Objective To establish the method for isolation and culture of urine-derived stem cells (USCs) from female patients with interstitial cystitis/bladder pain syndrome (IC/BPS).Methods The USCs were collected from fresh midstream urine samples from 6 female IC/BPS patients admitted to our hospital from June 2018 to December 2018.The 6 patients were 33-55 years old (average 41.5 years old),and their course of illness was 2-18 years (average 8 years).The USCs were isolated from the urine through times of centrifugation and cultured in specific medium.Growth curve and cell cycle of USCs were observed.The expression of surface markers of USCs was analyzed by flow cytometry and immunofluorescence,the smooth muscle and epithelial differentiation potential of USCs were detected by immunofluorescence staining of surface markers of smooth muscle cells and epithelial cells.Results USCs were successfully extracted from 3 of 6 female patients,and the success rate reached 50％ by once extraction.USCs showed a "rice-grain" spindle appearance and showed logarithmic growth.USCs expressed surface markers associated with mesenchymal stem cells (e.g.CD44,CD73,CD105,CD133) and embryonic stem cells [e.g.stage-specific embryonic antigen 4 (SSEA4)] and pericytes[e.g.CD146,platelet derived growth factor beta receptor (PDGFRB) and NG2],but didn't express hematopoietic stem cell surface markers(e.g.CD31,CD34 and CD45).When induced to smooth muscle cells or epithelial cells,the cells expressed the surface markers of smooth muscle cells [e.g.desmin,myosin,alpha-smooth muscle actin(otSMA) and vimentin] and epithelial cells(e.g.uroplakin 1A,uroplakin 3B,AE1/AE3 and cytokeratin 13).Conclusions The method of isolation and culture of USCs from female IC/BPS patients was successfully established,and it provides a preliminary technical method for exploring the application of USCs in the clinical study of IC/BPS patients with autologous treatment.

Objective To investigate the effects of leptin on Treg cells and the possible mecha-nism. Methods Leptin-deficient ( ob/ob) mice and homologous wild-type mice were used in this study. The percentages of Treg cells in spleen tissues and peripheral blood samples were measured by flow cytometry ( FCM) . Differences in Treg cell functionality were compared between the two groups. Splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by magnetic beads, were respectively cultured with leptin and anti-leptin neutralization antibody to evaluate the effects of leptin on Treg cells. Quantitative real-time PCR was performed to analyze the expression of Treg cell-related cytokines at transcriptional level. The levels of IL-10 and TGF-β in the supernatants of CD4+T cell culture were measured with Luminex technolo-gy. Results Compared with the wild-type mice, the ob/ob mice showed higher percentages of Treg cells in both peripheral blood samples and spleen tissues [(11. 56 ± 0. 72)％ vs (5. 47 ± 0. 81)％, (10. 16 ± 0.93)％ vs (6.29±0. 69)％]. Treg cells isolated from the ob/ob mice had stronger immunosuppressive effects on the proliferation of effector T ( Teff) cells and the secretion of TNF-α and IFN-γ than those from the wild-type mice [TNF-α:(1. 6±0. 2)％ vs (2. 4±0. 5)％, IFN-γ:(4. 3±0. 3)％ vs (7. 2±1. 2)％]. The percentages of Treg cells were decreased from (12. 2±1. 8)％ to (7. 6±0. 9)％ upon the in vitro treat-ment of CD4+ T cells from the ob/ob mice with leptin and the immunosuppressive effects of Treg cells were also weakened. However, the percentages of Treg cells were increased from (7. 8±0. 85)％ to (13. 1± 1. 5)％ upon the in vitro treatment of CD4+T cells from the wild-type mice with anti-leptin antibody and the immunosuppressive effects of Treg cells were improved as well. Moreover, the expression of Foxp3, IL-10 and TGF-β at transcriptional level and the levels of IL-10 and TGF-β in the ob/ob group were higher than those in the wild-type group. Conclusions Leptin deficiency significantly promoted the generation of Treg cells in mice and resulted in an increased expression of Foxp3, IL-10 and TGF-βat mRNA level and elevat-ed levels of IL-10 and TGF-β. The treatment of CD4+T cells with leptin might inhibit the generation of Treg cells through down-regulating the transcription of Foxp3, IL-10 and TGF-β.

