OBJECTIVE To know the present situation of management of endoscope disinfection in order to improve the disinfection quality.METHODS A questionnaire and on-spot-survey were sent to 8 hospitals equipped with endoscope.RESULTS Eight different grade hospitals had a system of endoscope management established and integrated into their quantity management system.One hospital had isolated cleaning and sterilizing rooms,of which 3 hospitals were well ventilated;6 hospitals had special storage cabinets for endoscope,for endoscope as well as cleaning and sterilizing troughs.All of the hospitals used multienzymes and timers when doing cleaning.CONCLUSIONS There are some problems in the present situation.The way of management monitoring and occupational protection must be further improved to reduce the nosocomial infection.

Objective To validate a new standardized training program for urological surgeons to improve their laparoscopic surgical skills. Methods The laparoscopic surgical training program was carried out by using the traditional mechanical simulators and animal models. Thirty-three trainees participated the urological laparoscopic surgical training program. Initially, the novices were assigned to practise basic laparoscopic skills step by step on the simulator with fixed trocar positions. After a period of basic training, they were allowed to practise on animal models for some particular proce-dures. Results All trainees (33/33, 100.0%) participated were able to perform all basic techniques skillfully and completed laparoscopic anastomosis accurately after the training. The time required for performing the partial nephrectomy, dismembered pyeloplasty and ureteral reimplantation on animal models declined from 64.0±18.4, 127.54±17.5 and 75.84±11.6 min at the beginning to 30.94±3.8, 65.2±7. 5 and 37.7±7.2 min after practicing these procedures 8 times (P<0.01). They could un-derstand the crucial procedures of the laparoseopic surgeries after 6 to 8 special trainings on animal models. Fifteen trainees (15/33, 45.5%) had started to carry out laparoscopic surgeries after-finish- ing the training program. Conclusions Our program enables the participants to improve their techniques in complicated laparoscopic surgeries. The challenging parts of reconstructive laparoscopic surgeries such as laparoscopic dismembered pyeloplasty can be taught by using animal models. This program could be incorpo-rated easily by all urological departments developing a laparoscopie surgical training program.

Objective To investigate the causes of re-fractures of non fracture vertebral body after percutaneous kyphoplasty (PKP).Methods 512 patients (618 vertebral bodies) treated with PKP because of osteoporosis VCFs were recruited from June 2010 to June 2014.There were 107 males (121 vertebral bodies) and 405 females (497 vertebral bodies) with the mean age of 70.38±7.59 years old (51 to 91 years).There were 406 single segment fracture and 106 double segment fractures cases,and the fracture segments were T4 to L5.The T value of the patients' bone mineral density (BMD) was from-1.0 to-5.2 SD.The clinic characteristics of all the patients including age,sex,body weight,body height,body mass index (BMI),BMD score of the spine,volume of bone cement,restoration rate of anterior/middle vertebral height,postoperative complications (pulmonary embolism,bone cement leakage,nerve injury),and treated vertebral level were analyzed.Results 52 patients (10.16％,52/512) experienced refractures of non fracture vertebral body after kyphoplasty,and 4 experienced re-fracture of the fracture vertebral body after kyphoplasty.The average age of the 52 patients was 71.88±7.74 years old,meanwhile,the ratio of female was 94.23％ (49/52),the mean T value of BMD-4.03±0.60 SD,the ratio of initial double segment fractures 51.92％ (27/52).In addition,among the 456 cases with no fracture,the average age was 70.21±7.56 years,the ratio of female was 77.19％ (352/456);the mean T value of BMD was-2.89±0.55 SD;the ratio of initial double segment fractures was 17.32％(79/456).The data above (age,T value of BMD and initial double segment fractures) were all with statistical significant differences.Whereas the BMI,volume of bone cement,intervertebral disc leakage and restoration rate of anterior/middle vertebral height had no significant difference between the two groups.Furthermore,in the re-fracture of non fracture vertebral body group,32 cases (61.54％,32/52) were nonadjacent fractures,and 20 (38.46％,20/52) were adjacent fractures.Conclusion Osteoporosis degree,female and initial double segment fractures were major risk factors in the development of re-fracture of non fracture vertebral body after PKP.

