Identification of a basement membrane-related genes signature with immune correlation in bladder urothelial carcinoma and verification in vitro.

BACKGROUND: Bladder urothelial carcinoma (BLCA) is the most common genitourinary cancer and the prognosis of patients is often poor. However, studies of basement membrane-related genes (BM-related genes) in BLCA are less reported. Therefore, we established a BM-related genes signature to explore their functional and prognostic value in BLCA. METHODS: In this study, a BM-related genes signature was constructed by LASSO-Cox regression analysis, and then a series of bioinformatics methods was used to assess the accuracy and validity of the signature. We constructed a nomogram for clinical application and also screened for possible therapeutic drugs. To investigate the functions and pathways affected by BM-related genes in BLCA, we performed functional enrichment analyses. In addition, we analyzed the immune cell infiltration landscape and immune checkpoint-related genes in the high and low-risk groups. Finally, we confirmed the prognostic value of BM-related genes in BLCA in vitro. RESULTS: Combining multiple bioinformatics approaches, we identified a seven-gene signature. The accuracy and validity of this signature in predicting BLCA patients were confirmed by the test cohort. In addition, the risk score was strongly correlated with prognosis, immune checkpoint genes, drug sensitivity, and immune cell infiltration landscape. The risk score is an independent prognostic factor for BLCA patients. Further experiments revealed that all seven signature genes were differentially expressed between BLCA cell lines and normal bladder cells. Finally, overexpression of LAMA2 inhibited the migration and invasion ability of BLCA cell lines. CONCLUSIONS: In summary, the BM-related genes signature was able to predict the prognosis of BLCA patients accurately, indicating that the BM-related genes possess great clinical value in the diagnosis and treatment of BLCA. Moreover, LAMA2 could be a potential therapeutic target, which provides new insights into the application of the BM-related genes in BLCA patients.

ß-Cryptoxanthin Maintains Mitochondrial Function by Promoting NRF2 Nuclear Translocation to Inhibit Oxidative Stress-Induced Senescence in HK-2 Cells.

The mechanisms of acute kidney injury and chronic kidney disease remain incompletely revealed, and drug development is a pressing clinical challenge. Oxidative stress-induced cellular senescence and mitochondrial damage are important biological events in a variety of kidney diseases. As a type of carotenoid, ß-Cryptoxanthin (BCX) has various biological functions, which means it is a potential therapeutic candidate for the treatment of kidney disease. However, the role of BCX in the kidney is unclear, and the effect of BCX on oxidative stress and cellular senescence in renal cells is also unknown. Therefore, we conducted a series of studies on human renal tubular epithelial (HK-2) cells in vitro. In the present study, we investigated the effect of BCX pretreatment on H2O2-induced oxidative stress and cellular senescence and explored the potential mechanism of BCX action. The results showed that BCX attenuated H2O2-induced oxidative stress and cellular senescence in HK-2 cells. Moreover, BCX promoted NRF2 nuclear expression, maintained mitochondrial function, and reduced mitochondrial damage in HK-2 cells. In addition, silencing NRF2 altered the protective effect of BCX on mitochondria and significantly reversed the anti-oxidative stress and anti-senescence effects of BCX in HK-2 cells. We concluded that BCX maintained mitochondrial function by promoting NRF2 nuclear translocation to inhibit oxidative stress-induced senescence in HK-2 cells. In light of these findings, the application of BCX might be a promising strategy for the prevention and treatment of kidney diseases.

[Microwave ablation of the canine prostate: an experimental study].

OBJECTIVE: To investigate the effect of transrectal ultrasound-guided microwave ablation of canine prostate tissue. METHODS: Guided by transrectal ultrasound, we conducted microwave ablation on each side of the prostate in 12 male dogs, 6 at 40 W/ 120 s (group A) and the other 6 at 40 W/160 s (group B), and observed the changes in the thermal lesions using grayscale ultrasound. After thermal ablation, we measured the volume of the thermal lesions by contrast-enhanced ultrasound (CEUS). Then we harvested the whole prostate from the animals and determined the lesion volumes in the fresh tissue specimens. RESULTS: Grayscale ultrasound revealed an echogenic area at the initiation of the microwave ablation procedure, which was enlarged with the increase of ablation time. At the end of the procedure, the lesions appeared as an irregular heterogeneous echogenic area. CEUS showed oval non-perfused areas, which appeared as well-defined non-echoic areas in sharp contrast with the surrounding normal prostate parenchyma with bolus injection of contrast material (Sonovue, 2.4 ml), and that the thermal lesion volumes of groups A and B were (1.18 +/- 0.23) cm3 and (1.52 +/- 0.23) cm3, respectively. The thermal lesions of the gross specimen exhibited an elliptical shape, pale color and clear margin, and their volumes were (1.13 +/- 0.20) cm3 and (1.48 +/- 0.20) cm3, respectively, in groups A and B. CONCLUSION: Different combinations of time and power can produce coagulative necrotic lesions of different volumes in the local prostatic tissue. CEUS can accurately manifest the lesion area and thus avoid excessive or inadequate ablation treatment.

