Objective To understand and identify the molecules related to the natural resistance to Schistosoma japonicum infection in Mirotus fortis. Methods Sera from Mirotus fortis without schistosome infection were collected. The S.japonicum adult worm cDNA library was immunologically screened with the sera. The positive recombinants were identified, cloned, sequenced and analysed with software and internet. Results Seven genes encoding antigens relevant to sera antibodies in Mirotus fortis were cloned and sequenced. These antigens included glyceraldehyde 3-phosphate dehydrogenase (GAPDH), serine protease inhibitors(SERPIN), 70 kDa heat shock protein(HSP70), 22\^6 kDa membrane-associated antigen, paramyosin (Sj97), cytochrome C and cathepsin B. Conclusion Many protein molecules might have been involved in natural resistance to \{S.japonicum\} infection in Mirotus fortis. The above 7 kinds of molecules may be identified as new candidates of vaccine against \{S.japonicum\} infection.

Objective To assess the value of the urine flow acceleration(UFA)versus maximum urinary flow rate (Qmax) for diagnosis of bladder outlet obstruction (BOO) in benign prostate hyperplasia (BPH).Methods A total of 50 men with BPH and 50 normal men were included in this study.Urodynamic examinations were performed in all patients according to the recommendations of the International Continence Society.Prostate volume,UFA and Qmax of each patient were analyzed and the results were compared between two groups.Results The UFA and Qmax of BPH group were much lower than that of the control group [(2.05±0.85)ml/s2 vs.(4.60±1.25)ml/s2 ; (8.50±1.05)ml/s vs.(13.00±3.35)ml/s,P＜0.05].The prostate volume in BPH group was increased compared with control group [(28.6±9.8) ml vs.(24.2±7.6)ml,P＜0.05].As diagnosis standard of UFA＜2.05 ml/s2 and Qmax＜ 10 ml/s,the sensitivity and specificity of UFA and Qmax in diagnosing BOO were (88％,75 ％)vs.(81％,63％).While compared with the result of P-Q chart,the Kappa values in correspondence analysis were 0.55 vs.0.35.The sensitivity,specificity and Kappa value of UFA in diagnosing BOO in BPHs were slightly higher than that of Qmax in comparison with the gold standard (BOO diagnosed by P-Q figure).Conclusions The UFA is a useful urodynamics parameter in diagnosing BOO of BPH.

Objective To investigate the voiding patterns of term and preterm newborns and whether voiding in term and preterm neonates was accompanied by any cortical arousal. Methods Between May 2013 and September 2013,64 hospitalized newborns at Neonatal Intensive Cave Unit in the Frist Affiliated Hospital of Zhengzhou University were recruited in this study. In these patients,31 cases were term newborns(20 male,11 female)and 33 cases were preterm newborns(19 male,14 female). The term and preterm newborns gestational ages at birth were(38. 2 ± 1. 2) weeks and(32. 1 ± 1. 6)weeks,weighted(3. 3 ± 0. 4)kg and(1. 7 ± 0. 3)kg,respectively and postnatal ages at study were[4 - 16(10. 5 ± 3. 6)]days and[4 - 16(11. 2 ± 3. 1)]days. The voiding volume(VV),post - void residual volumes(PRV),body movement rate and voiding frequency(VF)in 4 hours as well as the volume of milk and liquid fed at the same time frame were recorded and analyzed,retrospectively. At the same time electrocardiogram(ECG)and electroencephalogram(EEG)were recorded. The changes of heart rate(HR),EEG frequency,respiratory frequency (RF)during the 5 s period and 30 s before and after voiding onset were compared respectively. For cortical arousal definition the recommendations of the International Pediatric Work Group on Arousals(2005)were used. Results A total of 184 times of voiding were recorded. In preterm newborns,the VV and body movements rate were significantly lower compared with the term newborns[(21. 8 ± 7. 9)mL and(41 ± 21)% vs(26. 4 ± 8. 7)mL and(62 ± 19)% , t = 3. 75,4. 20,all P ﹤ 0. 05]. However,the VF and PRV were significantly higher in preterm newborns[(1. 7 ± 0. 9) mL and(3. 2 ± 1. 1)times vs(1. 2 ± 0. 7)mL and(2. 6 ± 0. 9)times,t = 2. 47,2. 38,all P ﹤ 0. 05]. Bladder voiding in these infants happened only during QS. In term newborns,HR frequency was higher during the 5 s interval before and after voiding onset when compared with the 30 s period before voiding onset[(152 ± 6)times/ min and(152 ± 5) times/ min vs(147 ± 6)times/ min,t = 5. 30,5. 76,all P ﹤ 0. 05]and the EEG frequency[(2. 6 ± 0. 1)Hz and (2. 6 ± 0. 1)Hz vs(1. 5 ± 0. 1)Hz,t = 70. 0,70. 0,all P ﹤ 0. 05]. While the HR and EEG frequency of preterm neo-nate was not changed before and after bladder voiding onset. The RF of both term and preterm neonates were not changed before and after bladder voiding onset. Conclusions The voiding patterns between term and preterm were sig-nificantly different and cortical arousal was found only in term neonates,which indicate the term newborns have better mature bladder function and development of nervous system.