Aim To investigate the effect and mecha-nism of valproic acid on neuroglobin expression, and the neuroprotective role of valproic acid against H2 O2-induced neurotoxicity. Methods Western blot, RT-PCR and luciferase assay were used to detect the pro-tein levels, mRNA levels and promoter activity of mouse and human neuroglobin induced by valproic acid. Luciferase assay was used to investigate the role of transcription factor CREB in the up-regulation of neuroglobin by valproic acid. MTT assay was used to evaluate the effect of valproic acid against H2 O2-in-duced neurotoxicity. Results VPA treatment marked-ly increased the protein levels, mRNA levels and pro-moter activity of Ngb in mouse N2 a cells and human SKNSH cells. CREB specific inhibitor KG501 or CREB dominant negative mutant KCREB attenuated VPA-induced Ngb promoter activity. VPA could pro-tect N2a cells from H2 O2-induced neurotoxicity. Con-clusion CREB mediates VPA-induced Ngb up-regula-tion, which may contribute to the neuroprotective effects of VPA in oxidative stress in neurons.

Objective:To investigate the effect of rapamycin (Rapa) on apoptosis of acute myeloid leukemia THP-1 cells induced by idarubicin (IDA) and its molecular mechanism.Methods:The THP-1 cells were treated with 10, 20, 40 and 80 nmol/L Rapa for 1 h, and the cells without Rapa treatment were set up. Western blot was used to detect the conversion of autophagy marker LC3 protein in THP-1 cells (the ratio of LC3Ⅱ/LC3Ⅰ), flow cytometry was used to detect the apoptotic rate, and the pretreatment concentration of Rapa was determined. THP-1 cells were treated with different concentrations of IDA for 24 h, the cell proliferation inhibition rate of IDA for THP-1 cells was detected by CCK-8 method, and the half maximal inhibitory concentration ( IC50) was calculated. THP-1 cells with or without Rapa treatment were treated by IDA with the concentration of lower than IC50 for 24 h, CCK-8 method was used to detect cell proliferation inhibition rate, flow cytometry was used to detect cell apoptosis, real-time fluorescent quantitative polymerase chain reaction was used to detect the expression changes of autophagy-related genes Beclin-1, LC3 and p62, and Western blot was used to detect the conversion of autophagy marker LC3 protein. Results:The ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells treated by 20 nmol/L Rapa was higher than that in the untreated cells ( P=0.002 4). The apoptotic rate in THP-1 cells treated by 80 nmol/L Rapa was higher than that in the untreated cells ( P=0.007 3). According to the results of Western blot and flow cytometry, 20 nmol/L Rapa was selected as the pretreatment concentration. The IC50 of IDA for THP-1 cells treated with IDA for 24 h was 59.874 nmol/L. After treated with 50 nmol/L IDA for 24 h, the proliferation inhibitory [(69.67±5.03)% vs. (41.67±3.51)%] and apoptotic rates [(74.35±4.83)% vs. (41.25±5.24)%] in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells (both P<0.05); the Beclin-1 and LC3 mRNA expression levels and the ratio of LC3Ⅱ/LC3Ⅰ in THP-1 cells pretreated by Rapa were higher than those in the unpretreated cells, and the expression of p62 mRNA was lower than that in the unpretreated cells (all P<0.05). Conclusion:Rapa can enhance the apoptosis of THP-1 cells induced by a relative low dose of IDA, which may be achieved through inducing excessive autophagy in THP-1 cells.

The anti-tumor activity of curcumin against androgen-independent prostate cancer cells in vitro and the possible mechanism were investigated. After curcumin treatment, the effect of curcumin on the proliferation of prostate cancer PC-3 cells was assessed by CFSE staining. Flow cytometery (FCM) was performed to analyze the cell cycle and the induction of apoptosis of tumor cells. A luciferase reporter gene assay was used to determine the effects of curcumin on the activities of intracellular NF-κB and AP-1 signaling pathways. The results showed curcumin could effectively inhibit the proliferation of PC-3 cells in vitro (P<0.05). Cells were arrested at G(2)/M phase. After curcumin treatment, the percentage of apoptotic cells was significantly higher than in control group (P<0.05). The results of the luciferase assay revealed that curcumin selectively inhibited the activities of the NF-κB and AP-1 signaling pathways in PC-3 cells significantly. It was suggested that curcumin could exert anti-tumor activity against androgen-independent prostate cancer cells in vitro by inhibiting cellular proliferation and inducing apoptosis, which was probably contributed to the inhibition of transcription factors NF-κB and AP-1.

