Objective To investigate the proportion of follicular fluid CD56~+ natural killer (NK) cells to the total lymphocytes and the ac-tivated CD56~+ NK cells to the total CD56~+ NK cells. To provide an evidence for improving the clinical pregnancy rate in in vitro fertilization (IVF) treatment by regulating the function of NK cells. Methods Triple color flow cytometry was used to analyze the proportion of CD56~+ NK cells and activated CD56~+ NK cells in mature follicular fluid. The IVF treatment outcome was closely followed up. The relationship be-tween the proportion of CD56~+ NK cells and activated CD56~+ NK cells and the IVF treatment outcome was analyzed statistically. Results The proportion of CD56~+ NK cells and activated CD56* NK cells in mature follicular fluid of women who got pregnancy by IVF treatment was (15.57±3.10)% and (2.63±0.94)% respectively,while the proportion of the women who didn't get pregnancy was (19.12±5.37)% and (4.06±2.08)% respectively,which was significantly higher than the pregnant group(P< 0.05). Conclusion The women with lower propor-tion of CD56~+ NK cells and activated CD56~+ NK cells in mature follicular fluid of is easier to get pregnancy by IVF. An altered NK cells pro-file in follicular fluid could therefore influence fertility in IVF treatment outcome.

Objective To explore the etiological factors of primary pathological duodenogastric reflux (DGR) through investigating the relationship between severity of bile refulx,the changes of surface gastric electric rhythm and gastric emptying movement in primary pathological DGR patients.Methods From January 2007 to April 2008 in Qingdao Municipal Hospital,58 cases of outpatients diagnosed as primary pathological DGR and 21 healthy individuals (control group) were collected and underwent 24-hour gastric bilirubin monitoring,gastric endoscopy,gastric electric rhythm,and gastric emptying test.The relationship between gastric electric parameters and gastric emptying,bilirubin reflux,Hp infection was analyzed.Results 1.The main frequency in fasting and postprandial of primary pathological DGR patients [(1.94±0.04) cpm vs (2.93±0.07) cpm; (2.12±0.03) cpm vs (3.35 ±0.05) cpm],the percentage of normal gastric slow wave in fasting and postprandial (74.46± 0.56 vs 85.55 ± 1.06 ; 63.97 ± 0.64 vs 86.13 ± 1.49),and fasting/postprandial power ratio (PR) (1.68±0.02 vs 2.75±0.09) were all lower than those of control group (P＜0.05).The percentage of bradygastria in fasting and postprandial of DGR patients (18.04±0.36 vs 7.76±0.78;23.73±0.91 vs 8.47±0.55),the percentage of tachygastria in fasting and postprandial (8.93±0.26 vs 5.75±0.66;13.02±0.40 vs 7.66±0.27) were higher than that of control group (P＜0.05).2.The main frequency of severe reflux patients in fasting and postprandial [( 1.68 ± 0.07) cpm vs (2.13 ± 0.07)cpm; (2.18±0.09) cpm vs (2.76±0.06) cpm],the percentage of normal gastric slow wave in fasting and postprandial (69.71±0.43 vs 80.35±0.68; 56.36 ±0.85 vs 72.34±0.80),fasting /postprandial PR (1.47±0.04 vs 2.02±0.04) were lower than those of mild-reflux group (P＜0.05).The percentage of bradygastria in fasting and postprandial of severe reflux patients (22.94 ± 0.68 vs 13.47 ± 0.61; 29.61 ± 1.14 vs 17.55 ± 0.51) and the percentage of tachygastria in fasting and postprandial (9.94 ± 0.54 vs 7.02 ± 0.42 ; 17.04 ± 0.70 vs 10.71 ± 0.20) were higher than that of mild-reflux group (P＜0.05).3.There was no significant difference of gastric electrical parameters in fasting and postprandial between Hp-positive and Hp-negative groups (P＞0.05).4.The ratio of gastric emptying in DGR group was significantly lower than that of control group (37.9％ vs 90.5 ％,P＜0.05).The gastric emptying delay in DGR group significantly increased compared with control group (60.3％ vs 9.5％,P＜0.05).There was no significant difference in gastric emptying delay between severe-reflux group and light-reflux group (69.0％ vs 51.7％,P ＞ 0.05).Conclusions There is dysfuntion of gastric myoelectrical activity and gastric motility in primary pathological DGR patients,which may be an important mechanism in pathological DGR.