Objective To investigate the changes in intracellular Ca2+ in the ureter smooth muscle cells (USMC) of rats with neuropathic urinary tract dysfunction (NUTD) and their significance.Methods Forty-five rats were randomly and averagely divided into NUTD group,experimental control (EC) group and blank control (BC) group.The NUTD group was operated with a spinal cord transection at the first lumbar level and the sacral cord was destroyed;in EC group the spinal process was partly bitten at the same position,but the spinal cord was not transected;BC group was given no operations.One week later,the video-urodynamic was performed to observe the acontractile detrusor (ACD),vesicoureteral reflux (VUR) and urinary tract dysfunction in rats among the NUTD group,EC group and the BC group.Video-urodynamic assessment was performed at the sixth week after operation.Ureter smooth muscle cells (USMC) were obtained by collagenase digestion.Intracellular Ca2 + in the USMC were observed by laser scanning confocal microscope.Then the effects of Bay K8644(10-8 mol/L,10-7 mol/L,10-6 mol/L) on cytosolic Ca2+ concentrations([Ca2+] i) in NUTD group were studied by calculating the fluorescence intensity.Results ACD and no detrusor overactivity were found in all rats in NUTD group and without vesicoureteral reflux.Immunofluorescence method confirmed that the cells were USMC.Compared with BC group (31.44 ± 2.82) and EC group (32.06 ± 3.67),the fluorescence intensity (FI) of intracellular Ca2 + in USMC was much lower in the NUTD group (9.80 ± 1.11),and there was significant difference(P ＜ 0.05).Bay K8644 (10-8 mol/L,10-7 mol/L,10-6 mol/L) increased the FI of [Ca2 +] i in a concentration-dependent manner,which were 3.80 ± 1.30,10.04 ± 2.15,19.89 ± 2.06,respectively,and there was significant difference (P ＜ 0.05).Conclusions The decrease in Ca2 + concentration in the ureter smooth muscle cells may be one of the important factors for the primary ureteral dysfunction of NUTD.And calcium channel agonist can be meaningful for adjusting abnormal Ca2+ concentration in USMC of NUTD.