Objective:Mutation in the gap junction beta 6 (GJB6) gene has been reported to be associated with an autosomal dominant disorder hidrotic ectodermal dysplasia (HED), characterized by congenital nail clubbing, alopecia and palmoplantar keratoderma. The aim of this study is to investigate relationship between genetic mutation in GJB6 and HED in an affected Chinese family.
Methods:We selected a Chinese HED family consisting of a total of 17 individuals including 8 HED patients (5 males and 3 females). The whole coding region of GJB6 was amplified by polymerase chain reaction and sequenced.
Results:Sequence analysis identified a heterozygous missense mutation c.31G>A (p.G11R) in GJB6 gene of affected individuals, but not in healthy individuals.
Conclusion:A c.31G>A (p.G11R) missense mutation in GJB6 gene is the genotypic characteristic for HED in Chinese population.

Objective To compare broth microdilution and agar dilution methods for in vitro testing of activities of fluconazole,ketoconazole and itraconazole against clinical Malassezia isolates.Methods Broth microdilution and agar dilution methods were used to determine the minimal inhibitory concentration(MIC)of fluconazole,ketoconazole and itraconazole for 27 clinical strains(5 species)of Malassezia.Results The minimal inhibitory concentration(MIC)ranges of fluconazole,ketoconazole and itraconazole were 0.25-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.125 mg/L respectively as shown by broth microdilution method,2-≥64 mg/L,≤0.03-0.5 mg/L and ≤0.03-0.25 mg/L respectively as revealed by agar dilution method.Both methods demonstrated that itraconazole possessed the strongest activity against Malassezia species,followed by ketoconazole and fluconazole.The agreement rate in MICs between the two methods was 78.8％,85.2％ and 88.9％,respectively for fluconazole,ketoconazole and itraconazole,with the intraclass correlation coefficients (ICCs)being 0.88,0.80 and 0.76 respectively.Conclusions Fluconazole,ketoconazole and itraconazole are highly active against Malassezia species in vitro,and itraconazole is the most active.Broth microdilution and agar dilution method coincide well in,and are applicable for,the antifungal susceptibility testing of Malassezia species in vitro.

The aims of this research were to construct prokaryotic expression vector containing the gene of porcine urate oxidase (pUOX), optimize the conditions of the expression of pUOX in recombinant Escherichia coli BL21(DE3), and analyze the in vitro activity and the enzymological properties of pUOX. The pUOX gene was amplified by RT-PCR from the extracted total RNA of porcine liver, and was inserted into the prokaryotic expression vector pET30a(+) to construct a recombinant expression vector pET30a(+)/pUOX. We identified the recombinant vector by endonuclease digestion and sequence analysis. The pUOX gene was amplified and cloned into the vector pET30a(+) successfully. And then the recombinant vector was transformed into E. coli BL21(DE3). The expression of pUOX with a molecular of approximately 41 kD was induced by IPTG. We also optimized the expression conditions of the recombinant protein. The recombinant protein was mostly located in the cytoplasm and it was insoluble. After the inclusion body was solved in 8 mol/L urea and refolding in 2 mol/L urea, the recombinant protein was collected and purified by Ni2+-NTA column. This recombinant protein had a specific activity of 50.61 IU/mg and showed similar properties of optimum temperature and thermal stability, base on the enzymatic assay and analysis of enzymological properties. These results would help to analyze the in vivo activity by testing animal.
Animals ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; Swine ; Urate Oxidase ; biosynthesis ; genetics ; isolation & purification

Objective To investigate the in vitro effects of bladder cancer cell proliferation after silencing Notch1 gene. Methods The siRNA eukaryotic expression vector of Notch1 (psiRNA1)was constructed and transfected into bladder cancer cell lines T24 and BIU-87. Methabensthiazuron (MTT) and flow cytometry (FCM) assays were used to detect bladder cancer cells line growth, cell cycle and apoptosis after the transfection. RT-PCR and Western blotting were used to determine the expression changes of Notch1 in these cell lines. Results After transfection for 72 h, the rate of G0/G1 phase cells inceased from (23.89±1.32) % to (80.13±2.69)% in T24 cell line, and increased from (24.63±1.68)% to (69.44±2.41)% in BIU-87 cell line (both P<0.05). In addition, apop-totic cell index in T24 and BIU-87 cell lines increased from (1.28±0.14)% to (13.75±1.23)%, from (1.01±0.27)% to (8.72±1.01)%, respectively(both P<0.05). The growth of T24 and BIU-87 cell lines was obviously inhibited 24 h after the transfection, and the inhibitory effects lasted until 96 h after the transfection. Notch1 mRNA and protein significantly downregulated after transfection compared to the control(P<0.05). Conclusions Silencing Notch1 expression can inhibit the prolif-eration of bladder cancer cell lines. Notch1 gene might act as a tumor gene in bladder cancer.