beginning at 10 weeks. Pale bodies were seen in some basophilic cells. Small cell changes appeared starting at 12 weeks. Some of these cells containing droplets like lipid vacuoles, invaded into the surrounding liver tissues. Both basophilic cell lesions and small cell changes were all positive for proliferating cell nuclear antigen (PCNA).

CONCLUSIONDevelopment of foci of basophilic small cells, with cells containing lipid vacuoles and pale bodies, and invading into the surrounding liver tissues are the changes highly suggestive of an early hepatocellular carcinoma transformation.

Animals ; Basophils ; pathology ; Carcinoma, Hepatocellular ; pathology ; Hepatocytes ; pathology ; Liver Neoplasms, Experimental ; pathology ; Male ; Precancerous Conditions ; pathology ; Rats ; Rats, Sprague-Dawley