Ginkgetin improved experimental colitis by inhibiting intestinal epithelial cell apoptosis through EGFR/PI3K/AKT signaling.

Excessive apoptosis of intestinal epithelial cells leads to intestinal barrier dysfunction, which is not only one of the pathological features of inflammatory bowel disease (IBD) but also a therapeutic target. A natural plant extract, Ginkgetin (GK), has been reported to have anti-apoptotic activity, but its role in IBD is unknown. This study aimed to explore whether GK has anti-colitis effects and related mechanisms. An experimental colitis model induced by dextran sulfate sodium (DSS) was established, and GK was found to relieve colitis in DSS-induced mice as evidenced by improvements in weight loss, colon shortening, Disease Activity Index (DAI), macroscopic and tissue scores, and proinflammatory mediators. In addition, in DSS mice and TNF-α-induced colonic organoids, GK protected the intestinal barrier and inhibited intestinal epithelial cell apoptosis, by improving permeability and inhibiting the number of apoptotic cells and the expression of key apoptotic regulators (cleaved caspase 3, Bax and Bcl-2). The underlying mechanism of GK's protective effect was explored by bioinformatics, rescue experiments and molecular docking, and it was found that GK might directly target and activate EGFR, thereby interfering with PI3K/AKT signaling to inhibit apoptosis of intestinal epithelial cells in vivo and in vitro. In conclusion, GK inhibited intestinal epithelial apoptosis in mice with experimental colitis, at least in part, by activating EGFR and interfering with PI3K/AKT activation, explaining the underlying mechanism for ameliorating colitis, which may provide new options for the treatment of IBD.

Prevalence and risk factors of pre-frailty and frailty in maintenance haemodialysis patients in China: A cross-sectional Study.

AIMS: To examine the prevalence and risk factors of pre-frailty and frailty in maintenance haemodialysis patients in China. DESIGN: A cross-sectional study. METHOD: From January to July 2017, using the convenience sampling method, a total of 503 maintenance haemodialysis patients from six hospitals in Lianyungang, China, were recruited for this study. The participants' socio-demographic, lifestyle factors and health information were assessed using a general information questionnaire. Frailty was evaluated based on the Fried frailty phenotype. Multi-categorical logistic regression was performed to examine factors associated with pre-frailty and frailty in this population, including age, sex, living alone, employment, educational level, body mass index, per capita monthly household income, smoking status, exercise status, primary diagnosis, dialysis age, frequency of dialysis, vascular access, congestive heart failure, other cardiac diseases, cerebrovascular disease, peripheral blood diseases, pain, albumin level and haemoglobin level. RESULTS: Among the 503 participants with an average age of 53.02 years (standard deviation 14.99), 178 had pre-frailty (35.3%) and were mostly young and middle-aged. The prevalence of pre-frailty among participants <60 years old was more than 40%. Regression analysis showed that lack of exercise, dialysis age ≤12 months, congestive heart failure and other cardiac diseases were positively associated with pre-frailty. Two hundred and eighteen participants were frail (43.3%), most of whom were aged ≥60. The prevalence of frailty in participants ≥60 was 71.4%. Regression analysis showed that advanced age, being female, obesity, low per capita monthly household income, lack of exercise, diabetes as the primary disease, dialysis age ≤12 months, congestive heart failure, other cardiac diseases, pain and low albumin level, were positively associated with frailty. In addition, more than half of the participants hardly exercised (64.6%), while lack of exercise was a risk factor for pre-frailty and frailty. A third of the participants had pain (33.4%), while pain was an independent risk factor for pre-frailty and frailty in these participants. CONCLUSION: Pre-frailty and frailty are common in patients with maintenance haemodialysis. Most of the elderly maintenance haemodialysis patients are frail, and most of the young and middle-aged patients are pre-frail. Clinicians should actively screen the pre-frailty and frailty among patients with maintenance haemodialysis, especially those with dialysis age ≤12 months. Many factors affect pre-frailty and frailty in this population. Tailored intervention measures should be designed and implemented based on these factors, giving priority to exercise guidance and pain management for patients to help them prevent or reverse pre-frailty and frailty.

Development of a Practical Screening Tool to Predict Sarcopenia in Patients on Maintenance Hemodialysis.

