Sulforaphane effectively inhibits HBV by altering Treg/Th17 immune balance and the MIF-macrophages polarizing axis in vitro and in vivo.

BACKGROUND: Hepatitis B virus (HBV) infection is a major public health problem. After HBV infection, viral antigens shift the immune balance in favor of viral escape. Sulforaphane (SFN) is a traditional Chinese medicine.It regulates multi-biological activities, including anti-inflammation, anticancer, and antiviral. However, few studies reported that SFN can inhibit HBV infection before. METHODS: An immunocompetent HBV CBA/CaJ mouse model and a co-culture model were used to explore the effect of SFN on HBV and whether SFN altered the immune balance after HBV infection. RESULTS: We found that SFN was able to reduce HBV DNA, cccDNA, HBsAg, HBeAg, and HBcAg levels in serum and liver tissues of HBV-infected mice. In vitro and in vivo experiments showed that SFN could significantly increase the expression of Cd86 and iNOS and inhibit the expression of Arg1 on macrophages after HBV infection. After SFN administration, Th17 markers in liver tissue and serum were significantly increased. There was no significant changes in the proportion of Treg cells in peripheral blood, but a significant increase in the proportion of Th17 cells and decrease of the Treg/Th17 ratio. Using a network pharmacology approach, we predicted macrophage migration inhibitory factor (MIF) as a potential target of SFN and further validated that MIF expression was significantly increased after HBV infection and SFN significantly inhibited MIF expression both in vitro and in vivo. There was an upward trend in HBV markers (p>0.05) after MIF overexpression. Overexpression of MIF combined with the use of SFN resulted in a significant reversion in the expression of HBV markers and polarization of macrophages towards the M1 phenotype. CONCLUSION: Our results indicated that immunocompetent HBV CBA/CaJ mouse model is a good model to evaluate HBV infection. SFN could inhibit the expression of HBV markers, promote polarization of macrophages towards the M1 phenotype after HBV infection, change the proportion of Treg and Th17 cells. Our findings demonstrate that SFN inhibit HBV infection by inhibiting the expression of MIF and promoting the polarization of macrophages towards the M1 phenotype, which illustrates a promising therapeutic approach in HBV infection.

Nectar robbing by bees affects the reproductive fitness of the distylous plant Tirpitzia sinensis (Linaceae).

Nectar robbing can affect plant reproductive success directly by influencing female and male fitness, and indirectly by affecting pollinator behavior. Flowers have morphological and chemical features that may protect them from nectar robbers. Previous studies on nectar robbing have focused mainly on homotypic plants. It remains unclear how nectar robbing affects the reproductive success of distylous plants, and whether defense strategies of two morphs are different. Nectar-robbing rates on the long- and short-styled morph (L-morph, S-morph) of the distylous Tirpitzia sinensis were investigated. We compared floral traits, the temporal pattern of change in nectar volume and sugar concentration, nectar secondary metabolites, and sugar composition between robbed and unrobbed flowers of two morphs. We tested direct effects of nectar robbing on female and male components of plant fitness and indirect effects of nectar robbing via pollinators. Nectar-robbing rates did not differ between the two morphs. Flowers with smaller sepals and petals were more easily robbed. The floral tube diameter and thickness were greater in L-morphs than in S-morphs, and the nectar rob holes were significantly smaller in L-morphs than in S-morphs. Nectar robbing significantly decreased nectar replenishment rate but did not affect nectar sugar concentration or sugar composition. After robbery, the quantities and diversity of secondary compounds in the nectar of S-morphs increased significantly and total relative contents of secondary compounds in L-morphs showed no obvious changes. Nectar robbing could decrease female fitness by decreasing pollen germination rate and thus decreasing seed set. Nectar robbing had no significant effects on male fitness. Robbed flowers were less likely to be visited by hawkmoth pollinators, especially in S-morphs. These results suggest that nectar robbing could directly and indirectly decrease the female fitness of T. sinensis, and different morphs have evolved different defense mechanisms in response to nectar-robbing pressure.

Quantitative Analysis on Molecular Characteristics Evolution of Gastric Cancer Progression and Prognosis.

