Objective To prepare recombinant human adenovirus type 3 expressing Norovirus cap-sid protein gene(Noro-orf2). Methods The cDNA for Noro-orf2 was amplifed by RT-PCR from stool of in-fantile gastroenteritis and cloned into the adenovirus shuttle vector pBSE3CMV-egfp. The vector pBSE3CMV-Nor was linearized with EeoR Ⅴ and Not Ⅰ, and transformed into E. coil BJ5183 with lined edenovirus ge-nomic DNA pLasmid pBRAdv3 by Rsr Ⅱ. The identification of recombinant adenovirus plasmid pBRAdv3E3dNor was performed by PCR, enzyme digestion and DNA sequencing. Then pBRAdv3E3dNor was digested with AsiS Ⅰ and transfeeted into Hep-2 cells with LipofectAMINETM 2000 to package recombi-nant adenovirus particles. Results Noro-orf2 was successfully inserted into the shuttle vector. The recombi-nant adenoviral plasmid pBRAdv3E3dNor was generated by homologous recombination in E. coil BJ5183 and confirmed by PCR and enzyme digestion. The recombinant adenovirus was successfully packaged and puri-fied. Norovirus eapsid protein gene expression was confirmed in Hep-2 cells by immunecytochemistry assay. Conclusion The recombinant type 3 adenovirus expressing Norovirus eapsid protein gene was successfully constructed. This study laid a foundation for developing vaccine against Norovirus.

Objective: To evaluate the accuracy of automated classification of ICD-O-3 morphology code from pathology reports by text-mining and support vector machine ( SVM ) , in order to provide basis for automated tumor coding in Chinese. Methods: The tumor report cards of Zhejiang residents from 2017 to 2019 were collected from Chronic Disease Surveillance Information Management System of Zhejiang Province. According to ICD-O-3, the keywords of the pathology reports were extracted, and SVM was used for automatic classification. The classification results were compared with those of 16 professionals with more than two years of experience in tumor coding, and the accuracy rate, recall rate and F-score were calculated for effect evaluation. Results: Totally 83 082 cases from 2017 to 2019 were included and were categorized into 17 morphological classifications, with 52 877 ( 63.65% ) cases of adenocarcinoma, squamous carcinoma and transitional cell carcinoma. A total of 1 090 keywords were enrolled into main corpus. The total F-score, accuracy rate and recall rate are 85.69, 77.20% and 96.27%, respectively. Conclusion Text-mining combined with SVM can improve the efficiency of ICD-O-3 morphology coding; however, the accuracy needs to be further improved.

Objective To identify the prevalence and etiology of kidney disease and the related risk factors in type 2 diabetic patients in rural Shanghai.Methods A cross-sectional study in type 2 diabetic patients was conducted in a community of Shanghai.Questionnaire,clinical examination and laboratory tests were completed to collect the information about sociodemographic and healthcare characteristics.Results A total of 1421 eligible patients with complete information were screened from 1487 type 2 diabetic patients between November 2008 and March 2009.Of them,40.75％ were men,59.25％ were women,aged 37-86 (61.33 ± 9.65 ) years old,with diabetic duration of 0.25-43.92 (7.85 ± 6.34) years.Among them,43.42％ had diabetic retinopathy,21.18％ had neuropathy; 69.95％ met the screening definition for hypertension,76.07％ for hyperlipidemia,15.55％ for hyperuricemia and 23.65％ for cardiovascular disease.The control rates of fasting blood glucose,glycosylated hemoglobin,blood pressure and serum cholesterol were 57.71％,33.99％,14.22％ and 2.46％,respectively.The prevalence of kidney disease,diabetic nephropathy and non-diabetic renal disease was 41.31％,18.51％ and 13.44％,respectively; and 9.36％ were diagnosed as renal insufficiency of unknown reasons.Age,diabetic duration,hyperuricemia,diabetic retinopathy and poor control of blood pressure were independently associated with kidney disease;age and poor control of blood pressure were independently associated with diabetic nephropathy; age and hyperuricemia were independent risk factors of renal insufficiency in patients with diabetic nephropathy.Conclusions Although the diabetic duration of these subjects is relatively short,the prevalence of complications including diabetic nephropathy is high.The high prevalence of non-diabetic renal disease shows the importance of further screening and diagnoses for prevention.Strict control of blood glucose,blood pressure,serum cholesterol and serum uric acid are key points of cutting down the prevalence of diabetic nephropathy and chronic kidney disease.

OBJECTIVETo study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments.

METHODSFecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced.

RESULTSTotally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March).

CONCLUSIONNV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain.

Caliciviridae Infections ; epidemiology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Feces ; virology ; Humans ; Infant ; Molecular Epidemiology ; Norovirus ; classification ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo study the possible differences in the interferon gamma receptor (IFN-gamma R1) response among a variety of clinical types in patients with chronic hepatitis B (CHB) and implications in pathogenesis.

