Objective:To explore the possible mechanism of astragaloside involved in the mouse podocytes injury induced by TGF-β1 in vitro.Methods:Mouse podocytes were cultured in vitro and then all cell were divided into 5 groups:normal control group , TGF-β1 treatment group ,TGF-β1 treatment +astragaloside low dose group ,TGF-β1 treatment +astragaloside middle dose group and TGF-β1 treatment +astragaloside high dose group.The proliferation rate of each group was investigated by MTT assay ,the expression of TRPC6 protein and mRNA were measured by Western blot and RT-PCR respectively after 48 hours.Results:TGF-β1 can significantly inhibit the proliferation of podocytes ( P<0.05) ,fusions of foot processes or even effaced of podocytes were observed .TGF-β1 could also increase the expression of TRPC6.Astragaloside could reduce the inhibition of TGF-β1 to the proliferain of podocytes significantly ,make the cell shape tend to be normal,and reduce the expression of TRPC6 mRNA and protein with dose-effect relation.Conclusion:TRPC6 play an impor-tant role in the TGF-β1 induecd podocytes injury .Astragaloside can alleviate podocytes injury by reduce the expression of TRPC 6.

Cannula ; Catheters ; Drainage ; Duodenal Diseases ; Humans ; Intestinal Fistula/surgery*

COP9 Signalosome Complex ; Carcinoma, Hepatocellular ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; chemistry ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins ; chemistry ; genetics ; metabolism ; Peptide Hydrolases ; chemistry ; genetics ; metabolism ; Phosphorylation ; Phosphoserine ; metabolism

Carcinoma, Hepatocellular ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Phosphorylation ; S-Phase Kinase-Associated Proteins ; metabolism

OBJECTIVETo establish a high-sensitivity, high-specificity and low-cost hydrogel chip platform for the clinical screening of Y chromosome microdeletions.

METHODSSite-specific extended primers with a common sequence at the 5' end were used for hybridizing with the target. The Cy5-dUTP was incorporated into the products by primer extension, and the products were labeled with fluorescence. Then the extended products were added to the chip for hybridizing with acrylamide-modified common probes immobilized on the chip. After removal of the free Cy5-dUTP by electrophoresis, the signals were obtained by fluorescence scanning. And the detecting conditions of this method were optimized.

RESULTSSY254 of 9 samples was successfully detected with the hydrogel chip. The results showed that 3 were normal and the other 6 with microdeletions (1 female sample as a negative control), which coincided with the results of conventional multiplex PCR-electrophoresis.

CONCLUSIONThe hydrogel chip platform we established has provided a new technique for the detection of Y chromosome microdeletions, and is beneficial to the diagnosis and treatment of male infertility.

Carbocyanines ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Deoxyuracil Nucleotides ; Humans ; Hydrogels ; Infertility, Male ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; diagnosis ; genetics

OBJECTIVETo investigate the effect of Xiaoyu Zhixue Tablet (XYZXT) on the expression of platelet membrane glycoprotein (GP) Ib/IX/V complex and GP I b alpha in patients with chronic renal failure (CRF) in early metaphase.

METHODSFifty-one patients with CRF in early metaphase (treated group) were treated with XYZXT, 3 months as the course of treatment for 2 courses. The previous therapies remained unchanged. Flow cytometry was used to assess the expression of platelet GP Ib/IX/V complex and GP Ib alpha in patients with CRF, and turbidity method was used to determine the platelet maximum aggregation rate (MAR), meanwhile the renal function was measured. The final data were compared with those before the treatment, and with those in the normal control group (31 healthy subjects).

RESULTSCompared with the normal control group, expressions of GP I b/IX/V complex and GP I b alpha, and platelet MAR in CRF patients were significantly lower (P=0.007, P=0.001, P=0.009) before the treatment; after the treatment with XYZXT, the above indexes in CRF patients were remarkably increased (P=0.033, P=0.026, P=0.045), but still lower than those in the normal control group, however, it was not statistically significant.

CONCLUSION(1) The expression of GP I b/IX/V complex in CRF patients of early metaphase was decreased, which lead to platelet aggregation dysfunction. This might be one of the reasons for the hemorrhagic trend in CRF. (2) XYZXT was able to upgrade expressions of GP I b/IX/V complex and GP I b alpha in CRF patients, improve platelet function and down-regulate platelet activation in patients with CRF.

