Abstract:

Abstract:As an innovation in minimally invasive surgery, transumbilical laparoendoscopic single-site surgery has emerged recently; it follows the concept of natural orifice transluminal endoscopic surgery (NOTES) and has gained rapid progress in urology, which has promoted the advancement of concept and technique of urological surgery and yielded some valuable experience. This paper discusses the retrospects and prospects of the technique by introducing the related animal experiments, application statuses home and abroad, difficulties and advantages, future trend, and so on, hoping to guide the clinical practice.

Abstract:Laparoendoscopic single-site surgery (LESS) is technically challenging; it can reduces instrument triangulation and robust retraction and is associated with a steep learning curve. The instruments of the daVinciR○ surgical system (Intuitive Surgical) are designed with seven degrees of motion mimicing the dexterity of the human hand and wrist. This inherent feature of the robotic arm provides superior ergonomics when performing LESS, especially for complex reconstructive surgery. This review analyzes the evidence supporting current and future application of robotic technology in the field of urologic LESS.

Abstract:Laparoendoscopic single-site surgery (LESS) and natural orifice transluminal endoscopic surgery (NOTES) are gaining the momentum of mini-invasive surgery, but many technical problems remain to be solved; difficulties in learning the technique hamper its popularization. A satisfactory training system can greatly promote the development of the new technique. In this paper, we introduce the current status of training for LESS and NOTES, and also introduce some valuable experience in our training courses.

Abstract:Objective To use Spider system, a novel laparoendoscopic single-site (LESS) platform, for partial nephrectomy in pigs, so as to evaluate the feasibility and ergonomic features of the Spider system. Methods A 2 cm incision was made on the lateral border of the rectus muscle; the subcutaneous tissues and muscle layer were dissected bluntly by the Kelly clamp. With the outside sheath the Spider system was introduced into the peritoneal cavity. After establishment of pneumoperitoneum, a 5 mm scopy was introduced. The whole Spider platform was fixed using the suspension system on the bedside. The procedures of partial nephrectomy were performed according to the conventional steps. Renal reconstruction was done by the 2-0 V-loc barbed suture using the knotless closure technique. Results The procedure of partial nephrectomy was successfully performed in 2 cases, with the operation time being 100 and 102 min, the docking time being 7 and 11 min, and the time periods of warm ischemia being 23 and 28 min. The V-loc barbed suture was used to close the defect of kidney without bleeding after unclamping the renal artery. Conclusion This novel Spider LESS platform is safe and feasible for partial nephrectomy in pigs, which better solves the problem of “cross manipulation” in single-port laparoscopy, avoiding external instrument clashing; but it still has many drawbacks like the lack of strength and precise maneuverability of the tips, which may hamper the visual field in the operation.

Abstract:Objective To make attempt for “no scar” cystectomy by pure LESS (laparoendoscopic single-site) surgery and transurethral NOTES (natural orifice transluminal endoscopic surgery) in pigs, so as to assess the feasibility and summarize the experience. Methods Female pigs weighing 30 kg were given general anesthesia. A 2 cm incision was made through the right lateral border of the rectus muscle, and the subcutaneous tissues and muscle layer were dissected bluntly with the Kelly clamp. The SILS port was introduced into the peritoneal cavity and the pneumoperitoneum was established. The bladder was dissected by the flexible forceps and scalpel. And the ureters and urethra were divided. Under the guidance of ureteroscopy, a 12 mm trocar was introduced into the urethra. Through the transurethral access, the small intestine was divided by two Endo GIAs. The lateral-lateral anastomosis of intestine and the left side of ureter-intestinal anastomosis were performed with assistance of the transurethral access. After the specimens were retracted, the urethra was closed by 2-0 suture. After the SILS port was removed, the right ureter and the small intestine for the stoma were pulled out of the peritoneal cavity. The right side of ureter-intestinal anastomosis was done outside of the body. Then the stoma was performed. Results Three cases were performed in a mean of (275±63) min (ranging from 210-335 min) without additional trocars. Except for the stoma,there were no other incisions on the surface of the abdominal wall. Conclusion This “no scar” LESS radical cystectomy can be performed successfully with assistance of the transurethral access in pigs, but the operation is technically difficult.