The dynamic changes of key biological characteristics from gastric low-grade intraepithelial neoplasia (LGIN) to high-grade intraepithelial neoplasia (HGIN) to early gastric cancer (EGC) are still unclear, which greatly affect the accurate diagnosis and treatment of EGC and prognosis evaluation of gastric cancer (GC). In this study, bioinformatics methods/tools are applied to quantitatively analyze molecular characteristics evolution of GC progression, and a prognosis model is constructed. This study finds that some dysregulated differentially expressed mRNAs (DEmRNAs) in the LGIN stage may continue to promote the occurrence and development of EGC. Among the LGIN, HGIN, and EGC stages, there are differences and relevance in the transcription expression patterns of DEmRNAs, and the activation related to immune cells is very different. The biological functions continuously changed during the progression from LGIN to HGIN to EGC. The COX model constructed based on the three EGC-related DEmRNAs has GC prognostic risk prediction ability. The evolution of biological characteristics during the development of EGC mined by the authors provides new insight into understanding the molecular mechanism of EGC occurrence and development. The three-gene prognostic risk model provides a new method for assisting GC clinical treatment decisions.

Combined influence of soil moisture and atmospheric humidity on land surface temperature under different climatic background.

Soil moisture (SM) and atmospheric humidity (AH) are crucial climatic variables that significantly affect the climate system. However, the combined influencing mechanisms of SM and AH on the land surface temperature (LST) under global warming are still unclear. Here, we systematically analyzed the interrelationships among annual mean values of SM, AH, and LST using ERA5-Land reanalysis data and revealed the role of SM and AH on the spatiotemporal variations of LST through mechanism analysis and regression methods. The results showed that net radiation, SM, and AH could well model the long-term variability of LST well and explain 92% of the variability. Moreover, SM played an essential and different role under the different LST backgrounds. The AH always displayed a greenhouse effect on the LST. This study provides essential insights into the global climate change mechanism from the surface hydrothermal processes perspective.

The association between fear of future workplace violence and depressive symptoms among nurses based on different experiences of workplace violence: a cross-sectional study.

BACKGROUND: Fear of future workplace violence (FFWV) has a negative impact on individuals' health. However, no study has investigated the association between FFWV and depressive symptoms. Nurses with different experiences of workplace violence may have different levels of FFWV and differences in mental health. This study explored the association between FFWV and depressive symptoms among Chinese nurses with different experiences of workplace violence. METHODS: A cross-sectional study was conducted involving 1888 Chinese nurses from 12 tertiary hospitals in Shandong Province. The Fear of Future Violence at Work scale was used to measure FFWV. Depressive symptoms were assessed using the 10-item Center for Epidemiologic Studies Depression scale. Multiple logistic regression analysis was used to examine the association between FFWV and depressive symptoms. RESULTS: The prevalence of depressive symptoms was 45.9% (no aggression group: 24.3%; non-physical violence group: 46.1%; physical violence group: 63.7%), and 72.8% of nurses had high levels of fear of future workplace violence (no aggression group: 60.2%; non-physical violence group: 75.6%; physical violence group: 70.8%). Compared with low levels of FFWV, high levels of FFWV were associated with more depressive symptoms among nurses in the no aggression group (odds ratio [OR] = 3.269, 95% confidence interval [CI]: 1.102-9.695) and in the non-physical violence group (OR = 2.338, 95% CI: 1.385-3.945). CONCLUSION: Nurses who had experienced physical violence had the most depressive symptoms and nurses with experience of non-physical violence had the greatest FFWV. Our findings suggested that there was a significant association between FFWV and depressive symptoms among Chinese nurses in the no aggression and non-physical violence groups. Hospital administrators need to address FFWV needs when developing strategies to reduce depressive symptoms among nurses.

Development of a Pancreatic Fistula Prediction Model After Pancreaticoduodenectomy Based on a Decision Tree and Random Forest Algorithm.

