Objective To explore the expressions of histamine receptor subtypes (H1, H2, H3, H4 receptor) in children's mid-urethral striated muscles and during mouse C2C12 striated myogenesis.Methods Children's mid-urethral striated muscle samples were paraffin embedded and tissue sections were made, then immunohistochemical staining was used to check H1, H2, H3, H4 receptors.C2C12 myogenesis was induced, the early differentiation early markers of desmin, middle marker of myogenin, late marker of myosin heavy chain and histamine 4 receptor subtype mRNA were checked by quantitative real-time polymerase chain reaction.Immunofluorescence staining was done to check 3 differentiation markers and histamine H3 receptor protein.Results During myogenesis, the expression of desmin mRNA in the differentiation of 2,4,6 days were 12,68,60 times as many as that of the undifferentiated myoblasts;the expression of myogenin mRNA in the differentiation of 2,4,6 days were 631,1 408,914 times as many as that of the undifferentiated myoblasts;the expression of myosin heavy chain mRNA in the differentiation of 2,4,6 days were 7 718,9 448,286 288 times as many as that of the undifferentiated myoblasts.The expression level of H1 receptor mRNA in the differentiation of 6 days was about 25％ to undifferentiated cells;the expression of H2 receptor mRNA in undifferentiated cells and differentiated cells groups had no significant difference (F =1.47, P ＞ 0.05);the expression of H3 receptor mRNA in the differentiation of 2,4,6 days was 28,103,198 times to undifferentiated cells;H4 receptor mRNA was not detected.In immunofluorescence staining, H3 receptor protein staining intensity increased with the differentiation.Immunohistochemistry of pediatric urethral striated staining indicated that H1, H2, H3 receptor staining was positive,H1 receptor showed strong positive staining, H3 receptor moderate positive staining,and H2 receptor showed weak positive staining.Conclusions Histamine receptor subtypes of H1 receptor, H2 receptor and H3 receptor were found during mouse striated myogenesis and in the children's mid-urethral striated muscles.The increasing expression of H3R with myogenesis might indicate it plays a role in mature striated muscle cells.

Objective: To study the infectious state of 3 kinds of viruses (CVB, CMV and EBV) in respiratory tract infection and its significance. Methods: Among 290 patients studied, there were 158 boys and 132 girls, aged from 6 months to 12 years. The antigen of coxsackievirus B (CVB-Ag) and antibody of coxsackievirus B (CVB-IgM), antibody of cytomegalovirus (CMV-IgM) and antibody of EB virus (EBV- IgM) were detected by ELISA method. Results: (1) The positive incidence of upper respiratory tract infection, bronchitis, pneumonia was higher than that of noninfectious disease group (P<0.01). The positive incidence of CVB was higher than that of CMV and EBV(P＜0.01). (2)There were cross viruses positive, especially in pneumonia group (P<0.05), usually CVB cross other virus infection. (3) The course of pneumonia was long in single CVB (P<0.01) and longer in cross infection of viruses (P<0.01). Conclusion: CVB infection is the most common one in the 3 viruses. Attention should be paid to multiple viruses infections in pneumonia children.

Object To optimize the best extraction of the total saponins in the root of Ophiopogon japonicus (Thunb.) Ker-Gawl. (RO). Methods The extract condition of total saponins in RO was observed by orthogonal design with ophiopogonis D as index. Results The optimum extract condition of RO was established. The more effective way was to smash RO to 5-10 mesh and extract total saponins in RO with ten times amount of 80% alcohol thrice by refluxing, one hour each time. Conclusion The total saponins in RO could be fully extracted in this condition.