METHODSThe serum level of IFN-gamma and the expression level of IFN-gamma R1 in peripheral leucocytes, from 53 CHB patients, were examined by ELISA and flow cytometry respectively, which were compared with the baseline levels of 15 healthy controls and were performed correlation analysis with alanine aminotransferase (ALT), total bilirubin (TBil) in serum and morphological change in hepatic tissues.

RESULTSThe results showed that the level of IFN-gamma R1 (28.89% 11.77%) expressed on the membranes of lymphocytes in CHB patients, which correlated with liver inflammation (r=0.621, P<0.01) and serum TBil level (r=0.575, P<0.01), was much higher than that (9.23% 1.30%) of the healthy controls (Z=3.988, P<0.05), and no obvious difference on the membranes of leucocytes. The serum levels of IFN-gamma in patients with cirrhosis and severe hepatitis were higher than those of healthy controls. And the two was no difference from each other, but the standard deviation in each group was relatively large.

CONCLUSIONThese findings suggest that the IFN-gamma signal transduction pathway is modulated through up-regulating the expression of IFN-gamma R1 on the membranes of lymphocytes, which takes part in the immuno-pathogenesis in CHB patients.

Adult ; Aged ; Female ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; blood ; Lymphocytes ; chemistry ; Male ; Middle Aged ; Receptors, Interferon ; blood

OBJECTIVETo investigate the mechanism underlying the inhibitory effect of tacrolimus (FK506) against acute liver graft rejection.

METHODSRat models of orthotopic liver transplantation were divided into 3 groups, namely the tolerance group with Brown Norway (BN) rats as the donors and Lewis rats as the recipients, rejection group with Lewis rats as donors and BN rats as recipients, and FK506 group with the same donor-recipient pair as in the rejection group and FK506 treatment. The recipients were sacrificed 7 days after the transplantation, and the hepatic histology, cytokine levels, and glucocorticoid-induced tumor necrosis factor-related protein ligand (GITRL) expression in the liver and Kupffer cells were observed and detected.

RESULTSCompared with the tolerance group, the rejection group showed increased GITRL expressions in the liver and Kupffer cells (P<0.05), which was significantly lowered by FK506 treatment (P<0.05). Acute liver graft rejection caused significantly elevated interferon-γ (IFN-γ) levels and decreased interleukin-10 (IL-10) levels in the plasma and Kupffer cells (P<0.05), and these changes were obviously attenuated by FK506 treatment (P<0.05).

CONCLUSIONThe effect of FK506 in suppressing acute liver graft rejection is probably associated with down-regulated GITRL expression in the liver and Kupffer cells.

Animals ; Carrier Proteins ; metabolism ; Graft Rejection ; prevention & control ; Kupffer Cells ; metabolism ; Liver ; metabolism ; Liver Transplantation ; Male ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Tacrolimus ; pharmacology

OBJECTIVETo observe the effect of Chuanhuang No.1 Recipe (CHR) on renal function and micro-inflammation in phase 3 chronic kidney disease (CKD) patients.

METHODSTotally 60 phase 3 CKD patients were randomly assigned to the treatment group (treated by CHR) and the control group (treated by Losartan Potassium), 30 in each group. All patients received basic treatment. Patients in the treatment group took CHR decoction, 400 mL each time, one dose per day, while those in the control group took Losartan Potassium, 50-100 mg per day. All medication lasted for 24 weeks. Changes of serum creatinine (SCr), blood urea nitrogen (BUN), estimated glomerular filtration rate (eGFR), serum uric acid (UA), 24 h urinary protein excretion (24 h U-pro), urinary microalbumin (U-Alb), high-sensitivity C-reactive protein (hs-CRP), serum tumor necrosis factor (TNF)-alpha, and serum IL-6 were detected and compared before and after treatment. Efficacy was also compared.

RESULTSCompared with before treatment, SCr and BUN significantly decreased in the treatment group (P<0.05, P<0.01); eGFR in- creased (P<0.05). Only UA obviously decreased in the control group (P<0.05), but with no obvious change in SCr, BUN, or eGFR. Compared with before treatment, 24 h U-pro decreased after treatment in the treatment group (P<0.05), but with less decreased level when compared with the control group. U- Alb was also significantly decreased in the control group (P<0.01). There was statistical difference in 24 h U-pro and U-Alb between the two groups after treatment (P<0.05). Compared with before treatment, hs-CRP obviously decreased after treatment in the two groups, but serum levels of TNF-alpha and IL-6 obviously decreased only in the treatment group (P<0.05). The total effective rate was obviously higher in the treatment group than in the control group (70.00% vs. 43.33%, P<0.01).

CONCLUSIONCHR could efficiently improve the renal function of phase 3 CKD patients and alleviate the micro-inflammation.

