AIM:To analyze zebrafish embryos and to identify erythropoietin (Epo) as an active renal anti-apoptotic factor in an Epo receptor (EpoR)-dependent manner. METHODS:For transient knockdown of Epo and EpoR in renal Tg (wt1b:EGFP) zebrafish reporter lines, the morpholino antisense oligonucleotide technique was used. Morphant zebrafish embryos were phenotypically analyzed using fluorescence microscopy. Apoptosis was determined by TUNEL assay and Annexin V staining. Western blot was used to identify Akt phosphorylation in Epo and EpoR knockdown zebrafish. RE-SULTS:Epo and EpoR zebrafish morphants exhibited pathophysiological changes within the pronephros, adversely affecting pronephric structure. Zebrafish embryos upon silencing of Epoa and EpoR showed a significant increase in the apoptosis within the zebrafish pronephros, consequently leading to renal pathophysiological effects. Decreased p-Akt was identified in Epo and EpoR knockdown zebrafish embryos. CONCLUSION:Epo is an essential regulator of renal development and function by interacting with its receptor EpoR and thereby repressing apoptosis, mechanistically by promoting Akt phospho-rylation.

BACKGROUNDData on the epidemiology of hypertension in Chinese non-dialysis chronic kidney disease (CKD) patients are limited. The aim of the present study was to investigate the prevalence, awareness, treatment, and control of hypertension in the non-dialysis CKD patients through a nationwide, multicenter study in China.

METHODSThe survey was performed in 61 tertiary hospitals in 31 provinces, municipalities, and autonomous regions in China (except Hong Kong, Macao, and Taiwan). Trained physicians collected demographic and clinical data and measured blood pressure (BP) using a standardized protocol. Hypertension was defined as systolic BP ≥ 140 mmHg and/or diastolic BP ≥ 90 mmHg, and/or use of antihypertensive medications. BP < 140/90 mmHg and < 130/80 mmHg were used as the 2 thresholds of hypertension control. In multivariate logistic regression with adjustment for sex and age, we analyzed the association between CKD stages and uncontrolled hypertension in non-dialysis CKD patients.

RESULTSThe analysis included 8927 non-dialysis CKD patients. The prevalence, awareness, and treatment of hypertension in non-dialysis CKD patients were 67.3%, 85.8%, and 81.0%, respectively. Of hypertensive CKD patients, 33.1% and 14.1% had controlled BP to < 140/90 mmHg and < 130/80 mmHg, respectively. With successive CKD stages, the prevalence of hypertension in non-dialysis CKD patients increased, but the control of hypertension decreased (P < 0.001). When the threshold of BP < 130/80 mmHg was considered, the risk of uncontrolled hypertension in CKD 2, 3a, 3b, 4, and 5 stages increased 1.3, 1.4, 1.4, 2.5, and 4.0 times compared with CKD 1 stage, respectively (P < 0.05). Using the threshold of < 140/90 mmHg, the risk of uncontrolled hypertension increased in advanced stages (P < 0.05).

CONCLUSIONSThe prevalence of hypertension Chinese non-dialysis CKD patients was high, and the hypertension control was suboptimal. With successive CKD stages, the risk of uncontrolled hypertension increased.

Adult ; Aged ; Awareness ; Female ; Humans ; Hypertension ; complications ; epidemiology ; therapy ; Male ; Middle Aged ; Prevalence ; Renal Insufficiency, Chronic ; complications