Objective: To understand the relationship between coxsackievirus B and pediatric diseases. Methods: The infectious state of coxsackievirus B in hospitalized children were studied. Among 796 children studied, there were 218 upper respiratory tract infection cases, 179 pneumonia, 106 asthma, 155 myocarditis, 19 allergic purpura and 89 other diseases. The antigen (CVB-Ag) and IgM (CVB-IgM) were detected using ELISA method. Results: (1)There were 47% positive of CVB in upper respiratory tract infection and 48% positive of CVB in pneumonia(no difference between them, P>0.05). (2) There were 62% positive of CVB in asthma, 61% positive of CVB in myocarditis and 68% positive of CVB in allergic purpura(no difference among them, P>0.05)； But the positive rate of CVB in asthma, myocarditis and purpura were higher than in upper respiratory tract infection and pneumonia, (P<0.05). (3) There were lower positive rate of CVB in other kinds of diseases (16%) and in healthy children (3%)(no difference between them, P>0.05). Conclusion: CVB infection was related to several kinds of diseases, the relationship between CVB infection and diseases such as asthma, myocarditis, and allergic purpura should be further studied.

Objective To investigate the effect of early application of clean intermittent catheterization(CIC) in infants with neurogenic bladder(NB).Methods Eighty-seven children with NB diagnosed in our urodynamic center were less than 1 year old when they first came to hospital from January 2007 to January 2010, and CIC was carried out at different age.Sixty-four patients were followed up for a long time and divided into early CIC group(less than 1 year old children) and late CIC group(more than 3 years old children) according to the treatment time.Early CIC group included 29 patients [19 boys and 10 girls with the mean age of (7.5 ±2.8) months].And 4 cases were suffering from postoperative spina bifida manifesta;22 cases with spina bifida occulta;2 cases with sacral dysplasia;1 case with meningitis.Late CIC group included 35 patients [20 boys and 15 girls with the mean age of (8.0 ±2.9) months].2 cases were suffering from postoperative spina bifida manifesta;28 cases with spina bifida occulta;4 cases with sacral dysplasia;1 case with postoperative pelvic surgery.Before the treatment, there were no significant differences of the bladder compliance (BC), the maximum cystometric capacity (MCC) and the safety bladder capacity (SBC) between two groups.Urodynamic parameters and complications of 64 patients who were successfully followed up for 6 years were compared.Results After 3 years follow up, BC, SBC and MCC in early CIC group [(8.5 ± 1.9) ml/cmH2O, (140 ±25) ml, (142 ±29) ml]were significantly higher than those of late CIC group [(7.0 ± 2.2) ml/cmH2O, (110 ± 31) ml, (120 ± 28) ml;all P ＜ 0.05].After 6 years follow up, BC, SBC and MCC in early CIC group [(12.0 ±2.5) ml/cmH2O, (210 ±26) ml, (230 ±30) ml] were significantly higher than those of late CIC group [(9.3 ± 2.3) ml/cmH2O, (192 ± 31) ml, (205 ± 35) ml;all P ＜ 0.05], and the vesicoureteral reflux rate [24.1％ (7/29)] in early treatment group was significantly less than that in late treatment group [54.3％ (19/35), P ＜ 0.05].Increases in BUN and serum creatinine were found in 6 cases (20.7％) in early CIC group and 17 cases (48.6％) in late CIC group, the difference was significant (P ＜ 0.05).Conclusion For NB patients, the effect of early CIC is better than that of late CIC.

Ras-associated domain family 1A (RASSF1A) genes are members of the RASSF family, which bind to Ras in a guanosine triphosphate(GTP)-dependent manner and then induce Ras-mediated apoptosis. The protein encoded by the RASSF1A gene is similar to the Ras effector protein, which can interact with DNA repair protein XPA, and can also inhibit the accumulation of cyclin D1, thereby inducing cell cycle arrest. The deletion or abnormal expression of RASSF1A gene is related to the pathogenesis of various malignant tumors, indicating that it has tumor suppressor function. RASSF1A gene methylation has been found in at least 37 tumors, and RASSF1A gene may be the most frequently described methylated gene in human cancers. In this paper, the abnormal methylation of RASSF1A gene in different malignant tumors was introduced, and the research progress of its related effects and mechanisms in malignant tumors of the respiratory system, digestive system, genitourinary system, and nervous system in recent years was reviewed, with a view to malignant tumors early diagnosis, individual molecular targeted therapy and prognostic evaluation provide important guidance.

