Wstępne badania dotyczące zróżnicowania ekspresji receptorów kortykotropiny w gruczolakach nadnerczy

Introduction: The ACTH receptor (ACTHR) is primarily expressed in the adrenal cortex. Previous studies focused on the regulatory function of ACTHR in glucocorticoid secretion, but research on adrenal tumours is rare. The aim of this study was to evaluate ACTHR expression in common adrenal adenomas and investigate its influence on adrenal tumorigenesis using adrenocortical H295R cells. Materials and methods: Real-time polymerase chain reaction and western blot were used to detect the expression of ACTHR in 18 aldosterone-producing adenomas, 16 cortisol-producing adenomas, 9 non-functional adenomas, and 12 normal adrenal samples. Lentiviral vector pLVX-mCMV-ACTHR-ZsGreen was transfected into the H295R cells to increase ACTHR expression. WST-1 and cell count were applied to evaluate cell proliferation at different ACTHR levels. TUNEL staining was used to measure cell apoptosis. Results: Compared with normal adrenal samples, the aldosterone-producing adenoma samples had higher ACTHR mRNA and protein levels. However, the mRNA and protein levels of ACTHR in non-functional adenomas and cortisol-producing adenomas were lower than those in the normal adrenal samples. Proliferative activity in the experimental cells was higher than that in the control cells in the first three days. The proliferative activity peaked in the second day. However, this trend was reversed in the fourth day and became more apparent with time. By contrast, TUNEL staining showed that ACTHR overexpression did not induce a significant difference between the two groups. Conclusions: Differential ACTHR expression may be determined by different types of adrenocortical tumours. ACTHR is more likely to have induced the proliferation rather than the apoptosis of H295R cells.Wstęp: Receptor kortykotropiny (adrenocorticotropic hormone receptor, ACTHR) występuje głównie w korze nadnerczy. Prowadzono wcześniej badania dotyczące regulacyjnej funkcji ACTHR w wydzielaniu glikokortykoidów, lecz rzadko obejmowały one chorych z guzami nadnerczy. Badanie przeprowadzono w celu oceny ekspresji ACTHR w najczęstszych gruczolakach nadnerczy i zbadania jej wpływu na rozwój tych nowotworów w komórkach kory nadnerczy ludzkich linii H295R. Materiał i metody: Do wykrycia ekspresji ACTHR w 18 gruczolakach wydzielających aldosteron, 16 gruczolakach wydzielających kortyzol, 9 gruczolakach nieczynnych hormonalnie i 12 próbkach prawidłowych nadnerczy użyto reakcji łańcuchowej polimerazy w czasie rzeczywistym oraz metody Western blot. W celu zwiększenia ekspresji ACTHR transfekowano wektor lentiwirusowy pLVX-mCMV-ACTHR-ZsGreen do komórek H295R. Proliferację komórek przy różnych poziomach ekspresji ACTHR oceniono na podstawie WST-1 oraz liczby komórek. W celu zmierzenia apoptozy komórek zastosowano barwienie metodą TUNEL. Wyniki: W porównaniu z próbkami prawidłowych nadnerczy w próbkach gruczolaków wydzielających aldosteron stwierdzono wyższą ekspresję mRNA i białka ACTHR. Jednak ekspresja mRNA i białka ACTHR w gruczolakach nieczynnych hormonalnie oraz w gruczolakach wydzielających kortyzol była niższa niż w próbkach prawidłowych nadnerczy. W ciągu 3 pierwszych dni aktywność proliferacyjna w komórkach eksperymentalnych była wyższa niż w komórkach kontrolnych. Aktywność proliferacyjna osiągnęła maksymalną wartość w drugim dniu. Jednak w czwartym dniu ta tendencja uległa odwróceniu, co było wyraźniejsze w miarę upływu czasu. Z kolei barwienie metodą TUNEL wykazało, że nad ekspresja ACTHR nie spowodowała istotnej różnicy między grupami. Wnioski: Zróżnicowanie ekspresji ACTHR może wynikać z różnych typów guzów nadnerczy ocenianych w badaniu. Wydaje się, że ACTHR pobudza proliferację, a nie apoptozę komórek H295R

