Objective To establish PKD2 gene recombinant lentivirus and to investigate its restorative effects on polycystin-2 and Wnt/β-catenin signaling pathways in Pkd2-null cell lines.Methods From August 2016 to February 2017,PKD2 gene was cleaved from the pcDNA3.1-TM-PKD2 plasmid and was inserted into the pLV-sfGFP 2A Puro by restriction enzymes Xba Ⅰ and Xho Ⅰ.The recombinant pLV-sfGFP-PKD2 plasmid was sequenced to verify a correct construction.Then we obtained recombinant lentiviruses by co-transfecting 293T cells with recombinant plasmid and packaging plasmids.B3/D3 (Pkd2 +/-) and B2/E8 (Pkd2-/-) cell lines were used to evaluate the effectiveness of lentivirus,they were divided into experimental group,control group,blank group and treated with pLV-sfGFP-PKD2 virus,pLV-sfGFP virus or culture media,respectively.The expression of PC2 was detected by Western blotting and immunofluorescence staining.Finally the cell proliferation was evaluated by detecting of proliferating cell nuclear antigen.The changes of Wnt/β-catenin signaling pathway were evaluated by real-time quantitative PCR.Results Enzyme digestion analysis and DNA sequencing showed that the recombinant plasmid pLV-sfGFP-PKD2 was constructed successfully.After infected with pLV-sfGFP-PKD2 virus,the expression of PC2 in the experimental group B2 and E8 cells(0.668 ±0.013,0.763 ±0.021) was restored to the normal level,compared with control group B3 and D3 cells,respectively (0.687 ± 0.015,P =0.164;0.776 ± 0.008,P =0.409).The proliferative activity in experimental group B2 cells(0.573 ±0.010) was significantly lower than that in control group B2 cells (0.848 ±0.031,P ＜0.01),and was returned to the level of blank group B3 cells (0.585 ±0.017,P =0.369).Reexpression of PKD2 in experimental group B2 cells also reduced the expression of Wnt7a,β-catenin,back to blank group B3 cells' level(0.037 ±0.005 vs.0.037 ±0.004,P=0.969;0.554 ±0.008 vs.0.571 ±0.013,P =0.64).Conclusions The recombinant pLV-sfGFP-PKD2 lentivirus has been constructed successfully.The lentivirus could rectify the absence of PC2 in PKD2-null cell lines,by which the hyperactivated Wnt/β-catenin signaling pathway and the abnormal cell proliferation caused by PC2 deficiency could be also restored to normal levels.

Objective To investigate the change of intracellular calcium ion concentration in prostate smooth muscle cells of SD rats with chronic abacterial prostatitis under high potassium solution.Methods SD rats were divided into experiment group and control group.The CP model was set up by castration and estradiol injection.The PSMC was cultured and purified in vitro.Laser confocal scanning microscope was used after the ceils were incubated with Quest Fluo-8TM.The cells were treated with high potassium solution,and the change of fluorescence intensity was observed.Results The pathologic specimens of the experiment group showed typical pathologic characteristics of chronic prostatitis under light microscope,the control group without inflammation performance.Using immunocytochemistry method confirmed that the experiment group and the control group were prostate smooth muscle cells.The change of fluorescence intensity of [Ca2+] i in the experiment group and control group in the high potassium solution was 27.86 ± 9.88 and 7.61 ± 4.31.There were statistically significant differences between the two groups (P ＜ 0.01).Conclusions High potassium solution cause intracellular calcium ion concentration increased.

Objective To investigate the prevalence and relevant factors of psychological disorders in patients with premature ejaculation (PE). Methods From September 2009 to October 2010,we employed the Self- Rating Anxiety Scale (SAS) and Self-Rating Depression Scale (SDS),Chinese Index of Sexual Function for Premature Ejaculation ( CIPE-5 ),NIH-Chronic Prostatitis Symptom Index (NIH-CPSI),International Index of Erectile Function (IIEF-5) and self-designed questionnaire to investigate the psychological status of 1164 patients with PE.The relevance between scores of SAS,SDS and such parameters as NIH-CPSI score,CIPE-5 score,erectile function,age,disease duration,occupation,education,personality characteristics and other factors were evaluated. Results The SAS and SDS scores of 1,164 cases with PE were 43.87 ± 10.53,44.05 ±9.81,respectively.If the cut-off points were set at SAS ≥ 50 and SDS ≥53,341 (29.3％) of cases had symptoms of anxiety and 217( 18.6％ ) patients had symptoms of depression.The SAS,SDS scores and detection rate of anxiety and depression were correlated with the CIPE-5 score,NIH-CPSI score,erectile function and duration of disease,personality characteristics ( P ＜ 0.05),while they were not correlated with age,occupation,and education. Conclusions Psychological disorders widely exist in patients with PE and may have a close relationship with a variety of factors.