BACKGROUND: The incidence of postoperative pancreatic fistula (POPF) after pancreaticoduodenectomy (PD) is high. We sought to develop a POPF prediction model based on a decision tree (DT) and random forest (RF) algorithm after PD and to explore its clinical value. METHODS: The case data of 257 patients who underwent PD in a tertiary general hospital from 2013 to 2021 were retrospectively collected in China. The RF model was used to select features by ranking the importance of variables, and both algorithms were used to build the prediction model after automatic adjustment of parameters by setting the respective hyperparameter intervals and resampling as a 10-fold cross-validation method, etc. The prediction model's performance was assessed by the receiver operating characteristic curve (ROC) and the area under curve (AUC). RESULTS: Postoperative pancreatic fistula occurred in 56 cases (56/257, 21.8%). The DT model had an AUC of .743 and an accuracy of .840, while the RF model had an AUC of .977 and an accuracy of .883. The DT plot visualized the process of inferring the risk of pancreatic fistula from the DT model on independent individuals. The top 10 important variables were selected for ranking in the RF variable importance ranking. CONCLUSION: This study successfully developed a DT and RF algorithm for the POPF prediction model, which can be used as a reference for clinical health care professionals to optimize treatment strategies to reduce the incidence of POPF.

Escitalopram-induced sinus bradycardia in coronary heart disease combined with depression: a case report and review of literature.

For patients with cardiovascular disease, using the antidepressant escitalopram may lead to unexpected adverse events. Here, a rare repeated sinus bradycardia event due to escitalopram is first reported. In an 82-year-old female patient with cardiac dysfunction using digoxin, tachycardia (average heart rate of 93 beats/min) was demonstrated by electrocardiogram (ECG). She began to take escitalopram and lorazepam due to depression, but sinus bradycardia (93.7% heart rate was <60 beats/min) and sinus arrest were first detected after 3 months. Its proportion decreased to 0.1% after discontinuation of digoxin and escitalopram for 1 day, and the rhythm returned to normal 2 weeks later. After 2 months, escitalopram was prescribed again in combination with quetiapine; then, 17.1% heart rate was <60 beats/min. After escitalopram and quetiapine withdrawal, the ECG showed the heart rhythm had normalized again. No other drug changes were made during these periods. Escitalopram was deemed to be a highly possible cause of sinus bradycardia according to its Naranjo's Algorithm score. Furthermore, literature on escitalopram-mediated cardiovascular adverse events was reviewed and analyzed. Empirically, escitalopram should be discontinued immediately if iatrogenic causes cannot be ruled out. Furthermore, ECG monitoring in escitalopram-related cardiovascular adverse events is highlighted, especially in patients receiving certain drug classes simultaneously (i.e., sinoatrial node inhibitors, antipsychotics).

Hepatitis B Virus Infection Promotes M2 Polarization of Macrophages by Upregulating the Expression of B7x In Vivo and In Vitro.

Several studies have reported that hepatitis B virus (HBV) infection is mediated by macrophages and that the B7x (B7-H4, VTCN-1) protein plays an important role in immune regulation in HBV-associated hepatocellular carcinoma (HBV-HCC). However, the relationship among HBV, macrophages, and B7x has not been studied. In this study, HBV-infected mouse model and coculture of HBV cell lines and macrophages were used to observe the changes in macrophages and the role of B7x after HBV infection. The expression of HBV markers (HBeAg, HBsAg), negative regulator of immunity (B7x), T-helper 17 (Th17)/T-regulatory (Treg)-related cytokines, and macrophage markers, as well as changes in the apoptosis and cell cycle of macrophages were analyzed through reverse transcription quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, western blot, and flow cytometry. The expression of HBsAg, HBeAg, and B7x increased and the levels of macrophage surface marker and Treg cells secrete related cytokines (IL-10 and TGF-ß) were altered after HBV infection both in vivo and in vitro. Apoptosis of macrophages increased, and cell cycle arrest occurred in vitro. These effects, except those in the cell cycle, were reversed when B7x was knocked down. Thus, HBV infection can promote the expression of B7x, which in turn regulates the Th17/Treg balance and affects the expression of HBsAg and HBeAg. The mechanism used by B7x likely involves the promotion of macrophage polarization and apoptosis. These results suggest that B7x is a novel target for HBV immunotherapy.

