Testis cell pyroptosis mediated by CASP1 and CASP4: possible sertoli cell-only syndrome pathogenesis.

BACKGROUND: Sertoli cell-only syndrome (SCOS) is the most serious pathological type of non-obstructive azoospermia. Recently, several genes related to SCOS have been identified, including FANCM, TEX14, NR5A1, NANOS2, PLK4, WNK3, and FANCA, but they cannot fully explain the pathogenesis of SCOS. This study attempted to explain spermatogenesis dysfunction in SCOS through testicular tissue RNA sequencing and to provide new targets for SCOS diagnosis and therapy. METHODS: We analyzed differentially expressed genes (DEGs) based on RNA sequencing of nine patients with SCOS and three patients with obstructive azoospermia and normal spermatogenesis. We further explored the identified genes using ELISA and immunohistochemistry. RESULTS: In total, 9406 DEGs were expressed (Log2|FC|≥ 1; adjusted P value < 0.05) in SCOS samples, and 21 hub genes were identified. Three upregulated core genes were found, including CASP4, CASP1, and PLA2G4A. Thus, we hypothesized that testis cell pyroptosis mediated by CASP1 and CASP4 might be involved in SCOS occurrence and development. ELISA verified that CASP1 and CASP4 activities in the testes of patients with SCOS were significantly higher than those in patients with normal spermatogenesis. Immunohistochemical results showed that CASP1 and CASP4 in the normal spermatogenesis group were mainly expressed in the nuclei of spermatogenic, Sertoli, and interstitial cells. CASP1 and CASP4 in the SCOS group were mainly expressed in the nuclei of Sertoli and interstitial cells because of the loss of spermatogonia and spermatocytes. CASP1 and CASP4 expression levels in the testes of patients with SCOS were significantly higher than those in patients with normal spermatogenisis. Furthermore, the pyroptosis-related proteins GSDMD and GSDME in the testes of patients with SCOS were also significantly higher than those in control patients. ELISA also showed that inflammatory factors (IL-1 ß, IL-18, LDH, and ROS) were significantly increased in the SCOS group. CONCLUSIONS: For the first time, we found that cell pyroptosis-related genes and key markers were significantly increased in the testes of patients with SCOS. We also observed many inflammatory and oxidative stress reactions in SCOS. Thus, we propose that testis cell pyroptosis mediated by CASP1 and CASP4 could participate in SCOS occurrence and development.

Obstructive sleep apnea is related to alterations in fecal microbiome and impaired intestinal barrier function.

Obstructive Sleep Apnea (OSA) is related to repeated upper airway collapse, intermittent hypoxia, and intestinal barrier dysfunction. The resulting damage to the intestinal barrier may affect or be affected by the intestinal microbiota. A prospective case-control was used, including 48 subjects from Sleep Medicine Center of Nanfang Hospital. Sleep apnea was diagnosed by overnight polysomnography. Fecal samples and blood samples were collected from subjects to detect fecal microbiome composition (by 16S rDNA gene amplification and sequencing) and intestinal barrier biomarkers-intestinal fatty acid-binding protein (I-FABP) and D-lactic acid (D-LA) (by ELISA and colorimetry, respectively). Plasma D-LA and I-FABP were significantly elevated in patients with OSA. The severity of OSA was related to differences in the structure and composition of the fecal microbiome. Enriched Fusobacterium, Megamonas, Lachnospiraceae_UCG_006, and reduced Anaerostipes was found in patients with severe OSA. Enriched Ruminococcus_2, Lachnoclostridium, Lachnospiraceae_UCG_006, and Alloprevotella was found in patients with high intestinal barrier biomarkers. Lachnoclostridium and Lachnospiraceae_UCG_006 were the common dominant bacteria of OSA and intestinal barrier damage. Fusobacterium and Peptoclostridium was independently associated with apnea-hypopnea index (AHI). The dominant genera of severe OSA were also related to glucose, lipid, neutrophils, monocytes and BMI. Network analysis identified links between the fecal microbiome, intestinal barrier biomarkers, and AHI. The study confirms that changes in the intestinal microbiota are associated with intestinal barrier biomarkers among patients in OSA. These changes may play a pathophysiological role in the systemic inflammation and metabolic comorbidities associated with OSA, leading to multi-organ morbidity of OSA.

