Objective To determine chromosomal localization of the primary gout susceptibility gene in a pedigree.Methods The clinical data and the peripheral blood samples were collected in the pedigree members and the genomic DNA was extracted from peripheral blood.A genome-wide screening was performed using 400 micro-satellite DNA markers in this family,and linkage analysis was used to determine the chromosomal location of the primary gout susceptibility gene.Results Linkage analysis showed that the maximum LOD score reached 1.50 at marker D4S1572 (at recombination fraction?=0.00).Conclusion Since D4S1572 is localized at 4q25,the primary gout susceptibility gene of this pedigree is localized at 4q25.

Objective To study the expression of mdm2 genes in nephroblastoma in children and relationship between mdm2 gene and clinical pathological parameters.Methods The protein expressions of mdm2 in 24 cases of nephroblastoma were detected with S-P immunohistochemical method.The relationships between mdm2 expression and clinicopathological parameters were analyzed.Results The positive rates of mdm2 in 24 cases of nephroblastoma were correlated with lymph node metastasis,clinical stage and tumor differentiation.There was a positive relationship between mdm2 protein expression and clinicopathological parameters such as lymph node metastasis,clinical stage and degree of differentiation(P

The active ingredient of traditional Chinese medicine, silybin (SBN), can inhibit the proliferation of cancer cells and enhance the anticancer effect of doxorubicin (DOX). However, due to non-targeting and short half-life of SBN and DOX, as well as different administration routes and pharmacokinetic processes, this combination drug cannot act on the tumor in the set order, seriously eliminating the synergistic effect between them and limiting the effect in vivo. Therefore, we intended to construct a nano-delivery system based on molybdenum disulfide (MoS₂), modified by polyethylene glycol (PEG) and sialic acid (SA), and co-loaded with SBN and DOX. The system induced the release of combined drugs under the dual-stimulation of pH and near infra-red (NIR), increased the free concentration of intracellular drugs, so as to achieve the synergistic effect between them. The animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Fujian University of Traditional Chinese Medicine. MoS₂-PEG-SA-SBN/DOX circulated in vivo, and effectively accumulated at tumor sites through enhanced permeability and retention effect (EPR) and SA-mediated active targeting. Under near infrared light irradiation, MoS₂-PEG-SA-SBN/DOX realized the combination of synergistic chemotherapy and photothermal therapy for tumor, thus achieving excellent anti-tumor effect in vivo. This study can provide a new idea and strategy for the clinical treatment of lung cancer. Taken together, MoS₂-PEG-SA-SBN/DOX can offer a new idea and strategy for the clinical treatment of lung cancer.

OBJECTIVESTo investigate the cure effect of tumor antigen specific CTL on a model of human hepatocellular carcinoma in nude mice LCI-D20.

METHODSDendritic cells (DCs) were induced from peripheral blood mononuclear cells of healthy people in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4) and were pulsed with tumor antigen from hepatocellular carcinoma cell line MHCC97H. Then tumor antigen specific cytotoxic T lymphocytes (CTLs) were induced. By intraperitoneal injection of tumour antigen specific CTLs into the LCI-D20, the preventive and therapeutic effects of these CTLs to HCC in the LCI-D20 model were assessed. Cytokine-induced killer (CIK) cells and phosphate buffer solution were used as controls at the same time.

RESULTSThe weights of tumors in the tumor antigen specific CTL group, in the CIK cell group and in the blank group were (1.11+/-0.63), (1.12+/-0.36) and (2.68+/-0.53) grams respectively (t = 5.18, t = 6.06, P < 0.01). The amount of blood alpha fetal protein in the tumor antigen specific CTL and CIK groups were (52.1+/-9.7) microg/L and (48.6+/-5.2) microg/L, and was (82.2+/-7.2) microg/L in the blank group (t = 17.26, t = 22.07, P < 0.01 respectively). The metastasis rates in livers were 16.7%, 16.7% and 58.3% in the tumor antigen specific CTL, CIK cell and blank control groups respectively (chi2= 4.44, P < 0.01). The survival time of the mice in the tumor antigen specific CTL group was (79.0+/-5.02) days, (73.3+/-7.0) days in the CIK group, and (52.3+/-5.2) days in the blank group (t = 14.56, t = 17.54, P < 0.01).

