Objective To identify the therapeutic effect and safety of mini -percutaneous nephrolithotomy combined with antegrade modular flexible ureteroscope for the treatment of staghorn stone.Methods The clinical data of 116 patients with staghorn calculi who underwent mini -percutaneous nephrolithotomy combined with antegrade modular flexible ureteroscope were retrospectively analyzed.Of the 116 patients,63 cases were men,53 cases were women .Age ranged from 2 5 to 6 7 years .The diameter ofcalculi ranged from 4 6 to 9 8 (mean =4 6 )mm .There were 63 large complete staghorn renal calculi in these patients.Results All procedures were performed successfully using a single lithotripsy tract.The mean surgical time was (125.4 ±30.0)minutes.The initial stone -free rate was 81.03%(94 /116).Twenty -two cases had several residual calculi from 12 to 25mm.Post -procedure complications included hemorrhage in 9 patients,fever(>38.5 ℃)in four patients,and reactive pleural effusion in one patient. Blood loss requiring transfusion,sepsis,adjacent organ injury and kidney loss were not observed.Conclusion Mini -percutaneous nephrolithotomy combined with antegrade modular flexible ureteroscope has good therapeutic effect in treating renal staghorn calculi,since the technique in treating the renal staghorn calculi makes the operation period shorter,the rate of stone -free higher,the operation wounds smaller,the rate of complication lower,the rate of surrounding organ injury less.

Objective To evaluate the diagnostic value of immunohistochemistry (IHC) for the identification and localization of Treponema pallidum (TP).Methods Rabbit anti-human TP polyclonal antibody labeled IHC was used to detect 20 paraffin-embedded biopsy samples from lesions of 14 patients with syphilis or suspected syphilis in Sir Run Run Shaw Hospital of Zhejiang University from January 2004 to May 2012.Results TP was detected in 80％ of all the 20 samples by IHC assay,including 83.3％ (5/6) in patients with primary syphilis,100.0％ (10/10) in patients with secondary syphilis,and 25.0％ (1/4) in patients with tertiary syphilis,with a positive diagnostic accuracy of 100.0％.TP was mainly present in lower part of epidermis or perivascular,characterized by an endotheliotropic and epitheliotropic patterns or in the tissue of granulomatous inflammation.Besides,the density of TP was associated with types of lesions.There were more TP in the lesions of syphilis chancre,syphilis proctitis and condyloma latum,and fewer TP in the lesions of squamous erythema,greyish-black plaque,ulcer of chest wall from tertiary syphilis,and least in syphilitic lymphadenitis.There were no correlations between the quantity of TP and the rapid plasma regain (RPR) test titer (P＞0.05).Conclusions IHC for TP is of both high sensitivity and specificity for the diagnosis of syphilis,suggesting that TP-IHC is helpful for the diagnosis of syphilis,especially for the diagnosis of early suspected syphilis with negative serological results,systemic damage of syphilis,or syphilis in untypical locations and unusual lesions.It can serve as an alternative method for the diagnosis of syphilis.

Aim To screen the differentially expressed microRNAs ( miRNAs ) induced by hypoxia precondi-tioning ( HPC ) in adult rat cardiomyocytes, and pre-dict miRNAs-regulated target genes and their func-tions. Methods Cardiomyocytes were isolated from a-dult rat ventricular myocardium and cultured ( in vitro) . The cells were divided into 2 groups: control group ( CON ) and hypoxia preconditioning group ( HPC) . The cardiomyocytes in HPC group were sub-jected to 10 min hypoxia followed by 30 min reoxygen-ation, while the cells in CON group were cultured un-der normal condition. After that, total RNA was ex-tracted and then subjected to miRNA microarray to screen differentially expressed miRNAs. The microar-ray results were further validated by quantitative RT-PCR ( qRT-PCR ) . Bioinformatics analysis was per-formed to predict the miRNAs-regulated target genes and analyze the enriched gene ontology ( GO) and sig-naling pathway ( Pathway) . Results HPC caused sig-nificant changes in miRNAs expression in cardiomyo-cytes as compared to CON group. A total of 12 miR-NAs were up-regulated and 14 miRNAs were down-reg-ulated ( P <0. 01 , FDR <0 . 05 ) . The differentially expressed 7 miRNAs with a fluorescent signal intensity>500 were selected for further bioinformatics analysis. The expression of miR-133b-5p, miR-664-1-5p and miR-6216 detected by qRT-PCR exhibited the similar patterns of up or down regulation to those shown in mi-croarray results. Bioinformatics analysis revealed that miRNAs-regulated target genes were significantly en-riched in 27 GOs and 6 signal pathways. Conclusion
The expression profile of miRNAs in rat cardiomyo-cytes is significantly affected by HPC. These differenti-ally expressed miRNAs might participate in HPC-in-duced cardioprotection by regulating their target genes in rat cardiomyocytes.

