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【Objective】 To explore the safety and efficacy of flexible ureteroscope in the treatment of upper and middle ureteral calculi complicated with lower ureteral stricture after the failure of rigid ureteroscopy. 【Methods】 Clinical data of 36 patients with middle and upper ureteral calculi and lower ureteral stricture treated with rigid ureteroscopy but failed during Oct.2019 and Oct.2021 were retrospectively analyzed. The patients’ average age was (46.2±13.2) years, and the maximum diameter of calculi was (1.3±0.3) cm. The intraoperative, postoperative and follow-up data were recorded. 【Results】 All 36 patients successfully completed first-stage operation. Intraoperatively, the stenosis degree was F6-8 and could be dilated to F9-11. The mean length of stenosis was (1.1±0.34) cm. No serious postoperative complications such as infection or bleeding occurred. Two patients were lost and 34 patients were followed up. There was no obvious hydronephrosis on ultrasound examination. The stone removal rates were 76.5%, 88.2% and 97.1%, respectively, in months 1, 2 and 3 after operation. One patient with residual stones underwent secondary ureteroscopy in month 3 and large stones were removed with stone removal basket. 【Conclusion】 In patients with middle and upper ureteral calculi and lower ureteral stricture, after the failure of rigid ureteroscopy, flexible ureteroscope is safe and effective, and can significantly increase the success rate of first-stage surgery.
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Metabolism reprogramming plays an important role in the process of tumor occurrence and development, and provides the necessary material basis for tumor cells. It can change the metabolic patterns of amino acids, glucose and fatty acids in tumor cells, which is one of the hallmark features of tumors. At present, it is shown that most tumors tend to take advantage of glycolysis for energy resource. In contrast, studies have shown that prostate cancer cells dependent more on the fatty acid oxidation pathway for metabolic reprogramming to obtain energy substances. Therefore, it is of great significance to understand the relation between key enzymes of lipid metabolism and regulatory genes for early diagnosis, targeted treatment and better prognosis of prostate cancer.
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To determine the effect of a recombinant lentivirus containing human stem cell leukemia (SCL) gene on the expression of c-kit protein in damaged interstitial cells of Cajal (ICC) under high glucose condition. Methods: After isolation of ICC, the cells were cultured for 24 hours until the cells were adherent. After identification by inverted microscope and immunofluorescence, ICC cells were divided into two groups: A control group and a high glucose group. The control group was added with a medium containing 5 mmol/L of glucose. The high glucose group was added with a medium containing 20 mmol/L of glucose. After 48 h of continuous cultivation, the high glucose group was divided into 3 subgroups: A blank group, an empty lentivirus group, and an experimental group. The blank group, the empty lentivirus group, and the experimental group were added a medium containing PBS solution, empty lentivirus, and a recombinant lentivirus containing the SCL gene with a glucose concentration of 5 mmol/L, respectively. The cultures were incubated for 24 and 48 h. The expression of c-kit protein in ICC in each group was detected by Western blot. Results: After 24 or 48 h, the expression of c-kit protein in ICC was significantly lower in the blank group and the lentivirus group than that in the control group, and the expression of c-kit protein in ICC was significantly higher in the experimental group than that in the blank group and the empty lentivirus group, but it was still lower than that in the control group (all P<0.05). Conclusion: The recombinant lentivirus of SCL gene can up-regulate the expression of c-kit protein in functionally impaired ICC under high glucose condition.
