[The role of EBUS-TBNA in the systematic evaluation of lymph node staging and resectability analysis in non-small cell lung cancer].

Objective: To evaluate the role of endobronchial ultrasound guided transbronchial needle aspiration (EBUS-TBNA) in lymph node staging and resectability assessment of patients with non-small cell lung cancer (NSCLC). Methods: The clinical data of 154 patients with NSCLC who underwent EBUS-TBNA from March 2015 to December 2018 were collected. All accessible mediastinal and hilar lymph nodes were systematically explored and punctured using EBUS-TBNA. EBUS-TBNA and CT were used for preoperative staging and resectability evaluation. Results: The sensitivity, specificity and accuracy of EBUS-TBNA were 94.2%, 100.0% and 96.0%, respectively, while those of CT were 89.9%, 31.8% and 72.0%, respectively. The differences were statistically significant (P<0.05). The sensitivity, specificity and accuracy of EBUS-TBNA in lymph nodes with short diameter less than 15 mm were 92.4%, 100.0% and 96.0%, respectively, while those of CT were 80.7%, 34.8% and 60.1%, respectively, with statistical differences (P<0.05). The staging of 62 patients was changed, 27 cases were up-regulated and 35 cases were down-regulated. Among them, 32 cases had been changed to resectable. The evaluating resectability of EBUS-TBNA showed excellent consistency with that of pathological results (Kappa=0.95). The sensitivity and specificity were 100.0% and 97.2%, respectively. Conclusion: EBUS-TBNA can systemically evaluate the metastatic status of NSCLC patients and improve the accuracy of preoperative lymph node staging and resectability assessment.

[Analysis of influencing factors of ureteral anastomotic stenosis following urinary diversion after laparoscopic radical cystectomy].

Objective: To investigate the influencing factors of ureteroenteric strictures (UES) in patients undergoing laparoscopic radical cystectomy plus urinary diversion (UD). Method: A total of 412 patients who underwent UD after radical prostatectomy from January 2008 to December 2016 were retrospectively included in this study. Age, gender, body mass index (BMI), diversion type, time to diagnosis of UES, duration of ureteral stent, postoperative complications, including urinary tract infections, ureteroenteric leakage and UES were collected. Kaplan-Meier curves were used to describe time to developing UES. Prognostic factors of UES were analyzed using COX proportional hazard regression model. Result: Median follow-up time was 37 (IQR 17-120) months. A total of 59 patients (70 sides) developed UES, including 34 cases on the left side, 14 cases on the right side and 11 cases on both sides, following UD after radical cystectomy. The median time to diagnosis of UES was 7 (IQR 4-11) months. The total incidence of UES was 14.3%. The incidence of UES was 10.9%, 13.3% and 14.1% at 1, 3 and 5 years after UD, respectively. Cox proportional hazard regression model analysis demonstrated that BMI≥25kg·m(-2) (P=0.008), ureteroenteric leakage (P=0.001) and urinary tract infections (P=0.037) were the independent risk factors associated with UES following UD after radical cystectomy. Conclusion: The incidence rate of UES following UD after radical cystectomy was relatively high, which occurs more common on the left side. Obese patients, combined with ureteroenteric leakage, urinary tract infection after UD, are more likely to develop into UES.

Genetic instability and CpG methylation in the 5'-flanking region of the PAI-1 gene in Chinese patients with gastric cancer.