Objective To validate the hypothesis that the phenotype reversion occurs in the smooth muscle cells in the corpus cavernosum of hypertensive rats with selenium food and explore selenium′s impact on erectile function. Methods 25 14-week-old male spontaneously hypertensive rates (SHR) were divided into 2 groups (SHR-ED-Se group n = 8 and SHR-ED-N group n = 9). SHR-ED-Se rats were fed with selenium diet, SHR-ED-N rats and WKY rats with normal diet. And 10 syngeneic normotensive rats (WKY) were used as control group. All rats were killed after 4-week feeding. Immunohistochemical staining and color image analysis system were used to observe expression of α-actin, desmin and osteopontin (OPN) in rats corpus cavernosum of different group. Results 2 SHR-ED-N rats died. Expression of α-actin of smooth muscle in corpus cavernosum in SHR-ED-N group was the lowest in all groups (P = 0.000), so did erection frequency (P < 0.05). Expression of OPN in SHR-ED-N is the highest (P = 0.000). No significant difference existed in expression of desmin in all group (P > 0.05). Conclusions Phenotype modulation of smooth muscle in corpus cavernosum of SHR rats can be reversed by selenium which means the smooth cells can transform from synthetic phenotype into the contractile phenotype, resulting ultimately in improving erectile function.

Aim To evaluate the effects of morphine preconditioning ( MPC ) on the expression of microR-NAs ( miRNAs ) induced by hypoxia-reoxygenation (H/R) in H9c2 myocardial cells. Methods H9c2 cells were randomly divided into 3 groups ( n=4 each) as follows:control group ( CON) , hypoxia/ reoxygen-ation group ( H/R ) and morphine preconditioning group ( MPC+H/R) . The cells were cultured in nor-mal condition in CON group. The cells were subjected to 5 h hypoxia followed by 1 h reoxygenation in H/R group and MPC+H/R group. Specifically, the cells in MPC+H/R group were preconditioned with morphine with the final concentration of 1 μmol·L-1 for 10 min before H/R. After the treatment, CCK-8 was used to detect cell viability and chemical colorimetry was used to detect lactate dehydrogenase ( LDH ) activity in the culture medium. Cell apoptosis was assessed by An-nexin-V-FITC/PI flow cytometry. Relative expression of Fas protein was detected by Western blot. The ex-pression of miRNA in myocardial cells was analyzed by quantitative reverse transcription polymerase chain re-action ( qRT-PCR ) . Results Compared with CON group, the cell viability was significantly decreased, while the LDH activity, apoptotic rate and Fas protein expression were dramatically increased in group H/R (P<0. 01). However, MPC significantly increased the cell viability, whereas it decreased the LDH activity, apoptotic rate and Fas protein expression induced by H/R injury ( P < 0. 01 ) . The expressions of miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7 e-5 p were markedly down-regulated by H/R as compared to CON group ( P <0. 05 ) , while MPC inhibited these miRNAs which were significantly down-regulated by H/R group ( P <0. 01 ) . Conclusion Morphine preconditioning might protect H9 c2 myocar-dial cells against H/R injury by regulating the expres-sion of miRNAs such as miR-133a-5p, miR-133b-5p, miR-664-1-5p, miR-6216 and let-7e-5p.