Aim To investigate the protective effects of ginsenosides ( GS) on reactive oxygen species-induced oxidative damage in mouse spermatogonial cells. Meth-ods Mouse spermatogonial cell oxidative stress model was established and the attenuating effects of ginsen-osides on germ cell oxidative damage were evaluated by determination of cell viability,malondialdehyde( MDA) formation,superoxide dismutase ( SOD) activity and glutathione ( GSH) level. Results The exposure to hypoxanthine/xanthine oxidase ( HX/XO) induced an elevation in MDA,while a decrease in germ cell viability,SOD activity and GSH level. However,supplementation with GS ( 10 mg?L -1) restored HX/XO-induced decrease in cell viability,SOD activity and GSH level and HX/XO-induced increase in MDA formation. Conclusion GS may exert antioxidant activity to attenuate reactive oxygen species-induced oxidative damage in mouse spermatogonial cells.

Objective To investigate the status of geriatric doctors' acknowledge and treatment of outpatients with lower urinary tract symptoms in 15 Chinese cities.Methods The investigation was conducted in 15 Chinese cities and involved 1056 geriatric doctors from December 2013 to February 2014.The questionnaire survey was made in behavior,attitude,skill and knowledge(BASK)of LUTS.The geriatric doctors participated in the investigation by answering the questionnaires about acknowledge for definition,etiology,diagnosis and treatment of the LUTS,and in results discussion,which was aimed at improving participants' cognition for diagnosis and treatment of elderly LUTS and at strengthening the practice.The final objective was to promote diagnosis standardization and to improve the level of clinical diagnosis and treatment for the elderly 1UTS.Results 95.9％ of geriatric doctors considered that they knew well the main clinical manifestations of LUTS.However,34.3 ％ of geriatric doctors were able to classify the symptoms of LUTS correctly.More than 90.0 ％ of geriatric doctors considered that the causes of LUTS were variable and they mastered the common cause of LUTS.But,38.0％ of them were able to completely master the causes of LUTS and 35.5％ would actively ask outpatients about the symptoms.70.0％ of the doctors thought they were familiar with the initial manifestations of LUTS and benign prostatic hypertrophy(BPH).However,many Chinese geriatric doctors didn't follow up the guidelines and 30.0％ of them failed to evaluate the severity of LUTS via IPSS(International Prostatic Symptom Score).75.8％ of geriatric doctors considered that they would give the patients the best treatment options by comprehensive assessment.However,60.0％ of them didn't know that α1 receptor blocker was the first line of treatment option of patients with moderate severe LUST.More than half of them were unclear that the patients with prostate volume under 40 ml could be treated by α1 receptor blocker.82.3％ of geriatric doctors considered that LUTS and hypertension should be treated respectively.But,55.0％ of them didn't know that ALLHAT trial has proved that Doxazosin would increase the risk of cardiovascular event.Conclusions The geriatric doctors are now able to pay attention to outpatients with LUTS,but still should further strengthen their acknowledge on the definition,cause,diagnosis and treatment of LUTS.

Objective To investigate the relationship between prostate histologic inflammation classification and prostate cancer (PCa) in needle biopsy specimens with serum prostatic specific antigen (PSA) under 20 μg/L.Methods The clinical records of patients who underwent prostate biopsy were retrospectively analyzed in Beijing Hospital Urological Department from January 2011 to December 2013.The patients underwent prostate biopsy because of PSA raised and without pelvic radiotherapy.The needle biopsy specimen was evaluated by the pathohistologic criteria of location,extent and grade classification of prostate inflammation.Results PCa was detected in 84 (37.2％) biopsies and benign prostatic disease (BPD) in 142 (62.8％) biopsies.There were significant differences between PCa and BPD in inflammation classification (location:P＜0.001,extent:P＜0.001,grade:P＜0.001).On multivariable Logistic regression analysis,both location and extent classification of chronic histologic prostatitis were found tobe significantly associated with a lower risk of PCa in biopsy (location:P =0.001,OR =0.114,95％ CI 0.032-0.405 ; extent:P =0.021,OR =0.232,95％ CI 0.067-0.804).There was no relationship between grade classification and PCa (P=0.223,OR=1.805,95％CI 0.698-4.667).Under the evaluation of location and extent classification,it could differentiate PCa with BPD in biopsy with a sensitivity,specificity,positive predictive value and negative predictive value of 91.7％,50.7％,52.4％,91.1％,respectively.Conclusion The location and extent classification of chronic prostatitis is found to be associated with a lower risk of PCa independently.