5-Aminolaevulinic acid-based photodynamic therapy suppresses lipid secretion by inducing mitochondrial stress and oxidative damage in sebocytes and ameliorates ear acne in mice.

Acne is a chronic inflammatory skin disease with wide-ranging effects, involving factors such as Propionibacterium acnes (P. acnes) infection and sebum hypersecretion. Current acne treatments are challenged by drug resistance. 5-aminolaevulinic acid (ALA) -based photodynamic therapy (PDT) has been widely used in the clinical treatment of acne, however, the mechanism of its action remains to be elucidated. In this study, by constructing a mice ears model of P. acnes infection, we found that ALA-PDT inhibited the proliferation of P. acnes in vivo and in vitro, significantly ameliorated ear swelling, and blocked the chronic inflammatory process. In vitro, ALA-PDT inhibited lipid secretion and regulated the expression of lipid synthesis and metabolism-related genes in SZ95 cells. Further, we found that ALA-PDT led to DNA damage and apoptosis in SZ95 cells by inducing mitochondrial stress and oxidative stress. Altogether, our study demonstrated the great advantages of ALA-PDT for the treatment of acne and revealed that the mechanism may be related to the blockade of chronic inflammation and the suppression of lipid secretion by ALA-PDT.

Cyanidin-3-O-glucoside plays a protective role against renal ischemia/ reperfusion injury via the JAK/STAT pathway.

PURPOSE: To investigate the role of cyanidin-3-O-glucoside (C3G) in renal ischemia/reperfusion (I/R) injury and the potential mechanisms. METHODS: Mouse models were established by clamping the left renal vessels, and in vitro cellular models were established by hypoxic reoxygenation. RESULTS: Renal dysfunction and tissue structural damage were significantly higher in the I/R group. After treatment with different concentrations of C3G, the levels of renal dysfunction and tissue structural damage decreased at different levels. And its protective effect was most pronounced at 200 mg/kg. The use of C3G reduced apoptosis as well as the expression of endoplasmic reticulum stress (ERS)-related proteins. Hypoxia/reoxygenation (H/R)-induced apoptosis and ERS are dependent on oxidative stress in vitro. In addition, both AG490 and C3G inhibited the activation of JAK/STAT pathway and attenuated oxidative stress, ischemia-induced apoptosis and ERS. CONCLUSIONS: The results demonstrated that C3G blocked renal apoptosis and ERS protein expression by preventing reactive oxygen species (ROS) production after I/R via the JAK/STAT pathway, suggesting that C3G may be a potential therapeutic agent for renal I/R injury.

Fucoxanthin Attenuates Oxidative Damage by Activating the Sirt1/Nrf2/HO-1 Signaling Pathway to Protect the Kidney from Ischemia-Reperfusion Injury.

Oxidative stress is a key component of renal ischemia/reperfusion (I/R) injury. Fucoxanthin (Fx), a marine carotenoid with enhanced antioxidant capacity, acts as a ROS inhibitor in diseases such as ischemic stroke and acute lung injury. We hypothesized that fucoxanthin could attenuate renal I/R-induced oxidative damage. C57BL/6 mice (n = 30) were randomly assigned to sham, IR, IR + DMSO, and IR + Fx (25, 50, and 100 mg/kg) groups. The renal I/R injury was induced by clamping the left kidney nephron tip in mice. Fucoxanthin was injected intraperitoneally 24 hours before surgery. Compared with the IR group, pretreatment with fucoxanthin significantly improved renal dysfunction and tissue structural damage and inhibited ROS levels and apoptosis. Consistent results were observed in HK-2 cells. Besides, we found that renal I/R resulted in decreased expression of Sirt1, Nrf2, and HO-1, while fucoxanthin upregulated the expression of Sirt1, Nrf2, and HO-1. The protective effects of fucoxanthin were significantly reversed by EX527 (a selective inhibitor of Sirt1) or si-Sirt1. In conclusion, our study investigated the protective effect of fucoxanthin against renal I/R injury, and the underlying mechanism may be related to the activation of the Sirt1/Nrf2/HO-1 signaling pathway by fucoxanthin to attenuate oxidative stress-induced apoptosis.