BACKGROUND Sarcopenia is a common complication in maintenance hemodialysis (MHD) and can increase patient hospitalization and mortality. No simple and reliable tools to identify sarcopenia exist. We aimed to develop a screening tool to predict MHD patients at high risk for sarcopenia. MATERIAL AND METHODS This cross-sectional study included 589 and 216 MHD patients for training and validation sets, respectively. We used diagnostic criteria developed by the Asian Working Group on Sarcopenia to screen for sarcopenia. The risk prediction model was established by univariate and multivariate logistic regression analyses. We used the area under the receiver operating characteristic curve (AUROC), calibration curve, Hosmer-Lemeshow test, and decision curve analysis (DCA) to evaluate the model's discrimination ability, calibration ability, and clinical utility. RESULTS The incidence of sarcopenia was 17.1% in the training set and 18.1% in the validation set. We constructed prediction models applying age, body mass index, calf circumference, and serum creatinine and plotted a nomogram. The training set model had an AUROC of 0.922, sensitivity of 85.1%, specificity of 85.9%, and chi-square value (Hosmer-Lemeshow test) of 5.603 (P>0.05); the DCA diagram showed that when the threshold probability was 0 to 0.95, the model predicted a net benefit for sarcopenia in MHD patients. The validation set model had an AUROC of 0.913, sensitivity of 94.3%, specificity of 82.9%, and chi-square value (Hosmer-Lemeshow test) of 9.822 (P>0.05). CONCLUSIONS The screening tool has good discrimination ability, calibration ability, and clinical utility. It could help to identify MHD patients at a high risk for sarcopenia.

Micronutrient Level Is Negatively Correlated with the Neutrophil-Lymphocyte Ratio in Patients with Severe COVID-19.

Aim: To explore the potential relationship between NLR and micronutrient deficiency in patients with severe COVID-19 infection. Methods: Sixteen patients were categorized into the mild group (mild COVID-19) and severe group (severe COVID-19) based on the guideline of the management of COVID-19. The lactate dehydrogenase (LDH); superoxide dismutase (SOD), the inflammatory markers (neutrophil lymphocyte ratio (NLR)), erythrocyte sedimentation rate (ESR), c-reactive protein (CRP), selenium (Se), iron (Fe), zinc (Zn), nickel (Ni), copper (Cu), chromium (Cr), cadmium (Cd), arsenic (As), and manganese (Mn) were measured in the blood. Results: Compared to the mild group, the NLR (P < 0.05) and the level of Se (P < 0.01), Fe (P < 0.05), and Zn (P < 0.05) were significantly decreased in the severe group. The level of Se, Fe, and Zn was significantly correlated to NLR levels. Furthermore, close positive correlation was found between NLR and severity of COVID-19. Conclusion: The micronutrient deficiency in the blood is associated with NLR in the severity of COVID-19 patients.

An epigenome-wide DNA methylation study of patients with COVID-19.

In the early 2000s, emerging SARS-CoV-2, which is highly pathogenic, posed a great threat to public health. During COVID-19, epigenetic regulation is deemed to be an important part of the pathophysiology and illness severity. Using the Illumina Infinium Methylation EPIC BeadChip (850 K), we investigated genome-wide differences in DNA methylation between healthy subjects and COVID-19 patients with different disease severities. We conducted a combined analysis and selected 35 "marker" genes that could indicate a SARS-CoV-2 infection, including 12 (ATHL1, CHN2, CHST15, CPLX2, CRHR2, DCAKD, GNAI2, HECW1, HYAL1, MIR510, PDE11A, and SMG6) situated in the promoter region. The functions and pathways of differentially methylated genes were enriched in biological processes, signal transduction, and the immune system. In the "Severe versus Mild" group, differentially methylated genes, after eliminating duplicates, were used for PPI analyses. The four hub genes (GNG7, GNAS, PRKCZ, and PRKAG2) that had the highest degree of nodes were identified and among them, GNG7 and GNAS genes expressions were also downregulated in the severe group in sequencing results. Above all, the results suggest that GNG7 and GNAS may play a non-ignorable role in the progression of COVID-19. In conclusion, the identified key genes and related pathways in the current study can be used to study the molecular mechanisms of COVID-19 and may provide possibilities for specific treatments.