Hyperpolarization-activated cyclic nucleotide-gated (HCN) channels are confirmed to be expressed in bladder interstitial Cajal-like cells (ICC-LCs), but little is known about their possible role in cystitis-associated bladder dysfunction. The present study aimed to determine the functional role of HCN channels in regulating bladder function under inflammatory conditions. Sixty female wild-type C57BL/6J mice and sixty female HCN1-knockout mice were randomly assigned to experimental and control groups, respectively. Cyclophosphamide (CYP)-induced cystitis models were successfully established in these mice. CYP treatment significantly enhanced HCN channel protein expression and I(h) density and significantly altered bladder HCN1 channel regulatory proteins. Carbachol (CCH) and forskolin (FSK) exerted significant effects on bladder ICC-LC [Ca²⁺]i in CYP-treated wild-type (WT) mice, and HCN1 channel ablation significantly decreased the effects of CCH and FSK on bladder ICC-LC [Ca²⁺]i in both naive and CYP-treated mice. CYP treatment significantly potentiated the spontaneous contractions and CCH (0.001-10 µM)-induced phasic contractions of detrusor strips, and HCN1 channel deletion significantly abated such effects. Finally, we demonstrated that the development of CYP-induced bladder overactivity was reversed in HCN1 -/- mice. Taken together, our results suggest that CYP-induced enhancements of HCN1 channel expression and function in bladder ICC-LCs are essential for cystitis-associated bladder hyperactivity development, indicating that the HCN1 channel may be a novel therapeutic target for managing bladder hyperactivity.
Animals ; Carbachol ; Colforsin ; Cyclophosphamide ; Cystitis ; Female ; Humans ; Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels* ; Mice* ; Telocytes* ; Up-Regulation* ; Urinary Bladder*

Objective To investigate the effect of intervention nursing on the formation of pressure ulcer in hypoalbuminemia patients with renal transplantation.Methods From February 2011 to June 2015,the 134 cases of hypoalbuminemia patients with renal transplantation in our hospital were divided into the observation group (73 cases)and the control group (61 cases).The control group were given conven-tional nursing methods for prevention of pressure ulcers,and the observation group were given corresponding intervention nursing measures on the basis of pressure ulcer risk factors.Then assessed the risk factors of pressure ulcers,the occurrence of pressure ulcers,and the rate of nursing satisfaction.Results The incidence of pressure ulcer in the observation group was significantly lower than that in the control group (P <0.05).The rate of nursing satisfaction in the observation group were significantly higher than that in the control group (P <0.05).The risk factors for hypoalbuminemia patients with renal transplantation were decrease of sensory response,increase of moisture in the compressed part,and increase of friction and shear forces in the compressed part.The occurrence of these risks could be significantly decreased after inter-vention nursing.Conclusion Effective nursing care can significantly reduce the incidence of pressure sores in hypoalbuminemia patients with renal transplantation and improve the nursing satisfaction.This method can be recommended for clinical nursing care for prevention of periop-erative pressure ulcer.

Objective To test the influence of peripherally inserted central catheter ( PICC) phlebotomizing and venipuncture ( VP) phlebotomizing on detection of coagulation function, so as to comfirm the feasibility of PICC. Methods Took 2 blood samples from the same patient, and divided the blood samples in to the PICC group and the VP group according to the methods of phlebotomizing to adopt a parallel controlled trial. Pearson correlation analysis and Bland-Altman analysis were taken during the following research. Results The 5 items of blood coagulation indexes testing showed that the results of PICC group and VP group were highly correlated, and the correlation coefficient was 0. 990 to 0. 997. The mean biases obtained by Bland-Altman analysis were closer to 0. Conclusion PICC and traditional method of VP showed obvious correlation and accordance in terms of the 5 indexes of coagulation function detection, which indicated that PICC is feasible for coagulation function detection.

Objective To explore the influence of catheterization for urinary tract infections and the importance of humanistic nursing. Methods Totally 248 patients who were admitted into our hospital from February 2013 to April 2014 were randomly divided into two groups. Patients in the control group (117 cases) were given spray fungicide and regular disinfection of urethra by nursing staff, and patients in the observation group (131 cases) were nursed by themselves or their family after been taught the way of spraying fungicide and cleaning. Ana-lyzed the psychological status, routine urine, urinary tract infections, nursing job satisfaction of the patients, and the relevant data were ana-lyzed by SPSS 17. 0 software. Results The third day after placing urinary catheter urethra, the relevant nursing can reduce the rate of urina-ry tract infections in patients. the difference of the two groups was statistically significant (P<0. 05). The rate of urinary tract infections at the seventh day after placing urinary catheter urethra was 39. 69% in the observation group and 41. 03% in the control group, the difference of the two groups was not statistically significant (P >0. 05). Infection rate of male patients (29. 01%) is higher than female patients (10. 69%). The satisfaction rate of urethra care in the observation group was 87. 79%, and the satisfaction rate of other project were all higher than 92. 5%. The satisfaction rate of urinary tract nursing check in the control group was 79. 49%, and the satisfaction rate of nurse-patient communication was 83. 76%. It was found that the difference of communication between nurses and patients, service attitude and the urethra care were statistically significant. Conclusion It can reduce the rate of urinary tract infection after placing urinary catheter in pa-tients for 7 days and nursed by patients themselves with spraying fungicide and cleaning. Strengthen the humanistic nursing care can improve nursing satisfaction.