Comparison of Thulium Fiber Laser Versus Holmium Laser in Ureteroscopic Lithotripsy: A Meta-Analysis and Systematic Review

OBJECTIVES: To compare the efficacy and safety of thulium fiber laser (TFL) to holmium: YAG (Ho: YAG) laser in ureteroscopic lithotripsy for urolithiasis. METHODS: PubMed, Web of Science, Embase, CENTRAL, SinoMed, CNKI database, VIP and Wanfang Database were systematically searched for all relevant clinical trials until September 2023. References were explored to identify the relevant articles. Meta-analysis was carried out for the retrieved studies using RevMan5.4.1 software, and the risk ratio, mean difference and 95% confidence interval were expressed. Statistical significance was set at p \u3c 0.05. The main outcomes of this meta-analysis were stone-free rate (SFR), perioperative outcomes and intraoperative or postoperative complications. RESULTS: Thirteen studies, including 1394 patients, were included. According to the results of pooled analysis, TFL was associated with significantly higher stone-free rate (SFR) [0.52, 95% CI (0.32, 0.85), P = 0.009], shorter operation time [-5.47, 95% CI (-8.86, -2.08), P = 0.002], and less stone migration [0.17, 95% CI (0.06, 0.50), P = 0.001]. However, there was no significant difference in terms of the laser time, duration of hospital stay, drop of hemoglobin level, total energy, postoperative ureteral stenting, the incidence of intraoperative complications or postoperative complications between TFL and Ho: YAGs. CONCLUSION: The findings of this study demonstrated several advantages of TFL in terms of higher SFR, shorter operative time and less stone migration. TRIAL REGISTRATION: The protocol of this systematic review was listed in PROSPERO ( www.crd.york.ac.uk/PROSPERO ) (Protocol number: CRD42022362550)

Information on harm outcomes was insufficiently reported in systematic reviews and meta-analyses of individual participant data

Meta-analysis is an important tool for summarizing findings from scientific research. But for harm outcomes,issues may arise due to the distribution problems, zero-events problems, as well as low statistical power. Individual participant data (IPD) meta-analysis represents a viable approach to overcome these shortcomings because all eligible small-scale studies can be treated as one large trial with multiple clusters through a one-stage analytic. An increasing number of IPD meta-analyses of healthcare interventions are published over the past years.6 Findings from these IPD meta-analyses contribute to the evolving body of best available evidence for informed decision-making. However, quality of safety reporting by the emerging IPD meta-analyses remains to be ascertained. In this study, we appraised the reporting of IPD meta-analyses, with a special focus on safety outcomes.L.TZ is funded by a National Key Research and Development Plan of China (Grant No. 2016YFC0106300).Scopu

Comparison of Thulium Fiber Laser versus Holmium laser in ureteroscopic lithotripsy: a Meta-analysis and systematic review

Abstract Objectives To compare the efficacy and safety of thulium fiber laser (TFL) to holmium: YAG (Ho: YAG) laser in ureteroscopic lithotripsy for urolithiasis. Methods PubMed, Web of Science, Embase, CENTRAL, SinoMed, CNKI database, VIP and Wanfang Database were systematically searched for all relevant clinical trials until September 2023. References were explored to identify the relevant articles. Meta-analysis was carried out for the retrieved studies using RevMan5.4.1 software, and the risk ratio, mean difference and 95% confidence interval were expressed. Statistical significance was set at p < 0.05. The main outcomes of this meta-analysis were stone-free rate (SFR), perioperative outcomes and intraoperative or postoperative complications. Results Thirteen studies, including 1394 patients, were included. According to the results of pooled analysis, TFL was associated with significantly higher stone-free rate (SFR) [0.52, 95% CI (0.32, 0.85), P = 0.009], shorter operation time [-5.47, 95% CI (-8.86, -2.08), P = 0.002], and less stone migration [0.17, 95% CI (0.06, 0.50), P = 0.001]. However, there was no significant difference in terms of the laser time, duration of hospital stay, drop of hemoglobin level, total energy, postoperative ureteral stenting, the incidence of intraoperative complications or postoperative complications between TFL and Ho: YAGs. Conclusion The findings of this study demonstrated several advantages of TFL in terms of higher SFR, shorter operative time and less stone migration. Trial registration The protocol of this systematic review was listed in PROSPERO ( www.crd.york.ac.uk/PROSPERO ) (Protocol number: CRD42022362550)

