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

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 .

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

siRNA drug research and development is becoming one of the main objectives in the future. However, due to the in-stability of siRNA and the complex environment in vivo, the safe and effective delivery of siRNA is limited in vivo. Thus, special vectors are used to assist siRNA to express biological effects. This paper reviews the advances in non-viral vector for delivery of siRNA in vivo.

Objective To summarize the clinical features and treatment of male urethral duplication.Methods The clinical data of 2 cases treated in June 2011 and April 2014 were analyzed retrospectively.The first case was a 5-year-old boy presented with passages of urine from two orifices in the penis.The second case was a 15-year-old boy presented with dorsal chordee and a sinus on the dorsum of the penis.The patient had a small amount of watery discharge occasionally dripping out of the opening for 10 years.The 2 patients underwent retrograde urethrography, which revealed a complete duplicated urethra with the channel arising from the proximal prostatic urethra ( class ⅡA2 according to the classification of Effman) .The 2 patients underwent excision of the accessory urethra under general anesthesia.Results The pathology reports of the 2 cases were hyperplasia of squamous epithelium and urothelial mucosa.Pathological diagnosis was urethral duplication.The first case was followed up for 1 year with a satisfactory functional and cosmetic outcome.The second case was followed up for 6 months and no watery discharge noticed from the residual dorsal chordee.Conclusions Urethral duplication is a rare congenital anomality affecting mainly boys.Clinical presentation varies depending on the different anatomical patterns of the urethral anatomy. Surgical management must be evaluated for each different anatomical variation.

Purpose To evaluate the correlation between T2-FLAIR hyperintense vessel sign (HVS) and the stenotic degree of internal carotid artery (ICA) and assess the HVS changes after the carotid endarterectomy (CEA). Materials and Methods Fifty-one patients with CEA were retrospectively enrolled. The stenosis of the bilateral ICA were as:≥90%, and<90%. The distribution of HVS locations was classified as three regions:sylvian fissure, sulci of temporo-occipital lobe and other areas. The presence and the location of HVS were counted. The extrension of HVS on T2-FLAIR were graded as:I:the presence of HVS was<1/3 of the MCA territory, II:the presence of HVS was≥1/3 of the MCA territory.χ2-test was performed for correlation between HVS and ICA stenosis. The difference of HVS and stenosis of ICA and their effects on CEA was accessed. Results HVS was significantly higher in the ICA stenosis more than 90%group than in the less than 90% group (χ2=23.584, P<0.001). The frequencies of HVS were 12, 34 and 15 in sylvian fissure, sulci of temporo-occipital lobe and other area, respectively. The proportion of grade II HVS was higher in the ≥ 90% group than in the<90% group (χ2=8.395, P<0.05). After CEA, HVS on 29 affected hemispheres were showed to be disappeared (n=24) or remained (n=5) in the treated side. Conclusion The presence and the grade of HVS were correlated with the stenotic degree of ICA. In the patients with ICA stenosis, HVS was most frequently found in the sulci of temporal lobe and occipital lobe, and seldom found in sylvian fissure. HVS disappeared after CEA indicating that HVS can be considered as a marker for CEA treatment.