GATA binding protein 4 promotes the expression and transcription of hepatitis B virus by facilitating hepatocyte nuclear factor 4 alpha in vitro.

BACKGROUND: GATA binding protein 4 (GATA4) has been reported as a potential target of gene therapy for hepatocellular carcinoma (HCC). It is well known that the main cause of HCC is the chronic infection of hepatitis B virus (HBV). However, whether the effect of GATA4 on HBV has not yet been reported. METHODS: In this study, the regulation of GATA4 on HBV was analyzed in vitro. In turn, the effect of HBV on GATA4 was also observed in vitro, in vivo, and clinical HCC patients. Subsequently, we analyzed whether the effect of GATA4 on HBV was related to hepatocyte nuclear factor 4 alpha (HNF4α) in vitro. RESULTS: The results showed that GATA4 significantly promoted the secretion of HBV surface antigen (HBsAg) and HBV e antigen in the cell culture medium, improved the replication of HBV genomic DNA, and increased the level of HBV 3.5 kb pre-genomic RNA and HBV total RNA (P < 0.05). Moreover, it was showed that HBV had no significant effect on GATA4 in vitro and in vivo (P > 0.05). At the same time, GATA4 expression was decreased in 78.9% (15/19) of HCC patients regardless of the HBV and HBsAg status. Among them, there were 76.9% (10/13) in HBV-associated patients with HCC (HBV-HCC), and 83.3% (5/6) in non-HBV-HCC patients. In addition, the expression of HNF4α was also up-regulated or down-regulated accordingly when stimulating or interfering with the expression of GATA4. Furthermore, stimulating the expression of HNF4α could only alleviate the HBsAg level and HBV transcription levels, but had no significant effect on GATA4. CONCLUSIONS: In summary, this study found that GATA4 has a positive effect on HBV, and the potential pathway may be related to another transcription factor HNF4α that regulates HBV.

mRBioM: An Algorithm for the Identification of Potential mRNA Biomarkers From Complete Transcriptomic Profiles of Gastric Adenocarcinoma.

PURPOSE: In this work, an algorithm named mRBioM was developed for the identification of potential mRNA biomarkers (PmBs) from complete transcriptomic RNA profiles of gastric adenocarcinoma (GA). METHODS: mRBioM initially extracts differentially expressed (DE) RNAs (mRNAs, miRNAs, and lncRNAs). Next, mRBioM calculates the total information amount of each DE mRNA based on the coexpression network, including three types of RNAs and the protein-protein interaction network encoded by DE mRNAs. Finally, PmBs were identified according to the variation trend of total information amount of all DE mRNAs. Four PmB-based classifiers without learning and with learning were designed to discriminate the sample types to confirm the reliability of PmBs identified by mRBioM. PmB-based survival analysis was performed. Finally, three other cancer datasets were used to confirm the generalization ability of mRBioM. RESULTS: mRBioM identified 55 PmBs (41 upregulated and 14 downregulated) related to GA. The list included thirteen PmBs that have been verified as biomarkers or potential therapeutic targets of gastric cancer, and some PmBs were newly identified. Most PmBs were primarily enriched in the pathways closely related to the occurrence and development of gastric cancer. Cancer-related factors without learning achieved sensitivity, specificity, and accuracy of 0.90, 1, and 0.90, respectively, in the classification of the GA and control samples. Average accuracy, sensitivity, and specificity of the three classifiers with machine learning ranged within 0.94-0.98, 0.94-0.97, and 0.97-1, respectively. The prognostic risk score model constructed by 4 PmBs was able to correctly and significantly (∗∗∗ p < 0.001) classify 269 GA patients into the high-risk (n = 134) and low-risk (n = 135) groups. GA equivalent classification performance was achieved using the complete transcriptomic RNA profiles of colon adenocarcinoma, lung adenocarcinoma, and hepatocellular carcinoma using PmBs identified by mRBioM. CONCLUSIONS: GA-related PmBs have high specificity and sensitivity and strong prognostic risk prediction. MRBioM has also good generalization. These PmBs may have good application prospects for early diagnosis of GA and may help to elucidate the mechanism governing the occurrence and development of GA. Additionally, mRBioM is expected to be applied for the identification of other cancer-related biomarkers.