Prognostic Significance and Diagnostic Value of Overexpressed lncRNA PVT1 in Colorectal Cancer.

BACKGROUND: The aim of this study is to investigate the expression of lncRNA PVT1 in CRC tissue compared to adjacent normal tissues, and reveal the association between lncRNA PVT1 expression level and the clinicopathological characteristics of patients with CRC. METHODS: We detected the lncRNA PVT1 relative expression of cancerous tissues in 130 patients with CRC by using real-time quantitative polymerase chain reaction. At the same time, we collected the clinicopathological and prognostic information. RESULTS: IncRNA PVT1 was overexpressed in CRC tissues compared to paired-adjacent normal tissues and the high expression rate was 72.31%. High expression of lncRNA PVT1 predicts a later tumor stage (p = 0.001), poorer tissue differentiation (p = 0.019), and higher plasma CEA level (p = 0.043). Additionally, the lncRNA PVT1 expression was closely related to lymph node metastasis (N1/N2 vs. N0) and distant metastasis (M1 vs. M0) in CRC patients (p = 0.002; p = 0.003), but not to tumor T classification (p = 0.314). The result of prognostic analysis indicated that the 1-year and 3-year DFS of the lncRNA PVT1 low and high expression patients were 93.8% and 81.1%, 69.3% and 44.7%, respectively. The median DFS was 44 months in low expression group and 26 months in high expression group, with statistical significance (p = 0.021). COX multivariate analysis showed that TNM staging (III/IV vs. I/II: HR = 6.342, 95% CI: 2.994 - 13.433, p < 0.001) and the lncRNA PVT1 expression (high expression vs. low expression: HR = 3.744, 95% CI: 1.493 - 9.392, p = 0.005) was closely related to DFS in CRC patients. As with tumor TNM staging, lncRNA PVT1 expression was also an independent prognostic predictor of DFS. The proportion of lncRNA PVT1 high expression (fold change ≥ 1.725) was higher than that of elevated CEA ( > 5 ng/mL) in different CRC stages, especially, there was a significant difference in stage I patients (X2 = 41.717, p < 0.0001). CONCLUSIONS: The lncRNA PVT1 was over-expressed in CRC tissues, which indicated a poor prognosis. The lncRNA PVT1 expression is far higher than the plasma CEA level in the early stage patients, which has the potential diagnostic value for early stage CRC.

Increased Expression of PCNA-AS1 in Colorectal Cancer and its Clinical Association.

BACKGROUND: The aim is to study the expression of PCNA-AS1 in colorectal cancer (CRC) tissue and paired-adjacent normal tissue and the relationship between its expression level and clinical pathological features. METHODS: Using real-time qPCR, PCNA-AS1 expression levels were detected in 114 cases to establish the relationship between its expression and clinicopathologic features. Moreover, the expressions of PCNA-AS1 were investigated in CRC and normal colonic epithelial cell lines. RESULTS: PCNA-AS1 was upregulated in CRC patients. The difference was statistically significant (p < 0.001). The expression level was significantly correlated with the tumor invasion and TNM stage. The area under the receiver operating characteristic curve, sensitivity, and specificity were 0.824, 0.632, and 0.860, respectively. Moreover, PCNA-AS1 was up-regulated in CRC cell lines. CONCLUSIONS: PCNA-AS1 may function as a potential tumor biomarker for diagnosing CRC.

Long Noncoding RNA PVT1 as a Potent Predictor of Prognosis in Cancers: a Meta-Analysis.