Abstract:Objective To compare the therapeutic outcomes of laparoendoscopic single-site (LESS) radical nephrectomy and standard laparoscopy, so as to understand the clinical advantages of transumbilical LESS radical nephrectomy. Methods Between Sep. 2008 and Jan. 2011, a single urologist in our center performed 105 transperitoneal laparoscopic radical nephrectomies, including 21 transumbilical LESS radical nephrectomies (Group A, Sep. 2009 and Jan. 2011). This group was subsequently matched to 21 standard laparoscopic RN procedures (Group B, Sep. 2008 and Mar. 2010). The perioperative outcomes and follow-up information, including scores of cosmetic satisfaction, were retrospectively analyzed in the two groups. The two groups were matched in patient age, body mass index, tumor size and location, and surgical indication (T1 stage). Results The LESS group had significantly quicker bowel function recovery (\[31.6±17.98\] h vs \[42.3±19.94\] h, P<0.05), lower postoperative pain score (3.5±0.84 vs 4.1±1.06, P<0.05), and a better cosmetic satisfaction score (8.2±0.71 vs 7.3±0.85, P<0.05). Notably, the mean operation periods for the first 10 cases were significantly different between the two groups(P=0.030), but not significantly for the last 10 cases (P=0.495)and for the whole group (P=0.076). Both groups remained metastasis- and recurrence-free during mean follow-up periods of (8.4±4.41) and (20.0±3.95) months(P=0.000). Conclusion The transumbilical LESS radical nephrectomy offers a better postoperative pain control, quicker recovery of bowel function, and a better cosmetic satisfaction compared with standard nephrectomy, but its therapeutic effect needs to be verified by long-term follow-up. The learning curve of this procedure appears not so steep for an experienced laparoscopist following a strict specialized training course.

Abstract:Objective To treat benign prostate hyperplasia (BPH) by single-port transvesical enucleation of the prostate (STEP) and to report our preliminary experience. Methods We performed STEP in 11 BPH patients, who had a mean age of 72.3±4.61 (ranging 67-80) years and a mean body mass index (BMI) of 22±3.81 (ranging 18-29) kg/m2. They had a mean prostate volume of 74.3±27.9 (ranging 19.9-116.9) ml (by ultrasonography), a mean IPSS of 27.1±4.89 (20-35), and a mean postvoid residual of 308.3±283.6 (18 to 1 000) ml. The mean maximum urinary flow rate was 8.42±4.09 ml/s (ranging 3.5-15.7) in 6 patients (not measured in others due to acute urinary retention in 4 patients and chronic obstructive renal failure in 1 patient). The mean baseline prostate-specific antigen (PSA) level was 6.41±2.89 (ranging 2.31-12.15) ng/ml before operation; prostate biopsy showed that 6 patients with PSA level ≥4 ng/ml had BPH. Under general anaesthesia, a 2 cm skin incision was made just below the umbilicus, the white line was incised and the peritoneum was pushed up to expose the dome of the bladder. The novel single-port device was inserted percutaneously into the bladder with assistance of cystoscopy. After establishment of pneumovesicum, the prostate adenoma was enucleated along the inner margin of its surgical capsule with a flexible scissor and extracted via the port. After suturing the bladder fissure, a three-way Foley catheter and retropubic tube were left for drainage. Results Open conversion was necessary in one case due to failure of single-port device insertion. The STEP was completed in 10 cases, with the mean operative duration being 155±30.1 (ranging 120-210) min and the estimated intraoperative blood loss being 355±288.1 (ranging 50-900) ml. One case was transfused with an amount of 1 200 ml, without any other severe complications. The mean specimen volume was 36.8±20.2 (ranging 12-76) ml, and the specimens were identified as BPH by pathological examination. Continuous bladder irrigation was used in all cases, with a mean period of 2.7±0.95 (ranging 2-4 days). The patients had a mean retropubic drainage for 2.4±0.92 (ranging 1-4) days, a mean hospital stay of 7.1±1.73 (ranging 5-11) days, and a mean duration of catheterization of 2 weeks. All 10 patients (excluding the one converted to open prostatectomy) were voiding spontaneously after the catheter retraction. The patients were followed up for 1-3 months after surgery, and it was found that the mean maximum urinary flow rate was 23.2±5.59 (ranging 16.4-26.9) ml/s, with a postvoid residual of 38.3±13.7(ranging 10-67) ml and an International Prostate Symptom Score(IPSS) of 2.1±0.86 (ranging 1-4). No patient had developed urinary incontinence, retention or dysuria. Conclusion Our initial experience shows that STEP is a safe, effective and feasible procedure, with advantages of less trauma and more cosmetic benefit, but its clinical effect needs to be observed by prospective comparative studies with a longer follow-up.