Objective To establish simple,stable and reliable rat models of oxygen glucose deprivation/reoxgenation(OGD/R)in adult neural stem cells(NSCs)in vitro.Methods The NSCs from adult Fisher344 rats were cultured in serum-free medium and identified using nestin and DAPI immunofluorescent double staining.These cells were washed with a Earle′s balanced salt solution without glucose for 2 times,then,incubated for different periods(2,4,6,8 and 10 h)in a trigas incubator with an atmosphere of 1％ O2,5％CO2 and 94％ N2,98％ humidity at 37 ° C.And then,these cells were removed from the anaerobic incubator,washed,and added DEME/F12 containing bFGF supplement.A normoxic-normoglycemic control group was employed.Morphological assessment of NSCs was performed by light microscopy after re-oxgenation for 24 h; CCK-8 colorimetric method was used to determine the survival and proliferation of NSCs,and flow cytometry was employed to detect the apoptosis of NSCs.Results After the setting of oxygen glucose deprivation for 2 h,the OD value and the survival rate in the OGD cells were increased as compared with those in control group without significant difference(P＞0.05).While the morphological damage of NSCs aggravated gradually and the OD value decreased in OGD cells following the prolongation of times; under the setting of oxygen glucose deprivation for 6 h,the OD value in OGD cells obviously decreased as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the survival rate obviously decreased and the apoptosis rate significantly increased in OGD cells as compared with that in the control group(P＜0.05); under the setting of oxygen glucose deprivation for 6 h,the apoptosis rate of NSCs excessed to 50％.Conclusion By means oftrigas incubator,simple,stable and reliable models of OGD/R in NSCs in vitro can be successfully established.

OBJECTIVETo reassess the diagnostic value of 24 hour urinary copper excretion in children with Wilson disease (WD).

METHODSFrom July 2005 to June 2007, inpatients over three years old in a pediatric liver center were assigned into WD and non-WD group.

RESULTS94 patients, including 26 cases in WD and 68 in non-WD group, were enrolled in this study. The median of 24 h urinary copper excretion was 98.5 microg in WD group and 25.8 microg in the non-WD group (Z = -6.111, P equal to 0.000). The area under receiver operator curve (ROC) was 0.909 (95% CI: 0.839-0.979, P equal to 0.000). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 84.6%, 91.2%, 89.4%, 78.6% and 93.9% respectively using 52.0 ug as a cutoff value, and 50.0%, 97.1%, 84.0%, 86.7% and 83.5% using 100 microg as a cutoff value. The goodness of fitness of 52 microg criteria was significantly higher than 100 microg criteria (kappacoefficient 0.760, 0.541 respectively, P equal to 0.000).

CONCLUSIONComparing to 100, 52 microg of 24 h urinary copper excretion as a cutoff value significantly improves the sensitivity and accuracy for diagnosing WD in children.

Adolescent ; Age Factors ; Ceruloplasmin ; Child ; Child, Preschool ; Copper ; urine ; Female ; Hepatitis ; diagnosis ; pathology ; urine ; Hepatitis A ; diagnosis ; pathology ; urine ; Hepatolenticular Degeneration ; diagnosis ; pathology ; urine ; Humans ; Liver ; pathology ; Male ; Penicillamine ; Predictive Value of Tests ; ROC Curve ; Sensitivity and Specificity ; Time Factors

OBJECTIVETo observe the expression of protein arginine N-methyltransferase (PRMT) genes in the lung and spleen of E3 rats with acute asthma.

METHODSE3 rats with ovalbumin-induced pulmonary inflammation were divided into two groups (n=10), and the validity of the acute asthma model was evaluated by histological observation with HE and PAS staining and by measurement of NO production. Semi-quantitative RT-PCR was employed to detect the expressions of PRMT1-PRMT6 genes in the lung and spleen tissues of the rats.

RESULTSIn the lung tissue of the asthmatic rats, the gene expressions of PRMT1 (P<0.01), PRMT2 (P<0.01), PRMT3 (P<0.05) and PRMT5 (P<0.05) were significantly increased, but the expression of PRMT4 gene (P<0.05) was significantly decreased as compared with those in the control tissue. In the spleen tissue of the asthmatic rats, the expressions of PRMT2 (P<0.05) and PRMT5 genes (P<0.05) showed a significant increase as compared with those in the control rat tissue.

CONCLUSIONThe gene expressions of PRMTs vary significantly between asthmatic rats and control rats, suggesting that PRMTs play an important role in the post-translational modification process of asthma-related genes.

