Objective To explore the silence of B-cell specific Moloney murine leukaemia virus insertion site 1(Bmi-1)by RNA interference on the proliferation of human leukemia cell line K562 and its mechanisms.Methods Small interfering RNA(siRNA)targeting Bmi-1 gene was designed and double-stranded siRNA was chemically synthesized.After double-stranded siRNA was transfected into K562 cells with Lipofectamine 2000,the proliferation of K562 cells was detected by MTT colorimetry,cell cycle was determined by flow cytometry,and the expression of Bmi-1 and P16 were analyzed by Western blotting.Results The siRNA targeting human Bmi-1 gene effectively prolonged the double time of K562 cells,increased the percentage of cells at G1 phase,and the expression of Bmi-1 was significantly down-regulated but the expression of P16 was up-regulated.Conclusion The siRNA targeting human Bmi-1 gene inhibits the proliferation of K562 cells,and up-regulates the expression of P16 in the cells.

To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cell (HKC), in vitro cultured HKC cells were divided into 3 groups: negtive control, low dose CTGF-treated group (rh CTGF, 2.5 ng/ml) and high dose CTGF-treated (rhCTGF, 5.0 ng/ml). Then the expression of alpha-smooth muscle actin (alpha-SMA) were assessed by indirect immuno-fluorescence, and the percentage of alpha-SMA positive cells were assessed by flow cytometry. RT-PCR were also performed to examine the mRNA level of alpha-SMA. Upon the stimulation of different concentrations of rhCTGF, the expression of alpha-SMA were markedly stronger than that in negative controls. The percentages of alpha-SMA positive cells were significantly higher in the stimulated groups than that of negative controls (38.9%, 65.5% vs 2.4%, P<0.01). alpha-SMA mRNA levels were also up-regulated by the stimulation of rhCTGF (P<0.01). These results suggest that CTGF can promote the transdifferentiation of human renal tubular epithelial cells towards myofibroblast (Myo-F).
Actins/metabolism ; Cell Differentiation/*drug effects ; Cells, Cultured ; Epithelial Cells/*cytology ; Immediate-Early Proteins/*pharmacology ; Insulin-Like Growth Factor Binding Proteins/pharmacology ; Intercellular Signaling Peptides and Proteins/*pharmacology ; Kidney Tubules/*cytology

Objective To explore whether Cl2MDP-loaded gelatin particles can induce the depletion of macrophage or can depress the immunity of macrophage in SD rat models of immune thrombocytopenic purpura(ITP) to treat the ITP rats.Methods SD rats were intravenously injected with 150 ?L of anti-platelet serum once per 24 hr to induce the ITP model.The platelet count was persistently less than 50?109L-1 in these models.After intravenous injection of a group dose of Cl2 MDP-gelatin particles,the platelet counts of these models were examined at the time of 4 hr,24 hr,48 hr,72 hr and 96 hr,respectively.The pathological change of macrophages in spleen was observed by electron microscopy.Results Cl2MDP-loaded gelatin particles can significantly increase the platelet counts of ITP models from less than 50?109L-1 to mean 180?109L-1 in 24 hr after injection.The pathological changes of macrophages were seen under electron microscope,such as degeneration, necrosis and so on.Furthermore,rats pre-treated with Cl2 MDP-loaded gelatin particles can evade the decrease of platelet counts when they were injected anti-platelet serum.Conclusion Cl2 MDP-loaded gelatin particles can promptly,effectively restoreplatelet counts of ITP models to physiological level with a dose dependent manner.So,the targeting therapy of drug-loaded gelatin particles offers a new approach to treat ITP.

Objective:To investigate the value of 64-slice spiral computed tomography (64SCT) in preoperative evaluation of spinal vascular intervention.Methods: Seventeen patients with spinal vascular malformations(SVM) scheduled for spinal vascular interventional therapy in our hospital between January 2012-December 2014 were collected, all received 64-detector spiral CT angiography (64-MDCTA) before surgery, image postprocessing technologies such as volume rendering (VR), maximum intensity projection(MIP) and multiplanar reconstruction(MPR) were applied to conduct three-dimensional reconstruction, the traveling case of spinal cord feeding artery was analyzed and were compared with DSA test results.Results: All of the 17 patients clearly showed the abnormality of spinal cord blood vessels and lesions scope, the SVM classification and lesions scope were exactly the same as that of DSA and intraoperative performance, the coincidence rate was 100%;1 case of patient was not shown fistula, the rest of the patients clearly shown the draining veins around the spinal cord, open position and flow tendency of radiculomedullary artery, the coincidence rates were 100.00%, 94.12%%, 100.00%.Conclusion: 64-MDCTA is important for SVM patients to preoperative evaluation of spinal vascular intervention, for it can fully and accurately reflect the open position and flow tendency of radiculomedullary artery, spatial relationship with the surrounding vessels, spinal cord and vertebral situation, should be widely applied.

