Efficacy of Mitomycin C Combined with Direct Vision Internal Urethrotomy for Urethral Strictures: A Systematic Review and Meta-Analysis.

BACKGROUND: The high recurrence of a urethral stricture after direct vision internal urethrotomy (DVIU) has been a problem for years. Mitomycin C (MMC) is an excellent antifibrosis antigen that has been used in many fields, but its effect on a urethral stricture remains controversial. The purpose of this review was to investigate the effectiveness of MMC in reducing the recurrence rate of a urethral stricture after the first urethrotomy. METHODS: Common databases were searched for publications prior to November 30, 2020. Randomized controlled and cohort trials were all included. Recurrence and success rates after the first urethrotomy of the posterior urethra were the main outcomes. Revman 5.3 was used for statistical analysis. Two evaluation systems, the Cochrane risk of bias tool and the Newcastle Ottawa Scale, were used to examine the risk of bias for RCTs and all studies. The quality of evidence was assessed by the Grading of Recommendations, Assessment, Development, and Evaluation standard. RESULTS: Sixteen trials were included, the reporting quality of which was generally poor, and the evidence level was very low to moderate. The addition of MMC could significantly reduce the recurrence rate of urethral strictures (risk ratio [RR] = 0.42; 95% confidence interval [CI]: 0.26, 0.67; p = 0.0002; 9 trials; 550 participants). The results of the subgroup analysis suggested that the effect of MMC combined with DVIU was significant in short (≤2 cm) anterior urethral strictures (RR = 0.39; 95% CI: 0.20, 0.78; p = 0.008), >12-month follow-up (RR = 0.45; 95% CI: 0.26, 0.76; p = 0.003). It also increased the success rate of the first urethrotomy procedure for posterior urethral contracture (RR = 0.74; 95% CI: 0.65, 0.84; p < 0.00001; 7 trials; 342 participants). Low-dose local injection of MMC was the most commonly used method. CONCLUSION: MMC combined with DVIU is a promising way to reduce the long-term recurrence rate of a short-segment anterior urethral stricture. It also increases the success rate of the first urethrotomy of the posterior urethra. However, more high-quality randomized controlled trials are needed.

High expression levels of DEF6 predicts a poor prognosis for patients with clear cell renal cell carcinoma.

Clear cell renal cell carcinoma (ccRCC) is one of the most common types of malignant tumors and early detection contributes to a better prognosis. Finding new biomarkers for the diagnosis or treatment remains meaningful. DEF6 guanine nucleotide exchange factor (DEF6) is upregulated in ccRCC compared to normal controls, but the relationship between DEF6 expression and prognosis in ccRCC is unclear. Moreover, the potential biological functions of DEF6 in ccRCC remains unclear. In the present study, the Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO), TISIDB and the clinical database of the Peking University First Hospital were used to analyze DEF6 expression in ccRCC. Immunohistochemistry (IHC), western blotting and reverse transcription­quantitative PCR were used to examine the DEF6 protein and mRNA expression levels in cell lines and clinical samples. Subsequently, the Kaplan­Meier method and Cox regression analyses were used to determine the impact of DEF6 expression on the overall survival of patients alongside other clinical variables in both the TCGA database and the present clinical database. The results showed that both DEF6 mRNA and protein expression levels were upregulated in ccRCC compared to normal controls. The Kaplan­Meier survival analysis showed that patients with high DEF6 expression had poor prognoses from both the TCGA database and the present clinical database. Univariate survival analysis and multivariate survival analysis revealed that DEF6 could be an independent prognostic factor for ccRCC. Additionally, bioinformatics analysis indicated that differentially expressed genes related to DEF6 expression influenced ccRCC by regulating the tumor immune microenvironment. In conclusion, overexpression of DEF6 is significantly correlated with a poor prognosis for patients with ccRCC and DEF6 may influence the biological processes involved with ccRCC by regulating the immune microenvironment.

Prognostic Value and Potential Biological Functions of CLDN8 in Patients with Clear Cell Renal Cell Carcinoma.