BACKGROUND: Plasmacytoma variant translocation 1 (PVT1), an oncogenic long noncoding RNA located in a recognized cancer-risk gene region-8q24, is significantly overexpressed in various cancers. Many studies have found that high expression of PVT1 was correlated with poor prognosis. METHODS: This meta-analysis was performed by searching electronic databases Pubmed, Web of Science, Chinese National Knowledge Infrastructure, WanFang, and ChongQing VIP for eligible papers on the prognostic impact and clinicopathological characteristics of PVT1 expression in cancer from inception to January 31, 2017. The hazard ratio (HR) and odds ratio (OR) with 95% confidence intervals (CI) were computed to estimate the pooled effect of PVT1 on prognosis of cancers using Stata 12.0 version software. RESULTS: Thirteen studies were finally included in this review with a total of 1559 patients. The pooled result indicated that overexpressed PVT1 predicts a poorer prognosis of cancerous patients for overall survival (HR = 1.91, 95% CI: 1.61 - 2.26, p < 0.001) and disease-free survival (HR = 1.90, 95% CI: 1.46 - 2.48, p < 0.001) or recurrencefree survival (HR = 1.77, 95% CI: 1.24 - 2.52, p = 0.002) or progression-free survival (HR = 2.84, 95% CI: 1.67 - 4.82, p < 0.001). High expression of PVT1 was closely associated with tumor-node-metastasis (TNM) stage (III/IV vs. I/II: OR = 3.19, 95% CI: 2.43 - 4.18, p < 0.001), and the significant correlation between PVT1 expression and TNM stage is found in T classification (T3/4 vs. T1/2: OR = 6.48, 95% CI: 2.93 - 14.31, p < 0.001) and lymph node metastasis (present vs. absent: OR = 2.56, 95% CI:1.36 - 4.80, p = 0.003), but not in distant metastasis of patients with cancers (yes vs. no: OR = 2.50, 95% CI: 0.72 - 8.66, p = 0.15). Furthermore, the cancerous patients with high PVT1 expression had a worse histological differentiation than those with low PVT1 expression (undifferentiated/poorly vs. moderately/well: OR = 1.48, 95% CI: 1.02 - 2.14, p = 0.039). CONCLUSIONS: PVT1 could serve as a potent predicator of prognosis in different types of cancers.

A prospective comparison of short term results and functional recovery after laparoscopic subtotal colectomy and antiperistaltic cecorectal anastomosis with short colonic reservoir vs. long colonic reservoir.

BACKGROUND: To observe and compare the short term results and functional recovery of laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis (LSCACRA) in the treatment of Adult slow transit constipation (STC) with two different reservoir length: short colonic reservoir and long colonic reservoir. METHODS: All STC patients treated with LSCACRA between April 2007 and December 2011 at our institution were followed up. Patients with 2 cm to 3 cm ascending colon preserved above the ileocecal junction were designated as observation group, whereas those preserved by 10 cm to 15 cm were classified as control group. 41 cases in the observation group and 40 cases in the control group were enrolled. Preoperative and outcome parameters of patients were collected, including gender, age, body mass index, operative time , blood loss, first flatus time, hospital stay, postoperative complications, Wexner constipation scale(WCS), Wexner incontinence scale, gastrointestinal quality of life index(GIQLI), abdominal pain intensity scale(APIS), abdominal pain frequency scale(APFS) and abdominal bloating scale(ABS). RESULTS: Laparoscopic surgeries were successfully carried out for all patients, without any case transferred to laparotomy or death related to surgery. The operative time, blood loss, first flatus time, and days of hospital stay of the two groups did not show significant differences. We found no significant differences on complications (Clavien-Dindo grade > I) between the two groups. No patient exhibited anastomotic leak. No fecal incontinence occurred in both groups. On the 3(rd), 6(th) and 12(th) month after operation, the parameters of both groups significantly improved compared with the preoperative conditions (P < 0.05) except the APIS at 3(rd) and 6(th) month in control group. On the 3(rd), 6(th) and 12(th) month after operation, the Functional Recovery outcomes of WCS、GIQLI、APIS、APFS and ABS in the observation group were superior to those in the control group (P < 0.05). CONCLUSION: LSCACRA has a significant effect in the treatment of STC in adult. Postoperative outcomes can be optimized by shortening the length of the preserved ascending colon above the ileocecal junction, which promise better life quality of patients. TRIAL REGISTRATION: Chinese Clinical Trial Registry ChiCTR-OPC-14005280, 2014-09-29.