MicroRNA-328-3p facilitates the progression of gastric cancer via KEAP1/NRF2 axis.

Gastric cancer is a common lethal malignancy and causes great cancer-related mortality worldwide. MicroRNA (miR)-328-3p is implicated in the progression of various human cancers; however, its role and mechanism in the progression of gastric cancer remain unclear.Human gastric cancer cells were incubated with miR-328-3p mimic, inhibitor or the matched negative control. Cell viability, colony formation, migrative and invasive capacity, cell apoptosis and oxidative stress were measured. To clarify the involvement of nuclear factor-E2-related factor 2 (NRF2) and kelch-like ECH-associated protein 1 (KEAP1), small interfering RNA was used. miR-328-3p was upregulated in human gastric cancer cells and tissues, and its level positively correlated with the progression of gastric cancer. miR-328-3p promoted cell viability, colony formation, migration and invasion, thereby facilitating the progression of gastric cancer. miR-328-3p mimic reduced, while miR-328-3p inhibitor increased apoptosis and oxidative stress of human gastric cancer cells. Mechanistically, miR-328-3p upregulated NRF2 via targeting KEAP1to attenuate excessive free radical production and cell apoptosis. miR-328-3p functions as an oncogenic gene and inhibiting miR-328-3p may help to develop novel therapeutic strategies of human gastric cancer.

Differences in the clinical characteristics and outcomes of COVID-19 patients in the epicenter and peripheral areas of the pandemic from China: a retrospective, large-sample, comparative analysis.

BACKGROUND: There is limited information on the difference in epidemiology, clinical characteristics and outcomes of the initial outbreak of the coronavirus disease (COVID-19) in Wuhan (the epicenter) and Sichuan (the peripheral area) in the early phase of the COVID-19 pandemic. This study was conducted to investigate the differences in the epidemiological and clinical characteristics of patients with COVID-19 between the epicenter and peripheral areas of pandemic and thereby generate information that would be potentially helpful in formulating clinical practice recommendations to tackle the COVID-19 pandemic. METHODS: The Sichuan & Wuhan Collaboration Research Group for COVID-19 established two retrospective cohorts that separately reflect the epicenter and peripheral area during the early pandemic. The epidemiology, clinical characteristics and outcomes of patients in the two groups were compared. Multivariate regression analyses were used to estimate the adjusted odds ratios (aOR) with regard to the outcomes. RESULTS: The Wuhan (epicenter) cohort included 710 randomly selected patients, and the peripheral (Sichuan) cohort included 474 consecutive patients. A higher proportion of patients from the periphery had upper airway symptoms, whereas a lower proportion of patients in the epicenter had lower airway symptoms and comorbidities. Patients in the epicenter had a higher risk of death (aOR=7.64), intensive care unit (ICU) admission (aOR=1.66), delayed time from illness onset to hospital and ICU admission (aOR=6.29 and aOR=8.03, respectively), and prolonged duration of viral shedding (aOR=1.64). CONCLUSIONS: The worse outcomes in the epicenter could be explained by the prolonged time from illness onset to hospital and ICU admission. This could potentially have been associated with elevated systemic inflammation secondary to organ dysfunction and prolonged duration of virus shedding independent of age and comorbidities. Thus, early supportive care could achieve better clinical outcomes.

Impaired immune and coagulation systems may be early risk factors for COVID-19 patients: A retrospective study of 118 inpatients from Wuhan, China.

The coronavirus disease 2019 (COVID-19) outbreak has become a global health threat and will likely be one of the greatest global challenges in the near future. The battle between clinicians and the COVID-19 outbreak may be a "protracted war."The objective of this study was to investigate the risk factors for in-hospital mortality in patients with COVID-19, so as to provide a reference for the early diagnosis and treatment.This study retrospectively enrolled 118 patients diagnosed with COVID-19, who were admitted to Eastern District of Renmin Hospital of Wuhan University from February 04, 2020 to March 04, 2020. The demographics and laboratory data were collected and compared between survivors and nonsurvivors. The risk factors of in-hospital mortality were explored by univariable and multivariable logistic regression to construct a clinical prediction model, the prediction efficiency of which was verified by receiver-operating characteristic (ROC) curve.A total of 118 patients (49 males and 69 females) were included in this study; the results revealed that the following factors associated with in-hospital mortality: older age (odds ratio [OR] 1.175, 95% confidence interval [CI] 1.073-1.287, P = .001), neutrophil count greater than 6.3 × 10 cells/L (OR 7.174, (95% CI 2.295-22.432, P = .001), lymphocytopenia (OR 0.069, 95% CI 0.007-0.722, P = .026), prothrombin time >13 seconds (OR 11.869, 95% CI 1.433-98.278, P = .022), D-dimer >1 mg/L (OR 22.811, 95% CI 2.224-233.910, P = .008) and procalcitonin (PCT) >0.1 ng/mL (OR 23.022, 95% CI 3.108-170.532, P = .002). The area under the ROC curve (AUC) of the above indicators for predicting in-hospital mortality were 0.808 (95% CI 0.715-0.901), 0.809 (95% CI 0.710-0.907), 0.811 (95% CI 0.724-0.898), 0.745 (95% CI 0.643-0.847), 0.872 (95% CI 0.804-0.940), 0.881 (95% CI 0.809-0.953), respectively. The AUC of combined diagnosis of these aforementioned factors were 0.992 (95% CI 0.981-1.000).In conclusion, older age, increased neutrophil count, prothrombin time, D-dimer, PCT, and decreased lymphocyte count at admission were risk factors associated with in-hospital mortality of COVID-19. The prediction model combined of these factors could improve the early identification of mortality risk in COVID-19 patients.

