Objective To investigate the regulative effect of interferon gamma (INF?) on the expression of platelet-derived endothelial cell growth factor / thymidine phosphorylase (PD-ECGF/TP) and its relation with the anti-cancer effect of 5′-deoxy-5-fluorouridine (5'DFUR) and 5-fluorouracil (5FU) in T24 bladder cancer cells. Methods PD-ECGF/TP mRNA and protein expressions were determined by RT-PCR and Western blot,respectively.Cytotoxicity of 5′DFUR,5FU or mitomycin C (MMC) against T24 cells was evaluated by MTS assay. Results The expressions of PD-ECGF/TP mRNA and protein were concomitantly elevated in T24 cells after INF? treatment.And INF? decreased IC 50 s of 5′DFUR [from (16.0? 3.6 )mmol/L to (6.2?0.9)mmol/L,( P

Objective To establish a method for rapidly detecting the G-protein ?3 subunit (GNB3) 825 site single nucleotide polymorphism (SNP) and to analyse the relationship between GNB3 825 site gene SNP and obesity. Methods The real-time fluorescent PCR was employed to analyse the GNB3 825 site gene SNP of 420 samples from 21 provinces and the the frequencies of genotypes were compared with those detected by gene sequencing. GNB3 825 site genotype, body weight, body mass index (BMI) and fat content were examined from 207 subjects and the correlation between GNB3 825 site gene SNP and obesity was analysed. Results The result by real-time fluorescent PCR showed that the frequencies of 825T and 825C haploid were 46.90% and 53.10%, respectively, and the frequencies of 825TT, 825TC and 825CC genotype were 22.38%, 51.42% and 28.10%, respectively, with no other genotype detected, which was consistent with the result by gene sequencing. BMI and fat content were significantly higher in subjects with GNB3 825TT than in subjects with other genotypes. Body weight was much higher in subjects with GNB3 825TT genotype than in subjects with 825CC genotype, but not significantly different with 825CT genotype. Conclusion A new rapid method for the detection of GNB3 825 site SNP has been successfully established. There existed significant correlation between GNB3 825TT genotype and obesity.

Objective To evaluate the nutritional status of iodine of pregnant and lactating women in coastal and inland areas of Zhejiang province,and to provide scientific basis for iodine supplementation.Methods In 2009,five coastal cities,five coastal counties and three inland areas from Zhejiang province were selected by the cluster sampling method,and one sub-district(township) was randomly selected in each city (county),then one community (village) was randomly selected in each sub-district (township).Thirty pregnant women,thirty lactating women and fifty children aged 8 - 10,were randomly selected in each community(village) as the investigation objects to collect instant urine samples to detect urine iodine,and to collect thirty resident edible salt and some water samples to detect salt iodine and water iodine,if the numbers of water source were more than 5 then 1 water sample was collected by their location of east,south,west,north and center in each community(village),otherwise,the numbers of water samples collected were equal to the number of community (village).Results The medians of salt iodine of coastal city,coastal county and inland area were 28.42,25.29,and 33.24 mg/kg,respectively,and the medians of water iodine correspondingly were 5.41,11.52,and 2.80 μg/L,respectively.The medians of urinary iodine of pregnant and lactating women in Zhejiang province were 148.72 and 161.34 μg/L,respectively,which were less than that of children aged 8 - 10(231.02 μg/L,Z =8.57,8.24,all P ＜ 0.017).The medians of urinary iodine in pregnant women from different areas were ranked in order as inland areas(211.42 μg/L),coastal counties (150.51 μg/L) and coastal cities (123.33 μg/L,Z =2.80,5.31,2.42,all P ＜ 0.017),and in lactating women,the medians of urinary iodine in inland areas( 197.43 μg/L) was higher than that in coastal cities( 139.64 μg/L,Z =4.03,P ＜ 0.017).Conclusions The overall levels of iodine nutrition in lactating and pregnant women in Zhejiang province are in the appropriate level,but that of the pregnant women in the coastal cities is inadequate.

Objective To evaluate the nutritional status of vitamin A among urban residents. Methods Using stratified random sampling method,a total of 400 residents from two cities in Zhejiang were selected. They were interviewed by questionnaire,also, the dietary survey, physical measurement and blood sample testing were used. According to Recommended Nutrient Intake( RNI)of vitamin A,to evaluate the intake of Vitamin A and analysis the nutritional status and influenc factors. Results The median of vitamin A concentration was 0. 463( inter-quartile range is 0. 213)μg/mL, and the percentage of severe vitamin A deficiency( VAD ),VAD,insufficiency and adequate serum vitamin A were 0. 25%,0. 50%,6. 75% and 92. 50% respectively. Among 299 individuals who participated in the dietary survey,the median of daily vitamin A intake was 350. 5( inter-quartile range was 351. 5)μgRE. Only 25. 42% of participants' dietary intake of vitamin A met the level of 80% RNI,while 61. 87% of peoples' daily intake of vitamin A under 60%RNI. Male, from the bigger city,older age,high level of triglyceride and more intake of cholesterol may contribute to a higher level of serum vitamin A. Conclusion The majority of urban residents had adequate vitamin A . But the intake of dietary vitamin A still should be paid more attention. Serum vitamin A was associated with age,gender and district.