Mitochondria-Targeted Antioxidant Mitoquinone Maintains Mitochondrial Homeostasis through the Sirt3-Dependent Pathway to Mitigate Oxidative Damage Caused by Renal Ischemia/Reperfusion.

Mitochondrial dysfunction is a critical factor contributing to oxidative stress and apoptosis in ischemia-reperfusion (I/R) diseases. Mitoquinone (MitoQ) is a mitochondria-targeted antioxidant whose potent anti-I/R injury capacity has been demonstrated in organs such as the heart and the intestine. In the present study, we explored the role of MitoQ in maintaining mitochondrial homeostasis and attenuating oxidative damage in renal I/R injury. We discovered that the decreased renal function and pathological damage caused by renal I/R injury were significantly ameliorated by MitoQ. MitoQ markedly reversed mitochondrial damage after I/R injury and inhibited renal reactive oxygen species production. In vitro, hypoxia/reoxygenation resulted in increased mitochondrial fission and decreased mitochondrial fusion in human renal tubular epithelial cells (HK-2), which were partially prevented by MitoQ. MitoQ treatment inhibited oxidative stress and reduced apoptosis in HK-2 cells by restoring mitochondrial membrane potential, promoting ATP production, and facilitating mitochondrial fusion. Deeply, renal I/R injury led to a decreased expression of sirtuin-3 (Sirt3), which was recovered by MitoQ. Moreover, the inhibition of Sirt3 partially eliminated the protective effect of MitoQ on mitochondria and increased oxidative damage. Overall, our data demonstrate a mitochondrial protective effect of MitoQ, which raises the possibility of MitoQ as a novel therapy for renal I/R.

Energy-Stress-Mediated AMPK Activation Promotes GPX4-Dependent Ferroptosis through the JAK2/STAT3/P53 Axis in Renal Cancer.

Energy stress is an unfavorable condition that tumor cells are often exposed to. Ferroptosis is considered an emerging target for tumor therapy. However, the role of ferroptosis in energy stress in renal cancer is currently unknown. In this study, we found that glucose deprivation significantly enhanced GPX4-dependent ferroptosis through AMPK activation. Further, AMPK activation suppressed GPX4 expression at the transcriptional level through the upregulation of P53 expression. Additionally, the inactivation of JAK2/STAT3 transcriptionally promoted P53 expression, thereby promoting AMPK-mediated GPX4-dependent ferroptosis. In conclusion, energy stress promotes AMPK-mediated GPX4-dependent erastin-induced ferroptosis in renal cancer through the JAK2/STAT3/P53 signaling axis.

[Ultrasound-guided transrectal microwave ablation of the prostate in dogs].

OBJECTIVE: To investigate the feasibility and safety of ultrasound-guided transrectal microwave ablation in reducing the prostate volume. METHODS: Ultrasound-guided transrectal microwave ablation of both sides of the prostate was conducted on experimental dogs with the output volume of 30W for 120 seconds. The dogs were sacrificed on the very day of the ablation, and the prostate and its surrounding tissues were excised for observation of the thermal lesions and pathological examination. RESULTS: A total of 12 thermal lesions were achieved on the two sides of the prostate. The ultrasonogram manifested dense echo and increasing extent in the ablated area, and then an irregular heterogeneous echogenic area and clearly differentiated margin. Pathological examination of the gross specimen showed a little stagnant blood under the rectal mucous, the urethra and bladder not injured, and the thermal lesions elliptical, clearly margined and with the mean size of (0.94 +/- 0.30) cm3. CONCLUSION: Ultrasound-guided transrectal microwave ablation of the prostate can effectively cause coagulative necrosis of the local tissue without inflicting thermal injury upon the surrounding tissues. Conventional grayscale ultrasound can give a real-time'display of the extent of thermal lesion and the whole process of the ablation.