Objective To evaluate the role of microRNA-133b-Sp (miR-133b-Sp) in apoptosis hypoxia/reoxygenation (H/R) induced by in rat cardiomyocytes.Methods Rat myocardial cell line H9c2 was cultured in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum.The cells were seeded in 96-well or 6-well plates and randomly divided into 4 groups (n=64 wells each):control group (group C);group H/R;miR-133b-5p mimic + H/R group (group M+H/R);miR-133b-Sp negative control + H/R group (group NC+H/R).The cells were exposed to 95％ N2-5％ CO2for 5 h at 37 ℃ followed by 1 h reoxygenation in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum in all the groups except group C.The cells were cultured in normal culture atmosphere in group C.In M+H/R and NC+H/R groups,the cells were transfected with miR-133b-5p mimic (final concentration 30 nmol/L) and miR-133b-5p negative control (final concentration 30 nmol/L),respectively,for 24 h before H/R.Total RNA was extracted from cells to detect the expression of miR-133b-5p using quantitative real-time PCR.The cell viability (by CCK-8) and lactic dehydrogenase (LDH) activity in the culture medium were detected.Cell apoptosis was assessed by Annexin V/PI flow cytometry.Apoptotic rate was calculated.Result Compared with group C,the cell viability was significantly decreased,and the LDH activity and apoptotic rate were increased in H/R,M+H/R and NC+H/R groups,the expression of miR-133b-5p was down-regulated in H/R and NC+H/R groups,and the expression of miR-133b-Sp was up-regulated in group M+H/R.Compared with group H/R,the cell viability was significanttly increased,the LDH activity and apoptotic rate were decreased,and the expression of miR-133b-5p was up-regulated in group M+H/R,and no significant change was found in the parameters mentioned above in group NC+H/R.Conclusion H/R in rat cardiomyocytes can induce cell apoptosis possibility through down-regulating the expression of miR-133b-5p

Objective To evaluate the effect of morphine preconditioning on the expression of miR-133b-Sp and Fas in rat cardiomyocytes subjected to hypoxia/reoxygenation (H/R).Methods Cardiomyocytes were isolated from healthy adult male Sprague-Dawley rats by using Langendorff perfusion.The cells were seeded into 24-well plates or 60 mm diameter dishes and randomly divided into 3 groups (n =24 each) using a random number table:control group (group C),group H/R,and morphine preconditioning group (group MPC).The cells in group C were cultured in normal culture atmosphere.In H/R and MPC groups,the cells were exposed to 95％ N2-5％ CO2 for 90 min followed by 120 min reoxygenation.In group MPC,the cells were cultured for 10 min in serum-free DMEM liquid culture medium containing morphine 1 μmol/L,and then were cultured for 30 min in morphine-free DMEM liquid culture medium before hypoxia.At 120 min of reoxygenation,the cells in 24-well plates were selected to detect the cell viability (by MTT),lactate dehydrogenase (LDH) activity in the culture medium,and cell apoptosis (by Hoechst 33234 staining).Apoptosis rate was calculated.Total RNA and protein were extracted from the cells in 60 mm dishes to detect the expression of miR-133b-5p and Fas mRNA (by quantitative real-time PCR) and Fas protein (by Western blot).Results Compared with C group,the cell viability was significantly decreased,LDH activity and apoptosis rate were increased,the expression of miR-133b-Sp was down-regulated,and the expression of Fas mRNA and protein was up-regulated in H/R group.Compared with H/R group,the cell viability was significantly increased,LDH activity and apoptosis rate were decreased,the expression of miR-133b-5p was up-regulatcd,and the expression of Fas mRNA and protein was down-regulated in MPC group.Conclusion The mechanism by which morphine preconditioning reduces H/R injury to rat cardiomyocytesis related to up-regulation of the expression of miR-133b-Sp and down-regulation of the expression of Fas.

Objective: To describe characteristics of cardiometabolic risk factors of children and adolescents aged 6-17 years in 7 cities in China from 2013 to 2015. Methods: Data was from the China Child and Adolescent Cardiovascular Health (CCACH) study. 12 590 children and adolescents were selected from 24 schools (3 kindergartens, 7 primary schools, 7 junior high schools and 7 senior high schools) in seven cities (Changchun, Yinchuan, Beijing, Jinan, Shanghai, Chongqing and Tianjin) during 2013-2015 by using a stratified cluster random sampling method. The demographic characteristics, e.g. birth date, feed status and history of disease, were collected by questionnaires. Anthropometric measurements, i.e. weight, height, waistline, blood pressure, total cholesterol, fasting plasma glucose, triglyceride and high-density lipoprotein, were also collected. The detection rate of metabolic syndrome was calculated respectively according to "international diabetes federation standard " and "definition and prevention of metabolic syndrome in Chinese children and adolescents " . Results: The prevalence of cardiometabolic risk factors such as obesity, hypertension, hyperglycemia and dyslipidemia was 12.0%(1 497/12 491), 18.2%(2 193/12 035), 24.4%(3 028/12 422) and 15.8%(1 977/12 490), respectively. The prevalence of these four cardiometabolic risk factors in males was significantly higher than that in females (all P values<0.05). The prevalence of metabolic syndrome was 3.3%(272/8 328) with international diabetes federation 2007 definition and 5.4% (453/8 325) with Chinese definition among children above 10 years old. The prevalence of hypertension, hyperglycemia, hypertriglyceridemia, high total cholesterol, low high-density lipoproteincholesterol and dyslipidemia increased with the change of obesity type from non-obesity to complex obesity (all P values<0.001). Conclusion The prevalence of cardiometabolic risk factors was still high in children and adolescents, which has become an important factor threatening the healthy growth of children and adolescents.