PURPOSE: Clear cell renal cell carcinoma (ccRCC) is among the most common malignant tumors worldwide, with a high incidence rate and poor prognosis. Currently, there are no biomarkers that can accurately guide prognostic evaluation and therapeutic strategy for ccRCC. The prognostic value and potential biological function of claudin-8 (CLDN8), a critical component of tight junctions in ccRCC, remain unclear. METHODS: Sequencing data were obtained from The Cancer Genome Atlas, International Cancer Genome Consortium, and Gene Expression Omnibus databases. R packages were used to explore CLDN8 mRNA expression levels and analyze differentially expressed genes. Results were validated in clinical specimens and cell lines, and bioinformatics analyses were conducted to explore the potential biological functions of CLDN8. Finally, functional analyses were carried out using 786-O ccRCC cell line. RESULTS: Both CLDN8 mRNA and protein expression levels were significantly lower in ccRCC compared with the normal control tissues. Kaplan-Meier analyses showed that low CLDN8 expression levels were associated with the poor overall survival, while univariate and multivariate Cox regression indicated that CLDN8 could serve as an independent prognostic factor in patient with ccRCC. Bioinformatic and Western blot analyses showed that CLDN8 suppressed proliferation, migration, and invasion of 786-O ccRCC cells through the epithelial-mesenchymal transition and AKT pathways. CONCLUSION: CLDN8 could serve as an independent prognostic factor in ccRCC, in which it suppresses 786-O proliferation, migration, and invasion through EMT and AKT pathways.

Overexpression of P4HB is correlated with poor prognosis in human clear cell renal cell carcinoma.

Prolyl 4-hydroxylase, beta polypeptide (P4HB) protein has been found to be associated with tumorigenesis in many types of tumor, However, the relationship between P4HB and clear cell renal cell carcinoma (ccRCC) has not been clarified. In this study, we focus on the correlation between P4HB expression and ccRCC. Through the Cancer Genome Atlas (TCGA) database, Gene Expression Omnibus (GEO) database, our database and immunohistochemical (IHC) staining. Compared with adjacent normal tissues, both the mRNA and protein levels of P4HB in ccRCC tissues were enhanced. The Kaplan-Meier survival analysis showed that high expression of P4HB is correlated with poor prognosis in both TCGA database and our own database. Multivariate survival analysis and Univariate analysis showed that P4HB expression and age are significantly correlative with poor prognose. All the results indicated that P4HB is correlated with poor prognosis in human clear cell renal cell carcinoma.

Inflammatory Stimuli Significantly Change the miRNA Profile of Human Adipose-Derived Stem Cells.

BACKGROUND: Human adipose-derived stem cells (hASCs) have been shown to have immunoregulatory properties in many studies. However, the mechanisms remain unknown. miRNAs are associated with many cellular processes, including immune responses. Thus, we hypothesized that miRNAs act as immunoregulators when hASCs are stimulated by inflammatory environments. METHODS: A set of cytokines was used to stimulate the hASCs in the cytokine group, while no cytokines were used to stimulate the cells in the normal group. A microarray was used to obtain the miRNA expression profile of hASCs, and RT-PCR was used to validate the miRNAs that were differentially expressed between the two groups. Target genes were predicted using online databases, and KEGG analysis was performed to identify the pathways enriched by the target genes of all the differentially expressed miRNAs. RESULTS: Five miRNAs were significantly upregulated, and 2 miRNAs were downregulated in the cytokine group compared with the normal group. We identified several immune-related pathways that are targeted individually or collectively by those miRNAs. CONCLUSION: Inflammatory stimuli changed the miRNA expression profile of hASCs. miRNAs may play a pivotal role in the immune response in hASCs and may be targets through which the immunoregulatory functions of hASCs can be enhanced.

Identification of three novel CTL epitopes within nucleocapsid protein of Hantaan virus.

Hantaan virus (HTNV) is a member of the Hantavirus genus that causes human hemorrhagic fever with renal syndrome (HFRS) in humans. The CTL response seems to play a key role in control of viral infection, but only a few HTNV epitopes recognized by the CTLs have been reported. Herein, we screened a panel of overlapping peptides covering the HTNV nucleocapsid protein by ELISPOT assays for those that can elicit IFN-γ production in vitro. Three novel CD8(+) CTL epitopes, N197-205 (RYRTAVCGL), N245-253 (KLLPDTAAV), and N258-266 (GPATNRDYL), were defined on the nucleocapsid protein and were found to be restricted by various HLA alleles including A11, A24, and B7. The epitopes were highly conserved among the reported HTNV strains and other hantanviruses, including Dobrava-Belgrade virus and Seoul virus, supporting their potential use in vaccine designs.

Keratinocyte growth factor gene therapy ameliorates ulcerative colitis in rats.