Objective To investigate the effect of erythropoietin(EDO)on the expression and function of aqua_porin_1( AQD _1)in the kidney of young male SD rats after release of bilateral ureter obstruction( BUO _ R). Methods Porty_eight young SD rats were randomly divided into bilateral ureteral complete obstruction(BUO)group (n﹦6),BUO_R group(n﹦12),BUO_R﹢EDO group( n﹦12)and Sham group( n﹦18). The BUO model was built through bilateral ureteral ligation. BUO group were killed after 24 h,and BUO_R group and BUO_R﹢EDO group were relieved after obstruction of 24 h. EDO(500 U∕kg)was given to BUO_R﹢EDO rats at 1 h after release of BUO,and then repeated 1 d,3 d and 5 d 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 and blood samples were collected on 3 d and 7 d(24 h,3 d,7 d for Sham group)after release of BUO. The u_rine samples were collected by using metabolic cage before death. The plasma osmotic pressure,creatinine(Cr)and u_rea nitrogen(BUN)in the plasma of young rats were detected. The expression of AQD_1 protein in all groups of kidney tissues was detected by adopting immunohistochemistry and Western blot. Results On day 3 after release of BUO,24 h water intake and urine volume of BUO_R﹢EDO group were higher than those of Sham group,but lower than those of BUO group(P〈0. 05),the urine osmotic pressure of BUO_R﹢EDO group was higher than that of BUO group,but lower than that of Sham group(P〈0. 05),while plasma osmotic pressure,Cr and BUN of BUO_R﹢EDO group were higher than those of Sham group,but lower than those of BUO group(P〈0. 05),and they were all of lower than BUO group( P 〈0. 05). On day 7 after release of BUO,there was no obvious change in Sham group,and the indexes of BUO_R group and BUO_R﹢EDO group gradually recovered,but they still did not reach the normal level(P〈0. 05). The difference between BUO_R group and BUO_R﹢EDO group was statistically significant(P〈0. 05). The immuno_histochemical results showed that the expression of AQD_1 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﹢EDO group than that of Sham group(P〈0. 05). Compared with 3 days after release of BUO,the staining intensity of BUO_R﹢EDO group and BUO_R group was enhanced,but still lower than that of the Sham group. These results were further confirmed by adopting Western blot,and BUO group was also the lowest of the four groups,and BUO_R﹢EDO group was higher than that of BUO group,but lower than that of Sham group( P〈0. 05). Conclusion EDO can promote not only the recovery of AQD_1 protein expression but also the recovery of renal function in young BUO_R rats.

Objective To explore the expression of aquaporin-2 (AQP-2) in human fetus kidney and amniotic fluid at different stages of pregnancy.Methods Twenty-two cases of aborted fetuses' kidneys were collected.They were divided into 3 groups according to the pregnancy age:8 cases in 17-23 + 6 weeks,8 cases in 24-31 +6 weeks,and 6 cases in 32-38 +6 weeks.Western blot was used to examine the expression of AQP-2 in the kidney.Twenty-four cases of the amniotic fluid were collected,and they were divided into 3 groups according to the pregnancy age:10 cases in 17-23 +6 weeks,6 cases in 24-31 +6 weeks,and 8 cases in 32-38 +6 weeks.Eight cases of healthy adult morning urine were collected as positive controls.The AQP-2 protein in the amniotic fluid was detected with the method of enzyme-linked immunosorbent assay (ELISA) and the osmotic pressure of amniotic fluid at different stages of the pregnancy was measured with the freezing point osmometer.Results The expression of AQP-2 was increased with the extending of pregnancy age,and the AQP-2 expressions in fetus kidney of 17-23 +6 weeks,24-31 + 6 weeks and 32-38 +6 weeks were 0.986 ± 0.335,1.566 ± 0.272,and 2.080 ± 0.246,respectively,and the difference was significant (P ＜ 0.05).The AQP-2 detected from amniotic fluid was positively correlated with the result of AQP-2 in the kidney(r =0.985,P ＜ 0.05),and the AQP-2 expression also increased with the extending of pregnancy age:17-23 +6 weeks,24-31 +6 weeks,32-38 +6 weeks and adult urine was (30.253 ±5.843) mg/L,(35.103 ±7.271) mg/L,and (42.580 ± 1.230) mg/L and (46.493 ± 0.450) mg/L,respectively.The osmolality of the amniotic fluid of 17-23 +6 weeks,24-31 +6 weeks,32-38 +6 weeks was (272.600 ± 4.827) mmol/L,(252.00 ± 15.360) mmol/L,and (261.750 ±5.560) mmol/L,respectively,and the difference was significant(P ＜0.05).Conclusions The AQP-2 expression in human fetus kidneys has good correlation with amniotic fluid,which indicates that the level of AQP-2 of the amniotic fluid may reflect the expression of AQP-2 in the fetus kidney.