Objective To investigate the effect of exogenous vascular endothelial growth factor (VEGF) on bone activity of rabbit heterotopic allograft decalcified bone.Methods 140 adult healthy China white rabbits were selected,no limitation with sex,20 rabbits as the donor preparation of allogenic decalcified bone,according to the random number table,the rest was divided into the experimental group (allograft decalcified bone ± VEGF) and the control group (Allograft decalcified bone),each group contained 60 rabbits.For the experimental group,the prepared 1.5 cm long homologous decalcified tibia was placed in rabbit right thigh of rectus femoris and vastus medialis muscle gap near by saphenous artery,and fixed on the femur with two 0.8 mm Kirschner wire.In the vicinity of the skin,implanted an osmotic pump which contain the VEGF solution 200 μl with concentration was 0.5 μg/ml.In the control group,implanted the isometric allograft decalcified bone in rabbit right thigh corresponding parts with the same method.Each group respectively at 0,2,4,6,8,10 weeks to death 10 white rabbits,By specimen observation,HE dyeing observation and detection of type Ⅰ glue protein fluorescence intensity,Analysis the bone activation degree of two groups of bone allograft decalcified.Results Experimental allograft decalcified bone gradually wrapped by connective tissue membrane,its surface appear different size of the pits and gradually increased and become deep,while the control group pits relatively little and shallow.In the experimental group and control group,the fluorescence intensity of type Ⅰ collagen reached its peak respectively at 8 weeks (47.57 ±3.50) and 10 weeks (45.07±6.02),with no statistically significant (P ＞ 0.05).Conclusion Rabbit allograft decalcified bone implanted in the muscle clearance with abundant blood supply can be transformed into activated bone after 10 weeks,and after applying exogenous VEGF,allograft decalcified bone can be transformed into activated bone after 8 weeks,the bone activation process obviously speed up.The reaults confirmed the exogenous VEGF can obviously promote the ectopic rabbit bone allograft decalcified bone activation process.

Objective: To investigate the expression of voltage-dependent potassium channel 1.3(Kv1.3) mRNA and protein during human monocyte-derived macrophage differentiation into foam cells and its function in foam cell formation. Methods: Human peripheral blood monocytes were isolated from healthy male volunteers by density gradient centrifugation and then by adherent method. The obtained monocytes were cultured for 5 days to differentiate into macrophages. Based on establishment of the human macrophage-derived foam cell model, the expression of Kv1.3 channel was investigated by immunocytochemical staining, reverse transcription-polymerase chain raction (RT-PCR) and Western blot. Furthermore, the effects of rMargatoxin, a Kv 1.3 channel-specific inhibitor, on cholesterol metabolism in macrophages incepting oxidized low density lipoprotein (OxLDL) were studied. Results: After the macrophages co-incubated with 30 mg/L OxLDL at 37 ℃ for 60 hours, the cellular volume obviously enlarged and many red lipid granules were deposited in cytoplasm. The total amount of cholesterol (TC),free cholesterol ( FC ) and cholesterol ester ( CE ) in cells markedly increased and the ratio of CE/TC rose from ( 14.4±6.8) % to (57.9±3.5) % (n=7,P＜0.05). However, the expression of Kv1.3 channel had no significant change. r Margatoxin (0.1 nmol/L and 10 nmol/L) markedly reduced the contents of TC, FC and CE in macrophages and the ratios of CE/TC decreased to (42.8±11.6) % and (22.6±8.0)% , respectively (n=7, P＜0.05). Meanwhile, the red lipid granules deposited in the cytoplasm of macrophages also decreased. Conclusion: These data clearly show that the expression of Ky1.3 channel does not change obviously during human monocyte-derived macrophage differentiation into foam cells and the blocking of it would prevent foam cell formation.

Objective To explore the relationship between congenital vesical ureteral reflux(VUR) and bladder dysfunction in children through videourodynamic examination.Methods Sixty-seven children with congenital VUR in the First Affiliated Hospital of Zhengzhou University from Apr.2011 to Jul.2013 were included,and their clinical information of urnary tract infection,detrusor activity,dysfunctional voiding and grade of VUR were recorded.All the children were categorized as normal,isolated detrusor overactivity (DO)and dysfunctional voiding (DV) (with or without DO) according to the manifestation of urodynamic patterns,who were also divided into groups of low grade (Ⅰ-Ⅱ) VUR or high grade (Ⅲ-Ⅴ) VUR.Data of video-urodynamic examination,urinalysis,and voiding cystourethrogram were collected to investigate the relationship between bladder dysfunction,sides and grade of VUR and urinary tract infection.Results Totally 73.1％ (49/67 cases) of children with VUR were found having bladder dysfunction,which consisted of 49.3％ (33/49 cases) of DO,23.8％ (16/49 cases) of DV.Children with isolated DO tended to manifest unilateral,low grade reflux (grade Ⅰ-Ⅱ) with less urinary infection.However,children with DV,isolated or combined with DO manifest bilateral,high grade reflux(grade Ⅲ-Ⅳ),and often with urinary infection.Conclusions Video urodynamic study is useful for evaluation of bladder function in children with VUR,which is important in management of VUR.