Cyanidin-3-O-glucoside plays a protective role against renal ischemia/ reperfusion injury via the JAK/STAT pathway

Purpose: To investigate the role of cyanidin-3-O-glucoside (C3G) in renal ischemia/reperfusion (I/R) injury and the potential mechanisms. Methods: Mouse models were established by clamping the left renal vessels, and in vitro cellular models were established by hypoxic reoxygenation. Results: Renal dysfunction and tissue structural damage were significantly higher in the I/R group. After treatment with different concentrations of C3G, the levels of renal dysfunction and tissue structural damage decreased at different levels. And its protective effect was most pronounced at 200 mg/kg. The use of C3G reduced apoptosis as well as the expression of endoplasmic reticulum stress (ERS)-related proteins. Hypoxia/reoxygenation (H/R)-induced apoptosis and ERS are dependent on oxidative stress in vitro. In addition, both AG490 and C3G inhibited the activation of JAK/STAT pathway and attenuated oxidative stress, ischemia-induced apoptosis and ERS. Conclusions: The results demonstrated that C3G blocked renal apoptosis and ERS protein expression by preventing reactive oxygen species (ROS) production after I/R via the JAK/STAT pathway, suggesting that C3G may be a potential therapeutic agent for renal I/R injury.

[Comparison of open versus laparoscopic surgeries for adrenal tumor: a meta-analysis].

OBJECTIVE: To systematically review the effectiveness and safety of open and laparoscopic surgeries for treatment of adrenal tumors. METHODS: The online databases including CNKI, PUBMED, SinoMed, EBSCO, Springerlink, WanFang Data, and VIP were searched for clinical trials published from 1999 to 2016. A meta-analysis was performed using RevMan 5.2 software. RESULTS: A total of 2340 patients in 25 trials were included. The results of meta-analysis showed that laparoscopic surgery was better than open surgery in terms of intestinal function recovery time (OR=-0.96, 95%CI [-1.22, -0.70] P<0.000 01), hospitalization time (OR=-3.48, 95%CI [-4.13, -2.78], P<0.000 01), complications (OR=0.22, 95%CI [0.14, 0.35], P<0.0001), and volume of blood loss (OR=-104.77, 95%CI [-138.95, -70.60], P<0.000 01). There was no significant difference in the surgery cost between open and laparoscopic surgeries. CONCLUSION: Laparoscopic surgery is superior to open surgery for treatment of adrenal tumors for shorter intestinal function recovery time, surgery duration, and hospitalization time and less complications and blood loss.

Discriminating Bipolar Disorder From Major Depression Based on SVM-FoBa: Efficient Feature Selection With Multimodal Brain Imaging Data.

Discriminating between bipolar disorder (BD) and major depressive disorder (MDD) is a major clinical challenge due to the absence of known biomarkers; hence a better understanding of their pathophysiology and brain alterations is urgently needed. Given the complexity, feature selection is especially important in neuroimaging applications, however, feature dimension and model understanding present serious challenges. In this study, a novel feature selection approach based on linear support vector machine with a forward-backward search strategy (SVM-FoBa) was developed and applied to structural and resting-state functional magnetic resonance imaging data collected from 21 BD, 25 MDD and 23 healthy controls. Discriminative features were drawn from both data modalities, with which the classification of BD and MDD achieved an accuracy of 92.1% (1,000 bootstrap resamples). Weight analysis of the selected features further revealed that the inferior frontal gyrus may characterize a central role in BD-MDD differentiation, in addition to the default mode network and the cerebellum. A modality-wise comparison also suggested that functional information outweighs anatomical by a large margin when classifying the two clinical disorders. This work validated the advantages of multimodal joint analysis and the effectiveness of SVM-FoBa, which has potential for use in identifying possible biomarkers for several mental disorders.

Enhancement of germ cell apoptosis induced by ethanol in transgenic mice overexpressing Fas Ligand.

It was suggested that chronic ethanol exposure could result in testicular germ cell apoptosis, but the mechanism is still unclear. In the present study, we use a model of transgenic mice ubiquitously overexpressing human FasL to investigate whether Fas ligand plays a role in ethanol-induced testicular germ cell apoptosis. Both wild-type (WT) mice and transgenic (TG) mice were treated with acute ethanol (20% v/v) by introperitoneal injection for five times. After ethanol injection, WT mice displayed up-regulation of Fas ligand in the testes, which was shown by FITC-conjugated flow cytometry and western blotting. Moreover, TG mice exhibited significantly more apoptotic germ cells than WT mice did after ethanol injection, which was demonstrated by DNA fragmentation, PI staining flow cytometry and TUNEL staining. In addition, histopathological examination revealed that degenerative changes of epithelial component of the tubules occurred in FasL overexpressing transgenic mice while testicular morphology was normal in wild-type mice after acute ethanol exposure, suggesting FasL expression determines the sensitivity of testes to ethanol in mice. In summary, we provide the direct evidences that Fas ligand mediates the apoptosis of testicular germ cells induced by acute ethanol using FasL transgenic mice.