Abstract:Objective To present our initial experience of single-port multi-channel laparoscopic urologic surgery for the transumbilical laparoscopic implantation of long-term peritoneal dialysis catheters. Methods On May 17th and August 18th ,2009, 2 patients underwent single-port multi-channel laparoscopic surgery for chronic renal failure (CRF) due to benign arteriolonephrosclerosis and type 2 diabetes mellitus(DM). The flexible laparoscopic instrumentation with a homemade single port and multichannels was used for the transperitoneal approach. Results The 2 cases were completed without conversion to standard laparoscopic or laparotomy. The operation time for implantation was 15-25 min, and the estimated blood loss was about 20 ml. The patients were discharged 1 day after operation and were followed up for 6-8 months. Conclusion Single-port multi-channel laparoscopy is feasible, safe, and effective for transumbilical laparoscopic implantation of long-term peritoneal dialysis catheters, with the advantages of minimal incision and better cosmetic benefit.

Abstract:Objective To present our experience on the first 100 cases receiving urological laparoendoscopic single-site surgery(LESS). Methods Between Dec. 2008 and May 2011, a cohort of 98 patients received a total of 100 TriPortTM LESS urologic surgeries for various indications, including 2 patients undergoing bilateral surgeries. The proceducres included radical nephrectomy (n=24), partial nephrectomy (n=3), simple nephrectomy (n=10), kidney cyst excision (n=21), adrenlectomy (n=12), transvesical simple prostatectomy (n=9), ureterolithotomy (n=6), and others (n=15).The demographics, perioperative outcomes, and follow-up data were prospectively analyzed. Results Of the 100 surgeries, 87 were smoothly completed. A 5 mm ancillary trocar was required in 9 cases. Conversion to standard laparoscopy and open surgery were needed in 1 and 3 patients, respectively. The overall complication rate was 15%, with 6% intraoperatively and 9% postoperatively. The complication rate was 5% for the first 50 cases and 10% for the second 50 cases. The patients were followed up for 9.9±5.61 (ranging 3-24) months. At the latest follow-up, all patients remained clinically healthy with a delightedly hidden scar. No evidence of recurrence or metastasis was detected in patients receiving tumor resection. Conclusion The LESS is safe and technically feasible for urological surgery, with minimal invasiveness and better cosmesis. Emphasis should be made on patient selection and indication criteria. Improvement in instrumentation, accumulation of clinical practice, and well-designed multi-center prospective clinical trials will promote the use of LESS surgery for treatment of urologic diseases.

