Objective To evaluate the clinical effectiveness of the Invance sling in treating children incontinence caused by neurogenic sphincteric incompetence. Methods Two children were treated with Invance sling for their neurogenic sphincteric incompetent incontinence because of congenital myelodysplasia.Both were male,aged of 9 and 10,respectively. Results Two patients were followed up of 12 and 14 months.Both patients micturated once each 2～4 hours with 1～2 episode of incontinence by chance in the daytime and without urinary pad.The maximum bladder capacities were 163 ml and 223 ml pre-operatively and those were 164 ml and 230 ml post-operatively,the retrograde maximum urethral pressures or retrograde leak point pressures were 37 cmH_2O and 27 cmH_2O pre-operatively while those were 45 cmH_2O and 37 cmH_2O post-operatively,maximum urethral closing pressures were 37 cmH_2O and 26 cmH_2O pre-operatively while those were 40 cmH_2O and 37 cmH_2O post-operatively,the functional urethral lengths were 2.5 cm and 3.0 cm pre-operatively while those were 3.5 cm and 4.0 cm post-operatively,the post-voiding residual urine volumes were 40 ml and 30 ml pre-operatively while those were 6 ml and 25 ml post-operatively.Post-operative voiding cystourethrography demonstrated that it was pressed to narrow at the suspended in the bulbous urethra segment. Conclusions The sling in treating children neurogenic sphincteric incompetent incontinence is simple,mini-invasive.It can increase the urethral resistance and continence better.But it has specific indication and should be applied prudently.

Objective To evaluate the value of detecting chlorophyll related genes of plankton in the diagnosis of drowning. MethodsEighteen rabbits were divided randomly into three groups: death by drowning (n=10), postmortem submersion (n=6) and control (n=2). The heart blood, lung, liver, kidney and brain tissues were taken from every rabbit. After isolated plankton from tissues with percoll and extracted their DNA, the chlorophyll-related genes, including EG (EG1 and EG2) and SK (SK1 and SK2), were detected using PCR technique. Meanwhile, diatom test was also performed from lung and liver tissues by nitric acid digestion method. ResultsFor the drowning group, the specific amplification products for EG1 were detected from 9 samples in heart blood, 10 samples in lung, 9 samples in liver, 7 samples in kidney and 8 samples in brain. The products for EG2 were detected from 8 samples, 10 samples, 7 samples, 5 samples and 7 samples accordingly. There were a small number of positives in heart blood, lung and kidney with SK1 and SK2 (≤2). For the postmortem submersion group, only one case was positive from heart blood and lung tissue respectively for EG1. No amplified product was detected for EG1 and EG2 in various tissues in control group, and also no product was detected for SK1and SK2 in other groups. In addition, diatoms were detected from 9 lung and 3 liver tissues in drowning group with the nitric acid digestion, and only one sample of lung was positive in the postmortem submersion group. ConclusionThe detection rate of the chlorophyll-related gene EG with PCR method was higher than that of diatom with nitric acid digestion method in drowning victims, and it can be used as a potentially useful tool for diagnosing drowning.