Sophoricoside improved Crohn's disease-like colitis by inhibiting intestinal epithelial cell apoptosis through PI3K/AKT signaling.

BACKGROUND AND AIMS: Increased apoptosis of intestinal epithelial cells (IECs) is a significant cause of intestinal barrier dysfunction in Crohn's disease (CD). Sophoricoside (SOP) is an isoflavone glycoside known for its anti-apoptotic properties. The aim of this study was to investigate the effects of SOP on mice with CD-like colitis and to understand the underlying mechanisms. METHODS: Mice treated with 2,4,6-trinitrobenzene sulfonic acid (TNBS) were used to examine the therapeutic effect of SOP on CD-like colitis and intestinal barrier damage. To further explore SOP's impact on IECs apoptosis and intestinal barrier protection, an in vitro colonic organoid apoptosis model induced by TNF-α was utilized. Network pharmacology was employed to predict the relevant pathways and molecular processes associated with SOP in the treatment of CD. RESULTS: Treatment with SOP significantly improved colitis symptoms in TNBS mice, as demonstrated by reductions in the Disease Activity Index (DAI), weight loss, colon shortening, macroscopic scores, colonic tissue inflammatory scores, and the expression of pro-inflammatory factors. Our experiments confirmed that SOP protects the intestinal barrier by counteracting IECs apoptosis. Additionally, this study established that SOP reduced IECs apoptosis by inhibiting the PI3K/AKT signaling pathway. CONCLUSIONS: SOP can reduce IECs apoptosis through the inhibition of the PI3K/AKT signaling pathway, thereby protecting the intestinal barrier. This study is the first to illustrate how SOP ameliorates colitis and protects the intestinal barrier, suggesting SOP has potential clinical application in treating CD.

Intestinal Adipocytes Transdifferentiate into Myofibroblast-like Cells and Contribute to Fibrosis in Crohn's Disease.

BACKGROUND AND AIMS: Intestinal fibrotic stenosis is a major reason for surgery in Crohn's disease [CD], but the mechanism is unknown. Thus, we asked whether intestinal adipocytes contribute to intestinal fibrosis. Adipocytes were found to transdifferentiate into myofibroblasts and confirmed to be involved in mesenteric fibrosis in our recent study. Here, we investigated the role and possible mechanisms of intestinal adipocytes in intestinal fibrosis in CD. METHODS: The intestinal tissue of patients with CD with or without fibrotic stenosis [CDS or CDN] and normal intestinal tissue from individuals without CD were obtained to assess alterations in submucosal adipocytes in CDS and whether these cells transdifferentiated into myofibroblasts and participated in the fibrotic process. Human primary adipocytes and adipose organoids were used to evaluate whether adipocytes could be induced to transdifferentiate into myofibroblasts and to investigate the fibrotic behaviour of adipocytes. LPS/TLR4/TGF-ß signalling was also studied to explore the underlying mechanism. RESULTS: Submucosal adipocytes were reduced in number or even absent in CDS tissue, and the extent of the reduction correlated negatively with the degree of submucosal fibrosis. Interestingly, submucosal adipocytes in CDS tissue transdifferentiated into myofibroblast-like cells and expressed collagenous components, possibly due to stimulation by submucosally translocated bacteria. Lipopolysaccharide [LPS]-stimulated human primary adipocytes and adipose organoids also exhibited transdifferentiation and profibrotic behaviour. Mechanistically, TLR4-mediated TGF-ß signalling was associated with the transdifferentiation and profibrotic behaviour of intestinal adipocytes in CDS tissue. CONCLUSIONS: Intestinal adipocytes transdifferentiate into myofibroblasts and participate in the intestinal fibrosis process in CD, possibly through LPS/TLR4/TGF-ß signalling.

The gut microbiota from maintenance hemodialysis patients with sarcopenia influences muscle function in mice.

