Recent researches have revealed that iron-overload could lead to osteoporosis and bone morphogenetic protein 6(BMP-6)could enhance the expression of hepcidin. Therefore, it is believed that BMP-6 might play a key role in the treatment of such kind of osteoporosis. This paper focuses on the newest papers regarding the relationship between BMP-6 and iron-overload osteoporosis in order to lay a foundation for further research on the osteoporosis induced by iron overloading.

Objective: To investigate the cytotoxicity of ICE gene transfection in Combination with Antitumor Chemicals killing Hepatocellular Carcinoma Cells in vitro.Methods: The recombinant plasmid pLXSN-hICE was transferred to virus packing cell PA317 by electroporation method. And then the retrovirus containing human ICE cDNA generated by these PA317 cells were used to transfect human hepatocellular carcinoma cell line HepG2. The apoptosis of transferred cells were examined by gel electrophoresis. The influence of chemotherapeutic drug Carbo-platin to the proliferation of hepatocellular carcinoma cell line SMMC7721 and its derivative cells(SMMC7721-hICE,SMMC7721-antisence hICE,SMMC7721-neo)was observed by incorporation of 3H-TDR. Results: Electrophoresis of DNA displayed the apoptosis ladder of HepG2 transfected by ICE gene. The proliferation of SMMC7721-hICE was significantly suppressed in vitro induced by Carbo-alatin compared to the other three cell lines. Conclusion: ICE gene transfection could greatly increase the susceptibility of SMMC7721 cells to apoptotic cell death following chemotherapy. These findings suggest that combining ICE gene transfection with utilizing antitumor drugs would represent a novel approach for the effective treatment of hepatocellular carcinoma.

BACKGROUND: One unit of umbilical cord blood does not have a sufficient number of peripheral blood stem cells to meet the requirements of transplantation in adults. One solution of this problem is their ex vivo expansion, which requires not only a longer time and higher culture conditions, but also easily leads to the differentiation of stern cells, thus affecting the effects of transplantation. OBJECTIVE: To transduce human interleukin-3(IL-3) gene into umbilical cord blood CD34" cells and to observe IL-3 expression. DESIGN, TIME AND SETTING: A cell-genomics in vitro experiment was performed in the Chengde Medical College in 2008. MATERIALS: Human peripheral blood mononuclear cells (PBMC) of healthy adult were provided by Chengde Blood Center, and umbilical cord blood was provided by Chengde Maternal and Child Care Hospital. Written informed consent was obtained from each donor. METHODS: Peripheral blood mononuclear cells were isolated by Ficoll gradient centrifugation and umbilical blood CD34+ cells were isolated using immunomagnetic beads method. IL-3 mRNA was extracted. IL-3 cDNA was synthesized by RT-PCR, and IL-3 cDNA was subcloned into eukaryotic expressing vector pcDNA3. In the experimental group, pcDNA3/IL-3 vectors were transduced into umbilical cord blood CD34+ cells, while in the control group, transfection was not performed. MAIN OUTCOME MEASURES: Detection of IL-3 level in umbilical cord blood CD34" cells suspension using ELISA kits. RESULTS: Theoretically, the amplified IL-3 cDNA was 616 bp, and actually, after agarose gel electrophoresis, the PCR products exhibited a strip with expected size under ultraviolet ray. Extraction of IL-3mRNA was successful and reversely transcripted cDNA was complete. A 616-bp inserted fragment was observed by agarose gel electrophoresis after double digestion with BamH Ⅰ and Xba Ⅰ, and it was the same as IL-3 sequence. Within 1-7 days after transfection, IL-3 level in the umbilical cord CD34+ cells suspension was significantly higher in the experimental group than in the control group (t = 3.46, P < 0.05). CONCLUSION: IL-3 cDNA was successfully cloned, and eukaryotic expressing plasmid pcDNA3/IL-3 that could be effectively expressed within short term in umbilical cord CD34+ cells was successfully constructed.

Objective To study the feature of MRI in ureter transitional cell carcinoma,to evaluate the diagnostic value in transitional cell carcinoma of ureter with MRI.Methods Heavily T_2-weighted fast spin echo pulse sequence,fat suppression pulse and MR urography(MRU)were performed.The MRI finding of the ureter transitional cell carcinoma were anlysed in 32 cases and were discssed with the review of literature.Results Fifteen lesions were located at the upper portionof the ureter,7 at mid portion and 10 at lower portion.Each case presented urinary obstruction,distention and uretal hydrocele.21 retrograde urleropyelogrhpy of nodular shaperal irregular,11 irregular the ureteral wall,10 dilate the ureter in 21 cases,11 infitrative lesion to grow in location,9 lymphanode to enlarge in surrounding of major arterial of abdominal and renal out in 11 cases.17—72 mm length the lesion,39 mm average,6—50 mm width the leion,17 mm average.Hypointense on T_1 WI and hyperintense on T_2 WI image in 23 cases,hyperintense on both T_1 WI and T_2 WI image in 5 cases,hypointense on T_1 WI and isointense on T_2 WI image in 2 case, slightly hypointense on both T_1 WI and T_2 WI images in 2 case.Ninteen homogeneous and 13 non homogeneous of signal in lesion,22 reliable and 5 suspicious diagnosis and 5 misdiagnosis in MRI. Conclusion The location,the shape,the spectrum of the tumor and change of surrounding tiessue were clear cuted in MRI,but further research in confirmation of the diagnosis.

AIMTo establish a method for detection of plasma total homocysteine with HPLC.

METHODSThe chromatography analysis was carried out using a Symmetry Shield RP18. The mobile phase was sodium acetate (0.08 mol/L) and methanol (1%) and we utilized a HPLC system with fluorescence detection of plasma homocysteine derivatized from reaction with 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate (SBD-F).