Objective To investigate the inhibitory effect on human ACHN cell line and its mice xenograft by using interferon α-1b combined with cyclooxygenase-2 inhibitor and the relevant mechanism in vitro and vivo experiment .Methods ACHN cell and the xenograft mice were devided into 4 groups(IFN-α1b,NS398,IFNα-1b+NS398 and control group).The inhibitory effects were tested by CCK8(Cell Counting Kit 8)assay after AHCN were treated for 24 h and 48 h.The expression of bcl-xl and COX-2 were detected by Western blot .The vol-ume of the xenografts of ACHN cell line and testing the expression of VEGF in xenografts were measured by immunohistochemistry assay .Re-sults Both IFNα-1b and NS398 exerted inhibitory effects on ACHN and this effects showed a rising trend with a increasing concentration of drugs.The combined group was more significant than monotherapy group (P<0.05).Western blot assay showed that IFNα-1b and NS398 downregulated the expression of bcl-xl and COX-2 in ACHN.The combined group was more significant than monotherapy group (P<0.05). The combined group has the greatest inhibitory effects on the xenografts of ACHN cell line compared with monotherapy group and control group(P<0.05).The expression of VEGF in tumor was obiviously inhibited in combined group compared with monotherapy group and con -trol group (P<0.05).Conclusion IFNα-1b combined with NS398 can inhibit the proliferation of ACHN and suppress the tumor growth .

Objective To investigate the effects of leptin on Th17 cells and the possible mechanism. Methods The leptin-deficient ( ob/ob) mice and their homologous wild-type mice were used in the study.The percentages of Th17 cells in peripheral blood samples, spleen tissues and lymph nodes were measured by flow cytometry ( FCM) analysis.The splenic CD4+T cells, separated from the ob/ob mice and the wild-type mice by using magnetic beads,were respectively cultured with leptin at various concentrations and with anti-leptin neu-tralization antibody to evaluate the effects of leptin on Th17 cells.The quantitative real-time PCR was performed to analyze Th17 cell-related cytokines at transcriptional levels.The levels of IL-6 and IL-17A in the supernatants of CD4+T cell culture were measured with Luminex technology.Results Compared with the wild-type mice, the ob/ob mice showed lower percentages of Th17 cells in both peripheral blood samples and spleen tissues (0.49%±0.03%vs 1.29%±0.1%, 1.56%±0.22%vs 2.47%±0.11%).There was a decrease in the percentages of Th17 cells upon the in vitro treatment of CD4+T cells from wild-type mice with anti-leptin antibody.The per-centages of Th17 cells were increased in a dose-dependent manner upon the in vitro treatment of CD4+T cells from ob/ob mice with leptin.Moreover, the levels of IL-17A and IL-6 and the transcriptional levels of RORγt, IL-17A and IL-6 in leptin deficiency group were lower than those of wild-type group, but were increased upon the treatment with leptin.No significant difference with the transcriptional levels of TGF-βand IL-23 was ob-served between the two groups with and without intervention.Conclusion Leptin deficiency seriously hampered the generation of Th17 cells in mice and resulted in a decreased expression of RORγt, IL-17A and IL-6 at mRNA level.The treatment of CD4 T cells with leptin might promote the generation of Th17 cells through up-regulating the transcription of RORγt and IL-6.

Objective To investigate the efficacy of tamsulosin in the distal ureter calculi expelling after ureteroscopic lithotripsy. Methods A total of 120 patients with distal ureteral calculi were enrolled and randomly divided into the observation group and the control group with 60 patients in each group. Patients in the observation group received tamsulosin and traditional treatment ( including liquid impact therapy,anti-inflammatory and analgesia) ,while patients in the control group received placebo and traditional treatment after operation. The number of colic episodes,lower urinary tract symptoms,analgesic dosage,and time of calculus removing were recorded and all the data were statistically analysed through software SPSS 13. 0. Results There was no singinificant difference between the two groups in the rate of calculus removing (P>0. 05), but it took less time in the observation group (P<0. 05). In addition, it was of less low urinary tract symptoms, low-er rate of analgesic usage and colic episodes in the observation group with statistically significant differences compared with the control group (P<0. 05). Conclusion The application of tamsulosin after ureteroscopic lithotripsy can reduce the rate of colic episodes and analgesic dosage and time of calculus removing.

Objective To investigate effects of hyperpolarization-activated cyclic nucleotide-gated cation nonselective channel in the hu-man ureter on the spontaneous contraction of smooth muscles. Methods Four HCN subtypes were detected in human ureteral tissue using reverse transcription polymerase chain reaction,Western blotting and immunohistochemical. ZD7288,the HCN blocker, was used to observe the changes of ureteral muscle contraction amplitude and frequency by applying the ureteral smooth muscle strip test in vitro. Results HCN1-4 isoforms were all identified in human ureter using reverse transcription-polymerase chain reaction and Western blotting. Through the immunohistochemical,HCN channel was found mostly in the urothelium layer and muscular layer of human ureteral wall. ZD7288 significantly decreased the bladder excitation. Conclusion All 4 HCN channel hypotypes exist in the human ureter, and affect the ureteral excitation.