Objective To understand the gene characteristics of envelope region of human T lymphotropic virus type Ⅰ(HTLV Ⅰ)from Shantou region of China. Methods The gene of proviral envelope region of HTLV Ⅰ Shantou strain (named HTLV Ⅰ WHP)was amplified by PCR,using 4 designed primers,and then was sequenced,meanwhile it was compared with that of various HTLV Ⅰ representative strains from all around the world,and subsequently it was analyzed to construct its phylogenetic tree.Results The nucleotide sequence of complete envelope region of HTLV Ⅰ WHP provirus we have obtained was 1466 bp.There no deletion and insertion in the deducted amino acid sequence.The results of phylogenetic tree analysis showed that the HTLV Ⅰ WHP strain was most closed to some of the strains from Japan and Caribbean,and like the latter,belonged to the subtypea of cosmopolitan group.Conclusion The HTLV Ⅰ strains either from the coastal region of Southeast China or from Japan,Caribbean and so on have the common origin of evolution.

10 U/ml was determined as positive value.Urinary NMP 22 protein was elevated in 22 cases.Bladder cancer was diagnosed in 11 cases.The sensitivity and specificity of the NMP 22 test were 100%(11/11) and 81%(46/57),respectively.Cystoscopy alone identified 35% of the cancers (4/11).Among 22 cases with elevated NMP 22,1 case was dignosized as bladder cancer during 1 year visit. Conclusions Urine NMP 22 is a new useful marker in early diagnosis of bladder cancer.

Objective To construct a prokaryotic expression vector of cystatin C (Cys C), purify Cys C protein produced by the expression system, and prepare its antiserum. Methods Total RNA was isolated from HL-60 cells, and human Cys C gene was amplified with RT-PCR. The cDNA fragment was cloned into pMD18-T vector and which was confirmed by sequencing. The enzyme-digested target fragment was cloned into PET-32(a) expression vector and transfected into E.coli. BL 21(DE3), in which Cys C expression was induced. After the inclusion body protein was purified through Ni2+ affinity chromatography, processed by dialysis, identified by Western blotting, a rabbit was immunized with the fusion protein, and the antiserum was obtained. Results The result of DNA sequence analysis showed that the cloned Cys C gene sequence was completely corresponding to GenBank data. SDS-PAGE and Western blotting showed that the expressed Cys C fusion protein was about 35?103, mainly existing in the inclusion body of E.coli., that could be purified through Ni2+ affinity chromatography. The titer of the antiserum to the purified protein was 1∶8 000 by ELISA, and Western blotting confirmed that the antiserum reacted specifically to the Cys C protein. Conclusion A recombinant Cys C protein and the specific polyclonal antibody have been obtained, which provides a basis for establishment of immunoassays of human Cys C.

Objective:To study the correlation between responses to Hepatitis B and HLA DRB1*07 DRB1*04 DRB1*1001. DQB1*0401 genes in Han population in Ningbo.Methods:A total of 240 Han people living in Ningbo received the routine vaccination of recombinant hepatitis B vaccine.The serum levels of anti-HBs antibody were examined and the subjects were divided into 2 groups according to tile results:negative responses group(n=120.anti-HBs D

Objective To prospectively assess the nutritional status in the patients with alimentary tract malignancy,and to elucidate the factors related to malnutrition.Methods The nutritional status of 328 patients with newly diagnosed alimentary tract malignancy was assessed using subjective global assessment(SGA)and serum levels of prealbumin and albumin.And the factors influencing the nutritional status of the patients with alimentary tract malignancy in different locations were analyzed.Results The prevalence of malnutrition was 64.43% in all,75.81% in colon cancer,63.24% in esophageal cancer,62.40% in gastric cancer and 60.27% in rectal cancer.The changes of nutritional status mainly manifested weight loss with the incidence of 67.39%,serum prealbumin level under 200 g/L with the incidence of 24.1% and serum albumin level less than 35 g/L with the incidence of 31.70%.And there was significant difference in weight loss and serum levels of prealbumin and albumin among the patients with different nutritional status(P=0.000).The factors that influence the nutritional status of the patients with alimentary tract malignancy include the location and TNM staging of tumors,and the age,appetite and digestive symptoms of the patients.Conclusion The patients with alimentary tract malignancy are susceptible to malnutrition due to the multiple factors such as the tumor location and metabolic impacts of tumor on host.Nutritional screening,assessment and early intervention should be emphasized in the inpatients with alimentary tract malignancy.

Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P

To observe the effects of Danshen-containing serum on SuFu and DYRK2 expression in the HSCs stimulated by leptin. SD rats (n = 60) were used to make danshen-containing serum by gastric perfusion for ten days with Danshen water decoction, normal saline and colchicine. The HSCs that were cultured in vitro would be stimulated for 24 hours by leptin (100 μg x L(-1)) except blank control group, after being intervened, the drug serum in each group would be cultured at 37 degrees C in 5% incubator. The cells would be collected after 24 hours, then the effects of danshen-containing serum on the proliferation of HSCs were detected by MTT, the expression of SuFu mRNA and DYRK2 mRNA were detected by RT-PCR, the expression of SuFu and DYRK2 proteins were tested by Western blot. Compared with blank control group, the expression of DYRK2 mRNA and DYRK2 proteins were enhanced obviously after stimulated the HSCs of rats by leptin (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, after cyclopamine group (Hh pathway inhibitor), Danshen-containing serum and colchicine were interfered, the expression of DYRK2 mRNA and DYRK2 proteins were decreased clearly (P < 0.01), but the expression of SuFu mRNA and SuFu proteins were increased significantly (P < 0.01 or P < 0.05). Compared with model group, adding purmorphamine (Hh pathway agonist) to model group and making it activate could increase the expression of DYRK2 mRNA and DYRK2 proteins, but the expression of SuFu mRNA and SuFu proteins were decreased significantly (P < 0.01). Compared with the model group, using the Danshen-containing serum to interfere the purmorphamine group could make the expression of DYRK2 mRNA and DYRK2 proteins decrease and the expression of SuFu mRNA and SuFu proteins increase significantly (P < 0.01). Danshen-containing serum would inhibition the activation and increment of HSCs by interfering the expression of SuFu and DYRK2 which were induced by leptin.
Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Liver Cirrhosis ; drug therapy ; genetics ; metabolism ; Male ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Repressor Proteins ; genetics ; metabolism ; Salvia miltiorrhiza ; chemistry

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective To observe the sensitivity of myocardium to ischemia/reperfusion (I/R) injury in mechanical trauma rats.Methods The mechanical trauma was established by Noble-Collip drum in the rats that were completely randomized into five groups:sham trauma group,trauma group,sham trauma and sham surgery group,sham trauma and I/R group,trauma and I/R group.The rats were subjected to 30 minutes of ischemia and one hour of reperfusion one week after trauma.The left ventricular systolic pressure (LVSP) and the left ventricular maximum rate of pressure rise and fall ( ± dp/dtmax ) were recorded with BL-410 biological signal recording and analysis system.The levels of serum creatine kinase isoenzyme MB (CK-MB) and cardiac troponinⅠ (cTnI) were detected by double antibody sandwich ABC-ELISA technique.At the end of reperfusion,the heart was excised and stained with Evan' s blue dye and triphenyltetrazolium chloride (TTC) to measure the infarct region with Image-Pro Plus 6.0 image analysis software.Results Compared with the sham traumatic I/R group,the cardiac function in vivo was significantly decreased in the traumatic I/R group ( P ＜ 0.01 ).While the serum CK-MB [ (4 960 ± 588 ) ng/ml:(2 925 ± 426) ng/ml,P ＜ 0.01 ],cTnI [ ( 18.10 ± 3.06 ) ng/ml:( 6.67 ±1.57 ) ng/ml,P ＜ 0.01 ] levels and myocardial infarct size [ ( 36.70 ± 7.42 ) ％:( 22.27 ± 4.54 ) ％,P＜0.01] were obviously higher in the traumatic I/R group compared with the sham traumatic I/R group.Conclusion Mechanical trauma increases the sensitivity of myocardium to I/R injury in rats.

Objective To observe the effects of selective α2-adrenergic receptor agonist alpha-Methylnore-pinephrine(α-MNE) as a vasopressin agent on hemodynamics, troponin T(cTnT) and myocardium in the rabbit cardiopulmonary resuscitation. Method Eighteen health rabbits, weighing 2.5 - 3.5 kg, both male and female,were provided by Lanzhou institute of veterinary medicine. After setting up rabbit model of cardiopulmonary resuscitation, 18 rabbits were randomly divided into three groups. The rabbits in group A as a operation-control group were processed with anesthesia, endotracheal intubation, and surgery without ventricular fibrillation induced. The rabbits in group B as a epinephrine group were administered with 30 ug/kg epinephrineduring CPR. The rabbits in group C as a MNE group were administered with 100 ug/kg α-MNE during CPR. The left ventrictdar end-diastolic pressure(LVEDP), left ventricular pressure rise and fall rate(± dp/dt) and serum concentrations of BNP were measured. Statistic package of SPSS 10.0 was used for the data analysis and significant differences between means were evaluated by ANOVA analysis. Results Compared with group A, LVEDP of other two groups gradually increased respectively(P < 0. 01), and peak ± dp/dt decreased in other two groups(P<0.01). Increase in LVEDP in group C was less than that in group B(P<0.05), whereas peak ± dp/dt in group C were higher than that in group B(P<0.05), at the same stage. Compared with group A, the cTnT of the remaining two groups increased(P<0.01, respectively),and reached peak at 30 minutes. In group C, the elevation of cTnT was less than that in group B(P<0.05) during the same period. In group B and C, myocardial injury was seen under a light microscope, but the injury in group C was lighter than that in group B. Conclusion The methylnorepinephrine can lessen the myocardial dysfunction after CPR.