Objective To investigate the effect of erythropoietin(EPO)on the expression of aquaporin - 2 (AQP2)in the kidney of young SD rats after release of bilateral ureter obstruction(BUO - R). Methods Thirty - two young SD rats were equally divided into 4 groups randomly(BUO group,BUO - R group,BUO - R ﹢ EPO group and Sham group,8 rats in each group). The BUO model was built through bilateral ureteral ligation. EPO(500 U/ kg)was given to BUO - R ﹢ EPO rats at 2 h after release of BUO,and then repeated 6 h,12 h,24 h and 36 h thereafter and the same volume of 9 g/ L saline was simultaneously given to BUO - R rats. The Sham group was prepared in parallel by laparotomy and free dissection of bilateral ureters but not ligated. Both side kidneys were harvested 48 h(72 h for Sham group)after release of BUO to examine the effect of EPO on the expression of AQP2 in inner medulla by immunohisto-chemistry,Real - time PCR and Western blot. The urine samples were collected by using metabolic cage before death. Results The osmotic pressure of BUO - R ﹢ EPO group was higher than that of BUO - R group,but lower than that of Sham group(P ﹤ 0. 05). Immunohistochemistry showed that the collecting duct wall thinned and lumen enlarged. After the pictures were analysized by using Image - Pro Plus software,it showed that the expression of AQP2 in collecting duct in BUO group was significantly down - regulated compared with that in Sham group,whereas,it was slightly weaker in BUO - R group and BUO - R ﹢ EPO group than Sham group(P ﹤ 0. 05). These results were further confirmed by a-dopting Western blot,and the relative quantity of AQP2 in BUO group was also the lowest of the four groups(P ﹤0. 05). Real - time PCR showed that the level of AQP2 mRNA in Sham group was(24. 30 ± 1. 03)folds of BUO group,(10. 60 ± 1. 05)folds of BUO - R group and(5. 70 ± 1. 01)folds of BUO - R ﹢ EPO group,respectively. Conclusion EPO could promote not only the recovery of AQP2 mRNA and protein expression but also the recovery of AQP2 function in young BUO - R rats.