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Humanos , Glucose , Células Intersticiais de Cajal , Lentivirus , Leucemia Mieloide Aguda , Proteínas Proto-Oncogênicas c-kitRESUMO
Objective @# To the effects and potential mechanisms of ST3GAL5 on biological behaviors of Bladder Urothelial Carcinoma(BLCA) . @*Methods @# Differentially expressed genes related to bladder cancer were identified using microarray analysis . Suitable bladder cancer cell lines were then screened . In vitro experimental measurements , including CCK8 , EdU , colony formation assays , transwell migration , flow cytometry apoptosis experiments , scratch assay , were used to evaluate the effects of ST3GAL5 on biological behaviors of BLCA . ST3GAL5 gene Kyoto Encyclopedia of Genes and Genomes ( KEGG) , gene set enrichment analysis ( GSEA) were analyzed using The Cancer Genome Atlas (TCGA) database . Finally , Western blot technology was used to verify the classical proliferation and metastasis related pathway factors . @*Results @# The combination of bioinformatics analyses and experimental measurements demonstrate that ST3GAL5 expression is aberrantly down⁃regulated in human cell lines of BLCA . Through Cancer Cell Line Encyclopedia (CCLE) database , HT⁃1376 cell lines were successfully screened for vitro test . Upregulation of ST3GAL5 was found to suppress the malignant biological behaviour of bladder cancer. GSEA enrichment analyses exhibited that ST3GAL5 and its co⁃expressed genes inhibited cell proliferation , invasion and metastasis of bladder urothelial carcinoma by activation of the PPAR pathway and inhibition of the PI3K/AKT pathway . The results of Western blot experiments verified that the key proteins of the PPAR signaling pathway showed a significant increase and the key proteins of the PI3K/AKT signaling pathway showed a significant decrease ( P <0. 05) after ST3GAL5 overexpression in bladder cancer. @*Conclusion @#ST3GAL5 gene might act as an oncogenic suppressor gene in bladder cancer , possibly inhibit the proliferation , invasion and metastasis of bladder cancer cells by activating the PPAR signaling pathway and inhibiting related molecules in the PI3K/AKT signaling pathway .
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Objective @#To investigate the effect of the combination of stem cell leukemia(SCL)recombinant lentivirus and connexin 43 ( Cx43 ) recombinant lentivirus on the function of diabetic cystipathy ( DCP) in guinea pigs.@*Methods @#After 90 healthy guinea pigs were fed normally for 1 week, streptozotocin(STZ)was injected intraperito⁃ neally at 200 mg/kg in a single dose, and random blood glucose was monitored weekly. After 12 weeks of normal feeding, 40 guinea pigs meeting the criteria were screened by urodynamic examination and randomly divided into 4 groups: diabetic group, SCL group, Cx43 group, and SCL + Cx43 group. In the diabetic group, 0. 2 ml of empty lentivirus without gene was instilled into the bladder through the urethra, in the SCL and Cx43 groups, 0. 2 ml of SCL lentivirus and Cx43 lentivirus were instilled using the same method, in the SCL + Cx43 lentivirus group, 0. 2 ml of each SCL and Cx43 lentivirus was instilled through the urethra. After 14 days of transfection, urodynamic examination was performed, and then the guinea pigs were executed after the examination, and the bladders were quickly removed for frozen bladder sections and fluorescent double staining. @*Results @#There were no differences in urodynamic examinations between the SCL and Cx43 groups compared to the diabetic group( P > 0. 05 ) . Urodynamic examination in the SCL + Cx43 group showed improvement in detrusor pressure, abdominal pressure and bladder pressure compared to the diabetic group(P < 0. 05) . In laser confocal experiments, the number of interstitial cells of Cajal (ICC) was reduced in the diabetic group, and the spindle structure and cell protrusions were obviously destroyed and appeared in a state of cell lysis. In the SCL and Cx43 groups, there was an improvement in the spindle structure and cell protrusion of ICC⁃like cells, and there was no significant change in the number of cells. In the SCL + Cx43 group, there was an increase in the number of ICC⁃like cells, a significant improvement in the spindle structure and cell protrusion, and the formation of ICC⁃dimers.@*Conclusion @#Transurethral co⁃infusion of SCL and Cx43 gene recombinant lentivirus can be successfully transfected in guinea pig DCP bladder, which can restore the number and structure of damaged ICC cells and form ICC dimer structure, improve the pressure of diabetic bladder forced urinary muscle, improve the weakness of urination, ventral pressure voiding and other characteristics. It provides a new direction for the treatment of DCP.