Abstract:Objective To investigate the toxic affects of 25 nm TiO2 nanoparticles in anatase crystal against human lung cancer cell line A549 after different exposure periods. Methods A549 cells were incubated with 100 μg/ml TiO2 nanoparticle suspension for 1 h, 4 h, 8 h and 24 h at 37℃ and 5%CO2. Then the damage degrees of cell membrane caused by TiO2 nanoparticle were assessed by LDH assay. ATP assay was used to evaluate the influence of TiO2 nanoparticle on mitochondria. SOD activity test was used to detect the oxidative damage of the cells caused by TiO2 nanoparticles. The cellular ultrastructure changes of A549 cells were observed by transmission electron microscopy. CCK-8 assay was performed to evaluate the viability of TiO2-treated cells after 1 h, 4 h, 8 h and 24 h exposure. Results With the prolongation of exposure time, the activity of extracellular LDH was increased and that of SOD was decreased; the concentration of intracellular ATP was reduced and the survival rate of cells was significantly decreased(P<0.05). The mitochondria and endoplasmic reticulum were swollen and expanded to different degrees. Conclusion TiO2 nanoparticles can result in oxidative damage and inhibit the growth of A549 cells, and the effects are in a time-dependent manner.

Abstract:Objective To observe the effects of magnetic stimulation on the expression of synaptophysin (SYN), growth associated protein43(GAP43), brain-derived neurotrophic factor (BDNF) in primary mouse hippocampal neurons, so as to explore the possible mechanism of magnetic stimulation in mediating synaptic plasticity. Methods Primary mouse hippocampal neurons were randomly divided into control group, sham stimulation group, and magnetic stimulation groups with various output intensities: 40% (1 Hz, 0.76 T), 60% (1 Hz, 1.14 T), and 80% (1 Hz, 1.52 T) . Cells were stimulated from the 2nd d to the 6th d after plantation; cells in the sham stimulation group was placed in the same environment, but did not accept magnetic stimulation, and cells were drawn at the same time on 7th d. Scanning electron microscopy, immune cell fluorescence, Western blotting analysis and RT-PCR Methods were used to study the morphological changes of neurons and the expressions of SYN, GAP43, and BDNF at protein and mRNA levels. Results In the 40% intensity group the neurite length of neurons and neural connections between cells were both increased, and the SYN-ir, BDNF-ir COD values and protein expressions were significantly higher than those in control group(P<0.05, P<0.01), but GAP43-ir COD value and protein level were not significantly increased; SYN, GAP43, and BDNF mRNA expressions were also markedly higher than those in the control group(P<0.05). In the 60% intensity group, the length and number of neuritis (woven into dense network) were also increased significantly compared with those in the control group (P<0.01) ; COD values of immune reactivity for SYN, GAP43, and BDNF, and their expressions at protein and mRNA were all significantly higher (P<0.01). The neuritis in the 80% intensity group had increased length and number, but with cell damage; GAP43-ir and BDNF-ir COD values and protein levels were also significantly increased (P<0.05, P<0.01), but SYN-ir COD value and protein expression were not significantly different from those of the control group; and SYN, GAP43, and BDNF mRNA expressions were significantly increased compared with control group (P<0.05, P<0.01). Conclusion Magnetic stimulation may increase SYN, GAP43 expression by promoting BDNF synthesis and secretion, and thus plays a very important role in affecting neuronal morphology, regulation of synaptic plasticity, promoting neural network construction; magnetic stimulation at different parameters Results in different stimulus Results .

Abstract:Objective To establish a renal carcinoma cell line A498 with CXCR4 stably silenced by using RNAi technique. Methods We designed three sequence-specific small interfering RNAs (siRNA) targeting CXCR4 gene and transfected siRNA into renal carcinoma cell line A498; the change of CXCR3 gene expression was observed by RT-PCR. The effective siRNA sequence was used to construct the recombinant plasmid, which was used to transfect A498 cells by liposome. Then the stably CXCR4 silenced cell lines were screened by using G418. RT-PCR and Westen blotting analysis were used to determine CXCR4 mRNA and protein expression. Flow cytometry was employed for determination of apoptosis in A498 cells. The invasion ability of cells was detected by transwell assay. Results Green fluorescence was seen in A498 cells transfected with recombinant shRNA plasmid. G418 screening yielded stably CXCR4-silenced A498 cell lines; RT-PCR and Western blotting analysis revealed that CXCR4 expression in CXCR4-shRNA group was significantly lower than that in the control group(P<0.05). The early apoptotic rate, late apoptotic rate, and total apoptotic rate in the CXCR4-shRNA group were significantly higher than those in the control group(P<0.05); transwell assay showed that the cell invasion ability in the CXCR4-shRNA group was significantly decreased compared with that in the control group (P<0.05). Conclusion We have successfully established an A498 cell line with CXCR4 stably silenced; the cell line has a higher apoptotic rate and lower invasion ability, which paves a way for future research.