Penetration of erythromycin through Staphylococcus epidermidis biofilm.

BACKGROUND: The catheter related infection caused by Staphylococcus epidermidis biofilm is increasing and difficult to treat by antimicrobial chemotherapy. The properties of biofilms that give rise to antibiotic resistance are only partially understood. This study aimed to elucidate the penetration of erythromycin through Staphylococcus epidermidis biofilm. METHODS: The penetration ratio of erythromycin through Staphylococcus epidermidis biofilms of 1457, 1457-msrA, and wild isolate S68 was detected by biofilm penetration model at different time points according to the standard regression curve. The RNA/DNA ratio and the cell density within the biofilms were observed by confocal laser microscope and transmission electromicroscope, respectively. RESULTS: The penetration ratios of erythromycin through the biofilms of 1457, 1457-msrA, and S68 after cultivation for 36 hours were 0.93, 0.55 and 0.4, respectively. The erythromycin penetration ratio through 1457 biofilm (0.58 after 8 hours) was higher than that through the other two (0.499 and 0.31 after 24 hours). Lower growth rate of the cells in biofilm was shown, with reduction of RNA/DNA proportion observed by confocal laser microscope through acridine orange stain. Compared with the control group observed by transmission electrmicroscope, the cell density of biofilm air face was lower than that of agar face, with more cell debris. CONCLUSIONS: Erythromycin could penetrate to the Staphylococcus epidermidis biofilm, but could not kill the cells thoroughly. The lower growth rate of the cells within biofilm could help decreasing the erythromycin susceptibility.

Efficacy and safety of micafungin for invasive candida infections: a meta-analysis of randomized controlled trials.

BACKGROUND: Invasive fungal infections such as candidiasis and mold infections cause significant morbidity and mortality in seriously ill patients. Micafungin is an echinocandin antifungal agent with potent activity against most species of Candida and Aspergillus. We did this meta-analysis to clarify whether micafungin offers superior efficacy and safety compared with other antifungal agent for treating infections associated with invasive candidiasis. METHODS: We did a meta-analysis of randomized controlled trials to examine whether micafungin has superior efficacy and safety compared with other antifungal agents recommended by the treatment guidelines for fungal infection. Seven trials involving 2913 patients were included in this analysis. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULTS: Micafungin was associated with significantly better treatment success compared with the comparator antifungal agents (modified intention to treat, 2851 patients; random-effects model, OR 1.20, 95%CI 1.00 - 1.45, P = 0.0487). In addition, micafungin was more effective than the comparators for antifungal prophylaxis of neutropenic patients undergoing hematopoietic stem cell transplantation (OR 1.47, 95%CI 1.08 - 2.00, P = 0.01). Although there was no significant difference between the compared regimens in terms of the incidence of adverse drug effects (OR 0.94, 95%CI 0.77 - 1.11), fewer patients treated with micafungin withdrew from the studies because of adverse events (OR 0.64, 95%CI 0.44 - 0.94). CONCLUSIONS: Micafungin has a good safety and tolerability profile, with an efficacy at least comparable to the other antifungal agents. Micafungin offers advantages over other agents for antifungal prophylaxis. Micafungin offers an appropriate alternative for antifungal prophylaxis rather than the treatment of invasive candida infections.

Clinical study of distribution and drug resistance of pathogens in patients with severe acute pancreatitis.