Objective To obtain alpaca single domain antibody targeting Her2.Methods An alpaca was immunized with human recombination Her2 protein mixed with Freund's adjuvant.Total RNA was extracted from the alpaca's blood and was used to synthesize first strand cDNA.Single domain antibody variable region (VHH) gene of the alpaca was amplified by PCR and cloned into pMES4 vector for library construction.After screening, E.coli BL21 (DE3) was transformed with selected clones and was induced with IPTG for the expression of recombinant proteins.The nanobody was purified by nickel ion affinity chromatography column.The affinity of the nanobodies to Her2 was tested.Results After the second round of screening, two antibody clones were selected, H3 and H5.The affinity of H5 was 8.106×10-10mol/L.Histochemistry results showed that H5 could recognize Her2 antigen in breast tumor tissue.Conclusion An Her2 specific nanobody derived from alpaca is obtained through phage display library screening, which can recognize human Her2 antibody in human breast tumor tissue.

Objective To investigate the feasibility and clinical effect of the tubeless minipercutaneous nephrolithotomy (PCNL) in treatment of upper ureteral calculi.Methods From March 2014 to March 2015,all the patients with upper ureteral calculi except for those with severe infection,pyonephrosis or renal cortex less than 5 mm were randomized into two groups,the standard PCNL group (24 F nephrostomy tube) and the mini-PCNL group (18 F nephrostomy tube).After PCNL,all the patients received ultrasound examination to check residual stones,perforation and urine leakage.DJ tube was placed and the channel of PCNL was packed with hemostatic sponge without nephrostomy tube.There were 26 patients in standard group and 28 in mini-group.The operation time,postoperative hemoglobin change,postoperative visual analogue pain score (VAS),the time when urine turned clean,postoperative urinary extravasation,hydrothorax,fever and the stone-free rate were compared between two groups.Particularly,these data were compared in those aged＞65 years.Results The operation time in the standard PCNL and the mini-PCNL group was (58.3 ±21.8) and (86.4±23.3) minutes respectively,and had a significant difference (t=10.836,P＜0.05).The decrease in hemoglobin level was (8.3±5.8) g/L and (7.7±0.5.4) g/L,the VAS scores was (3.8±0.8) and (3.6±1.1),the time when urine turned clean was (11.9±4.7) h and (9.6±5.6) h,the postoperative hospital stay was (5.1±0.8) d and (4.8±1.2) d,and the stone free rate was 92.3％ and 89.2％ in standard PCNL and mini-PCNL group respectively(P＞ 0.05).No significant difference were found between two groups (all P＞0.05).There was one patient who got fever more than 38.5℃ in the standard group and 2 cases in the mini-group.Each group had 1 slight hydrothorax,and no blood infusion and perinephric urinary extravasation were found.The application of packing hemostatic sponge in the nephrostomy channel was feasible and suitable for both the standard and tubeless mini-PCNL groups.Conclusions The application of packing hemostatic sponge in the nephrostomy channel is feasible and suitable in both the standard and tubeless miniPCNL.It is safe for the treatment of renal and ureteral calculi,and it can decrease the hemorrhage and urine leakage,which works for the elderly patients too.