OBJECTIVETo investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.

METHODSThe Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.

RESULTSThe copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).

CONCLUSIONThere was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.

Actins ; Antioxidants ; metabolism ; Carcinoma, Hepatocellular ; blood ; genetics ; Case-Control Studies ; DNA Copy Number Variations ; DNA, Mitochondrial ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Neoplasms ; blood ; genetics ; Reactive Oxygen Species ; metabolism

OBJECTIVETo investigate the relationship between mRNA expression of platelet-derived endothelial cell growth factor (PD-ECGF) and invasion of bladder transitional cell carcinoma (BTCC).

METHODSThe mRNA expression of PD-ECGF in BTCC was detected by RT-PCR. The target PCR bands were analyzed by NIH Image 1.62 software.

RESULTSThe mRNA level of PD-ECGF in BTCC was 3.86 times as high as that of normal bladder mucosa (t = 2.36, P < 0.05). The expression level of stage Ta, T1 and T2-4 tumor was 1.33, 4.02 and 7.59 times as high as that of normal bladder mucosa, respectively. That of Grade 3 tumor was 2.27 times as high as that of Grade 1 - 2 tumor (t = 3.52, P < 0.01).

CONCLUSIONThe mRNA expression of PD-ECGF was positively correlated with the invasiveness and grade of BTCC. The results suggest that the mRNA level of PD-ECGF might be used as an indicator of tumor progression and a guide for clinical treatment of bladder transitional cell carcinoma.

Adult ; Aged ; Carcinoma, Transitional Cell ; metabolism ; pathology ; Cell Differentiation ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; RNA, Messenger ; analysis ; Thymidine Phosphorylase ; genetics ; Urinary Bladder Neoplasms ; metabolism ; pathology

OBJECTIVETo investigate the therapeutic efficacy of standard antiviral therapy applied after interferon (IFN) treatment failure in patients with chronic hepatitis C (CHC).

METHODSCHC patients who completed a 48-week course of IFN therapy (pegylated (Peg)-IFNa-2a at 180 mug, qw, ih with or without ribavirin (RBV) at 15 mg/kg/w) in our hospital between January 2009 and June 2012 but who showed no response (at week 48) or who relapsed (at week 72) were enrolled in the study. Prior to initiating the 48-week course of retreatment therapy (Peg-IFNa-2a plus RBV as above), the hepatitis C virus (HCV) genotype was detected and the viral load measured (baseline) by PCR of HCV RNA. Each patient's response to therapy was classified as follows: baseline vs. week 4 (rapid virological response, RVR), vs. weeks 12 and 24 (early virological response, EVR), vs. week 48 (end of treatment virological response, ETVR) and vs. week 72 (sustained virological response, SVR).

RESULTSOf the total 235 cases administered retreatment therapy, 60.0% (n = 140) achieved RVR, 77.4% (n = 182) achieved EVR, 83.8% (n = 197) achieved ETVR, 68.0% (n = 68%) achieved SVR, and 15.7% (n = 37) relapsed. Stratification analysis of recurrence (n = 158) and non-responsive (n = 77) sub-groups showed that the recurrence group experienced significantly higher rates of RVR, EVR, ETVR and SVR, but a significantly lower rate of relapse. Stratification analysis of genotype 1b carrier (n = 206) and non-1b carrier (n = 29) sub-groups showed that the 1b carriers had significantly lower rates of RVR, EVR, ETVR and SVR, but a significantly higher rate of relapse. Finally, the patients who achieved RVR (vs. non RVR, n = 95) and EVR (vs. non-EVR, n = 53) showed higher rates of SVR and ETVR.

CONCLUSIONCHC patients who fail to respond to the initial course of standard IFN-based therapy may achieve SVR upon retreatment, especially those infected with the HCV genotype 1b.

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Female ; Genotype ; Hepacivirus ; genetics ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; administration & dosage ; therapeutic use ; Interferons ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; administration & dosage ; therapeutic use ; Recombinant Proteins ; administration & dosage ; therapeutic use ; Retreatment ; Ribavirin ; administration & dosage ; therapeutic use ; Treatment Failure

OBJECTIVETo construct a single-chain human anti-EGFR antibody (scFv) and truncated protamine (tP) fusion protein, ScFv/tP, carrying small interfering (si)RNA directed against the heat shock protein Hsp47, a collagen-binding glycoprotein, in order to evaluate the role Hsp47 in apoptosis of hepatic stellate cells.

METHODSA single chain of the human variable fragment was obtained by phage display and fused with the tP gene and with or without (negative control) the Hsp47 siRNA sequences. Following expression and purification of the scFv/tP fusion protein and the scFv/tPHsp47 siRNA fusion protein, internalization capabilities were tested in isolated human hepatic stellate cells and the QSG-7701 human hepatocyte cells with visualization by immunofluorescent staining. The DNA binding ability of the fusion proteins were verified by gel shift assay.Following ScFv/tP-Hsp47 siRNA fusion protein transfection into the human hepatic stellate cells, the levels of Hsp47 mRNA and protein expression were tested by RT-PCR and Western blotting; in addition, effects of siRNA-mediated silencing of Hsp47 on cell proliferation and apoptosis were analyzed by the cell counting kit (CCK)-8, flow cytometry and Western blot detection of the apoptosis marker poly (ADP-ribose) polymerase (PARP).