BACKGROUND: Previous researches about necrotic pancreatic tissue infections are numerous, but the study on systemic infection related to the severe acute pancreatitis (SAP) treatment period is limited. This study aimed to investigate the distribution and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP during the past three years. METHODS: A retrospective study was conducted on the distribution, category and drug resistance of pathogenic bacteria in patients who had hepatobiliary surgery for SAP from 2008 to 2011. RESULTS: A total of 594 pathogenic bacteria samples were isolated. Among them 418 isolates (70.4%) were Gram bacteria negative, 142 isolates (23.9%) were Gram bacteria positive, and 34 isolates (5.7%) were found fungi. The most common Gram negative bacteria were Escherichia coli (19.8%), and the dominant Gram positive pathogenic bacteria were Enterococcus faecium. The distribution of SAP-related infectious pathogens was mainly in peritoneal drainage fluid, sputum, bile, and wound secretions. Almost all the Gram negative pathogenic bacteria were sensitive to carbapenum. Extended-spectrum ß-lactamases (ESBLs) producing strains were more resistant to penicillins and cephalosprins than the ESBLs non-producing strains. Staphylococcus was sensitive to vancomycin and linezolid. The drug resistance of meticillin-resistant staphylococcus (MRS) to commonly used antibiotics was higher than meticillin-sensitive streptococcus (MSS). Enterococcus sp. exhibited lower drug-resistance rates to vancomycin and linezolid. CONCLUSIONS: Gram negative bacteria were the dominant SAP-related infection after hepatobiliary surgery. A high number of fungal infections were reported. Drug resistant rates were high. Rational use of antibiotics according to the site of infection, bacterial species and drug sensitivity, correctly executing the course of treatment and enhancing hand washing will contribute to therapy and prevention of SAP-related infection and decrease its mortality.

[Placement of dual double-J stents following high-pressure balloon angioplasty for treatment of ureter-ileum bladder anastomosis stricture].

OBJECTIVE: To evaluate the benefit of placement of dual double-J stents following high-pressure balloon angioplasty for treatment of ureter-ileum anastomosis stricture after total bladder resection. METHODS: Seventeen patients (11 males and 6 females, mean age 56.65±6.28 years, 23 sides) undergoing total bladder resection were included in this study. Unilateral and bilateral ureteral stricture occurred postoperatively in 11 and 6 patients, respectively; 13 patients had ureter-ileum bladder anastomosis stricture after ileal bladder substitution, and 4 patients had ureter-ileum stricture after orthotopic construction of ileal neobladder. The control group consisted of 21 patients undergoing open surgery. RESULTS: In the double-J stenting group, the effective rate was 82.6% (19/23), similar to that of 85.7% (18/21) in the control group (P>0.05). Compared with the control group, the stenting group showed a significantly reduced mean time of operation (87.42±10.35 min vs 34.12±7.52 min, P<0.05), intraoperative blood loss (203.16±32.67 ml vs 21.54±6.15 ml, P<0.05), and mean postoperative hospital stay (10.12±1.19 vs 3.24±0.35 days, P<0.05). CONCLUSION: As a safe and minimally invasive approach to the management of ureter-ileum bladder anastomosis stricture, placement of dual double-J stents following high-pressure balloon angioplasty produces a effect comparable with that of open surgery.

Treatment of benign ureteral stricture by double J stents using high-pressure balloon angioplasty.

BACKGROUND: Balloon dilatation angioplasty is a minimally invasive surgery for treating benign ureteral stricture. The aim of this study was to investigate the effect of placing double J (D-J) stents using high-pressure balloon angioplasty in treating benign ureteral stricture. METHODS: A total of 42 patients (48 cases) with benign ureteral stricture (42 had benign ureteral stricture) were investigated by inserting dual D-J stents using high-pressure balloon angioplasty. The control group contained 50 patients (57 cases) employing the conventional balloon angioplasty with a single D-J stent inserted for comparison. RESULTS: The overall effective rate of the treated and control groups was 87.8% (36/41) and 62.7% (32/51), respectively (P < 0.05). CONCLUSION: This new approach produces a better curative effect than the conventional balloon angioplasty with a single D-J stent insertion in treating benign ureteral stricture.

[Construction of eukaryotic expression vector for VEGF165 gene and its expression in rat bladder smooth muscle cells].

OBJECTIVE: To construct an eukaryotic expression vector for vascular endothelial growth factor (VEGF) 165 gene and obtain VEGF expression in rat bladder smooth muscle cells. METHODS: VEGF165 cDNA was cloned into the eukaryotic expression vector pcDNA3.1(-), and the resultant recombinant vector pcDNA3.1(-)/VEGF165 was transfected into the rat bladder smooth muscle cells by electroporation. VEGF expression in the cells was determined by RT-PCR and immunofluoresence assay, and the biological activity of VEGF in the supernant of the transinfected cell culture was tested by MTT assay. RESULTS AND CONCLUSION: VEGF expression was obtained in the transinfected cells, and the supernant of the transinfected cell cultures stimulated the proliferation of the endothelial cells.