RESULTSThe average recoveries were 95.8 - 100.8% and the relative standard deviations were 1.2-2.0%.

CONCLUSIONThe results showed it to be a rapid and accurate method for the determination of homocysteine level in plasma.

AIM: To observe the effects of Decoction of Bu-Shen-Huo-Xue-Xie-Zhuo (BSHXXZ) on focal segmental glomerulosclerosis (FSGS) in rats. METHODS: Unilateral nephrectomy was adopted firstly, one week later adrimycin was intravenously adminstered to 20 rats twice in a 23-day period to establish an advanced focal segmental glomerulosclerosis model in the rat. Changes in urine protein, blood chemistry, and histology of the kidney were investigated for 110 days after unilateral nephrectomy. RESULTS: The BSHXXZ decoction reduced edema, proteinuria, hyperlipidemia, azotemia and ascites, and increased albumin in blood. Light microscopic, electron microscopic, immunohistochemical and polarizing microscopic examination all showed that the pathologic changes in the treatment group were less than that of the model group. CONCLUSION: Decoction of BSHXXZ could markedly improve renal function and delay the progression of glomerulosclerosis and renal interstitial fibrosis.

Objective To investigate the effect of fluvastatin (FLV) on the expression of β1 integrin in puromycin aminonucleoside (PAN)-treated podocytes and its mechanism.Methods Cultured human podocytes were divided into PAN,different concentrations of fluvastatin (1 × 10-8 to 1 × 10-5 mol/L),SOD,H2O21 groups respectively.Expressions of β1 integrin and reactive oxygen species (ROS) in podocytes were detected by Western blotting and DCFHDA (2' 7'-Dichlorofluoresecein 3' 6'-diacetate) respectively.The viability of podocyte was determined by MTT colorimetry.Results PAN and H2O2 significantly decreased the expression of β1 integrin and increased the synthesis of ROS in podocytes (P＜0.05respectively).Lower concentration fluvastatin or SOD treatment up-regulated β1 integrin and downregulated ROS of podocytes induced by PAN (P＜0.05 respectively).MTT revealed that lower podocyte viability was found in higher concentration fluvastatin,PAN and H2O2 groups.Lower concentration fluvastatin and SOD could protect podocytes against PAN.Conclusion Fluvastatin attenuates the injury of podocyte induced by PAN and increases the expression of β1 integrin,whose mechanism may be associated with the inhibition of the ROS activity.

Objective To explore the effect and mechanism of fluvastatin on the expression of fibronectin(FN) in human peritoneal mesothelial cells (HPMCs) induced by high-glucose peritoneal dialysate (HGPDS).Methods Cultured HPMCs were randomly divided into control,HGPDS,HGPDS plus GSK650394 10-5 mol/L (the competitive inhibitor of SGK1),different concentrations of fluvastatin,fluvastatin 10-6 mol/L and GSK650394 10-5 mol/L alone.The morphology change of HPMC was observed by light microscopy.The cellular viability was detected by MTT colorimetry.The mRNA and protein expressions of serum and glucocorticoid-inducible kinase 1 (SGK1) and FN were detected by RT-PCR,Western blotting or ELISA.Results After incubation with HGPDS,the cell morphology changed from typical cobblestone-like appearance to fibroblast-like appearance,and the cell viability was inhibited significantly (P＜0.05).Fluvastatin 10-6mol/L and GSK650394 could improved the cell morphology and the cell viability injured by HGPDS (P＜0.05).Compared with the normal control group,the mRNA and protein expressions of SGK1 and FN increased significantly in HPMC treated with HGPDS(P＜0.05).GSK650394 significantly decreased the high expression of SGK1 and FN (P＜0.05),also the fluvastatin had same effects as GSK650394 in dose-dependent manner (P＜0.05).Conclusions High-glucose peritoneal dialysate can increase FN expression in human peritoneal mesothelial cells,which can be attenuated by fluvastatin.The protective role of fluvastatin in HPMC may be partially achieved through the signal pathway of SGK1.

Objective To compare the saccharometabolism with the pancreatic islets functions and insulin resistance index in children with severe stress. Methods Thirty children with severe stress and 30 healthy children in control group were tested. The levels of fasting blood glucose (FBG), fasting insulin (FINS) and fasting C - peptide (FCP) were detected by radioimmunoassay respectively and insulin sensitivity index (ISI), insulin resistance index (IR) and fasting blood cell function index (FBCI) were calculated statistically. Results There were significant differences between the children with severe stress and the normal controls in the levels of FINS, FCP and FBG,(all P0.05). Conclusion There is insulin resistance with the significant decrease in the insulin sensitivity index and significant increase in insulin resistance index in the children with severe stress, which may cause the disorder in glucose metabolism in children with severe stress.

Non-small-cell lung carcinoma (NSCLC) is one of the most frequently diagnosed malignancies worldwide.Previous studies have shown that microRNA-449b (miR-449b) functions as a tumor suppressor in many cancers.However,the role of miR-449b in NSCLC is still unknown.In the present study,miR-449b was significantly downregulated in NSCLC samples and cell lines.Bioinformatics analysis revealed that 3'-UTR region of leucine rich repeat containing G protein-coupled receptor 4 (LGR4) mRNA had putative complementary sequences to miR-449b,which was further confirmed by the luciferase assay.Western blotting showed that restoration of miR-449b in NSCLC cells decreased the expression of LGR4.Interestingly,over-expression of miR-449b inhibited growth and invasion of NSCLC cells in vitro.Furthermore,ectopic expression of LGR4 reversed miR-449b-suppressed proliferation and invasion of NSCLC cells.Therefore,the data of the present study demonstrate that miR-449b inhibits tumor cell growth and invasion by targeting LGR4 in NSCLC.