RESULTSIndirect immunofluorescence revealed that the ScFv/tP fusion proteins were internalized into human hepatic stellate cells but not into the QSG-7701 cells.The ScFv/tP-Hsp47 siRNA fusion protein caused reduced expression of Hsp47 mRNA and protein expression in the human hepatic stellate cells, as well as increased the cells' apoptosis remarkably.

CONCLUSIONThe ScFv/tP fusion protein can be used as a transfection reagent to deliver Hsp47 siRNA into hepatic stellate cells and to mediate apoptosis via blockade of Hsp47 expression.

Apoptosis ; Cell Proliferation ; HSP47 Heat-Shock Proteins ; genetics ; Hepatic Stellate Cells ; cytology ; Humans ; Protamines ; metabolism ; RNA, Messenger ; RNA, Small Interfering ; Receptor, Epidermal Growth Factor ; immunology ; Single-Chain Antibodies ; Transfection

OBJECTIVETo investigate the renal protective activity of Hsian-tsao Mesona procumbens Hemsl. water extracts in diabetic rats.

METHODSThirty Sprague-dawley female rats were randomly divided into three groups (n = 10 each), "control group" with intraperitoneal saline injection, "diabetic group" with 60 mg of intraperitoneal streptozotocin injection per kg of body weight and "Hsian-tsao group" with intragastric administration of Hsian-tsao extraction everyday for 4 weeks after intraperitoneal streptozotocin injection. The body weight and blood sugar were measured before and after model induction in the three groups. Thrombospondin-1 (TSP-1) expressions in the kidney were monitored by immunohistochemistry. Kidney ultrastructural changes were also analyzed by using transmission electron microscopy.

RESULTSBefore diabetic model induction, there were no significant differences among the three groups in body weight and blood sugar. Four weeks after the induction of diabetes, the differences became statistically significant. Electron microscopy also revealed disruption of the foot processes of the podocytes and other damages in diabetic group. These damages were significantly less severe in Hsian-tsao group when compared with the diabetic group. TSP-1 expressions in the kidney were significantly increased in both the diabetic group and Hsian-tsao group, but it was relatively lower in Hsian-tsao group than in diabetic group.

CONCLUSIONOur results showed that Hsian-tsao treatment in the diabetic rats effectively prevented the pathological alterations in the kidney and decreased the TSP-1 expression. It was suggested that Hsian-tsao had protective effect on the kidneys of the diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; pathology ; Diabetic Nephropathies ; metabolism ; pathology ; prevention & control ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Kidney ; drug effects ; metabolism ; pathology ; ultrastructure ; Lamiaceae ; chemistry ; Rats ; Rats, Sprague-Dawley ; Thrombospondin 1 ; metabolism

OBJECTIVETo investigate the mRNA expression of platelet-derived endothelial cell growth factor (PD-ECGF) in superficial bladder cancer and its significance.

METHODSPD-ECGF mRNA expressions were determined by RT-PCR in 28 cases of superficial bladder cancers and 6 cases of normal bladder mucosa. The relation between PD-ECGF mRNA expression and tumor invasion to lamina propria or recurrence after transurethral resection was also analyzed.

RESULTSSome degree of PD-ECGF mRNA expression was present in all the samples. The PD-ECGF mRNA level was 3.1-fold higher in pT(1) tumors than in normal bladder mucosa (t = 2.13, P < 0.05) and 2.2-fold higher in pT(1) tumors than in pT(a) tumors (t = 2.66, P < 0.05); G(3) tumors expressed 3.3-fold higher PD-ECGF mRNA than normal bladder mucosa (t = 2.44, P < 0.05) and 2.5-fold higher than G(1 - 2) tumors (t = 3.36, P < 0.01). Eleven cases recurred during the mean follow-up period of 18 months. Three-fold higher PD-ECGF mRNA expression was showed in cases who recurred after transurethral resection than that in cases who did not recur (t = 4.49, P < 0.01). The specificity and sensitivity of predicting tumor recurrence were 82.4% and 81.8% respectively using 0.095 as a cutoff value of PD-ECGF mRNA level in this group of superficial bladder cancer.

CONCLUSIONPD-ECGF mRNA expression correlates with tumor dedifferentiation and plays an important role in the early invasion in superficial bladder cancer. To analyze the PD-ECGF mRNA level contributes to the evaluations of tumor differentiation and invasion to lamina propria as well as recurrence prediction in superficial bladder cancer.

Carcinoma, Transitional Cell ; metabolism ; pathology ; Follow-Up Studies ; Humans ; RNA, Messenger ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Thymidine Phosphorylase ; genetics ; metabolism ; Urinary Bladder Neoplasms ; metabolism ; pathology