ObjectiveTo define the difference of urine analysis result,α1-microglobulin (α1-MG) and β2-microglobulin ( β2-MG) between those patients suffering from calcium oxalate stone,non-calcium oxalate stone and non-urolithiasis controls at the same time period.MethodsData from 100 patients admitted to the Department of Urology,First Affiliated Hospital of Anhui Medical University from July,2010 to September,2010 were reviewed.66 patients (45 men,21 women) suffered from urolithiasis,and 34 patients (22 men,12 women) were non-urolithiasis.Patients' ages in urolithiasis group varied from 13 to 78 years and the male to female ratio was 2.1∶1.0.The patients in non-urolithiasis controlgroup aged from 12 to 80 years and the male to female ratio was 1.8∶1.0.Blood and urine were taken from the patient the next morning after admission.The biochemistry from blood and 24 h urine were measured by automatic biochemistry analyzer.The α 1-MG and β2-MG content were measured by radioimmunoassay.The stone compositions were analyzed by infrared spectroscopy.ResultsThere was difference in the levels of serum creatinine and blood urea nitrogen among three groups ( P ＜ 0.05).In controls,those with calcium oxalate stone had higher level of urinary α1-MG and β2-MG,but there were no differences in the urinary electrolyte levels.Group of non-calcium oxalate stones urinary uric acid levels were higher than calcium oxalate and control groups,the difference was statistically significant.ConclusionsIn the formation of uric acid stones,uric acid increased as independent risk factors.α1-MG,β2-MG may promote the formation of calcium oxalate stones.

Objective To discuss the clinical features of bilateral multiple renal angiomyolipomas (RAML) with tuberous sclerosis complex (TSC).Methods Clinical data of 10 cases of bilateral multiple RAML associated with TSC treated from Jan.2003 to Dec.2010 were retrospectively reviewed.There were 4 males and 6 females in the study group,aged from 18 to 45 years,with an average age of 34 years.Three cases were treated urgently due to RAML hemorrhage.Five cases were treated due to back pain.One case was treated for hematuria.One case was diagnosed during physical examination.Facial angiofibroma occurred in 7 cases and facial flat speckle in 1 case.Epilepsy was indentified in 6 cases.Four cases had hypophrenia and 2 cases presented with symptoms of the classic triad:epilepsy,facial angiofibroma and hypophrenia.One case underwent urgent selective transcatheter arterial embolization (SEA).Eight cases (9 sides) underwent renal sparing operation.Conservative therapy was adopted in 2 cases.ResultsThe SEA was done successfully and bleeding ceased.AⅡ surgeries were completed successfully.The diameter of the biggest tumor resected was 8.0 cm.The number of tumors resected was 2 - 6,with an average of 3.5.The mean operative time was 70 min (range 50 to 120 min).The mean time of controlling the renal artery was 26 min.The mean intraoperative hemorrhage was 150 ml.One case required a blood transfusion.None of the cases re-hemorrhaged after operation.Pathology revealed RAML.There was no recurrence in 8 cases and follow-up ranged from 3 to 60 months,with an average of 25 months.A hemorrhage did occur on the oppositer side in 1 case 4 months after surgery.In 2 cases,a conservative therapy of regular monitoring and follow-up at 24 and 36 months was chosen.The serum creatinine of all the patients was in a normal range.Conclusions The bilateral multiple RAML associated with TSC must be diagnosed as soon as possible and treated depending on the size,location and developing circumstance of the tumor in a timely fashion.Renal sparing surgery and SEA are effective therapies.

Objective To analyze the diagnosis and treatment for complications of renal caliceal diverticulum with calculi or infection. Methods A retrospective investigation was performed on 29 cases with renal caliceal diverticulum. The 29 cases included 11 males and 18 females aged 18 to 61 years. Among the study group, 3 cases were simple renal caliceal diverticulum, 12 cases were diagnosed as diverticular calculi and 14 cases presented recurrent urinary tract infections including 3 cases with urinary fistula after unroofing and decompression as renal simple cyst from another hospital. Ten cases underwent an open operation that unroofed and decompressed the cyst, and sutured the diverticular neck. Eight cases underwent laparoscopic operation similar to the open operation, including lithotomy in caliceal diverticulum in 2 cases. Eleven cases diagnosed with caliceal diverticular calculi were taken one-stage percutaneous nephrolithotomy including dilating the diverticular neck, remaining the nephrostomy catheter and Double-J ureteral stents, and 1 case was transferred to open operation.Results The open and laparoscopic operations were performed successfully. One case was cured by Double-J ureteral stenting after postoperative urinary leakage. One case was transferred to open operation for the failure of percutaneous puncturation. X-ray examination revealed that there were no remaining stones after the operation. All the patients were followed up for 6 to 24 months without calculi and infection recurrence. Conclusions Stones and infection are common that complications of renal caliceal diverticulum. Percutaneous nephrolithotomy, laparoscopy and other operations were effective and feasible treatment options for cases with complications of renal caliceal diverticulum. Exact diagnosis was very important for treatment of renal caliceal diverticulum before operation.

