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 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.

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

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 observe the preventive and therapeutic effect of Chinese herbal medicine film µwave irradiation on mammary hyperplasia in rats. Methods Rat models of experimental mammary hyperplasia were established by intramuscular injection of estrogen and a small quantity of progestin. Two weeks after modeling, the rats were treated with Chinese herbal medicine film µwave irradiation. Before and after the treatment, mammary diameter of the second nipples were measured, serum concentrations of estrogen (E2), progesterone (P) and prolactin (PRL) were determined by radioimmunoassay,pathological changes of mammary hyperplasia were examined under microscope,and the rate of mammary hyperplasia was measured. Results Chinese herbal medicine film markedly inhibited experimental mammary hyperplasia in rats,and the rate of mammary hyperplasia were 23.4 %in medicine film group and 20.9 %in medicine film µwave irradiation group respectively. Medicine film regulated the concentrations of estrogen and progestin,reduced the level of PRL and alleviated the pathological severity of mammary hyperplasia obviously. Conclusion Chinese herbal medicine film µwave irradiation have certain preventive and therapeutic effect on mammary hyperplasia in rats.

Background and purpose:Now 3-dimensional conformal radiotherapy(3DCRT)and intensitymodulated radiotherapy(IMRT)are widely used in the treatment of head and neck tumor.For the parotid,this target area is located on the side of the head and the tumor has a concave shape.What kind of radiation method can be used more eriectively to achieve dose uniformity and protection organs at risk is the topic of much discussion.The postoperative irradiation of parotid tumor is varied in the techniques used.In this study,3-dimensional conformal radiotherapy (3DCRT),intensity-modulated radiotherapy(IMRT)and simplified forward planned multi-segment radiotherapy (MSRT)were compared to conventional planning techniques in order to investigate the potential advantages of these new treatments.Methods:The conventional planning included the large opposed lateral fields with 2 or 3 weight ratio at the target lateral(2F-2D)and the unilateral field with mixture of 6 MV photon and electron beams(X+E).The 3D techniques included 3DCRT,MSRT and IMRT.Their dose distributions were calculated and compared for 8 patients treated in our center.Different beam arrangements were used for 3D techniques.In each case.the dose of PTV was prescribed to 60 Gy.All plans were compared using dose-volume histogram data.The conformity index(CI)and heterogeneity index(HI)of dose were used to evaluate the dose coverage of the target volume.Dose sparing of brain stem.spinal cord and the contra lateral parotid was also compared.To compare IMRT and MS RT,the timing ofplanning and radiation delivery was recorded.Results:Compared to conventional planning,the 3DCRT,MSRT and IMRT plans produced adequate target coverage,and the CI showed 3DCRT plans(0.78)produced poorer target coverage than MSRT(0.81)and IMRT(0.85).MRST and IMRT plans showed a significant reduction in maximum dose to the spinal cord,brainstem and the contra lateral parotid,compared to the conventional plans,while the 3DCRT plan did not show significant sparing of these structures.MSRT and IMRT plans produced better dose coverage among all the techniques.The efficacy of beam delivery comparing between two modulated planning showed MSRT was better.Conclusion:For postoperative irradiation of parotid cancer,3D planning techniques generated better target dose-coverage,without compromising the dose-sparing advantages of important structures.A satisfactory dose distribution can be obtained using MSRT and such a simple technique may be suitable for replacing IMRT.

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.