Objective To observe the changes of the blood culture results before and after implementing the quality control on blood culture .Methods The blood culture results in 1 year before implementing the quality control and in 1 year after implementing the quality control were performed the contrastive analysis .Results After implementing the quality control on blood culture ,the positive rate of blood culture was elevated by 2 .4% and the pollution rate was decreased by 8 .6% after the quality control .Conclu‐sion Implementing the total quality control on blood culture can improve the positive rate of blood culture and reduces the pollu‐tion rate of blood culture .The determination of contamination bacteria must combine the laboratory detection data with the clinical situation for ensuring to provide accurate and effective antimicrobial agents for clinic .

Objective To investigate the role of connective tissue growth factor (CTGF) in transdifferentiation of human renal tubular epithelial cells (HKC).Methods Cultured HKC were divided into 3 groups: (1) negative control; (2) low dose CTGF treated (rhCTGF 2. 5 ng/ml); (3) high dose CTGF treated (rhCTGF,5. 0 ng/ml). Expression of ?-smooth muscle actin (?-SMA) and fibronectin(FN) mRNA were measured by RT-PCR. Indirect immunofluorescence and flow cytometry methods were used to assess the level of intracellular ?-SMA protein. Concentration of FN secreted into the media was determined by ELBA. Results Upon the stimulations of different concentrations of rhCTGF,expression of ?-SMA and FN mRNA increased markedly(P

OBJECTIVETo discuss the applicatioen of flap for the urethroplasty in the distal penile segment in the staged correction of severe hypospadias.

METHODSFrom Oct. 2004 to Aug. 2014, 25 cases with severe hypospadias were treated by staged urethroplasty for urethra reconstruction. The urethral meatus were located at peroscrotum in 4 patinets, at scrotum in 8 patients and at perineum in 13 cases. At the first stage, the urethral plate was divided and chordee was corrected. Then tubularized transverse island flap was used to prefabricate partial distal urethra. The defective urethra was repaired by using the Thiersch-Duplay principle at the second stage.

RESULTSAll patients completed both stages of the operation. The follow-up duration was 6-72 months (average, 24 months). In the first-stage, the modified tabularized transverse preputial island flap was performed on 10 patients, whereas the modified preputial double-faced island flap was performed on the other 15 patients. All of the prefabricated partial distal urethras had no evidence of stenosis or scarring. The result of the second-stage procedure was a complete penis with integrated urethral. All patients were satisfied with cosmetic and functional results. Neither stricture nor diverticula was observed. A good urinary stream during the urination was achieved in 19(19/ 25, 76%) patients. Five cases (5/25, 25%) developed urethrocutaneous fistula and one cases developed meatal stenosis (1/25, 4%) after the second stage repair.

CONCLUSIONSStaged urethroplasty using flap is a good choice for severe hypospadias. The successful rate is relatively high with good cosmetic and functional results.

Cicatrix ; Follow-Up Studies ; Humans ; Hypospadias ; surgery ; Male ; Penis ; surgery ; Perineum ; Reconstructive Surgical Procedures ; methods ; Scrotum ; Surgical Flaps ; Time Factors ; Urethra ; abnormalities ; surgery ; Wound Healing