AIM: To investigate the effect of keratinocyte growth factor (KGF) gene therapy in acetic acid-induced ulcerative colitis in rat model. METHODS: The colitis of Sprague-Dawley rats was induced by intrarectal infusion of 1 mL 5% (v/v) acetic acid. Twenty-four hours after exposed to acetic acid, rats were divided into three experimental groups: control group, attenuated Salmonella typhimurium Ty21a strain (SP) group and SP strain carrying human KGF gene (SPK) group, and they were separately administered orally with 10% NaHCO(3), SP or SPK. Animals were sacrificed and colonic tissues were harvested respectively on day 3, 5, 7 and 10 after administration. Weights of rats, colonic weight/length ratio and stool score were evaluated. Histological changes of colonic tissues were examined by hematoxylin and eosin (HE) staining method. The expression of KGF, KGF receptor (KGFR) and TNF-α were measured either by enzyme-linked immunosorbent assay or Western blotting. Immunohistochemistry was used to detect the cellular localization of KGFR and Ki67. In addition, superoxide dismutase (SOD) activity and malondialdehyde (MDA) contents in the homogenate were measured. RESULTS: Body weight and colonic weight/length ratio were declined in SPK group compared with SP and control groups (body weight: 272.78 ± 17.92 g vs 243.72 ± 14.02 g and 240.68 ± 12.63 g, P < 0.01; colonic weight/length ratio: 115.76 ± 7.47 vs 150.32 ± 5.99 and 153.67 ± 5.50 mg/cm, P < 0.01). Moreover, pathological changes of damaged colon were improved in SPK group as well. After administration of SPK strain, KGF expression increased markedly from the 3rd d, and remained at a high level till the 10th d. Furthermore, KGFR expression and Ki67 expression elevated, whereas TNF-α expression was inhibited in SPK group. In the group administered with SPK, SOD activity increased significantly (d 5: 26.18 ± 5.84 vs 18.12 ± 3.30 and 18.79 ± 4.74 U/mg, P < 0.01; d 7: 35.48 ± 3.35 vs 22.57 ± 3.44 and 21.69 ± 3.94 U/mg, P < 0.01; d 10: 46.10 ± 6.23 vs 25.35 ± 4.76 and 27.82 ± 6.42 U/mg, P < 0.01) and MDA contents decreased accordingly (d 7: 7.40 ± 0.88 vs 9.81 ± 1.21 and 10.45 ± 1.40 nmol/mg, P < 0.01; d 10: 4.36 ± 0.62 vs 8.41 ± 0.92 and 8.71 ± 1.27 nmol/mg, P < 0.01), compared with SP and control groups. CONCLUSION: KGF gene therapy mediated by attenuated Salmonella ameliorates ulcerative colitis induced by acetic acids, and it may be a safe and effective treatment for ulcerative colitis.

Keratinocyte growth factor (KGF) gene therapy mediated by an attenuated form of Salmonella typhimurium ameliorates radiation induced pulmonary injury in rats.

The aim of this study is to investigate the effect of KGF (Keratinocyte growth factor) gene therapy mediated by the attenuated Salmonella typhimurium Ty21a on radiation-induced pulmonary injury in rats model. Sprague-Dawley rats were divided into three groups: TPK group (treated with TPK strain, attenuated Salmonella typhimurium Ty21a-recombined human KGF gene); TP group (treated with TP strain, attenuated Salmonella typhimurium Ty21a-recombined blank plasmid); and Saline group (treated with saline). After intraperitoneal administration for 48 h, the thoraxes of the rats were exposed to X-ray (20 Gy), and the rats were administered again two weeks after radiation. On the 3rd, 5th, 7th, 14th and 28th day after radiation, the rats were sacrificed and lung tissues were harvested. Histological analysis was performed, MDA contents and SOD activity were detected, mRNA levels of KGF, TGF-ß, SP-A and SP-C were measured by Real-time RT-PCR, and their concentrations in the BALF were quantified with ELISA. Administration of TPK strain improved the pathological changes of the lung on the 28th day. In the TPK group, KGF effectively expressed since the 3rd day, MDA contents decreased and SOD activity increased significantly, on the 7th day and 14th day respectively. SP-A and SP-C expression elevated, whereas TGF-ß expression was inhibited in the TPK group. These results suggest that this novel gene therapy of KGF could ameliorate radiation-induced pulmonary injury in rats, and may be a promising therapy for the treatment of radiative pulmonary injury.

[Modification and activity observation of hepatocyte growth factor gene on bone marrow mesenchymal stem cells].