Objective: To assess the effect of viral macrophage inflammatory protein (vMIP) -Ⅱ on the expression of apolipoprotein B mRNA-editing enzyme-catalytic polypeptide-like 3G (APOBEC3G) , and to explore the mechanisms. Methods: A recombinant plasmid pEGFP-N3-K4 (vMIP-Ⅱ plasmid group) and an empty plasmid pEGFP-N3 (empty plasmid group) were separately transfected into 293T cells, and quantitative PCR and Western blot analysis were performed to evaluate the effect of transfection with vMIP-Ⅱ gene on the APOBEC3G expression in 293T cells. Some 293T cells in the empty plasmid group and vMIP-Ⅱ plasmid group were treated with 1 000 IU/ml interferon (IFN) -α for 36 hours, and then Western blot analysis was conducted to determine the APOBEC3G expression in the empty plasmid group and vMIP-Ⅱ plasmid group with or without IFN-α treatment. Some 293T cells transfected with vMIP-Ⅱ plasmids were treated with 75 μmol/L AG490 (a JAK/STAT signaling pathway inhibitor) and 20 μmol/L U0126 (an ERK signaling pathway inhibitor) separately; after 24 hours, total protein was extracted from 293T cells, and Western blot analysis was conducted to determine the expression of APOBEC3G. A recombinant plasmid containing APOBEC3G promoter was constructed by using a luciferase reporter gene, and the promoter fragment included the full-length promoter sequence (POS) of APOBEC3G, sequences with the lengths of 1 560, 960, 720, 480, 420, 360, 330 and 240 bp, and the regulatory element-free region (NEG) of APOBEC3G, separately. Some 293T cells were co-transfected with the recombinant plasmid carrying luciferase reporter gene and vMIP-Ⅱ plasmid (experimental group), or the recombinant plasmid and empty plasmid (control group). Subsequently, the activity of the APOBEC3G promoter was evaluated, and the key promoter region through which the transcriptional activity of APOBEC3G was regulated by vMIP-Ⅱ was analyzed. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference (LSD) -t test. Results: The mRNA and protein expression of APOBEC3G was significantly higher in the vMIP-Ⅱ plasmid group (2.500 ± 0.013, 1.472 ± 0.013 respectively) than in the control group (1, 0.364 ± 0.030 respectively; t = 6.22, 6.54 respectively, both P < 0.05) . The APOBEC3G expression significantly differed among the empty plasmid group, vMIP-Ⅱ plasmid group, empty plasmid + IFN-α group and vMIP-Ⅱ plasmid + IFN-α group (1, 2.030 ± 0.108, 2.700 ± 0.081 and 2.600 ± 0.099 respectively; F = 67.026, P < 0.001) , but there was no significant difference between the vMIP-Ⅱ plasmid group and empty plasmid + IFN-α group (t = 3.46, P > 0.05) . The APOBEC3G expression also significantly differed among the vMIP-Ⅱ plasmid group, vMIP-Ⅱ plasmid + AG490 group and vMIP-Ⅱ plasmid + U0126 group (0.617 ± 0.025, 0.179 ± 0.061, 0.359 ± 0.012 respectively; F = 70.019, P < 0.001) , and was significantly lower in the vMIP-Ⅱ plasmid + AG490 group and vMIP-Ⅱ plasmid + U0126 group than in the vMIP-Ⅱ plasmid group (t = 9.66, 11.836 respectively, both P < 0.01) . Luciferase activity assay showed that the promoter activity significantly differed among the vMIP-Ⅱ plasmid groups transfected with POS, 1 560-, 960-, 720-, 480-, 420-, 360-, 330-, 240-bp or NEG sequences (F = 81.092, P < 0.001) , and the APOBEC3G promoter activity decreased greatly from the 720-bp group to 480-bp group. Conclusion vMIP-Ⅱ upregulates the expression of APOBEC3G, likely through the JAK/STAT signaling pathway or the key promoter region regulating the transcriptional activity of APOBEC3G.

Objective To study the prevalence,risk factors of overactive bladder (OAB) in middle-aged and senior residents in Zhengzhou China.Methods A randomized,community-based,crosssectional study was performed on 10 160 residents aged 40 or older in urban area of Zhengzhou by using a stratified system sampling approach.A questionnaire including the subjects' basic information,previous history,present history,the Chinese overactive bladder symptom score (OABSS) was filled on site.The diagnostic criteria for OAB was 'an urgency score for Question 3 of 2 or more,and an OABSS of 3 or more'.Chisquare test was used to determine the differences of prevalence between genders,age groups,BMI and people with and without diabetes mellitus (DM).A pairwise comparison was conducted between different age,BMI group by using Bonferroni method.Results A total of 10 160 residents were investigated and finally 9805 (96.5％) were qualified for final statistical analysis.The mean age was (57.9 ± 9.7) years.The overall prevalence of OAB was 2.1％ (209/9805),of which,with OABdry 1.0％,and OABwet 1.1％.Male subjects were more likely suffered from OAB than female,with 2.7％ (84/3129) versus 1.9％ (125/6676).The prevalence of OAB in both male and female increased with age.There was no significant difference in the prevalence of male and female before the age of 60 years (1.2％ versus 1.4％,P ＞ 0.05) and more common in men than in women after the age of 60 years (4.6％ versus 2.6％,P ＜ 0.05).The prevalence of the subjects with DM was significantly higher than those without DM (P ＜ 0.05).The subjects with BMIs of 30 or more were nore likely to have OAB (3.2％ versus 1.8％,P ＜ 0.05).Conclusions The prevalence of OAB increases with advancing age.The prevalence of male is higher than female after the age of 60 years.The diabetics and obese people are more likely to have OAB.