Objective To investigate the value of detrusor isovolumetric test(DIT)on female SUI with sus-pected detrusor contraction weakness. Methods A total of 86 SUI patients with suspected detrusor contraction weakness diagnosed by preoperative bladder cystometry(CMG)were enrolled in this study.DIT method was used to evaluate the bladder function. The difference of first sensation cystometry capacity, maximum cystometric capacity, bladder compliance and maximum detrusor pressure were compared between the two methods.Patients in group Ⅰwhowere diagnosed with normal detrusor function with DIT method underwent TOT surgery. Ninety normal SUI patients were selected as control group Ⅱ,who were diagnosed with normal detrusor function with CMG method. The surgery effect was compared between the two groups. Results By using DIT detection,there were 65 cases showed normal detrusor function of the 86 cases(75.6%).The maximum detrusor pressure with DIT was(31.2 ± 12.9)cm H2O versus(11.7 ± 3.1)cm H2O with cystometry(P＜0.05).No significant differences were observed in the first sensation cystometry capacity,maximum cystometric capacity and bladder compliance.The effective rate of TOT in both group Ⅰ and group Ⅱ was more than 95%,with no significantly difference between the two groups (P＞0.05). Conclusions DIT is a valuable method to evaluate the bladder function of female SUI patients with suspected detrusor contraction weakness.

Objective:To investigate the expression of IKBKE and NF-κB in pancreatic cancer, and to explore the effect of IKBKE on pancreatic cancer proliferation and migration.Methods:Immunohistochemistry staining was used to study the expression of IKBKE and NF-κB in tissues of 61 pancreatic cancer patients admitted to the Second Hospital of Tianjin Medical University from January 2012 to January 2017 and 13 normal pancreatic tissues. The correlations between those expression to clinicopathological features were analyzed. Lentivirus mediated RNAi was transfected into pancreatic cancer cells to block IKBKE. Western blot was performed to test the silencing effeciency; CCK-8 and plate clone and scratch assays were used to investigate the proliferation and migration of pancreatic cancer.Results:Immunohistochemical staining showed that 60 (98.4%) of IKBKE staining were weakly positive, positive, and strongly positive in pancreatic cancer tissues, which were significantly higher than normal pancreatic tissues(76.9% cases were weakly positive and the rest were negative), and the differences were statistically significant ( P<0.05). All cases of NF-κB exhibited weakly positive expression and above in pancreatic cancer tissues, which was markedly higher than normal tissues (30.8% cases were weak positive and the rest were negative staining), statistically significant ( P<0.05). Survival analysis showed that patients with high level of IKBKE showed a shorter overall survival ( P<0.05). CCK-8, plate cloning and scratch assays showed that the proliferation and migration of were significantly decreased in IKBKE knocking down group ( P<0.05). Conclusions:IKBKE and NF-κB are highly expressed in pancreatic cancer, and IKBKE is correlated with NF-κB in pancreatic cancer. Blocking of IKBKE could distinctly inhibit the proliferation and migration of pancreatic cancer.