Objective To investigate the effect of reptroperitoneal laparoscopic operation on the parameters of platelet, D-dimer and thrombomodulin(TM). Methods Forty cases were divided into two groups according to the operative way, retroperitoneal laparoscopic operation (n= 20) and open operation (n=20). Blood samples were taken preoperatively and at the end of the surgery. The following parameters were measured and compared within each group and between groups: platelet count (PLT), mean platelet volume (MPV), platelet distributionwidth(PDW), D-dimer, TM. ResultsThere were no significant differences for the PLT, PDW, MPV, TM and D-dimer between before and after operation in each group. There was no difference between 2 groups either for all these indicators.No patients from either group suffered thrombosis or abnormal bleeding as a pastoperative complication. Conclusion Compared with the conventional operation, retroperitoneal laparoscopic operation dioesd not induce more change on parameters of platelet, D-dimer and TM.

OBJECTIVETo investigate the prevalence of sexual dysfunction in chronic prostatitis patients and its relation with patients' age and the course of disease.

METHODSQuestionnaire surveys were completed by 2 000 subjects who met the criteria for chronic prostatitis after clinical examination and expressed prostatic secretion analysis. Included in the questionnaire were: (1) age, height, weight, occupation, course of disease and treatment received; (2) NIH-CPSI; and (3) IIEF-5.

RESULTSThe prevalence of sexual dysfunction in patients with chronic prostatitis was 49.0% : 26.2% of the subjects with premature ejaculation, 15.0% with erectile dysfunction and 7.7% with both premature ejaculation and erectile dysfunction.

CONCLUSIONThe prevalence of sexual dysfunction in patients with chronic prostatitis is high, which is negatively correlated with both the age of the patients (P < 0.001) and the course of the disease (P < 0.001).

Adult ; Age Factors ; Chronic Disease ; Humans ; Male ; Middle Aged ; Prostatitis ; complications ; Sexual Dysfunctions, Psychological ; epidemiology ; etiology

OBJECTIVETo explore the inhibitory effect of He-Ne laser irradiation on fibroblast growth of hypertrophic scars in culture.

METHODSHe-Ne laser with wavelength of 632.8 nm, power density of 50 mW/cm(2) and doses of 3 J/cm(2), 30 J/cm(2), 90 J/cm(2) and 180 J/cm(2) was used to irradiate human scar fibroblasts in culture 1, 3 and 5 times respectively, and then the cell count and cell cycle analysis were done.

RESULTSRepeated irradiation with He-Ne laser at dose of 180 J/cm(2) three and five times led to an evident decrease in total cell number compared with that of the control group and there was a significant difference (P<0.05). The cell cycle analysis showed after three and five times of irradiation with 180 J/cm(2) He-Ne laser the cell number in S-phase decreased from 51% to 20% and 14% respectively, the cell number in G(0)/G(1) phase increased from 28% to 55% and 60% respectively, and the cell percentage in Sub-G1 phase was 6.7% and 9.8% respectively.

CONCLUSIONSRepeated irradiation with 180 J/cm(2) He-Ne laser can inhibit scar fibroblasts growth in culture. It may be that He-Ne laser irradiation causes cell stagnation in G(0)/G(1) phase and apoptosis.

Cell Division ; radiation effects ; Cells, Cultured ; Cicatrix ; pathology ; Dose-Response Relationship, Radiation ; Female ; Fibroblasts ; cytology ; radiation effects ; Helium ; Humans ; Lasers ; Male ; Neon

Objective　To establish animal model for hypertrophic scar and study the characters of its morphology and collagen metabolism. Methods　A total of 64 round wounds (diameter of 6 mm each) with total skin loss were made on the ventral side of rabbit ear using a trephine. Morphology and collagen metabolism of scar wounds were studied at 14,21,35,70 and 98 days after operation, respectively. Results　There were 76% elevated scars developed (45/59 wounds) on the ventral side of rabbit ear at 21 days and 46% elevated scars disappeared (11/24) at 98 days after operation. There were numerous fibroblast proliferation and whorl-arranged collagen fibers at 21 and 35 days. The number of fibroblast decreased, but irregular-arranged fibers still presented in the elevated scars at 70 and 98 days after operation. Hydroxyproline content in elevated scars at 21 days was higher than that in normal skin (P<0.05), and at 35 days was 3 times as that in normal skin and at 98 days was also markedly higher than that in normal skin (P<0.05). Conclusion　Excessive deposition of collagen is a characteristic of hypertrophic scar in rabbits. The conversion of normal scarring to hypertrophic scarring in rabbits occurs at 14～21 days after operation. Both development and regression of hypertrophic scar in rabbit are quicker than that in human.