We explored a possible correlation of genetic instability and CpG methylation in the 5'-flanking region of the PAI-1 gene with clinicopathologic features of gastric cancer in Chinese patients and looked for molecular markers for diagnosing gastric tumor development. Microsatellite instability and loss of heterozygosity of the PAI-1 gene locus D7S515, D7S471 and pai-1 in 50 specimens of gastric cancer and relevant pericancerous tissues were detected by PCR-single strand conformation polymorphism (PCR-SSCP) with sliver staining. Methylation-specific PCR was used to detect CpG methylation in the 5'-flanking region of the PAI-1 gene. Microsatellite instability was significantly more common in the negative than in the positive serosa infiltration group of gastric cancer (42.86 vs 2.33%). The frequency of microsatellite instability was significantly lower in the cases with lymph node metastasis than in those without metastasis (18.18 vs 2.56%); however, it was significantly higher in the low differentiation group than that in the middle or high differentiation groups (21.05 vs 0.00%). CpG methylation in the 5'-flanking region of the PAI-1 gene did not differ significantly. Microsatellite instability and loss of heterozygosity of the PAI-1 gene apparently regulates the development of gastric cancer through different pathways. Microsatellite instability could be used as a molecular marker for the development of gastric cancer. CpG methylation in the 5'-flanking region of the PAI-1 gene appears not to be involved in the development of gastric cancer.

Prediction of the prognosis of patients with acute-on-chronic hepatitis B liver failure using the model for end-stage liver disease scoring system and a novel logistic regression model.

The objective of this study was to determine the predictive value of the model for end-stage liver disease (MELD) scoring system in patients with acute-on-chronic hepatitis B liver failure (ACLF-HBV), and to establish a new model for predicting the prognosis of ACLF-HBV. A total of 204 adult patients with ACLF-HBV were retrospectively recruited between July 1, 2002 and December 31, 2004. The MELD scores were calculated according to the widely accepted formula. The 3-month mortality was calculated. The validity of the MELD model was determined by means of the concordance (c) statistic. Clinical data and biochemical values were included in the multivariate logistic regression analysis based on which the regression model for predicting prognosis was established. The receiver-operating characteristic curves were drawn for the MELD scoring system and the new regression model and the areas under the curves (AUC) were compared by the z-test. The 3-month mortality rate was 57.8%. The mean MELD score for the patients who died was significantly greater than those who survived beyond 3 months (28.7 vs 22.4, P = 0.003). The concordance (c) statistic (equivalent to the AUC) for the MELD scoring system predicting 3-month mortality was 0.709 (SE = 0.036, P < 0.001, 95% confidence interval 0.638-0.780). The independent factors predicting prognosis were hepatorenal syndrome (P < 0.001), liver cirrhosis (P = 0.009), HBeAg (P = 0.013), albumin (P = 0.028) and prothrombin activity (P = 0.011) as identified in multivariate logistic regression analysis. The regression model that was constructed by the logistic regression analysis produced a greater prognostic value (c = 0.891) than the MELD scoring system (z = 4.333, P < 0.001). The MELD scoring system is a promising and useful predictor for 3-month mortality of ACLF-HBV patients. Hepatorenal syndrome, liver cirrhosis, HBeAg, albumin and prothrombin activity are independent factors affecting the 3-month mortality. The newly established logistic regression model appears to be superior to the MELD scoring system in predicting 3-month mortality in patients with ACLF-HBV.

Inhibition of autophagy by 3-MA promotes hypoxia-induced apoptosis in human colorectal cancer cells.

OBJECTIVE: Cell autophagy reduces the sensitivity of cancer cells to therapeutic reagents in various types of human cancer. Therefore, the aim of our study was to use human colorectal cancer HCT116 cells to explore whether inhibition of autophagy by 3-Methyladenine (3-MA, an autophagy inhibitor) is able to enhance hypoxia-induced apoptosis in vitro. MATERIALS AND METHODS: HCT116 cells were treated with 3-MA, hypoxia, or 3-MA plus hypoxia, and the autophagy, apoptosis and proliferation of the HCT116 cells were investigated. Western blot analysis was used to detect autophagy specificity protein microtubule-associated protein light chain 3 (LC3) expression. Effects on apoptosis were evaluated by using flow cytometry (JC-1 staining to measure mitochondrial membrane potential) and annexin V-propidium iodide (PI) staining. RESULTS: The results showed that the treatment of HCT116 cells in vitro with hypoxia alone increased autophagy as well as apoptosis, whereas combination treatment with 3-MA and hypoxia markedly inhibited hypoxia-induced autophagy, but increased hypoxia-induced cell apoptosis. CONCLUSIONS: Autophagy might play a role as a self-defense mechanism in hypoxia-treated colon cancer cells, and its inhibition could be a promising strategy for the adjuvant chemotherapy of colon cancer.

