Bacillus mycoides JA20-1 was screened and identified as a biocontrol bacterium with a high capacity for producing volatile organic compounds(VOCs) in the laboratory. This strain had significant inhibitory effects on various postharvest disease pathogens in crops, such as Botrytis cinerea, as well as soil-borne disease pathogens in ginseng, such as Sclerotinia ginseng. In order to accelerate its industrialization process, in this study, single-factor experiments and response surface optimization methods were used. The fermentation medium and fermentation conditions in the shake flask of strain JA20-1 were systematically optimized by using cell production volume as the response variable. Meanwhile, the biocontrol effect of JA20-1 on B. cinerea of ginseng during the storage period was evaluated by using the method of fumigation in a dry dish in vitro. The results indicated that the optimal fermentation medium formulation for strain JA20-1 was as follows: 1% yeast paste, 1% soluble starch, 0.25% K_2HPO_4·3H_2O, and 0.2% NaCl. The optimal fermentation conditions in the shake flask were vaccination size of 3%, culture volume of 50 mL in a 250 mL Erlenmeyer flask, pH of 6.2, fermentation temperature of 34 ℃, shaking speed of 180 r·min~(-1), and incubation time of 18 hours. The bacteria count in the fermentation broth under these conditions reached 2.17 × 10~8 CFU·mL~(-1), which was 6.58 times higher than before. The average control efficacy of the fermentation broth on Botrytis cinerea of ginseng under in vitro fumigation reached 61.70% and 84.04% respectively, when 20 mL and 30 mL per dish were used. The research provided theoretical support and technical foundation for the development and utilization of Bacillus mycoides JA20-1 and the biocontrol of soil-borne diseases in ginseng and postharvest diseases in crops.
Botrytis/drug effects* ; Fermentation ; Panax/microbiology* ; Plant Diseases/prevention & control* ; Volatile Organic Compounds/metabolism* ; Bacillus/physiology* ; Pest Control, Biological/methods* ; Biological Control Agents/metabolism* ; Culture Media/chemistry*

BACKGROUND: Blood glucose and serum albumin have been associated with cardiovascular disease prognosis, but the impact of admission-blood-glucose-to-albumin ratio (AAR) on adverse outcomes in critical ill coronary artery disease (CAD) patients was not investigated. METHODS: Patients diagnosed with CAD were non-consecutively selected from the MIMIC-IV database and categorized into quartiles based on their AAR. The primary outcome was 1-year mortality, and secondary endpoints were in-hospital mortality, acute kidney injury (AKI), and renal replacement therapy (RRT). A restricted cubic splines model and Cox proportional hazard models assessed the association between AAR and adverse outcomes in CAD patients. Kaplan-Meier survival analysis determined differences in endpoints across subgroups. RESULTS: A total of 8360 patients were included. There were 726 patients (8.7%) died in the hospital and 1944 patients (23%) died at 1 year. The incidence of AKI and RRT was 63% and 4.3%, respectively. High AAR was markedly associated with in-hospital mortality (HR = 1.587, P = 0.003), 1-year mortality (HR = 1.502, P < 0.001), AKI incidence (HR = 1.579, P < 0.001), and RRT (HR = 1.640, P < 0.016) in CAD patients in the completely adjusted Cox proportional hazard model. Kaplan-Meier survival analysis noted substantial differences in all endpoints based on AAR quartiles. Stratified analysis and interaction test demonstrated stable correlations between AAR and outcomes. CONCLUSIONS The results highlight that AAR may be a potential indicator for assessing in-hospital mortality, 1-year mortality, and adverse renal prognosis in critical CAD patients.

This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.
Male ; Animals ; Mice ; Cisplatin/pharmacology* ; Carcinoma, Hepatocellular/genetics* ; MAP Kinase Signaling System ; Beclin-1 ; Apoptosis ; Liver Neoplasms/genetics* ; Necrosis ; Proto-Oncogene Proteins c-bcl-2/metabolism* ; Cell Line, Tumor ; RNA, Messenger/metabolism* ; Autophagy