Adult ; Aged ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Fluorescence ; Gene Expression Regulation ; drug effects ; Humans ; Kidney Failure, Chronic ; drug therapy ; physiopathology ; Kidney Function Tests ; Male ; Middle Aged ; Platelet Aggregation ; drug effects ; Platelet Glycoprotein GPIb-IX Complex ; metabolism ; Tablets

OBJECTIVETo study the effects of genistein on the proliferation, apoptosis induction and expression of related gene proteins of human colon cancer cells in vitro and in vivo, and its mechanisms of action.

METHODSMTT colorimetric assay was used to detect the effects of genistein on the proliferation of human colon adenocarcinoma SW480 cells. Light and transmission electron microscopy were used to study the histological and ultrastructural changes. Flow cytometry was used to determine the effects of genistein on cell cycle and apoptosis. Flow cytometry and immunohistochemistry were used to determine the effects of genistein on apoptosis induction and expression of related gene proteins of colon cancer cells.

RESULTSThe MTT colorimetric assay showed that genistein inhibited the proliferation of SW480 cells in a dose-dependent and time-dependent manner, and the highest inhibition rate was 60.2% after 80 microg/ml genistein treatment for 72 h. The light microscopy revealed that many genistein-treated cancer cells were shrunken, disrupted, or showing cytoplasmic vacuolization. The electron microscopic examination showed cell shrinkage, nuclear fragmentation and pronounced chromatin condensation, sometimes formed crescent chromatin condensation attached to the nuclear membrane. The results of flow cytometry showed that: after SW480 cells were treated with 0, 20, 40, 80 microg/ml genistein for 48 h, the FI values of PCNA were 1.49 +/- 0.02, 1.28 +/- 0.04, 1.14 +/- 0.03, and 0.93 +/- 0.08; the FI values of VEGF were 1.75 +/- 0.02, 1.34 +/- 0.06, 1.32 +/- 0.04, and 1.23 +/- 0.04; the fluorescence index (FI) values of p21 were 1.26 +/- 0.05, 1.36 +/- 0.06, 1.61 +/- 0.03, and 1.73 +/- 0.03, respectively. There were statistically significant differences between the control group and each treatment group (P < 0.05 or P < 0.01). The scores of immunohistochemical staining of PCNA and VEGF proteins were decreased, while p21 increased. There were statistically significant differences between the control group and each treatment group (P < 0.05 or P < 0.01).

CONCLUSIONGenistein can inhibit the growth of colon cancer cells via apoptosis induction and cell cycle arrest at G(2)/M phase. The anti-tumor mechanisms of genistein may be related with the down-regulation of expression of VEGF and PCNA, and up-regulation of the expression of p21.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Anticarcinogenic Agents ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Colonic Neoplasms ; metabolism ; pathology ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Dose-Response Relationship, Drug ; Gene Expression Regulation, Neoplastic ; Genistein ; administration & dosage ; pharmacology ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo explore the application value of detection of Hepcidin together with indicator of iron overload on clinical diagnosis and treatment of MDS with iron overload by measuring Hepcidin and iron load indices of transfusion dependent myelodysplastic syndrome (MDS) patients.

METHODSEnzyme-linked immunosorbent assay (ELISA), radioimmunoassay and colorimetry were used to determine the Hepcidin, serum ferritin (SF) and serum iron (SI) levels of 106 serum samples from 68 cases of transfusion dependent MDS patients, 30 serum samples of MDS patients without transfusion and 60 serum samples of controls.

RESULTSFor MDS group, Hepcidin level in blood transfusion < 9 U subgroup was significantly higher than that in control group \[(583 ± 50) µg/L vs (175 ± 35) µg/L\] and there was a strong positive correlation between Hepcidin levels and SF (r = 0.976), but no correlation between Hepcidin and SI (r = 0.284); Both Hepcidin and SF level in transfusion 9 ∼ 24 U subgroup was significantly higher than those in control group \[(665 ± 80) µg/L vs (175 ± 35) µg/L; (1445 ± 275) µg/L vs (112 ± 26)µg/L\]; whereas for SI level, there was no difference between transfusion 9 ∼ 24 U subgroup and the control group. Hepcidin did not correlate with SF or SI; For blood transfusion > 24 U group, all of Hepcidin, SF and SI levels were higher than those in control groups \[(703 ± 64) µg/L vs (175 ± 35) µg/L; (2587 ± 352) µg/L vs (112 ± 26)µg/L; (20 ± 4) µg/L vs (14 ± 4) µmol/L\], Hepcidin negatively correlated with SF and SI (r = -0.536; r = -0.456). Hepcidin levels of RARS patients were significantly lower than RAEB patients \[(260 ± 40) µg/L vs (442 ± 51) µg/L\], and there was no significant difference between RARS group and control group regardless of the number of blood transfusion.