[ABSTRACT]AIM:Toestablishatransgenicheterozygousmousemodelofprecancerouslesionsofcolorectal cancer with p110δmutation in the C57BL/6J background for serving the studies on colorectal cancer research mediated by p110δ.METHODS:The transgenic heterozygous mice were generated by crossing in p110δD910A/D910A mouse and ApcMin/+mouse, and the genotype was detected by PCR .Compared with ApcMin/+mice, transgenic heterozygous mice ( ApcMin/+;p110δD910A/D910A)were counted, and the number and size of intestine polyps were analyzed after methylene blue staining . The intestinal tissue structure was assessed by HE staining .RESULTS:The transgenic heterozygous mouse model of pre-cancerous lesions of colorectal cancer with p 110δmutation was established .The number and size of polyps in the transgenic heterozygous mice were declined .CONCLUSION: A transgenic heterozygous mouse model of precancerous lesions of colorectal cancer with p 110δmutation was successfully established .The initial phenotype of intestinal tumors in transgenic mice was observed .This model will greatly contribute to the related research of colorectal cancer in mice .

BACKGROUND:Stable and efficient labeling of dental pulp stem cel s in vitro is most important in tracer technique, which is also the basis of tooth regeneration in vivo.
OBJECTIVE:To determine the optimal condition and method for transfection of stem cel s derived from rat dental pulp with green fluorescent protein infection by lentiviral vector and to determine whether green fluorescent protein-labeled dental pulp stem cel s maintain their stem cel properties.
METHODS:Rat dental pulp stem cel s were obtained by modified enzyme digestion method, to identify the immune phenotype and differentiation potential. Dental pulp stem cel s were infected with green fluorescent protein by lentiviral vector for 24 and 48 hours at different multiplicity of infection (MOI) (5, 10, 25, 50 and 100). The infection efficiency and fluorescence intensity were analyzed by inverted fluorescent microscopy. The clonal and proliferation ability, cel cycle and the mineralization potential were compared before and after transfection. Based on those mentioned above, we could evaluate the influence of infection on their biological characteristics.
RESULTS AND CONCLUSION:Flow cytometry results showed that rat dental pulp stem cel s expressed STRO-1 and CD146 but not CD34 or CD45. The dental pulp stem cel s could differentiate into osteoblasts and
adipocytes when cultured in specific medium for each lineage differentiation. The highest efficiency of infection and strongest fluorescence expression appeared at 48 hours of infection and MOI 50. There were no significant differences in growth ability, cel colony formation rate and cel cycle before and after transfection (P>0.05). And the alkaline phosphatase expressed positively. Infection for 48 hours at MOI 50 is optimal for transfecting dental pulp stem cel s with green fluorescent protein by a lentiviral vector, thereby providing reliable tracer method for the study of rat dental pulp stem cel s in vivo.

Objective To translate the International Consultation on Incontinence Modular Questionnaire for Male Lower Urinary Tract Symptoms (ICIQ-MLUTS) and ICIQ-MLUTS long form (ICIQ-MLUTS LF) into Chinese and assess their metric properties and applicability.Methods After obtaining permission from the International Consultation on Incontinence Questionnaire (ICIQ),original ICIQ-MLUTS and ICIQ-MLUTS LF were translated into Chinese according to ICIQ validation protocol and cultural adaptation guideline.During November 2017 to August 2018,Chinese version of ICIQ-MLUTS and ICIQ-MLUTS LF were tested by administering them in 6 and 12 hospitals in China respectively.To validate the translated questionnaires,the following tests were undertaken.The content validity was determined by indepth interviews with participants and experts.The correlation coefficients of ICIQ-MLUTS and ICIQ-MLUTS LF with International Prostate Symptom Score (IPSS) were assessed to validate criterion validity.Cronbach's alpha test was used to explore internal consistency.And the test-retest reliability was evaluated by calculation of intraclass correlation coefficient.Results In total,Chinese ICIQ-MLUTS and ICIQ-MLUTS LF were administrated to 135 and 230 male patients with lower urinary tract symptoms respectively.Both questionnaires had good content validity and good criterion validity with IPSS (Pearson correlation 0.846 and 0.833 for ICIQ-MLUTS and ICIQ-MLUTS LF respectively,both P ＜ 0.001).The Cronbach's alpha coefficient was 0.797 for ICIQ-MLUTS,and 0.853 for ICIQ-MLUTS LF.Intraclass correlation coefficient was 0.986 and 0.985 respectively (both P ＜ 0.001),showing good test-retest reliability.Conclusions The Chinese version of ICIQ-MLUTS and ICIQ-MLUTS LF had good validity and reliability,which can be used to assess Chinese male patients with lower urinary tract symptoms.