CONCLUSIONTumor antigen specific CTLs may prevent metastasis in the LCI-D20 model and prolong the survival time.

Animals ; Antigens, Neoplasm ; immunology ; Carcinoma, Hepatocellular ; immunology ; pathology ; Cell Line, Tumor ; Dendritic Cells ; cytology ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Interleukin-4 ; pharmacology ; Liver Neoplasms ; immunology ; pathology ; Male ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Recombinant Proteins ; T-Lymphocytes, Cytotoxic ; immunology

Objective To measure albumin in urine by HPLC and conduct primary clinical application Methods Solvent gradient and appropriate wave length was optimized and performance of the HPLC method was evaluated.Urine albumin of 46 patients with diabetes was measured.Results In standard and urine,retention time of Alb was 13.1 min.The linear measuring range extends to 1 820 mg/L.The lower limit of measurment for Alb was 4.2 mg/L.The intra-assay CV and the inter-assay CV were 3.36%,4.12% and 1.93%,1.97% at 24.5 mg/L and 546.9 mg/L of Alb respectively.Analytical recovery rate were 96.3%,98.2% and 97.5%.Microalbuminuria rate was 54.3% by HPLC,26.1% by immunoassay in 46 patients with diabetes.Conclusions Measurement of Alb in urine by HPLC is feasible as routine method until quantifying urinary total Alb conveniently.HPLC is the same to suit research for diabetic nephropathy and so on.

OBJECTIVETo develop a human ovarian carcinoma SKOV3 model in severe combined immunodeficiency (SCID) mouse and to study its biologic characteristics.

METHODSHuman ovarian carcinoma SKOV3 cells were injected intraperitoneally into female SCID mouse to establish a transplantation model of human ovarian carcinoma. The biological characteristics, metastasis and morphology of transplanted tumors were studied.

RESULTAll tumors grew progressively with no sign of regression. The tumor cells spread around the peritoneal cavity and mainly on the diaphragm, mesentery, peritoneum and around the liver, which was confirmed by histopathology. The morphology, growth pattern and CA125 secretion of primary culture of transplanted cells remained as same as those of ovarian carcinoma cell line SKOV3.

CONCLUSIONAn intraperitoneal transplantation model of human ovarian carcinoma SKOV3 in SCID mice has been developed successfully, which can simulate the biological behavior of peritoneal metastasis of human ovarian carcinoma.

Animals ; Disease Models, Animal ; Female ; Humans ; Mice ; Mice, SCID ; Neoplasm Transplantation ; Ovarian Neoplasms ; pathology ; ultrastructure ; Peritoneal Neoplasms ; secondary ; Transplantation, Heterologous

OBJECTIVESTo investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.

METHODSOne hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.

RESULTS4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).

CONCLUSIONSIn ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.

Adaptor Proteins, Signal Transducing ; Base Pair Mismatch ; Carrier Proteins ; Chromosomal Instability ; Cystadenocarcinoma, Mucinous ; genetics ; DNA Methylation ; DNA Repair ; DNA, Neoplasm ; genetics ; DNA, Satellite ; Female ; Genes, Neoplasm ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics

OBJECTIVETo evaluate left ventricular function in newborn infants of mothers with gestational diabetes mellitus (GDM).

METHODSForty newborn infants of mother with GDM (GDM group) and forty normal newborn infants (control group) were enrolled in this study. Two-dimensional speckle tracking imaging was used to measure interventricular septal thickness, posterior left ventricular wall thickness and left ventricular ejection fraction in both groups. Left ventricular rotation and torsion were evaluated for all participants.

RESULTSInterventricular septal thickness in the GDM group was much higher than in the control group (0.45±0.06 mm vs 0.34±0.05 mm; P<0.05). Posterior left ventricular wall thickness in the GDM group was also higher than in the control group (0.45±0.17 mm vs 0.31±0.02 mm; P<0.05). There was no difference in the left ventricular ejection fraction between the two groups (P>0.05). Peak subendocardial rotation, peak subepicardial rotation, peak bulk rotation and peak mural torsion were higher in the GDM group than in the control group (P<0.05).