Abstract:Objective To evaluate the clinical value of modular flexible ureteroscope combined with Holmium laser for treatment of upper urinary calculi. Methods A total of 33 patients with upper urinary tract calculi, including 22 males and 11 females, aging 20-86, were recruited in the present study. Fourteen patients were with upper ureteral calculi, 1 with ureteral and upper calyx calculi, 1 with ureteral and middle calyx calculi, 1 with ureteral and lower calyx calculi, 1 with upper calyx calculi, 2 with middle calyx calculi, 5 with lower calyx calculi, and 8 with pelvis calculi. Twenty-six patients had left calculi and 7 had right calculi; one patient had negative calculi. The diameter of the calculi ranged 0.6-2.3 cm. The patients were under general or spinal anaesthesia and at cutting stone position. After dilating the ureter, we inserted the guide wire under the vision of F8/9.8 rigid ureteroscope, then F12/14 flexible ureteral access sheath. Finally, holmium laser lithotripsy was performed by F8.0 “Polyscope” modular flexible ureteroscope. The power of the laser was 1.0 J and frequency was 10-20 Hz. F6 D-J tube was regularly input for 4 weeks post-operation, and Forley-urethral tube was input for 1 to 7 days. KUB was performed 1 day post-operation to detect the result of lithotropsy and the position of Double-J tube (in some patients, KUB was performed 2-4 weeks post-operation). If more than 6mm residual stone was detected, ESWL was performed.Results Lithotripsy was performed successfully in 27 patients (27/33); the operation time was 25-75 min (averagely 38 min); and the bleeding in the operation was less. Two patients had low fever post-operation; there were no ureteral perforation, high fever or septicaemia. The success rate of one-time lithotripsy was 82%. In the other 6 patients (6/33), 1 patient was transferred to ESWL due to polypi around the stone; 1 was transferred to PCNL due to ureter stricture,2 were transferred to ESWL due to small angle of lower calyx and pelvis, and 2 were transferred to URS due to ureter stricture.Conclusion F8 modular flexible ureteroscope is safe, convenient, and effective for lithotripsy of the upper tract calculi, especially for upper ureteral, pelvis, middle calyx, upper calyx stones.

Abstract:Objective To establish equations for predicting the 24-hour urine protein (24-UP)excretion by analyzing the correlation of four parameters including urine protein/creatinine ratio (Upro/Ucr), urine albumin/creatinine ratio (Ualb/Ucr), urine protein/osmolality ratio (Upro/Uosm) and albumin/osmolality ratio (Ualb/Uosm) with 24-UP. Methods Totally 530 patients were randomly divided into two groups, with 300 for establishing equations and the other 230 for testing the performance of the equations. The equations were set up by linear regression model. The performance of the equations was assessed by Pearson correlation, Bland-Altman plot, calculating the difference and ROC analysis. Results The equations were as follows: E1-UP (mg/24 h)=86.1 (Upro/Ucr \[mg/mmol\]) 0.595, E2-UP (mg/24 h)=53.5 (Ualb/Ucr \[mg/mmol\])0.704, E3-UP (mg/24 h)=21.3(Upro/Uosm \[mg/osm\])0.598, and E4-UP (mg/24 h)=10.9(Ualb/Uosm \[mg/osm\])0.698. Equation based on Ualb/Ucr achieved a better dependence and accuracy compared with the other 3 equations. However, ROC analysis showed no significant difference in diagnostic performance among the 4 equations. We also noticed that excellent correlation was found between the predicted urine protein excretion and 24-hour protein excretion when the patients were at CKD 1 to 4 stages. When patients were at CKD 5 stage, the accuracy and dependence of the equations fell sharply. Conclusion Though the correlation and accuracy of Ualb/Ucr are better than the other 3 Methods , the overall performance is not significantly different among the 4 Methods .