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Objective @#To investigation of the interaction between interstitial cells of Cajal (ICCs) and connexin 43 ( Cx43) in bladder contraction and its significance.@*Methods @#Eighty male guinea pigs were randomly divided into blank control group,Glivec group,Gap27 group and Glivec + Gap27 group.Four groups of guinea pigs were per- fused with saline,Glivec,Gap 27,and Glivec + Gap 27 every morning for 2 months.Success of urodynamic testing model after 2 months.Bladder tissue was collected for an in vitro muscle strip test to detect muscle contraction in each group.Correlation between c-Kit and Cx43 was detected by immunofluorescence.The interaction between c- Kit and Cx43 in the bladder was further validated by qRT-PCR and Western blot.Ultrastructural changes in the muscle layer of the bladder were observed by electron microscopy. @*Results @#Urodynamics revealed increased blad- der compliance in the experimental group compared to the blank control group (P<0. 05) ; bladder compliance increased in the Glivec + Gap27 group compared to the Glivec and Gap27 groups (P<0. 01) .In vitro muscle strip experiments revealed that the frequency and tone of bladder muscle strip contractions were lower in the experimental group compared to the blank control group (P<0. 05) ,and that muscle strip contractions were weaker in the ex- perimental group after administration of acetylcholine (ACH) compared to the control group(P<0. 05) .Immuno- fluorescence showed that c-Kit was co-expressed with Cx43 on ICCs cells.qRT-PCR and Western blot suggested that the protein expression level and gene expression level of Cx43 in bladder tissues were lower after inhibition of c-Kit than in the blank control (P<0. 05) ; after inhibition of Cx43,the protein expression level and gene expres- sion level of c-Kit in bladder tissues the levels of c-Kit protein expression and gene expression in bladder tissues were lower than those in the blank control group (P<0. 05) .Electron microscopy revealed that the mitochondrial structure of bladder smooth muscle was disrupted after simultaneous inhibition of c-Kit and Cx43.@*Conclusion@#Cx43 is expressed on bladder ICCs and the two may be jointly involved in regulating bladder contractile function ; the joint reduction of Cx43 and c-Kit may have disrupted the mitochondria of bladder smooth muscle,affecting its function and consequently bladder contractile function.
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Objective To observe the effects of green fluorescence protein mediated by lentivirus in bladder, and to de?termine the amount of virus obtained good transfection effects. Methods lentivirus carring GFP gene was perfused using transurethral approach into bladder of guinea pigs. Samples of bladder, liver, kidney and lungs were collected for frozen sec?tions after feeding seven days. The distribution of green fluorescence was observed using laser confocal microscopy. Re?sults The titer of lentivirus was 4 × 108. GFP was found under the mucosa when the amount of lentivirus transurethral was 30μL. GFP was distributed widely in muscle layer using 40μL lentivirus. GFP was detected even stronger in muscle layer when the amount was 50μL. GFP was found in muscle layer when 25μL lentivirus was injected intravenously. GFP was not found in other tissues than in bladder via transurethral perfusion. There was higher GFP in liver, lungs and other organs than in bladder via intravenous injection. Conclusion Lentivirus can mediate GFP transfecting bladder of guinea pig successful?ly and escape the distribution of GFP all over the body intravenously, which will bring new research direction and method for clinical treatment of diseases in bladder.
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Objective To investigate the expressions of calcium sensitive receptor (CaSR) and tight junction protein (Claudin)-14 in renal calcium oxalate stone rat model. Methods Thirty male SD rats were randomly divided into control group (n=15) and model group (n=15). The rat model of renal calcium oxalate stone was established by gavaging 1%glycol (2 mL/d) and 2% ammonium chloride. The expression of CaSR protein was detected using immunohistochemical assay. RT-PCR was used to detect the Claudin-14 mRNA expression. The expression levels of CaSR and Claudin-14 protein were de-tected by Western blot assay respectively. The full automatic biochemical analyzer was used to detect rat renal function and changes of blood and urine biochemical indices. Results A large stone crystallization was observed under light microscope in model group. The serum levels of Cr, BUN, 24-h urine calcium and urine volume were significantly higher in model group than those of control group (P<0.01). There were no significant differences in serum calcium level and urinary pH value be-tween two groups. The expression levels of Claudin-14 mRNA and CaSR protein were significantly higher in model group than those of control group (P<0.01). The Claudin-14 protein expression was specifically higher in renal tissues of model group. There was no Claudin-14 protein expression in control group. There was a positive correlation between CaSR and Claudin-14 protein expression in renal tissues of model group. Also, CaSR and Claudin-14 protein expressions were posi-tively correlated with the 24-h urine volume. Conclusion The increased expressions of CaSR and Claudin-14 involved in the formation of kidney stone by increasing the urinary calcium excretion.