A potential pathological role of angiopoietins (Ang) in glomeruli following podocyte injury-induced progressive glomerulosclerosis was explored. Eighty male Wistar rats were randomly allocated into sham operation group (Sham, n = 25), Uninephrectomy group (UPHT, n = 25) and Uninephrectomy+Daunorubicin group (DRB, n = 30). In DRB group, daunorubicin (5 mg/kg) was injected via tail vein on the 7th and 14th day after uninephrectomy. At week 1, 2, 4, 6 and 8 respectively following establishment of the animal model, 5 rats in Sham group and UPHT group, and 6 in DRB group were taken respectively for determining 24-h urinary protein excretion rate (24hUPER), blood urea nitrogen (BUN) and serum creatinine (Scr). The sections of kidneys were examined by an electric microscope, PAS staining, immunohistochemical staining and in situ hybridization histochemistry. The results showed that 24hUPER, BUN and Scr in DRB group were more than those in Sham group and UPHT group at the same time points, and there was a trend towards an increase on level of GSI in DRB group from week 2 to week 8. Electric microscopy revealed that podocyte injury presented in DRB group. The expression of Ang1 mRNA and protein in glomeruli of DRB group was decreased, while the expression of Ang2 protein in glomeruli of DRB group increased. Meanwhile, the expression of Ang1 mRNA had a negative correlation with the expression of Ang2 mRNA, and the expression of Ang1 protein had a positive correlation with the expression of Ang1 mRNA, and had a negative correlation with 24hUPER, BUN, Scr, glomerular sclerotic index (GSI), the expression of Ang2 protein and CoIV protein. The expression of Ang2 protein had a positive correlation with the expression of Ang2 mRNA, and had a positive correlation with 24hUPER, BUN, Scr, GSI, the expression of CoIV protein. It was concluded that podocyte injury might lead to an alteration in the expression of Ang1 and Ang2 within glomeruli. Ang2 may get rid of inhibition from Ang1 for downregulation of the Ang1 expression, which facilitate upregulation of the Ang2 expression in glomeruli to promote progressive glomerulosclerosis in the rats.

To investigate the role and mechanisms of apoptosis and apoptosis signaling pathway in 5/6 nephrectomy rat model (SN(x)), the mRNA and protein levels of caspase-3, -8, -9 and apoptosis were detected by in situ end labeling (TUNEL), immunohistochemistry, RT-PCR, Western-blotting 1, 2, 4, 8, 12, 16, 26 and 40 weeks after 5/6 nephrectomy rat model was made respectively. The rats in the model group developed glomerular sclerosis and renal interstitial fibrosis. The number of the apoptototic cells in glomeruli, renal tubule and renal interstitium was remarkably higher in the model group than that in the control group (P < 0.05, P < 0.01). Changes of mRNA and protein level of caspase-3, -8, -9 had the same tendency and was up-regulated wavily in the rat model compared with the control group (P < 0.05). Peaks in model appeared on the 4th and the 40th week respectively. The growth amplitude of caspase-9 was remarkably higher than that of caspase-8. It is concluded that the development of 5/6 nephrectomy rat model was correlated with the apoptosis of glomeruli, renal tubule and renal interstitium. Both of death receptor and mitochondria signaling pathways are involved in the process and the latter might play a primary role.

The changes of Toll-like receptor (TLR) 2, 4 of peripheral blood mononuclear cells (PBMCs) in the acute abdomen patients associated with systemic inflammatory response syndrome (SIRS) and their potential significance were explored. A clinical study was performed on 103 acute abdomen patients in whom 65 were associated with SIRS. Forty healthy individuals served as normal controls. The mRNA expression of TLR2, 4 was detected by RT-PCR, and the expression of TNF-alpha and IL-6 by ELISA. The level of plasma endotoxin, hospital stay and mortality were measured. It was found that the endotoxin level was increased to varying degrees in all the acute abdomen patients, and the endotoxin level was and hospital stay longer in SIRS group than in non-SIRS group (P<0.01). TLR2 mRNA, TLR4 mRNA, IL-6 and TNF-alpha could be detected with low value in normal controls, but they were up-regulated markedly on the 1st day after admission. Then TLR4 mRNA, IL-6 and TNF-alpha were decreased gradually, but TLR2 mRNA maintained at a high level till the 5th day. These indexes above in SIRS group were higher than those in non-SIRS group (P<0.01). The results of correlation analysis revealed the expression of TLR2, 4 mRNA was positively correlated with the levels of TNF-alpha and IL-6, and the hospital stay. The results of Logistic regression demonstrated that overexpression of TLR2, 4 mRNA might result in higher risk of multiple organ dysfunction syndrome (MODS). It was concluded that in the acute abdomen patients associated with SIRS, the expression of TLR2, 4 in PBMCs was increased markedly, suggesting that TLR might play an important role in the pathogenesis of acute abdomen associated with SIRS.