Background Heavy metals may play an important role in environmental risk factors associated disorders of activities of daily living (ADL) in older adults. Objective To investigate the associations between plasma levels of six heavy metals (zinc, arsenic, cadmium, lead, manganese, and copper) and ADL disorders in older adults. Methods A cross-sectional survey was conducted from 2018 to 2019 among 1412 rural elderly people in Gongcheng Yao Autonomous County, Guangxi Zhuang Autonomous Region. The plasma metal concentrations were detected by inductively coupled plasma mass spectrometry (ICP-MS), and subsequently classified into three groups (T1-T3) based on tertiles, with the T1 group as the reference. The samples were assessed for ADL disorders using the ADL scale. Logistic regression and restricted cubic spline model (RCS) were used to estimate the odds ratio (OR) and 95% confidence interval (CI) to assess the associations between heavy metals and the prevalence of reporting ADL disorders. Results The mean age of the study population was (68.52 ± 5.92) years, 825 (58.4%) female and 587 (41.6%) male. Of these, 372 (26.34%) subjects reported ADL disorders, and most of them were female (74.30%). The results of logistic regression showed that the participants in the cadmium T2 group (0.15-0.25 µg·L⁻¹) had a higher risk of ADL disorders compared to the T1 group (≤0.15 μg·L⁻¹) (OR=1.552, 95%CI: 1.086, 2.134). After stratification by sex, the relative risk of ADL disorders was lower in the plasma copper T3 group (>1019.58 µg·L⁻¹) compared to the T1 group (≤868.12 μg·L⁻¹) in men (OR=0.481, 95%CI: 0.232, 0.998). The relative risk of ADL disorders was higher in the plasma cadmium T2 group (0.15-0.25 µg·L⁻¹) compared to the T1 group (≤0.15 μg·L⁻¹) in women (OR=1.758, 95%CI: 1.182, 2.616). The RCS results showed that the risk of ADL disorders in men was nonlinearly associated with copper (P_nonlinear=0.011, P_overall<0.05). Conclusion High levels of cadmium are positively associated with the risk of reporting ADL disorders, while high levels of copper are negatively associated with the risk of reporting ADL disorders in men.

Objective To observe the effects of short hairpin RNA (shRNA) targeting Her2 on its gene expression when the shRNA was stably transfected into human ovarian cell lines, SKOV3 and SKOV3. ipl, which have different extent of malignancy and investigate the changes of the biological characters of the two cell lines after the stable transfection. Methods The plasmids expressing shRNA targeting Her2 gene were transfected into SKOV3 and SKOV3. ipl cells. The stably transfected cells were gained by antibiotic screening. The expression of Her2 before and after the transfection was detected by RT- PCR and western blot. The transwell experiment was used to observe the invasion ability of the cancer cells before and after the transfection, and the parent and the transfected cells were injected into nude mice to observe the tumor growth. Results After the stable transfection with Her2 shRNA, mRNA and protein levels of Her2 gene in SKOV3 and SKOV3. ipl cells were remarkably reduced. The expression of mRNA were (68.0±3. 1) %, (40. 8±2. 0) %, (99. 9±1.3) %, (42. 4±2. 5) %. The expression of protein were (72. 1±3.4) %, (36. 4±1.5) %, (98.2±1.7) %, (40. 7±2. 1) %. The invasion ability into basilar membrane of the transfected cells was greatly reduced compared with the parent cells. The invasion cell numbers were 7.6±1.1, 1.8±0. 8, 36. 2±9.7, 15.7±7. 2. The growth rate of the planted tumors was lower in transfected groups than that of the parent groups. Conclusions (1) The expression of Her2 gene in SKOV3 and SKOV3. ipl cells was remarkably reduced by RNA interference targeting Her2. (2) The biological characters of SKOV3 and SKOV3. ipl cells are changed when the expression of Her2 gene is reduced by RNA interference.