Background: Sarcopenia is a common complication in patients undergoing maintenance hemodialysis (MHD). Growing evidence suggests a close relationship between the gut microbiota and skeletal muscle. However, research on gut microbiota in patients with sarcopenia undergoing MHD (MS) remains scarce. To bridge this knowledge gap, we aimed to evaluate the pathogenic influence of gut microbiota in the skeletal muscle of patients with MS, to clarify the causal association between gut microbiota and skeletal muscle symptoms in patients with MS and identify the potential mechanisms underlying this causal association. Methods: Fecal samples were collected from 10 patients with MS and 10 patients without MS (MNS). Bacteria were extracted from these samples for transplantation. Mice (n=42) were randomly divided into three groups and, after antibiotic treatment, fecal microbiota transplantation (FMT) was performed once a day for 3 weeks. Skeletal muscle and fecal samples from the mice were collected for 16S rRNA gene sequencing and for histological, real-time PCR, and metabolomic analyses. Results: Mice colonized with gut microbiota from MS patients exhibited notable decreases in muscle function and muscle mass, compared with FMT from patients with MNS. Moreover, 16S rRNA sequencing revealed that the colonization of MS gut microbiota reduced the abundance of Akkermansia in the mouse intestines. Metabolome analysis revealed that seven metabolic pathways were notably disrupted in mice transplanted with MS microbiota. Conclusion: This study established a connection between skeletal muscle and the gut microbiota of patients with MS, implying that disruption of the gut microbiota may be a driving factor in the development of skeletal muscle disorders in patients undergoing MHD. This finding lays the foundation for understanding the pathogenesis and potential treatment methods for sarcopenia in patients undergoing MHD.

Knockdown of GSG2 inhibits the development and progression of non-small cell lung cancer in vitro and in vivo.

Lung cancer has been recognized as the most common malignant neoplasm of the respiratory system with extremely high morbidity, among which non-small cell lung cancer (NSCLC) accounts for the majority. Many published literatures have revealed the roles of GSG2 in the progression of ovarian cancer, bladder cancer and breast cancer. However, there were no reports on the relationship between GSG2 and NSCLC. Herein, GSG2 was identified as a potential tumor promoter in NSCLC development, whose abundant expression was observed in NSCLC tissues compared with adjacent nonmalignant tissues and statistically correlated with more advanced tumor stage, more malignant grade and higher risk of lymphatic metastasis. Subsequent in vitro loss-of-function experiments indicated that GSG2 depletion could arrest cell cycle and suppress cell proliferation and migration while enhancing cell apoptosis. At the same time, the suppressive effects of GSG2 depletion on NSCLC development were verified by in vivo experiments. In conclusion, the current study identified GSG2 as a tumor promoter in development and progression of NSCLC, which could work as a novel therapeutic target for NSCLC.

Erratum: Targeting cysteine-rich angiogenic inducer-61 by antibody immunotherapy suppresses growth and migration of non-small cell lung cancer.

[This corrects the article DOI: 10.3892/etm.2018.6274.].

Characterization of the gut microbiota in hemodialysis patients with sarcopenia.

PURPOSE: Maintenance hemodialysis (MHD) patients are at high risk of sarcopenia. Gut microbiota affects host metabolic and may act in the occurrence of sarcopenia importantly. This study aimed to study the characterization of the gut microbiota in MHD patients with sarcopenia, and to further reveal the complex pathophysiology of sarcopenia in MHD patients. METHODS: Fecal samples and clinical data were collected from 30 MHD patients with sarcopenia, and 30 age-and-sex-matched MHD patients without sarcopenia in 1 general hospital of Jiangsu Province from December 2020 to March 2021. 16S rRNA sequencing technology was used to analyze the genetic sequence of the gut microbiota for evaluation of the diversity, species composition, and differential microbiota of the two groups. RESULTS: Compared to MHD patients without sarcopenia, the ACE index of patients with sarcopenia was lower (P = 0.014), and there was a structural difference in the ß-diversity between the two groups (P = 0.001). At the genus level, the relative abundance of Tyzzerella_4 in the sarcopenia group was significantly higher than in the non-sarcopenia group (P = 0.039), and the relative abundance of Megamonas (P = 0.004), Coprococcus_2 (P = 0.038), and uncultured_bacterium_f_Muribaculaceae (P = 0.040) decreased significantly. CONCLUSION: The diversity and structure of the gut microbiota of MHD patients with sarcopenia were altered. The occurrence of sarcopenia in MHD patients may be influenced by gut microbiota.