CONCLUSIONBoth Hepcidin and SF levels in MDS patients regardless of transfusion dependent or not, or the number of blood transfused were higher than those of normal controls, the increase of Hepcidin can not synchronize with the increase of SF level due to the increased blood transfusion, when blood transfusion > 24 U, Hepcidin level showed a negative relationship with SF and SI, reflecting the decreased ability of Hepcidin to inhibit body iron absorption during the increase of blood transfusion, which finally would lead to iron overload. We can predict the occurrence of iron overload in transfusion dependent MDS patients by dynamic monitoring concentration of Hepcidin.

Adult ; Aged ; Aged, 80 and over ; Antimicrobial Cationic Peptides ; blood ; Blood Transfusion ; Female ; Ferritins ; blood ; Hepcidins ; Humans ; Iron ; blood ; Iron Overload ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; therapy

OBJECTIVE: To investigate expression of ryanodine receptors (RyRs) in rabbit penile corpus cavernosum smooth muscle (CCSM) cells. METHODS: New Zealand White Rabbit CCSM cells were cultured by primary tissue culture. Using CCSM cells and fibroblast have different adherence velocity, CCSM cell can be purified. Identification of CCSM cell was by inverted microscope and immunofluorescence technique. The cDNA sequence of RyRs was found out by searching genebank. Three pair of primers were designed by Primer Premier 5.0. The RyRs subunits mRNA was detected by reverse transcription PCR in cultured CCSM cells. RESULTS: After 7d, we found growth of cultured cells. While 15 to approximately 20 d, cells filled the bottom of culture flask. They were identified by inverted microscope and immunofluorescence technique. After purification, purity of CCSM cells was near to 100%. It suggested only RyRs1 subunit was expressive in CCSM by RT-PCR. CONCLUSION RyRsl subunit is expressed in CCSM cells. It suggests that RyRs contribute to the regulation of Ca2+ in CCSM cells.
Animals ; Calcium/metabolism* ; Cell Culture Techniques/methods* ; Cells, Cultured ; DNA Primers ; Male ; Myocytes, Smooth Muscle/metabolism* ; Penis/cytology* ; RNA, Messenger/metabolism* ; Rabbits ; Reverse Transcriptase Polymerase Chain Reaction ; Ryanodine Receptor Calcium Release Channel/metabolism*

To perform a systemic review and meta-analysis of the diagnostic accuracy of PET (CT) and metaiodobenzylguanidine (MIBG) for diagnosing neuroblastoma (NB),electronic databases were searched as well as relevant references and conference proceedings.The diagnostic accuracy of MIBG and PET (CT) was calculated for NB,primary NB,and relapse/metastasis of NB based on their sensitivity,specificity,and area under the summary receiver operating characteristic curve (AUSROC) in terms of per-lesion and per-patient data.A total of 40 eligible studies comprising 1134 patients with 939 NB lesions were considered for the meta-analysis.For the staging of NB,the per-lesion AUSROC value of MIBG was lower than that of PET (CT) [0.8064±0.0414 vs.0.9366±0.0166 (P＜0.05)].The per-patient AUSROC value of MIBG and PET (CT) for the diagnosis of NB was 0.8771±0.0230 and 0.6851±0.2111,respectively.The summary sensitivity for MIBG and PET (CT) was 0.79 and 0.89,respectively.The summary specificity for MIBG and PET (CT) was 0.84 and 0.71,respectively.PET (CT) showed higher per-lesion accuracy than MIBG and might be the preferred modality for the staging of NB.On the other hand,MIBG has a comparable diagnosing performance with PET (CT) in per-patient analysis but shows a better specificity.