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Objective The calcium oxalate stone is the most common type of the kidney stones.By building the rat renal calcium oxalate stone model, preliminary study the function of CaSR-Claudin14 regulating pathways on renal calcium oxalate stone for-mation model in rats. Methods 30 Male S-D rats were randomly divided into control group (n=15) and model group (n=15). Adult male S-D rats were given ethylene glycol and ammonium chloride to induce urolithiasis.Application of full automatic biochemical analyzer to test rat renal function and the changes of urine biochemical index.Immunohistochemistry was used to detect the expression of CaSR protein;RT-PCR was used to detect the Claudin-14 mRNA expression;Western Blotting was used to detect the expression of CaSR and Claudin-14 protein respectively. Results By observing model group has large stones crystallization under light microsco-py;model group rats 24 h urine calcium are significantly higher than control group([9.66 ±1.10]mmol vs [3.26 ±0.60]mmol, P0.05 ) .Expression of Claudin-14 mRNA in the model group is significantly higher than normal control group([0.150 ± 0.004] vs [0.047 ±0.008], P<0.01); Expression of Claudin-14 protein in the model group is significantly higher than normal control group([1.526 ±0.089] vs 0, P<0.01).Expression of CaSR protein in the model group is significantly higher than normal control group([6.697 ±0.051] vs [5.016 ±0.053], P<0.05). Conclusion CaSR can raise the expression of Claudin-14, increase re-nal tubular urinary calcium excretion to promote renal calcium oxalate stone formation.
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Objective To study the beneficial effect and mechanism of Alcea rosea roots in nephrolithiasis model in-duced by 1%ethylene glycol in rats. Methods We randomly divided 60 male Wistar rats into six groups,including control group, model group, Alcea rosea roots lower-dose preventive group, Alcea rosea roots high-dose preventive group, Alcea ro-sea roots lower-dose curative group, Alcea rosea roots high-dose curative group. Control group was free to access food and water;model group was given 1%ethylene glycol drinking water and was fed with normal diet, preventive group was given 1%ethylene glycol drink and Alcea rosea roots in low (250 mg/kg) or high dose (500 mg/kg) each day, curative group re-ceived 1%ethylene glycol drink each day and Alcea rosea roots in low or high dose from day 15 to day 28. At the end of the experiment, various renal functional and injury markers such as urine volume, calcium, phosphate, magnesium, urea, creati-nine, and oxalate were evaluated using urine, serum, and kidney homogenates. The kidneys were removed and prepared for histological evaluation of calcium oxalate deposits. Results In model groups, urine output, urea, creatinine, 24 h urine Ca2+, and oxalate and MDA were increased compared to those in control group(P<0.05). GSH and SOD were increased in preventative and curative groups compared to those in the model group(P < 0.05). The urea, creatinine, 24 h urine Ca2+, urine oxalate, MDA were reduced in preventive and curative groups compared to those in the model group(P<0.05). The number and size of calcium oxalate crystal deposits were also less and smaller, and the kidney damage was less severe in pre-ventive and curative groups compared to in the model group. Conclusion The extract of Alcea rosea roots can prevent and treat calcium oxalate urinary stone formation in rats and protect renal function.
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Objective To discuss the types of ICC and characteristics of spontaneous Ca2+ waves of different types of ICC in the bladder of guinea pig. Methods Frozen-sections were made from the bladder of guinea pig and ICC were cultured in vitro. Cells were stained by indirect immunofluorescent method and detected by Laser scanning confocal microscope. The ICC cultured in vitro were divided randomly into 4 group: dimmer ICC,monomer ICC,dimmer ICC treated with 2-APB group and monomer ICC treated with 2-APB group according to the cell morphology and disrupted with 2-APB.The calcium concentration of ICC cultured in vitro were marked with Fluo-4 AM and disrupted by 2-aminoethoxydipheylbrate (2-APB, 100 μmol/L) in dimmer ICC treated with 2-APB group and monomer ICC treated with 2-APB group. The calcium oscillation function of ICC was observed under Laser scanning confocal microscope. Results For the monomer ICC and dimmer ICC in frozen sections and cultured in vitro,there were increased frequency (P<0.01) and amplitude (P<0.05) of spontaneous Ca2+ waves in dimmer ICC compared with the monomer monomer ICC. But after the cells disrupted by 2-APB after 15 min,There were decreased frequency (P<0.01) and amplitude (P<0.01) of spontaneous Ca2+ waves in the dimmer ICC treated with 2-ABP group compared with the dimmer ICC. The changes(P>0.05) of spontaneous Ca2+ waves was not statistical significance in monomer ICC treated with 2-ABP group compared with monomer ICC group. Conclusions The bladder of guinea pig may exist 2 different types of ICC, dimmer ICC and monomer ICC. The excitability of spontaneous Ca2+ waves of dimmer ICC could be higher than in monomer ICC. The special structure of dimmer ICC may contribute to the formation of high spontaneous Ca2+ waves.