Preliminary proteomic analysis of radiation response markers in rectal cancer patients.

OBJECTIVE: To provide biomarkers related to the radiation response of patients to avoid unnecessary side effects on those who were not sensitive to radiotherapy. PATIENTS AND METHODS: In the present study, we compared the different four proteins (PDIA3, Vimentin, Galectin3, Dhe3) patterns in rectal tumor tissue before and after radiation therapy by using 2-D PAGE, mass spectrometry, and bioinformatics analysis. RESULTS: The protein level of Galectin3 and PDIA3 were downregulated in rectal cancer patients before and after radiotherapy (1.42 folds); while Dhe3 protein and Vimentin were upregulated (1-2 folds), and we also revealed Vimentin as its role in the negative regulation of the well-known transcription factor ATF4. CONCLUSIONS: Our study showed that four candidate proteins, including PIDA3, Galectin3, Dhe3, and Vimentin, might be the potential biomarkers in the identification of radiation response in rectal cancer.

Study on in vitro sperm capacitation and egg-penetration of giant panda.

A preliminary report on in vitro sperm capacitation and egg-penetration of giant panda is briefly presented. The panda spermatozoon consists of head, neck and tail, just like the spermatozoa of other animals. Before capacitation sperm heads clustered together and dispersed after capacitation. They were then able to swim straight forward. During the time of in vitro capacitation the plasma membrane of the sperm head was first expanded to various degrees, then disintegrated, and finally became detached. The electro-dense material in the acrosome appeared in small clumps with high density. Extensive vesiculation occurred between the bi-layered acrosome membranes and thus led to disintegration. Vesiculation in panda sperm differs from that reported in hamsters. When the capacitated panda spermatozoa came into contact with the hamster eggs, the region between the acrosome collar and postacrosome cap first fused with the egg membrane followed by the penetration of the nucleus into the cortex of the egg. Some of the penetrating sperm nuclei became decondensed and some did not. The success of in vitro sperm capacitation and egg-penetration of giant panda is of great significance, suggesting that it is possible to carry out in vitro fertilization and embryo transplantation in this endangered species.

Salicylate, an aspirin metabolite, specifically inhibits the current mediated by glycine receptors containing alpha1-subunits.

BACKGROUND AND PURPOSE: Aspirin or its metabolite sodium salicylate is widely prescribed and has many side effects. Previous studies suggest that targeting neuronal receptors/ion channels is one of the pathways by which salicylate causes side effects in the nervous system. The present study aimed to investigate the functional action of salicylate on glycine receptors at a molecular level. EXPERIMENTAL APPROACH: Whole-cell patch-clamp and site-directed mutagenesis were deployed to examine the effects of salicylate on the currents mediated by native glycine receptors in cultured neurones of rat inferior colliculus and by glycine receptors expressed in HEK293T cells. KEY RESULTS: Salicylate effectively inhibited the maximal current mediated by native glycine receptors without altering the EC(50) and the Hill coefficient, demonstrating a non-competitive action of salicylate. Only when applied simultaneously with glycine and extracellularly, could salicylate produce this antagonism. In HEK293T cells transfected with either alpha1-, alpha2-, alpha3-, alpha1beta-, alpha2beta- or alpha3beta-glycine receptors, salicylate only inhibited the current mediated by those receptors that contained the alpha1-subunit. A single site mutation of I240V in the alpha1-subunit abolished inhibition by salicylate. CONCLUSIONS AND IMPLICATIONS: Salicylate is a non-competitive antagonist specifically on glycine receptors containing alpha1-subunits. This action critically involves the isoleucine-240 in the first transmembrane segment of the alpha1-subunit. Our findings may increase our understanding of the receptors involved in the side effects of salicylate on the central nervous system, such as seizures and tinnitus.