Comparison of the prevalence and associated factors of cognitive frailty between elderly and middle-young patients receiving maintenance hemodialysis.

PURPOSE: This study aimed at comparing the prevalence of cognitive frailty and explore the differences in the influencing factors between elderly and middle-young patients receiving maintenance hemodialysis (MHD). METHODS: In this cross-sectional study, the frailty phenotype, mini-mental state examination, and clinical dementia rating were used to assess the current status of cognitive frailty in 852 patients receiving MHD from four hospitals in Lianyungang City and Xuzhou City, Jiangsu Province, China; the influencing factors were then analyzed for statistical significance. RESULTS: Of the total 852 patients receiving MHD, 340 were classified into an elderly group (≥ 60 years) and 512 into a middle-young group (< 60 years). The prevalence of cognitive frailty was 35.9% and 8.8%, respectively. The results of multivariate logistic regression analysis showed that the independent factors of cognitive frailty were age (P < 0.001), education level (P = 0.010), nutritional status (P = 0.001), serum albumin level (P = 0.010), calf circumference (P = 0.024), and social support level (P < 0.001) in the elderly group and comorbidity status (P = 0.037), education level (P < 0.001), nutritional status (P = 0.008), serum creatinine level (P = 0.001), waist circumference (P < 0.001), and depression (P = 0.006) in the middle-young group. CONCLUSION: The prevalence of cognitive frailty was significantly higher in the elderly group than in the middle-young group, and the influencing factors differed between the two populations.

Clinical Features for Severely and Critically Ill Patients with COVID-19 in Shandong: A Retrospective Cohort Study.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a novel pathogen, has caused an outbreak of coronavirus disease 2019 (COVID-19) that has spread rapidly around the world. Determining the risk factors for death and the differences in clinical features between severely ill and critically ill patients with SARS-CoV-2 pneumonia has become increasingly important. AIM: This study was intended to provide insight into the difference between severely ill and critically ill patients with SARS-CoV-2 pneumonia. METHODS: In this retrospective, multicenter cohort study, we enrolled 62 seriously ill patients with SARS-CoV-2 pneumonia who had been diagnosed by March 12, 2020. Clinical data, laboratory indexes, chest images, and treatment strategies collected from routine medical records were compared between severely ill and critically ill patients. Univariate and multivariate logistic regression analyses were also conducted to identify the risk factors associated with the progression of patients with severe COVID-19. RESULTS: Of the 62 patients with severe or critical illness, including 7 who died, 30 (48%) patients had underlying diseases, of which the most common was cardiovascular disease (hypertension, 34%, and coronary heart disease, 5%). Compared to patients with severe disease, those with critical disease had distinctly higher white blood cell counts, procalcitonin levels, and D-dimer levels, and lower hemoglobin levels and lymphocyte counts. Multivariate regression showed that a lymphocyte count less than 109/L (odds ratio 20.92, 95% CI 1.76-248.18; p=0.02) at admission increased the risk of developing a critical illness. CONCLUSION: Based on multivariate regression analysis, a lower lymphocyte count (<109/L) on admission is the most critical independent factor that is closely associated with an increased risk of progression to critical illness. Age, underlying diseases, especially hypertension and coronary heart disease, elevated D-dimer, decreased hemoglobin, and SOFA score, and APACH score also need to be taken into account for predicting disease progression. Blood cell counts and procalcitonin levels for the later secondary bacterial infection have a certain reference values.

Effects of intradialytic resistance exercise on systemic inflammation in maintenance hemodialysis patients with sarcopenia: a randomized controlled trial.

PURPOSE: To investigate the effect of intradialytic resistance exercise on inflammation markers and sarcopenia indices in maintenance hemodialysis (MHD) patients with sarcopenia. METHODS: Forty-one MHD patients with sarcopenia were divided into an intervention group (group E, n = 21) and a control group (group C, n = 20). Group C patients only received routine hemodialysis care, whereas group E patients received progressive intradialytic resistance exercise with high or moderate intensity for 12 weeks at three times per week (using the weight of the lower limbs and elastic ball movement of the upper limb) on the basis of routine hemodialysis care. RESULTS: After 12 weeks, a significant difference in physical activity status (maximum grip strength, daily pace, and physical activity level), Kt/V, and C-reactive protein was found between groups E and C. Inflammatory factors (interleukin (IL)-6, IL-10, and tumor necrosis factor(TNF)-α) increased or decreased more significantly in group E than in group C. CONCLUSIONS: This study showed that intradialytic resistance exercise can improve physical activity effectively and reduce microinflammatory reactions even if this simple exercise does not affect the muscle mass in MHD patients with sarcopenia.