Acute Disease ; Animals ; Asthma ; enzymology ; Female ; Male ; Protein Processing, Post-Translational ; Protein-Arginine N-Methyltransferases ; classification ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Inbred Strains

OBJECTIVETo explore the efficacy of intraperitoneally injected epirubicin (EPI)-loaded poly (d, l)-lactic acid (PLA) microspheres (MS) alone or combined with free epirubicin (FEPI) in treating hepatocellular carcinoma (HCC) in mice.

METHODSMice that were transplanted with H22 ascites HCC were randomized into seven groups, which were intraperitoneally injected with blank microspheres, normal saline, three different doses of microspheres (9, 18, and 36 mg/kg EPI) , FEPI (9 mg/kg) , and the combination (microspheres with EPI 4.5 mg/kg + FEPI 4.5 mg/kg). The survival time of all animals was recorded. The rates of increase in life span of all the treatment groups were calculated.

RESULTSEPI-PLA-MS significantly prolonged the survival time of HCC mice in a dose-dependent manner, with a maximal tolerated dose (MTD) of 18 - 36 mg/kg. The combination group had the highest average survival time, median survival time, and rate of increase in life span, which were (40.0 +/- 16.9) days, 33.5 days, and 222.58%, respectively.

CONCLUSIONEPI-PLA-MS combined with FEPI is highly effective in treating HCC in mice when intraperitoneally injected.

Animals ; Delayed-Action Preparations ; Epirubicin ; administration & dosage ; therapeutic use ; Infusions, Parenteral ; Lactic Acid ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Microspheres ; Polyesters ; Polymers

OBJECTIVETo study the cytotoxic effect of allogenetic natural killer (NK) cells in vitro on human CD34+ acute myelogenous leukemia cells.

METHODSCD34 expression on acute myelogenous leukemia KG1a cells was detected by flow cytometry. KG1a cells were co-cultured at different effector-to-target (E:T) ratios with NK cells isolated from 5 healthy individuals using magnetic cell sorting. Lactate dehydrogenase (LDH) release assay was employed to examine the cytolysis of KG1a cells in the co-culture, and the inhibition rate of the KG1a cell colony formation in methylcellulose was determined with K562 cells sensitive to NK cells as the control.

RESULTSA expression rate as much as (98.0-/+1.1)% was detected for CD34 antigen on KG1a cells, and the isolated NK cells (CD3(-)CD16+CD56+ cells) had a purity of (93.2-/+3.7)% after magnetic cell sorting. Allogenetic NK cells exhibited obvious cytotoxicity and colony inhibition in vitro against KG1a cells at different E:T ratios, and the effects were significantly enhanced as the E:T ratios increased (P<0.05). At the same E:T ratio, the cytotoxicity and colony inhibition rate of allogenetic NK cells against KG1a cells was lower than those against K562 cells (P<0.05).

CONCLUSIONAllogenetic NK cells exhibit obvious cytotoxicity and colony formation against CD34+ acute myelogenous leukemia cells.

Antigens, CD34 ; immunology ; Coculture Techniques ; Cytotoxicity, Immunologic ; Flow Cytometry ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; Leukemia, Myeloid, Acute ; immunology

OBJECTIVETo observe the changes of serum free testosterone (FT) and testosterone secreting index (TSI) in ED patients, and to assess the contribution of these two indexes to the diagnosis of ED caused by endocrine factors.

METHODSWe studied 120 ED patients and 30 healthy men undergoing pre-marital medical check-up in Jiangsu Province Hospital of TCM by analyzing the scores on erectile function and desire domain in IIEF, testing the serum total testosterone, luteinizing hormone by chemiluminescent enzyme immunoassay (CLIA), measuring free testosterone by radioimmunoassay( RIA), and calculating TSI.