OBJECTIVE: To evaluate the transfection efficiency and expression level of hepatocyte growth factor (HGF) by transfecting a recombinant adenovirus carrying HGF gene (Ad-HGF) into bone marrow mesenchymal stem cells (BMSCs) and to explore the effect of the expression supernatant on BMSCs in vitro so as to lay a foundation for the manufacture of gene medicine which expresses efficient cell factors. METHODS: Rat BMSCs were isolated using Percoll density gradient method and cultured according to the adherent property of BMSCs. The expression of c-Met was detected by immunohistochemical examination. BMSCs were infected with a recombinant adenovirus carrying green fluorescent protein gene (Ad-GFP) at multiplicity of infection (MOI, 0, 25, 50, 100, and 200 pfu/cell). To select an optimal MOI, the transfection efficiency and the degree of cell damage were assayed by flow cytometry and MTT, respectively, at 48 hours after transfecting. The expression of HGF in BMSCs transfected with optimal MOI Ad-HGF was measured with ELISA assay. MTT method was used to evaluate the proliferation effect of HGF expression supernatant on BMSCs. RESULTS: Immunohistochemical staining showed that BMSCs expressed c-Met receptor for HGF. At 48 hours after transfecting with different MOI Ad-GFP (0, 25, 50, 100, and 200 pfu/cell), the transfection efficiencies were 0.34% +/- 0.04%, 40.72% +/- 0.81%, 61.72% +/- 1.04%, 85.33% +/- 0.83%, and 17.91% +/- 0.63%, respectively; and the highest transfection efficiency was observed at 100 pfu/cell MOI. The cell damage was obviously observed when MOI was 200 pfu/cell. The expression of HGF in BMSCs reached the highest level after being transfected with 100 pfu/cell MOI Ad-HGF for 48 hours. The expression product could stimulate the proliferation of BMSCs. The proliferation of BMSCs gradually rose with the increase of HGF protein, and reached the highest level at 10% (320 pg). CONCLUSION: BMSCs can be transfected efficiently with Ad-HGF and express HGF protein, which stimulates the proliferation of BMSCs. It suggests that BMSCs is an ideal repair cells with gene vector.

[The effects of lead exposure on primary cultured hippocampus neurons apoptosis and the expressions of Bcl-2, Bax, p53 and NOS1].

OBJECTIVE: To observe effects of plumbum on the apoptosis of primary cultured hippocampus neurons. METHODS: After primary cultured hippocampus neurons of rats treated with different concentration of Pb (C2H3O2) (10, 50, 100 and 200 microg/ml), to observe the morphological changes of the apoptosis of hippocampus cells by Giemsa dyeing, to detect the rate of apoptosis by flow cytometry, to observe the expression of Bcl-2, Bax and p53 by immunocytochemical stain and to analyze quantitatively the average fluorescence intensity of free Ca2+ in kytoplasm of each concentration group by laser scanning confocal microscope (LSCM). RESULTS: The results of Giemsa dyeing showed typically morphological changes of apoptosis after hippocampus neurons treated with certain concentration of Pb(C2H3O2), and had dose-dependent; The results of flow cytometry showed,after cells treated respectively in 50, 100, 200 microg/ml for 24h, that the rate of apoptosis increased and compared with the control group the difference of were all statistical significance (P < 0.01); The results of immunocytochemical stain showed that, with dose of Pb increasing, the expression of Bcl-2 protein decreased, the intensity of dyeing increasingly degraded and the expression of Bax and p53 was up-regulation, and compared with the control group, IOD value of cells had statistical significance (P < 0.01); The results of LSCM showed that the concentration of Ca2+ in kytoplasm was also increased with the dose of Pb (C2H3O2) increasing, and the average fluorescence intensity in groups of three concentration of Pb(C2H3O2) was all significantly higher than in the control group (P < 0.01) and had dose-response relation. CONCLUSION: Plumbum might lead to different degrees of the apoptosis of hippocampus neurons. It is supposed that Pb in certain concentration range might affect the normal function of hippocampus neurons in learning and memory, and so on.

[Study on bone marrow mesenchymal stem cells transfected with adenovirus hepatocyte growth factor gene promoting wounds repair in diabetic rats].

OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cells (BMSCs) transfected with adenovirus hepatocyte growth factor (Ad-HGF) on wound repair in diabetic rats. MRTHODS: BMSCs from male Wistar rats were isolated by density gradient centrifugation, cultured, and transfected with Ad-HGF. The multiplicity of infection was 100. Diabetic models were established in 20 female Wistar rats by diets in high fat and sugar plus intraperitoneal injection of streptozotocin (30 mg/kg). Then 2 full-thickness skin wounds (approximately 1.5 cm in diameter) were made on the dorsum. The rats were randomly divided into 4 groups (n = 5 rats). After wounding, the 0.3 mL suspensions of BMSCs (group A), Ad-HGF (group B), BMSCs transfected with Ad-HGF (group C), and PBS (group D) were injected directly into the dermal of wounds. The transverse diameter and longitudinal diameter of wound were measured at 21 days after treatment. At 7 days and 28 days after treatment, HE staining was performed to evaluate wound healing. The contents of hydroxyproline and advanced glycosylation end products (AGEs) in the wounds were measured by enzyme linked immunosorbent assay and fluorospectrophotometer, respectively, at 3, 7, 14, and 28 days after treatment. RESULTS: At 21 days after treatment, the wounds almost healed in group C, and the transverse diameter and longitudinal diameter were 0 and (0.110 +/- 0.024) cm, respectively. But the wounds healed partially in groups A, B, and D, and the transverse diameter and longitudinal diameter were (0.470 +/- 0.051) cm and (0.590 +/- 0.041) cm, (0.390 +/- 0.042) cm and (0.480 +/- 0.032) cm, and (0.700 +/- 0.068) cm and (0.820 +/- 0.068) cm, respectively. There were significant differences in wound healing between group C and groups A, B, and D (P < 0.05). The wound healing time of group C [(20.5 +/- 1.9) days] was significantly shorter (P < 0.05) than those of groups A, B, and D [(28.3 +/- 1.9), (25.9 +/- 2.3), and (36.6 +/- 5.1) days]. At 7 days, the HE staining showed that evident epidermis transportation, collagen formation, and leukocytes infiltration were observed in group C. At 28 days, the HE staining showed that the epidermis in group C was significantly thinner and more regular than those in other groups, and the decreased collagen and many small vessels were observed in group C. The content of hydroxyproline in group C was higher than those in groups A, B, and D at 7 days and 14 days (P < 0.05). The contents of AGEs in group C was lower than those in groups A, B, and D at 14 days and 28 days (P < 0.05). CONCLUSION Transplantation of BMSCs transfected with Ad-HGF can accelerate the wounds repair in diabetic rats.

[Human papillomavirus 16E6 gene mutation in cervical carcinoma tissues].

BACKGROUND AND OBJECTIVE: Human papillomavirus (HPV) 16 is the most common type of high-risk human HPVs. HPV16 E6 gene and its specific mutations are considered as risk factors causing cervical carcinoma (CC). This study was to investigate HPV16 E6 mutations in Lanzhou region and explore the relationship between HPV16 E6 mutations and the development of CC. METHODS: Tissue DNA was extracted from 23 patients operated on for CC and five normal cervical controls. The partial sequence of the HPV16 E6 gene (nucleotide 201-523) was amplified by PCR from the tissue DNA extracted from the samples. PCR fragments were sequenced and analyzed. RESULTS: The positive rates of HPV16 E6 in five normal cervical and 23 CC tissues were 0 (0/5) and 82.61% (19/23), respectively. Prototype HPV16E6 gene was found in six cases (33.33%) while mutation in the E6 gene was detected in 12 cases (66.67%), among which a 350G mutation was found in 11 cases (61.11%). Moreover, a 249G mutation was identified in one CC case (5.56%). CONCLUSIONS: There is a high HPV infection rate in CC tissues in Lanzhou region, and most of the HPV16E6 are mutated.

The oncoprotein LMO3 interacts with calcium- and integrin-binding protein CIB.

The protein LMO3 belongs to the LIM only (LMO) group of transcriptional regulators, which act as molecular adaptors for protein-protein interactions. However, little is known about its interactive proteins and functions. Evaluating LMO3 in a yeast two-hybrid screen, we identified the calcium- and integrin-binding protein CIB as an LMO3-binding protein, which binds via the second LIM domain (LIM2) of LMO3. Cotransfection of LMO3 and CIB resulted in a shift in LMO3 protein from the nucleus to the cytoplasm. In functional assays, LMO3 induced C8 astrocyte proliferation was suppressed by the overexpression of CIB. This study demonstrates one function for LMO3 in C8 cells and suggests that one role of the CIB/LMO3 complex is to inhibit cell proliferation.