Abstract:Objective To investigate the associated factors influencing the compliance to methadone maintenance treatment by analyzing the data of dropout patients in a single center.Methods The subjects were heroin addicts from Shanghai Hongkou District Mental Health Center who were on methadone maintenance treatment(MMT); they were divided into two groups: the study group(n=54) consisted of those who stopped methadone treatment within 3 months, the control group(n=107)were those who received treatment for more than 6 months. Factors such as social demographics, degree of drug dependence, methadone dose, coping style, and social support were collected, and their influences on dropout in methadone treatment were analyzed.Results Insufficient understanding, low positive response level, high negative response level, and social support were risk factors for the compliance to methadone maintenance treatment in heroin addicts.Conclusion Comprehensive interventions should be taken based on the risk factors, so as to reduce drop-out rate of MMT.

Abstract:Objective To investigate the association of serum Cystatin C level and mild cognitive impairment(MCI) in elderly people with type 2 diabetes mellitus(T2DM) and to analyze the risk factors for MCI. Methods A total of 156 elderly people with type 2 diabetes mellitus were divided into two groups: MCI and normal cognition(NC) group. All subjects were evaluated with cognition scores of Mini-mental state examination(MMSE), Montreal Cognitive Assessment(MoCA), Self-Rating Anxiety Scale(SAS), Center for Epidemiogical Studies Depression Scale(CES-D), Screening Scale for Parkinson disease (SSPD), Activity of Daily Living(ADL), and Global Deterioration Scale(GDS). CystatinC, HbA1c, triglyceride(TG), total cholesterol(TC), and blood pressure were examined. Independent t-test, correlations and logistic regression analysis were performed. Results The elderly diabetic patients complicated with MCI showed a higher level of serum Cystatin C than those in NC group (P=0.018). Cystatin C was negatively correlated with the score of MMSE, and score, visuospatial/executive, naming, attention, language and delayed recall of MOCA (P<0.001). Logistic regression analysis showed that diabetes duration, hypertension, HbA1c and Cystatin C were the variables of the equation. Conclusion Multiple risk factor including the higher level of serum Cystatin C may induce MCI in elderly T2DM people, but larger sample study is needed for further verification.

Abstract:Objective To identify, isolate, and evaluate cells with side population phenotype (SP, with the property of stem cells) from hepatocellular carcinoma. Methods Capable of pumping fluorochrome Hoechst33342 outside of cells, SP cells were isolated and purified by flow cytometry. Results We found that the ratios of SP cells in PLC,Huh7,Hep3B,SMMC7721 cell line and an HCC specimens were 0.8%, 0.3%,0.6%,0.3%,and 0.4%, respectively. We did not find SP cells in HepG2. NOD-SCID subcutaneous transplant experiments showed that SP cells had a high ability of tumorigenicity than non-SP cells. Conclusion Hepatocellular carcinoma cell lines and specimens contain a small number of SP cells, which have a higher tumorigenicity than non-SP cells; this finding paves a way for further analysis of the stem-like property of SP cells.

Abstract:Objective To explore the ultrasonic presentations of primary hepatic carcinoid tunors. Methods The ultrasonic data of 9 cases of hepatic carcinoid tumors, who were pathologically confirmed from September 2002 to November 2008 in our hospital, were retrospectively analyzed. Results Ultrasonic examination showed that the diameters of the 9 tumors were all >5 cm. The tumors had an irregular morphology and presented with strong or high echo with inhomogeneous echo inside. The tumors had no noticeable capsule and had clear borders and rich blood supply. Wide bleeding and necrosis of tumor could cause the cystic mass. There were no primary nidus or tumor emboli of portal veins. Conclusion Ultrasonic diagnosis of liver carcinoid tumors still lacks specific presentations, and efforts should be made to persistently summarize the experience to improve the diagnosis.