TGFBI promotes proliferation and epithelial–mesenchymal transition in renal cell carcinoma through PI3K/AKT/mTOR/HIF-1α pathway

Abstract Background Renal cell carcinoma (RCC) is presently recognized as the most prevalent kidney tumor. However, the role and underlying mechanism of action of the conversion factor-inducible protein (TGFBI), an extracellular matrix protein, in RCC remain poorly understood. Methods In this study, we employed Western blot, quantitative real-time polymerase chain reaction (qRT-PCR), and immunohistochemistry techniques to assess the expression of TGFBI in RCC tissues or cells. Furthermore, we analyzed the proliferation and migration of RCC cells using CCK8, cloning, scratching, and migration assays. Additionally, we examined apoptosis and cell cycle progression through flow cytometry, analysis. Lastly, we employed gene set enrichment analysis (GSEA) to investigate the biological processes associated with TGFBI, which were subsequently validated. Results The findings indicate that TGFBI exhibits significantly elevated expression levels in both renal cell carcinoma (RCC) tissues and cells. Furthermore, the knockdown of TGFBI in SiRNA transfected cells resulted in the inhibition of RCC cell proliferation, migration, invasiveness, apoptosis, and alteration of the cell cycle. Additionally, TGFBI was found to impede the epithelial-mesenchymal transition (EMT) process in RCC cells. Bioinformatics analysis suggests that TGFBI may exert its influence on various biological processes in RCC through the tumor immune microenvironment. Moreover, our study demonstrates that TGFBI promotes RCC progression by activating the PI3K/AKT/mTOR/HIF-1α. Conclusions Our research indicates that TGFBI exhibits high expression in RCC and facilitate RCC progression and metastasis through various molecular mechanisms. Hence, TGFBI has the potential to be a novel therapeutic target for the diagnosis and treatment of RCC in the future

UCHL5 Promotes Proliferation and Migration of Bladder Cancer Cells by Activating c-Myc via AKT/mTOR Signaling

Ubiquitin C-terminal hydrolase L5 (UCHL5) is a deubiquitinating enzyme (DUB) that removes ubiquitin from its substrates. Associations between UCHL5 and cancer have been reported in various tissues, but the effect of UCHL5 on bladder cancer has not been thoroughly investigated. This study investigates the expression and function of UCHL5 in bladder cancer. UCHL5 was shown to be abnormally expressed using IHC of tissue microarray and Western blotting. Several procedures were performed to assess the effect of UCHL5 overexpression or knockdown on bladder cancer, such as cell proliferation, colony formation, wound-healing, and Transwell assays. In addition, RNA-Seq and Western blotting experiments were used to verify the status of downstream signaling pathways. Finally, bladder cancers with knockdown or overexpression of UCHL5 were treated with either SC79 or LY294002 to examine the participation of the AKT/mTOR signaling pathway and the expression of downstream targets c-Myc, SLC25A19, and ICAM5. In contrast to adjacent tissue samples, we discovered that UCHL5 was substantially expressed in bladder cancer samples. We also found that UCHL5 downregulation significantly suppressed both tumor growth in vivo and cell proliferation and migration in vitro. According to RNA-Seq analyses and Western blotting experiments, the expression of c-Myc, SLC25A19, and ICAM5 was modified as a result of UCHL5 activating AKT/mTOR signaling in bladder cancer cells. All things considered, our findings show that increased UCHL5 expression stimulates AKT/mTOR signaling, subsequently triggering the expression of c-Myc, SLC25A19, and ICAM5, which in turn promotes carcinogenesis in bladder cancer. UCHL5 is therefore a potential target for therapy in bladder cancer patients