Objective To describe the experiences when different methods were used to treat early-onset antibody-mediated rejection (AMR) after kidney transplantation.Methods The clinical data of 42 recipients who experienced early-onset acute AMR after kidney transplantation in our department from Jan.2010 to Apr.2016 were retrospectively analyzed.The recipients were divided into 3 groups based on different strategies against AMR:group A (plasma exchange with intravenous immunoglobin);group B (bortezomib solo),and group C (combination of bortezomib and sirolimus).Results All the AMR episodes were diagnosed by kidney biopsy 9-27 days after transplantation.The AMR reversal rate in groups B and C was significantly higher than that in group A (100％ versus 60.00％,P=0.034;100％ versus 60.00％,P=0.007).The AMR recurrence rate in groups B and C was significantly lower than that in group A (0 versus 41.67％,P =0.035;0 versus 41.67％,P =0.007).The recipient survival rate was 100％ in all the three groups.There were 11 graft losses in group A,and none in group B or C.The graft survival rate in group B at 6 months,1 year and 3 years was significantly higher than in group A (100％ versus 60.00％,P =0.034;100％ versus 55.00％,P =0.021;100％ versus 50.00％,P =0.013).The graft survival rate in group C at 6 months and 1 year was significantly higher than in group A (100％ versus 60.00％,P =0.007;100％ versus 55.00％,P =0.003).There was no significant difference in AMR reversal rate,AMR recurrence rate and graft survival rate between groups B and C.There was no significant difference in incidences of infection,hyperlipidemia and bone marrow suppression among the three groups.The incidence of diarrhea in groups B and C was significantly higher than in group A (50.00％ versus 0,P =0.001;42.86％ versus 0,P =0.001).The incidence of peripheral neuritis in group B was significantly higher than in group A (25.00％ versus 0,P =0.02),but similar to group C.There was no significant difference in average serum creatinine level among three groups within 1 year after treatment (P＞ 0.05).Antibodies against human leukocyte antigen (HLA) and donor specific antibodies were detected in all the 42 recipients before treatment.The negative conversion ratio of panel reactive antibody (PRA) in group A was significantly lower than in groups B and C (10.00％ versus 87.50％,P＜ 0.001;10.00％ versus 92.86％,P ＜ 0.001).The PRA recurrence rate in group A was significantly higher than in groups B and C (85.00％ versus 37.50％,P＜0.001;85.00％ versus 0,P＜0.001),while that in group B was significantly higher than in group C (37.50％ versus 0,P =0.014).The ratio of Treg in peripheral blood at 3-12 month after treatment in group C was significantly higher than in groups A and B (P＜0.05).Conclusion Treatment for early-onset AMR after kidney transplantation based on bortezomib might be an effective and safe strategy.Graft longterm survival might benefit from the combination of bortezomib and sirolimus.

Objective To discuss the efficiency and safety of conversion from tacrolimus(Tac)to cyclosporine A(CsA) in patients with new onset diabetes after transplantation (NODAT).Methods The glucose metabolism parameters and related clinical indicators in 45 Tac treated renal transplantation recipients who developed NODAT were retrospectively analyzed.The oral immunosuppressive strategy was Tac + mycophenolate mofetil (MMF) + prednisone(Pred).Results 32 cases were converted to CsA whereas 13 patients stuck to Tac.After conversion,fasting plasma glucose (FPG)decreased from(8.2 ± 2.7)mmol/L to(5.9 ± 1.2)mmol/L(P ＜ 0.01)and HbA1c level decreased from (7.0 ± 0.9) ％ to (6.1 ± 0.7) ％ (P ＜ 0.05).The level of FPG and HbA1c was lower in the conversion group than in the control group(P ＜ 0.05).During the 1-year follow-up,the curative rate of NODAT was 53.1％ (17/32) in the conversion group while it was 0％ in the control group.No acute rejection happened after the conversion.There was no obvious change in renal function.The 1-year survival rate of patient and the transplanted kidney was 100％.Blood pressure and lipid levels were stable after the conversion.Conclusion Conversion from Tac to CsA is a simple and effective strategy to improve glucose metabolism in renal transplantation recipients with NODAT.

In the present study ,we aimed to observe the effect of curcumin on TSST-1-induced inflammatory cytokines in splenocytes of mouse and provide evidence for the further study on the effect of curcumin on inflammatory shock .Lactate de-hydrogenase (LDH) release assay was used to determine cytotoxicity of different doses of TSST-1 and curcumin .Inflammatory cytokines were determined by ELISA and flow cytometry .The doses of TSST-1 and curcumin we used in the present study did not cause significant cytotoxicity .TSST-1 induced higher level of IFN-γand IL-2 production but relatively lower level of TNF-α.The production of IL-1β,IL-6 and IL-12 was undetectable .TSST-1 induced Th1 cytokines (IFN-γand IL-2) were not IL-12-dependent which was different from LPS-induced IFN-γ.Curcumin significantly reduced IFN-γand TNF-αproduction at the concentration of 15 umol/L (P<0 .05) ,but had no effect on IL-2 production (P>0 .05) .It’s suggested that curcumin could significantly inhibit the production of IFN-γand TNF-αby splenocytes induced by TSST-1 ,but could not affect the prolifera-tion of T cells .