AIM: To explore the correlation between the expression levels of microRNA-377-3p(miR-377-3p)and microRNA-365-3p(miR-365-3p)in serum and aqueous humor and the degree of diabetes macular edema(DME).METHODS: A total of 60 DME patients(60 eyes)admitted to 363 Hospital from February 2021 to February 2022 were selected in this prospective study(the severe eye was selected if both eyes had DME, while the right eye was selected if the same degree of DME), including 24 mild eyes, 21 moderate eyes and 15 severe eyes. In addition, another 60 patients(60 eyes)with type 2 diabetes(without fundus disease)admitted to our hospital during the same period were selected as the control group. The basic clinical data of all subjects were collected, including body mass index(BMI), smoking history, drinking history, hypertension, hyperlipidemia, the course of diabetes, glycated hemoglobin levels, fasting blood glucose and homocysteine(Hcy); the expression levels of miR-377-3p and miR-365-3p were detected by real-time fluorescent quantitative PCR(qRT-PCR).RESULTS: The course of diabetes, glycosylated hemoglobin, fasting blood glucose and Hcy in DME group were obviously higher than those in control group(all P<0.05); the expression levels of miR-377-3p and miR-365-3p in serum of patients in DME group were lower than those in control group(all P<0.05); the expression levels of miR-377-3p and miR-365-3p in serum and aqueous humor in severe group were obviously lower than those in moderate group and mild group, and those in moderate group were obviously lower than those in mild group(all P<0.05); the expression levels of miR-377-3p and miR-365-3p in serum were negatively correlated with central macular thickness(CMT; r=-0.342, -0.374, all P<0.05), the expression levels of miR-377-3p and miR-365-3p in aqueous humor were negatively correlated with CMT(r=-0.425, -0.503, all P<0.05); the multivariate Logistic regression analysis showed that the course of diabetes, increased fasting blood glucose and Hcy were risk factors for DME in type 2 diabetes patients, and serum miR-377-3p and miR-365-3p were protective factors for DME in type 2 diabetes patients(P<0.05).CONCLUSION: The expression of miR-377-3p and miR-365-3p in serum and aqueous humor of patients with DME is low, which is negatively related to the severity of DME patients.

Literature regarding the impacts of heavy metal exposure on erectile dysfunction (ED) is scarce. We aimed to evaluate the correlation between 10 urinary metals and ED in a large, nationally representative adult male sample. The dataset was extracted from the National Health and Nutrition Examination Survey (NHANES) during the period of 2001-2002 and 2003-2004. Weighted proportions and multivariable logistic regression analysis adjusted for confounding variables were utilized to determine the relationship between metal exposure and ED. Weighted quantile sum (WQS) regression was utilized to evaluate the impact of a mixture of urinary metals on ED. A total of 1328 participants were included in our study. In multivariable logistic regression analysis, cobalt (Co) and antimony (Sb) were positively associated with ED (odds ratio [OR]: 1.36, 95% confidence interval [CI]: 1.10-1.73, P = 0.020; and OR: 1.41, 95% CI: 1.12-1.77, P = 0.018, respectively) after full adjustment. Men in tertile 4 for Co (OR: 1.49, 95% CI: 1.02-2.41, P for trend = 0.012) and Sb (OR: 1.53, 95% CI: 1.08-2.40, P for trend = 0.041) had significantly higher odds of ED than those in tertile 1. Furthermore, the WQS index was significantly linked with increased odds of ED after full adjustment (OR: 1.31, 95% CI: 1.04-1.72, P < 0.05). Our study expanded on previous literature indicating the possible role of heavy metal exposure in the etiology of ED. The evaluation of heavy metal exposure should be included in the risk assessment of ED.
Adult ; Humans ; Male ; United States ; Erectile Dysfunction/etiology* ; Nutrition Surveys ; Metals, Heavy ; Risk Assessment

OBJECTIVES: To investigate the clinical characteristics and risk factors for early-onset necrotizing enterocolitis (NEC) in preterm infants with very/extremely low birth weight (VLBW/ELBW). METHODS: A retrospective analysis was performed on the medical data of 194 VLBW/ELBW preterm infants with NEC who were admitted to Children's Hospital Affiliated to Zhengzhou University from January 2014 to December 2021. These infants were divided into early-onset group (onset in the first two weeks of life; n=62) and late-onset group (onset two weeks after birth; n=132) based on their onset time. The two groups were compared in terms of perinatal conditions, clinical characteristics, laboratory examination results, and clinical outcomes. Sixty-two non-NEC infants with similar gestational age and birth weight who were hospitalized at the same period as these NEC preterm infants were selected as the control group. The risk factors for the development of early-onset NEC were identified using multivariate logistic regression analysis. RESULTS: Compared with the late-onset group, the early-onset group had significantly higher proportions of infants with 1-minute Apgar score ≤3, stage III NEC, surgical intervention, grade ≥3 intraventricular hemorrhage, apnea, and fever or hypothermia (P<0.05). The multivariate logistic regression analysis showed that feeding intolerance, blood culture-positive early-onset sepsis, severe anemia, and hemodynamically significant patent ductus arteriosus were independent risk factors for the development of early-onset NEC in VLBW/ELBW preterm infants (P<0.05). CONCLUSIONS VLBW/ELBW preterm infants with early-onset NEC have more severe conditions compared with those with late-onset NEC. Neonates with feeding intolerance, blood culture-positive early-onset sepsis, severe anemia, or hemodynamically significant patent ductus arteriosus have a higher risk of early-onset NEC.
Child ; Infant ; Female ; Pregnancy ; Infant, Newborn ; Humans ; Infant, Premature ; Infant, Extremely Low Birth Weight ; Ductus Arteriosus, Patent ; Enterocolitis, Necrotizing/etiology* ; Retrospective Studies ; Infant, Newborn, Diseases ; Infant, Premature, Diseases/etiology* ; Risk Factors

Objective:To explore the association between atypical small acinar proliferation(ASAP)and subsequent diagnosis of intermediate and high risk prostate cancer(PCa),and analyze whether delaying repeat biopsy timing is safe and effective.Meth-ods:From June 2000 to June 2022,we retrospectively analyzed the clinical data of 276 patients accepting prostatic biopsy and diag-nosed with ASAP in China-Japan Union Hospital of Jilin University.54.7％(151/276)patients had a repeat biopsy.We used statis-tic methods to process the data.Results:25.2％(38/151)patients were diagnosed with PCa on repeat biopsy.Among them,78.9％(30/38)patients had Gleason score(GS)3+3 and 21.1％(8/38)had GS 3+4 disease.There were 4 and 6 patients got RP re-spectively in the two cohorts.Only 5.3％(8/151)of ASAP patients were diagnosed as intermediate risk PCa in repeated biopsy and specially,no high risk PCa was identified in our study.Conclusion:It was safe and valid to delay the repeat biopsy.