Abstract:Ionizing radiation (IR) can cause immune dysfunction, resulting in severe damages to human health and affecting the recovery of radiation injury. Dendritic cells (DCs) are the most potent antigen presenting cells (APCs), which are also the initiator and modulator of immune system. Studies on the radiation effects of DCs can not only help to understand the mechanism of radiation-induced immunosuppression, but also facilitate the research on DCs function for radioprotection. DC-based cancer vaccine combined with radiotherapy also provides new insights into cancer therapy. This paper reviews the influence of IR on the viability, phenotype, antigen presenting function and migration capacity, cytokine secretion of DCs, as well as the underlying mechanism. We also review some advances in tumor therapy by combining DC vaccine with radiotherapy, hoping to provide reference for studying radiation-induced immunosuppression and radioprotection.

Abstract:MicroRNA (miRNA) is a group of recently discovered small RNA molecules that have a significant role in regulating gene and protein expression. These small RNAs (-22 nt) can bind to 3′- untranslated regions (3′-UTRs) and induce mRNA degradation and repress protein expression. They play important roles in development and maintenance of normal cell function. More recently, a few studies have suggested that miRNA may also serve as an important regulator of circadian rhythmicity, which casts new lights on our understanding of biological clocks. Here we describe the function of miRNA in regulation of circadian system.

Abstract:G Protein-coupled receptors constitute a super family of cell surface proteins. β-arrestin 1 and 2 are widely expressed intracellular adaptor and scaffolding proteins that play important roles in G protein-coupled receptor desensitization, internalization, intracellular traf?cking and G Protein-independent signalling. Recent developments in bioluminescence resonance energy transfer （BRET）technology enable monitor the GPCR–β-arrestin complexes in live cells and real time. In concert with a range of experimental tools, investigators can elucidate the ever-expanding roles of β-arrestins in mediating GPCR function, which will be benefit for developing new generation drugs targeting GPCRs.

Abstract:Objective To observe the cleavage effect of Caspase-3 on PIAS1 protein in vitro. Methods PIAS1 protein was expressed in HEK293T cells. Co-immunoprecipitation (Co-IP) was used to observe the binding between PIAS1 protein and Caspase-3. The cleavage effect of Caspase-3 on PIAS-1 protein was detected by in vitro proteolytic cleavage test. Results Co-IP result showed that Caspase-3 bond to PIAS1 in vitro. Proteolytic cleavage test showed that PIAS1 was cleaved by active Caspase-3. Conclusion It has been shown that Caspase-3 can interact with PIAS1 protein in vitro, and PIAS1 can be cleaved by Caspase-3 in vitro.

Abstract:目的 构建Smad泛素调节因子1 (Smurf1) 基因的RNA干扰( RNAi) 慢病毒表达载体,观察其转染骨肉瘤U20S细胞后对Smurf1基因表达的抑制作用,为后续研究奠定基础。方法 构建Smurf1 基因过表达质粒,同时针对Smurf1基因序列设计合成4 条RNAi片段并构建RNAi 慢病毒载体。过表达质粒和RNAi 质粒共转染293T细胞后应用Western印迹法进行外源筛靶,筛靶出成功的干扰质粒PSC2939。用慢病毒包装PSC2939后感染U20S细胞,进行内源验证,荧光定量PCR和Western印迹检测U20S细胞中Smurf1 mRNA及蛋白的表达。结果 经PCR及测序证实,Smurf1基因过表达载体及4种干扰载体均成功构建；Western 印迹外源筛靶显示,PSC2939干扰载体能有效敲减目的基因的表达；慢病毒包装后的PSC2939干扰载体能有效抑制U20S细胞中Smurf1蛋白的表达,对Smurf1 mRNA的抑制率达到60%以上。结论 成功构建可供感染的Smurf1 基因RNAi 慢病毒载体,为进一步研究Smurf1基因在骨肉瘤细胞中的作用提供了有效的实验工具。