Adult ; Blood Urea Nitrogen ; C-Reactive Protein ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Inflammation ; Interleukin-6 ; metabolism ; Losartan ; therapeutic use ; Male ; Middle Aged ; Phytotherapy ; Renal Insufficiency, Chronic ; drug therapy ; Tumor Necrosis Factor-alpha ; metabolism ; Urea

OBJECTIVEObserve the effects of Goutengsan on SOD, MAO-B, GSH-PX, NO, LDH, index of brain, rate of death and so on in rats to study therapeutic effects and mechanism of Goutengsan on Alzheimer dementia (AD) model.

METHODOne hundred and twenty rats were randomly divided into 6 groups, 3 experimental groups of which were daily administrated with Goutengsan extract whereas the model and control groups were given NS (0.01 mL x g(-1)). Aniracetam at 0.1 g x kg(-1) served as a positive control. At the 5th day after administration, all groups except the control were administrated (ip) with AlCl3 (100 mg x kg(-1) ) for successive 50 days at 1 day interval. After administration, the death rate, body weight, training scores, brain index, MAO-B, SOD, GSH-Px in brain and NO, LDH in serum were determined.

RESULTThe brain index, SOD, GSH-Px activities as well as NO content of drug-treated groups were strikingly higher that of model group, and had not obvious difference from that of normal group except content of LDH was higher.

CONCLUSIONGoutengsan could increase the brain index, cut down the rate of death, stable increase of body weight, promote the endogenous antioxidant activity, enhance the clearance of lipid peroxide and other metabolic waste, inhibit the MAO-B activity, reduced the leakage of LDH and maintain the content of NO at a normal level. Therefore Goutengsan could protect cells, delay senile, improve symptoms of AD.

Aluminum Compounds ; pharmacology ; Alzheimer Disease ; chemically induced ; drug therapy ; metabolism ; Animals ; Body Weight ; drug effects ; Brain ; drug effects ; metabolism ; Chlorides ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Female ; Glutathione Peroxidase ; metabolism ; Lipid Peroxidation ; drug effects ; Male ; Malondialdehyde ; metabolism ; Memory ; drug effects ; Nitric Oxide ; metabolism ; Oxidative Stress ; drug effects ; Pyrrolidinones ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

OBJECTIVE: To develop a method to measure trihexyphenidyl, chlorpromazine and clozapine in human blood with GC-MS. METHODS: The specimens were alkalized (pH > 10) and extracted with V (benzene):V(ethyl acetate) = 1:1, and qualitatively analyzed using GC-MS-Full Scan with internal standard SKF525A. The specimens were alkalized (pH > 10) and extracted with V(benzene):V(ethyl acetate) = 1:1, and quantitatively analyzed using GC-MS-SIM with internal standard diazepam-d5. RESULTS: The lowest detection limits of trihexyphenidyl, chlorpromazine and clozapine were 0.3, 0.3 and 0.7 ng/mL (S/N > or = 3) respectively. The calibration curve in 20-10 000 ng/mL showed a good linear distribution. The recovery rate was 79.9% to 85.5%. The RSDs of intraday and interday were less than 5.1%. CONCLUSION The established method was simple, sensitive and accurate for simultaneous determination of trihexyphenidyl, chlorpromazine and clozapine in human blood, and can be applied in forensic toxicological cases.
Adult ; Antipsychotic Agents/poisoning* ; Chlorpromazine/blood* ; Clozapine/blood* ; Female ; Forensic Toxicology ; Gas Chromatography-Mass Spectrometry/methods* ; Humans ; Hydrogen-Ion Concentration ; Male ; Middle Aged ; Reproducibility of Results ; Sensitivity and Specificity ; Solvents/chemistry* ; Trihexyphenidyl/blood*

OBJECTIVETo explore the relationship between HBV core promoter mutations and liver damage or HBeAg status.

METHODSNested polymerase chain reaction (nPCR) was used for amplification of HBV DNA core promoter in 59 sera from patients with chronic hepatitis B in Guangxi, then the HBV DNA positive products were sequenced by direct sequencing.

RESULTSThe HBV DNA positive rate of was 59.3%(35/59). All the patients were infected by mutants. The commonest mutation was the double mutation (A --> T at nt1762 and G --> A at nt1764), counting for 57.1% (20/35). The next was C --> G at nt1799, counting for 54.4% (19/35), but this was no function. A --> G at nt1752 (resulting in isoleucine to valine) was seen in 37.1% (13/35) of the HBV DNA positive patients, and T --> C at nt1753 was seen in 20% (7/35). The significant difference in the frequency of T1762A1764 mutant was found between HBeAg positive patients (31.3%) and negative patients (79.0%).

CONCLUSIONSHBV core promoter mutations are common among patients with chronic hepatitis B in Guangxi. T1762A1764 mutant is associated with HBeAg status and chronic hepatitis.

Adolescent ; Adult ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; virology ; Humans ; Liver ; pathology ; Male ; Middle Aged ; Point Mutation ; Promoter Regions, Genetic ; genetics