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AIM: To determine the expression of c-kit mRNA and protein in the bladders in guinea pigs with diabetic cystopathy (DCP) and to explore the correlation and mechanisms between c-kit expression and DCP. METHODS: Sixty guinea pigs were divided randomly into control group (n=20) and experimental group (n=40). The guinea pigs in experimental group were injected with streptozotocin (STZ) to induce diabetes mellitus. After fed for 10 weeks, the animals in both groups were tested with urodynamics, and the guinea pigs in experimental group were divided into the subgroups of DCP and the diabetic no-cystopathy (NDCP) group according to the results of urodynamics. mRNA expression of c-kit was detected by reverse transcription polymerase chain reaction (RT-PCR) and protein expression of c-kit was tested and analyzed by laser scanning confocal microscope. RESULTS: Decreased expression of c-kit mRNA was observed in DCP group compared to control and the NDCP group. The ratio of c-kit mRNA and GAPDH was 5.66±0.54 in controls (P<0.05), 5.54±1.28 in NDCP group (P<0.05) and 4.65 ±0.47 in DCP group. c-kit protein expression significantly declined in DCP group. The mean value of fluorescence intensity was 856.52± 53.03 in control group, 844.67± 59.24 in NDCP group and 548.69± 48.51 in DCP (P<0.01).CONCLUSION: The declined expression of c-kit) gene at transcription and translation levels destroys the SCF/c-kit signal pathway, leading to the dysfunction of Cajal-like) cells in DCP guinea pig, so the abnormal expression of c-kit gene is involved in the pathogenesis of DCP.
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Objective To determine vascular endothelial growth factor(VEGF)-460 gene polymorphism in Uyghurs and its relationship to urolithiasis in south Xinjiang. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),gene sequencing and genetic analysis methods were used in 200 urolithiasis patients of Uyghurs, and 200 healthy Uyghurs. Results The distribution of genotype and allele had no significant difference between urolithiasis patients and normal controls (P>0. 05). The frequencies for the CC,TT and CT genotypes in patients with urolithiasis and normal controls were 1.5 %, 29.0 %, 69.5 % and 0. 5 %, 27.5 %, 72.0 %, respectively. The frequencies for C and T allele were 36.2%,63.7% and 36.9% ,63.1%, respectively. Conclusions The results of VEGF-460 gene polymorphisms indicate no significant relationship between patients with turolithiasis and normal controls in Uyghurs in south Xinjiang,which may not be urolithiasis susceptibility genetic locus.
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Purpose To clarify the morphologic changes and significance of interstitial cells of Cajal-like cells in the bladder of diabetic guinea pig.Methods The experimental guinea pigs were induced by a single intraperitoneal injection of streptozotocin(STZ).The diabetic guinea pig model, who were made succesful for 4 weeks, were detected through the urodynamic evaluation.The distribution of ICCs-like cells in the detrusor, which were taken from these guinea pigs,were quantitatively analyzed after frozen section and indirect immunofluorescence methods and observed by electron microscopy.Results In the diabetic group, the maximum detrusor pressure was signifently lower than controls (P<0.01), maximum bladder capacity, compliance, residual volume and leak point pressures was significently higer than controls (P<0.05 or P<0.01);In detrusor, compared with control, the ICCs-like cells of diabetes group showed significently decreased in quantity. In diabetic bladder, the quantity of gap junctions between ICCs-like cells and cells around,such as smoth muscle cells,terminal nerves and other ICCs-like cells,decreased significently.And more cavities and less organoids occurred in cytoplasm of diabetic ICCs-like cells. Mitochondria became swollen or even disapeared, accompanied with swollen endocytoplasmic reticulum.Conclusion The decrease and injures of ICCs-like cells in bladder may be one of the mechanisms resulting in urodynamic changes of diabetic cystopathy.
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Objective To investigate the expression level of cyclooxygenase-2 (COX-2) and its effects on cell proliferation and apoptosis in bladder cancer. Methods The immunohistochemical method was used to determine the expression of COX-2 and Ki67 in 54 cases of bladder cancer, 29 cases of adjacent tumor tissue and 10 cases of normal bladder specimen respectively, and the apoptotic levels of the obove three tissues were assessed by counting the number of the cells labeled by TUNEL method. Results The positive rate of COX-2, the mean labelling index of Ki67 and the cellular apoptosis index in the cancer tissues were significantly higher than those in adjacent tumor tissues and normal bladder specimens(P