CONCLUSIONSCardiac function may be impaired in newborn infants of mothers with GDM, with changes in left ventricular shape and abnormalities of left ventricular rotation and torsion. However, infants have a normal ventricular blood ejection under the cardiac compensation. Two-dimensional speckle tracking imaging technique can be used for early detection of left ventricular function.

Diabetes, Gestational ; physiopathology ; Female ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Stroke Volume ; Ventricular Function, Left

BACKGROUND: Adipose-derived mesenchymal stem cells (ADSCs) that are manufactured in good manufacturing practice (GMP) clean rooms should be made into stem cell preparations before administration. Low-temperature preparation has many advantages over cryopreservation preparation; however, little is reported on the effect of short-term low-temperature storage on the biological characteristics of stem cells. OBJECTIVE: To evaluate the effect of 24-hour low-temperature storage using multiple electrolytes containing 5% human serum albumin on the biological characteristics of ADSCs.METHODS: ADSCs at passages 3-6 at a concentration of 5×109/L were suspended in multiple electrolytes containing 5% human serum albumin. Cell suspension was transferred into cryogenic vials, and then these vials were placed in a cold chain shipping box for 2-8 ℃ low-temperature storage for 24 hours. Cell morphology, adhesion ability, cell viability, cell diameters and cell immunophenotyping before and after the storage were observed. RESULTS AND CONCLUSION: (1) After low-temperature storage of ADSCs for 24 hours, the number of dead cells increased. Although cell viability decreased significantly, it was still higher than 80%. Cell diameters of living cells increased significantly. (2) After low-temperature storage of ADSCs for 24 hours, few cells which were circle-shaped lost adhesion ability, and most cells could adherently grow, with the spindle-shaped morphology similar to the cells before preservation. (3) After low-temperature storage of ADSCs for 24 hours, HLA-DR, CD34 and CD45 were negatively expressed with a positive rate lower than 2%; CD29, CD73 and CD105 were positively expressed with a positive rate higher than 95%. However, the cell cluster was clearly divided into two parts after the preservation. Cells with enlarged diameters moved right in the FSC/SSC dot-plot. These results show that low-temperature preparation storage has no significant effect on the stemness of ADSCs, such as adhesion ability, cell viability and cell immunophenotype.

Objective To assess the diagnostic value of combined testing of urinary bladder cancer antigen(UBC),hyaluronic acid(HA)and survivin in the detection of bladder cancer.Methods This study included 64 bladder cancer patients and 20 urinary benign disease patients.The examinations of urine UBC by enzyme-linked immunosorbent assay(ELISA),HA by radioimmunology assay,survivin by RT-PCR and urine cytology were performed in them.Results The sensitivity of UBC(85.9%,55/64),HA (89.1%,57/64)and survivin(93.8%,60/64)was significantly higher than that of urine cytology (40.6%,P＜0.01).The specificity of UBC,HA,survivin and urine cytology was 85.0%(17/20),80.0% (16/20),95.0%(19/20)and 95.0%(19/20),respectively;there was no significant difference among these 4 methods(P＞0.05).The sensitivity of UBC,HA and survivin was also significantly higher than that of urine cytology in different histologic stages and grades(P＜0.05).The sensitivity of UBC and survivin was not significantly different among different histologie stages and grades(P＞0.05).With regard to HA test, the sensitivity in G_2 and G_3 groups was significantly higher than G_1 group(P＜0.01),but there was no differ- ence between G_2 and G_3 groups(P＞0.05);and no difference among different histologic stages(P＞0.05). However,the sensitivity of cytology was improved with the higher grade of bladder cancer(P＜0.01);there was no difference among histologic stages(P＞0.05),By combined use of UBC,HA and survivin,both the sensitivity and specificity were 100%.Conclusions The study indicates that UBC,HA and survivin are better diagnostic markers for the early detection of urinary bladder cancer.These tests are simple,feasible and noninvasive with higher sensitivity and specificity.In addition,combined use of them can improve the diag- nostic sensitivity and specificity.