RESULTSOf the 120 ED patients, 5% and 1538% were below the reference norm of TT and FT values respectively. TT, FT and TSI decreased with age, with statistical with FT and TSI, but not with TT. FT and TSI statistically declined with lower IIEF score on ED domain, but this was not the case with TT. There were no significant differences in TI, FT and TSI among different sexual desire groups the ED patients.

CONCLUSIONFT is much more valuable than TF in the diagnosis of ED with hypogonadism. Both FT and TSI are important parameters in assessing the severity of ED.

Adult ; Aged ; China ; Erectile Dysfunction ; blood ; metabolism ; Humans ; Immunoenzyme Techniques ; methods ; Luteinizing Hormone ; blood ; Male ; Middle Aged ; Radioimmunoassay ; Surveys and Questionnaires ; Testosterone ; blood ; secretion

OBJECTIVETo study the relationship between hepatitis B immunoglobulin (HBIG) injection before delivery and hepatitis B virus (HBV) S gene mutation.

METHODS18 neonates infected with HBV in uterus and their mothers were divided to a) HBIG group (8) in which their mothers received HBIG injection before delivery and b) control group (10) in which their mothers never received any treatment HBV DNA fragments were amplified by nest-PCR from sera of these neonates and their mothers. S gene region of these HBV DNA fragments were directly sequenced and data on mutations was analyzed.

RESULTSThere was no significant difference on nucleotide and amino acid changes in the S gene between the HBIG group and the control group. The majority HBV strains of newborn (17/18) were identical to their mother's dominant strains before delivery, including four mutation HBV strains. Among 18 newborns with HBV intrauterine infection, 12 were infected by B type (adw2), and 6 by C type (adrq+).

CONCLUSIONMothers who were asymptomatic HBsAg carrier and received injections ofHBIG before delivery would not be influenced by HBV S gene mutation. HBV intrauterine transmission with or without gene mutation might occur in the third-trimester of pregnancy. Gene mutation of HBV was not the main factor in intrauterine transmission of HBV.

Female ; Genes, Viral ; Hepatitis B ; prevention & control ; transmission ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; drug effects ; genetics ; Humans ; Immunoglobulins ; administration & dosage ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; prevention & control ; Mutation ; Pregnancy

OBJECTIVETo study the effectiveness of treating hepatocellular carcinoma (HCC) in mice with locally administered epirubicin-loaded poly( D, L) - lactic acid microspheres (EPI-PLA-MS ).

METHODSEPI-PLA-MS was prepared with double emulsion solvent evaporation technique. Five groups of mice (n = 8 in each group) were intraperitoneally injected with five different doses of free epirubicin (FEPI), and the maximum tolerated dose (MTD) was calculated. Then 15 mice with transplanted subcutaneous H22 HCC were divided into three groups (n = 5), which were respectively intratumorally injected with normal saline (NS), blank microspheres, and EPI-PLA-MS (with 9 mg/kg of EPI). After two weeks the tumors were excised and weighed. Another 15 mice with transplanted H22 ascites HCC were divided into three groups (n = 5), which were intraperitonealy injected with the same drugs, and the increased life span were registered exactly.

RESULTSThe MTD of intraperitoneally injected FEPI was 9 mg/kg. The tumour-inhibiting rates was 40.35% and 36.09% when EPI-PLA-MS were administered by intratumoral injection to the mice with subcutaneous H22 HCC. It significantly prolonged the survival time of mice with H22 ascites HCC and the increased life span by 153.49% and 142.22% when EPI-PLA-MS were intraperitoneally administered.

CONCLUSIONEPI-PLA-MS is a new sustained-release preparation with high-efficacy and low-toxicity in treating HCC and has shown promising prospects when administered locally.

Animals ; Antibiotics, Antineoplastic ; administration & dosage ; Delayed-Action Preparations ; Drug Carriers ; Epirubicin ; administration & dosage ; Injections, Intraperitoneal ; Lactic Acid ; pharmacology ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Mice ; Mice, Inbred Strains ; Microspheres ; Polyesters ; Polymers ; pharmacology