ObjectiveTo investigate the relationship of blood glucose,blood lipids,serum uric acid,fibrinogen level and prognosis of patients with acute lacunar cerebral infarction.MethodsClirical data of 127 cases with acute lacunar cerebral infarction were retrospectively analyed.Patients with normal blood glucose,blood lipid,uric acid and fibrinogen level were enrolled into A group of 64 cases,63 cases with abnormal levels were selected as group B,an average of 2 years of follow-up,observing the two groups of neurologic,re-infarction incidence and prevalence of vascular dementia were obsenved in two groups.ResultsAfter 4 weeks of treatment,A group of neural function defect score(9.6 ±7.3) was significantly lower than group B( 14.3 ±7.4) (t =3.8696,P ＜0.05) ;95.4％ total effective rate in A group was significantly higher than B group of 82.0％ ( x2 =5.8623,P ＜ 0.05 ).After an average follow-up of 2 years,A group of recurrent cerebral infarction rate( 18.8％ ),vascular dementia rates(7.8％ ),were significantly lower than group B(28.6％),17.5％(11/63)(x2 =6.5421,5.9623,P＜0.05).ConclusionA number of abnormal serum biochemical indexes had a significant impact on short and long-term prognosis of acute lacunar cerebral infarction.

Objective To study the type variation of microglial activation in spinal dorsal horn of rats after sciatic nerve injury.Methods Healthy adult male Sprague-Dawley rats were randomly divided into the control and experimental groups, 24 rats in each group.The experimental group underwent ligation of sciatic nerve trunk to generate nerve injury in the rats.The pain behavior in the rats was measured at the 1th, 7th and 14th postoperative days, and the changes of microglial activation in the rat lumbar spinal cord dorsal horn was detected by immunofluorescence staining.qRT-PCR assay was used to validate the activation trends of M1 and M2 types of microglia cells.Results No significant changes were found in the microglial cells in the spinal cord dorsal horn of rats in the sham-operation group during 14 days after operation.In the sciatic nerve ligation group at 1 day after operation, no significant change was observed in the number of microglial cells, but the expression of marker of M1 microglia was significantly increased.At 7 and 14 days after operation, the number of microglial cells and the expression of M1 microglia marker in the spinal cord dorsal horn were increased significantly.Conclusions Microglia activation in the spinal dorsal horn starts at the first day after sciatic nerve injury, and lasts at least for two weeks after the operation.M1 microglia activation dominates during this period.

Aim To study the effects of propofol on apoptosis of cortical astrocytes isolated from neonatal rats.Methods Pure astrocytes(AST)were obtained from the cultured cerebral cortical cells and identified by the GFAP stain technology in neonatal rats.AST cells were treated with different concentrations of propofol(0,10,30,90 μmol·L-1)for 8 hours.Cell viability was measured by MTT method,and the apoptosis was detected by Annexin V/PI double staining.Cytochrome C(cyt-C)leakage was detected by Western blot.Caspase-3 and caspase-9 activities were measured by spectrophotometry.Results AST cells were administered with different concentrations of propofol(0,10,30,90 μmol·L-1)for 8 hours.It decreased the survival rate in a concentration-dependent manner,induced the leakage of cyt-C in mitochondria,up-regulated the expression of pro-apoptotic protein caspase-3 and caspase-9,and induced AST apoptosis.Conclusion Propofol induces the apoptosis of astrocytes in neonatal rat cortex in vitro,which may be related to the activation of mitochondria apoptosis pathway induced by mitochondrial cyt-C release.