Abstract:目的 观察腺病毒(Adts)介导碱性成纤维细胞生长因子2（FGF-2）直接体内转基因对大鼠脊髓损伤的治疗作用。方法 将SD大鼠在T10水平制备脊髓损伤模型,随机分为体内转基因治疗组和损伤对照组,用携带FGF-2和绿色荧光蛋白(GFP)的Adts行直接体内转基因治疗；荧光显微镜观察体内转基因表达,后肢运动功能评分(BBB评分)检测大鼠运动功能恢复情况, EC染色观察损伤后脊髓灰质和白质残存组织的体积变化。结果 BBB评分检测到体内转基因治疗组大鼠后肢运动功能有明显恢复,与对照组相比,差异有统计学意义(P<0.05)；体内转基因治疗组脊髓灰质残存组织的体积明显增加,与对照组相比差异有统计学意义(P<0.05)。结论 由Adts介导FGF-2的直接体内转基因治疗能够明显促进损伤脊髓的功能恢复。

Abstract:目的 观察喉鳞癌组织中蛋白激酶C (protein kinase C,PKC)-δ的表达水平,探讨其可能的临床意义。方法 采用免疫组织化学法对60例喉鳞癌组织和20例正常喉黏膜 PKC-δ蛋白表达情况进行检测。结果 喉鳞癌组织中PKC-δ表达显著高于正常对照组织(P<0.01)。Ⅲ～Ⅳ期喉鳞癌组织 PKC-δ 表达明显高于Ⅰ～Ⅱ期（P<0.05）；伴有颈淋巴结转移的喉鳞癌组织PKC-δ表达明显高于无淋巴结转移组织（P<0.05）；PKC-δ表达阳性组与阴性组患者间生存率差异有统计学意义(P<0.05)。结论 PKC-δ 在喉鳞癌组织中高表达,并且其表达程度与喉鳞癌的分期及是否存在淋巴结转移有关,提示PKC-δ过表达可能与喉鳞癌的发生、发展相关。

Abstract:目的 观察人脑胶质瘤组织中MMP-7和整合素αvβ5的表达,并探讨其可能的临床意义。方法 采用免疫组织化学S-P法,检测38例不同病理分级脑胶质瘤和13例正常脑组织中MMP-7和整合素αvβ5的表达情况,探讨二者表达的相关性。结果 MMP-7和整合素αvβ5在脑胶质瘤中阳性表达均高于正常脑组织(P<0.01)。MMP-7蛋白表达水平与脑胶质瘤病理分级正相关（r=0.426,P<0.01）；整合素αvβ5蛋白表达与病理分级正相关（r=0.323,P<0.05）。人脑胶质瘤中MMP-7的表达水平与整合素αvβ5的表达水平正相关(r=0.578, P<0.01)。结论 MMP-7和整合素αvβ5表达水平的测定可能有助于早期识别具有高度恶性的人脑胶质瘤并判断其预后。

Abstract:目的 探讨人胰腺癌组织中BRAF和KRAS基因突变情况及两者的关系。方法 对36例胰腺癌组织及癌旁胰腺组织中BRAF基因15号外显子及KRAS基因12号密码子进行测序。结果 36例胰腺癌组织中,25例出现KRAS基因12号密码子突变(69.4%,25/36),3例出现BRAF基因V600E突变(8.3%,3/36),且均同时存在KRAS突变；11例KRAS野生型胰腺癌组织中未发现BRAF突变；癌旁胰腺组织中均未检出突变。结论 胰腺癌组织中BRAF基因突变可能并非主要突变,且常与KRAS基因突变同时存在。