XPG Asp1104His polymorphism and gastrointestinal cancers risk: a meta-analysis.

Several studies have reported the association between the Asp1104His polymorphism in xeroderma pigmentosum group G (XPG) gene and risk of gastrointestinal cancers. However, the results are inconsistent. This meta-analysis was performed to assess the association between XPG Asp1104His polymorphism and gastrointestinal cancers risk. Relevant studies were identified using PubMed, Web of Science, CNKI, WanFang and VIP databases up to July 22, 2014. The pooled odds ratio (OR) with a 95% confidence interval (CI) was calculated using the fixed- or random effects model. 13 case-control studies from twelve publications with 4275 patients and 5735 controls were included. Overall, a significant association was found between the XPG Asp1104His polymorphism and the risk of gastrointestinal cancers (dominant model: OR = 1.15, 95% CI: 1.05-1.26; His/His vs. Asp/Asp: OR = 1.15, 95% CI: 1.01-1.32). When the analysis was stratified by ethnicity, similar results were observed in Asians under homozygote model; in stratification analysis by cancer type, increased cancer risk was detected in colorectal and hepatocellular carcinoma, but not for other gastrointestinal cancers. Furthermore, in subgroup analysis by source of control, we failed to detect any association among population, hospital and family-based populations. This meta-analysis indicated that the XPG Asp1104His polymorphism may be a risk factor for gastrointestinal cancers, especially of colorectal cancer.

[Diagnosis of obscure gastrointestinal bleeding among different age groups by double balloon enteroscope].

OBJECTIVE: To investigate the diagnostic value of double balloon enteroscope (DBE) on obscure gastrointestinal bleeding(OGIB) and to analyze etiological characteristics among different age groups. METHODS: The clinical data of patients undergoing DBE due to OGIB in the Department of Gastroenterology in Renmin Hospital of Wuhan University from January 2007 to January 2012 were retrospectively analyzed and compared among different age groups. Patients were divided into the young group(age≤40, n=86), the middle age group(aged 41-59, n=81), and the elderly group (age≥60, n=49). The detection of bleeding origin by DBE was compared between different age groups. RESULTS: Diagnosis rates in young, middle age, elderly group were 83.7%(72/86), 87.7%(71/81), 81.6%(40/49) without statistical differences(P>0.05). Complication rates in the young, middle age, and elderly group were 1.2%(1/86), 2.5%(2/81), 2.0%(1/49) without statistic difference(P>0.05). The most common cause in young group was diverticulum/replica malformation while the most common location was ileum. The most common cause in both middle age and elderly group was tumor. CONCLUSIONS: DBE is an effective and safe method for diagnosis of OGIB among different age groups. Each age group has its etiological characteristics. Diagnosis and therapeutic strategy based on age-related characteristics is worthy of further investigation.

Functional genes based analysis of sulfur-oxidizing bacteria community in sulfide removing bioreactor.

Sulfur-oxidizing bacteria (SOB) are the main microorganisms that participate in the bioremediation of sulfide-rich wastewater. To reveal the SOB community structure and determine which members of SOB contribute to the sulfide oxidation in a sulfide-rich cloth printing and dyeing wastewater treatment plant, specific primer pairs dsrA 625F/877R, soxB 704F/1199R, and sqr 473F/982R based on the SOB functional genes encoding dissimilatory sulfite reductase, sulfate thioesterase/thiohydrolase, and sulfide: quinone oxidoreductase were designed. The restriction fragment length polymorphism analysis showed that the diversity indices and the abundance of each OTU have no significant changes after time, which suggested the SOB community in the sulfide removing bioreactor have high steady phylogenetic analysis of functional gene-based clone libraries detected the SOB from Chlorobia, α-proteobacteria, ß-proteobacteria, and γ-proteobacteria. The combined clone library showed the presence of dominant members of the SOB species closely related to families Halothiobacillaceae (17%), Hydrogenophilaceae (14%), and Rhodocyclaceae (13%), which may contribute to the sulfide oxidation in wastewater treatment process. This work provides a precise understanding of SOB microbial community within sulfide removing bioreactor, and the result gives assistance for the optimization of the treatment systems for sulfide biological degradation.