Objective To explore the role of spinal cord p300 in neuropathic pain induced by chronic constriction nerve injury (CCI) in rats.Methods Thirty two Sprague-Dawley (SD) rats were randomly divided into sham and CCI groups,14 days after surgery,immuno-fluorescence staining and Western blot were used to detect distribution and expression of p300 protein.After rats were successfully implanted with an intrathecal catheter and accepted CCI surgery,another 24 rats were randomly divided into three groups (n =8):dimethyl sulfoxide (DMSO) group,p300 acetyltransferase inhibitor C646 group,and control C37 group.Each rat were administered through the intrathecal catheter from day 7 to 14 and mechanical withdraw threshold were tested.Results (1) The p300 positive cells were detected mainly in neurons,and p300 protein in spinal cord of CCI group were significantly higher than sham group (P ＜0.05).(2) C646 alleviated significantly neuropathic pain in rats,without significant changes in pain threshold after injection of C37 and DMSO.Conclusions The p300 protein in spinal cord was involved in the development of neuropathic pain in rats,the mechanism may be referred to its acetyltransferase activity.

BACKGROUND: The investigation of culturing, passaging and establishing human limbal stem cells can strengthen the recognition of the stem cells and provide the enough cellular reserve for the basic and clinical research of limbal stem cell transplantation.OBJECTIVE: To explore a method of pessaging and establishing cell line of human limbal stem cells cultured in vitro.DESIGN: Randomized controlled observation.SETTING: Gannan Medical College.MATERIALS: The experiment was performed at Scientific Center of Gannan Medical College and the National Key Laboratory of Ophthalmology Hospital Affiliated to Sun Yat-Sen University from June 2003 to April 2004. Fresh human limbus corneae were isolated from two healthy donors. Procedures were performed according to the informed consent of the donors. Main reagents contained RPMI-1640 (Sigma R8755, containing L-glutamine) and 200 g/L fetal calf serum (FCS) (Gibco 16140-071). DMEM medium, chondroitin sulfatase and human epidermal growth factor (hEGF) were purchased from Sigma Co. USA; HEPES and DMSO were bought from Gibco, USA; 100% glycerinum was purchased from Yunjia Huangpu Pharmaceutical Product Limited Company, PRC; glutaraldehyde was bought from E.Merk, Germany; Alcohol, chlorhydric acid, acetone and methyl aldehyde were purchased from Beijing Chemical Agent Company, PRC; 0.25% parenzyme was bought from Shanghai Xinhua Pharmaceutical Factory, PRC.Above-mentioned reagents were analytical pure grade.METHODS: After digestion, human limbal tissues in limbal basilar part with an abundant pigment were cultured in the culture flask containing RPMI-1640 and 200 g/L FCS and in culture dish containing amniotic extracellular matrix (AECM) as the cultural supporter. Primary and passage cells were observed under light microscope and scanning electron microscope (SEM). The revival ratio of stem cell refrigeration of every generation was calculated by the trypanblau exclusion experiment.MAIN OUTCOME MEASURES: ① Observational results of limbal stem cells during the primary culture and serial subcultivation in vitro, and ② revival ratio of stem cell refrigeration.RESULTS: ①Findings of primary culture: Most limbal stem cells in the culture flask had the adherence and were arrayed uniformly sparsely to form monolayer and adhered to the bottom of culture flask under the inverted phase contrast microscope after 1-day culture. ② Findings of serial subcultivation: After human epidermal growth factor (hEGF) was added into the second passage, cells were scattered into the monolayer and adhered to grow quickly.Morphological variability of all the cells increased obviously when passage the 30th generation. The cellular volume was obviously increasing, and the round or irregular round cells gathered together. The 33rd generation human limbal stem cells still could vigorously differentiate, proliferate and grow in ACEM. ③ The revival ratio of stem cell refrigeration was 82.2%.CONCLUSION: The human limbal stem cell lines were preliminarily established by culturing and freezing the cells of 33 generations in vitro. The human limbal stem cell lines preferred to grow in the culture dish containing AECM as the cultural supporter.