Objective To investigate the effects of the aqueous extracts of ganoderma leucocontextum (GLAE) on cognitive decline of aging rats and possible regulation mechanism. Methods Fifty rats were divided into five groups, control group, model group, GLAE low-dose group, GLAE middle-dose group and GLAE high-dose group. Aging SD rat models were made by D-galactose, and then treated continuously with different doses (0, 50, 100, 200 mg/ kg) of GLAE. The novel object recognition and step down test were performed to detect the changes of rats cognitive function. The brain tissue was stained with toluidine blue, Giemsa and HE staining and observed. The cerebral cell DNA damage was detected by comet assay. Expressions of protein kinase A (PKA) / cyclic adenosine monophosphate (cAMP)-response element binding protein (CREB) signaling pathway related factors in brain were respectively detected by ELISA, Western blotting and Real-time PCR. Results Compared with the model group, administration of GLAE could obviously alleviate rats cognitive decline and pathological change. The levels of cell DNA damage reduced markedly (P<0. 05). The contents of cAMP, brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF), and also expression levels of mRNA and protein of PKA, BDNF, NGF, CREB in the brain increased significantly in each medicated group (P<0. 01, P< 0. 05). Conclusion GLAE can improve cognitive function, and its mechanism may be related to activation of brain PKA/ CREB signaling pathway, increase in neurotrophic factor content and inhibition of cell DNA damage.

Gut microbiome sequencing studies have great potential to translate microbial analysis outcomes into human health research. Sequencing strategies of 16S amplicon and whole-metagenome shotgun (WMS) are two main methods in microbiome research with respective advantages. However, how sample heterogeneity, sequencers and library preparation protocols affect the sequencing reproducibility of gut microbiome needs further investigation. This study aims to provide a reference for the selection of sequencing technologies by comparing differences in microbial composition from different sampling sites. The results of three widely adopted sequencers showed that the technical repetition correlation (r= 0. 94) was high in WMS method, while the biological repetition correlation (r = 0. 69) was low. Bray-Curtis distance identified that dissimilarity from biological replicates was larger than that of technical replicates (P<0. 001). In addition, dissimilarity and specific taxonomic profiles were observed between 16S and WMS datasets. Our results imply that homogenization is a necessary step before sample DNA extraction. The sequencers contributed less to taxonomic variation than the library preparation protocols. We developed an empirical Bayes approach that " borrowed information" in calculations and analyzed batch effect parameters using standardized data and prior distributions of (non-) parameters, which may improve population comparability between 16S and WMS and provide a basis for further application to fusion analysis of published 16S and microbial datasets.

There are two serious obstacles to tumor immunotherapy. Firstly, the immune response of the tumor is seriously reduced due to immunosuppressive tumor microenvironment (ITM) and low immunogenicity of tumor. The second obstacle is the dense and complex heterogeneous structures, which seriously prevent the nanoparticles (NPs) from penetrating deeper into tumor tissue. Immunogenic cell death (ICD) induced by doxorubicin (DOX) is an effective method to enhance tumor immune activity. However, interferon-γ (IFN-γ) secreted by cytotoxic T lymphocytes (CTL) after ICD induction would increase the expression of indoleamine 2,3-dioxygenase 1 (IDO1) and enhance ITM. IDO1 siRNA would reduce the expression of IDO1 protein, regulate the tumor immunosuppressive microenvironment and regulate ITM, so as to enhance the ICD effect of DOX. In this paper, a novel charge conversional, particle size reduction and highly penetrable NPs based on a pH sensitive copolymer poly(ethylene glycol)-poly-L-lysine-2,3-dimethylmaleic anhydride (mPEG-PLL-DMA, PLD) and polyamidoamine (PAMAM) dendrimers to achieve deep delivery of tumor tissue. DOX and IDO1 siRNA were encapsulated to achieve efficient tumor immunotherapy. Preparation and cell level experiments showed that PLD material had significant pH sensitivity. Results of 3D tumor penetrable experiment in vitro showed that adding the pH sensitive material PLD significantly improved the permeability of the preparation. In addition, 4T1 tumor model was established for BALB/c mice and all animal experiments were displayed in according with the requirements of the Animal Experiment Ethics Committee of Shenyang Pharmaceutical University. The results of in vivo efficacy experiments and tissue experiments evaluated that IDO1 siRNA significantly improved the ICD effect owing to DOX, so as to significantly inhibit tumor growth.