Comparison and characterization of microbial communities in sulfide-rich wastewater with and without propidium monoazide treatment.

A 16S rRNA gene-based culture-independent approach was used to study the bacterial and archaeal communities in a sulfide-rich wastewater. Propidium Monoazide (PMA) treatment was applied to limit the analysis to the fraction of viable cells in environment. A total of 104 and 68 clones respective from bacterial clone library and archaeal library were picked and analyzed by restriction fragment length polymorphism (RFLP). 35 RFLP patterns from bacterial clone library and 10 RFLP patterns from archaeal clone library were unique and the respective clones were selected for sequencing. BLAST analysis and RFLP analysis showed that the bacterial clone library mainly consisted of Gammaproteobacteria (73%), Anaerolineae (6%), Bacilli (5%), Deltaproteobacteria (7%), Clostridia (4%), Bacteroidetes (1%), and Chlorobia (1%); Methanomicrobia (99%) and Thermococci (1%) were the only two lineages of the archaeal domains. This study gave a first insight into the overall microbial structure in a cloth printing and dyeing wastewater treatment plant with high concentration of sulfide and increased knowledge on the applicability of the PMA treatment in combination with PCR-based molecular techniques to analyze only viable cells in microbial ecology.

Method to detect only viable cells in microbial ecology.

Propidium monoazide can limit the analysis of microbial communities derived from genetic fingerprints to viable cells with intact cell membranes. However, PMA treatment cannot completely suppress polymerase chain reaction (PCR) amplification when the targeted gene is too short. PMA treatment in combination with two-step nested PCR was designed to overcome this problem. Four experiments were performed to determine the limitation of PMA treatment and to evaluate the suitability of the method by applying the following samples: (1) pure cultures of Escherichia coli O157:H7, Enterobacter aerogenes, and Alcaligenes faecalis; (2) pond water samples spiked with heat-killed E. coli O157:H7 and E. aerogenes; (3) anaerobic sludge samples exposed to increasing heat stress; and (4) selected natural samples of estuarine sediment and lake mud. Results from the first two experiments show that PMA treatment cannot efficiently suppress dead cells from PCR amplification when the targeted gene is as short as 190 bp, however, the two-step nested PCR can overcome this problem. The last two experiments indicate the method that PMA treatment in combination with two-step nested PCR is useful for viable cells detection in microbial ecology.

RhoA and RhoC -siRNA inhibit the proliferation and invasiveness activity of human gastric carcinoma by Rho/PI3K/Akt pathway.

AIM: To evaluate the effects of adenovirus-mediated gene transfer of RhoA siRNA and RhoC siRNA on proliferation and invasion of SGC7901 cells by Rho/PI3K/Akt pathway. METHODS: Plasmid of RhoA siRNA and RhoC siRNA were constructed and transfected into SGC7901 cells. siRNA and LY294002 (PI3K inhibitor) were designed as the control group. The mRNA and protein expressions of RhoA and RhoC were respectively detected with RT-PCR and western blotting. In order to find out the changes of proliferation and invasion power of SGC7901 cell lines, we analyzed the data by MTT, Boyden chamber and evaluated apoptosis of cell with flow cytometry. We treated BALB/C nude mice with RhoA and RhoC-siRNA, and tumor control rate (%) in nude mice was calculated. RESULTS: RhoA and RhoC siRNA transfections specifically down-regulated the corresponding mRNA and protein levels in SGC7901 Cells. The experiment of permeated artificial basal membrane showed that the invasion power of SGC7901 cell lines are on the decline after treatment of Ad-RhoA and RhoC-siRNA (12.64 +/-3.27 vs 87.38 +/- 17.38, P < 0.05). The values of 490 nm wavelength light absorption were different in the five groups. The number of alive cells in the group of RhoA and RhoC-siRNA was lower than others in the 6(th) d (0.71 +/- 0.01 vs 3.82 +/- 0,11 P < 0.05). The apoptosis rate of transfected RhoA and RhoC-siRNA group with FACS were 19.07% +/- 1.78 and there were significant differences between treated and control groups (19.07 +/- 1.78% vs 1.23 +/- 0.11%, P < 0.01). The tumor transplantation experiment in BALB/C nude mice showed intratumoral injection of RhoA or RhoC siRNA can inhibit tumor growth. CONCLUSION: RhoA and RhoC siRNA gene therapy mediated by adenovirus may be useful for inhibiting growth and invasion of SGC7901 through a PI3K/Akt pathway. These results provide a novel therapeutic target in preventing gastric cancer cell invasion and metastasis.