Targeting cysteine-rich angiogenic inducer-61 by antibody immunotherapy suppresses growth and migration of non-small cell lung cancer.

Non-small cell lung cancer (NSCLC) is the most frequent type of human lung cancer; lung cancer is responsible for the highest rates of cancer-associated mortality in the world. Cysteine-rich angiogenic inducer-61 (CYR-61) has been identified as a tumorigenesis-, development- and metastasis-related gene, and is reported to enhance proliferation, migration and invasion through hepatocyte growth factor (HGF)-induced scattering and the metastasis-inducing HGF/Met signaling pathway in tumor cells and xenograft models. CYR-61 is a protein that promotes human lung cancer cell metastasis and is closely related to the patient's prognosis in NSCLC. The purpose of the present study was to investigate whether CYR-61 may serve as a dual potential target for gene therapy of human NSCLC. In the present study, an antibody targeted against CYR-61 (anti-CYR-61) was constructed and the therapeutic effects and underlying mechanism of this antibody in NSCLC cells and mice with NSCLC was investigated. It was observed that NSCLC cell viability, migration and invasion were inhibited while cell apoptosis was induced by the neutralization of CYR-61 protein by anti-CYR-61. Western blotting demonstrated that extracellular signal-regulated kinase (ERK) and protein kinase B (AKT) expression levels in NSCLC cells were decreased following treatment with anti-CYR-61. In addition, it was observed that inhibition of NSCLC cell viability was achieved by the suppression of the epithelial-mesenchymal transition signaling pathway. ERK and AKT phosphorylation levels were downregulated in NSCLC cells and tumors following anti-CYR-61 treatment. Analysis of a murine model indicated that tumor growth was inhibited and tumor metastasis was significantly suppressed (P<0.01) following anti-CYR-61 treatment for CYR-61. In conclusion, CYR-61 may serve as a potential target for gene therapy for the treatment of human NSCLC.

Scoring System to Predict the Risk of Surgical Site Infection in Patients with Esophageal Cancer after Esophagectomy with Cervical Anastomosis.

BACKGROUND: Surgical site infection (SSI) surveillance has become increasingly important during the peri-operative period of esophagectomy with cervical anastomosis (McKeown esophagectomy). This study sought to clarify the risk factors for SSI and to develop a stratification scoring system to predict SSI after esophagectomy with cervical anastomosis. PATIENTS AND METHODS: All patients who underwent elective esophagectomy with cervical anastomosis were studied between January 2010 and December 2016 in the Chinese Academy of Medical Sciences Cancer Hospital (CAMS). Univariable analysis and multivariable logistic regression were used to screen the independent risk factors. A risk stratification scoring system was developed based on multivariable logistic regression parameters. The model derivation set involved 711 consecutive cases, and the validation set involved 168 consecutive cases. RESULTS: In the model derivation set, there were 711 patients, of whom 146 were found to have SSI and the incidence rate was 20.53%. Multivariable analysis found that SSI was associated independently with the following adverse risk factors: peripheral vascular disease, prior chest surgery, no pre-operative surgical antibiotic prophylaxis (SAP) administration within 120 minutes prior to incision, low serum albumin, and low pre-albumin at post-operative day zero to three, respectively. Each of these factors contributed one point to the risk score and a risk stratification scoring system was established. The SSI rates were increased gradually in the low, intermediate, high, and extremely high-risk groups (p < 0.001). The area under the receiver operating characteristic (AUROC) curve was 0.706 for the logistic regression model and 0.704 for the scoring system. In the validation set, the model performed equivalently (AUC = 0.824). CONCLUSIONS: The validated stratification scoring system could predict accurately the risk of SSI after esophagectomy with cervical anastomosis. This could be helpful in the selection of high-risk patients requiring frequent monitoring and more aggressive interventions to decrease the incidence of SSI.