C2orf40 inhibits hepatocellular carcinoma through interaction with UBR5.

BACKGROUND AND AIM: Hepatocellular carcinoma (HCC) urgently needs a marker for early diagnosis and targeted treatment. C2orf40 has been identified as a tumor suppressor gene in many cancers. However, the precise role and regulatory mechanism by C2orf40 contribute to HCC remain elusive and merit exploration. METHODS: Reverse-transcription PCR, quantitative real-time PCR, and methylation-specific PCR were used to detect expression and methylation of C2orf40 in HCC cell lines or tissues. The effects of C2orf40 in liver cancer cells were examined via colony formation, CCK8, transwell, and flow cytometric assays. The effect of C2orf40 on tumorigenesis in vivo was determined by xenografts and immunohistochemical analysis. Western blot, indirect immunofluorescence, Co-IP, and cycloheximide (CHX) were used to further investigate the potential mechanism of C2orf40. RESULTS: The down-regulation of C2orf40 in hepatocellular cancer tissue samples is often related to the degree of methylation of its promoter CpG. The recovery of C2orf40 expression in HCC cell lines can induce G0/G1 phase arrest and apoptosis and also inhibit cell migration and invasion by reversing the epithelial-mesenchymal transition (EMT) process, both in vivo and in vitro. In addition, C2orf40 can increase the expression of p21 through interaction with UBR5. CONCLUSIONS: Low expression levels of C2orf40 are related to the hypermethylation of its promoter. C2orf40 can inhibit HCC through UBR5-dependent or p53-independent mechanisms. C2orf40 may be a diagnostic biomarker and a potential therapeutic target in HCC.

Aloperine inhibits hepatitis C virus entry into cells by disturbing internalisation from endocytosis to the membrane fusion process.

Aloperine, a natural alkaloid isolated from the Chinese traditional herb Sophora alopecuroides, is a broad-spectrum antiviral agent with anti-inflammatory activity. Here, we found that aloperine effectively inhibited hepatitis C virus (HCV) propagation in Huh7.5 cells and primary human hepatocytes without cytotoxicity, and it blocked HCV cell-to-cell viral transmission. The antiviral mechanism evidence demonstrated that aloperine inhibits HCV internalisation from endocytosis to the membrane fusion process, and the target may be associated with host factors. Aloperine additively inhibited HCV propagation with direct-acting antivirals (DAAs) and was effective against HCV variants resistant to known DAAs. Therefore, aloperine might be a natural lead compound for the development of innovative antivirals, and the combined use of aloperine with DAAs might contribute to eliminating liver diseases caused by HCV infection.

Erratum: Author correction to 'Up-regulation of glycolipid transfer protein by bicyclol causes spontaneous restriction of hepatitis C virus replication' [Acta Pharmaceutica Sinica B 9 (2019) 769-781].

[This corrects the article DOI: 10.1016/j.apsb.2019.01.013.].

Up-regulation of glycolipid transfer protein by bicyclol causes spontaneous restriction of hepatitis C virus replication.

Bicyclol is a synthetic drug for hepatoprotection in clinic since 2004. Preliminary clinical observations suggest that bicyclol might be active against hepatitis C virus (HCV) with unknown mechanism. Here, we showed that bicyclol significantly inhibited HCV replication in vitro and in hepatitis C patients. Using bicyclol as a probe, we identified glycolipid transfer protein (GLTP) to be a novel restrictive factor for HCV replication. The GLTP preferentially bound host vesicle-associated membrane protein-associated protein-A (VAP-A) in competition with the HCV NS5A, causing an interruption of the complex formation between VAP-A and HCV NS5A. As the formation of VAP-A/NS5A complex is essential for viral RNA replication, up-regulation of GLTP by bicyclol reduced the level of VAP-A/NS5A complex and thus inhibited HCV replication. Bicyclol also exhibited an inhibition on HCV variants resistant to direct-acting antiviral agents (DAAs) with an efficacy identical to that on wild type HCV. In combination with bicyclol, DAAs inhibited HCV replication in a synergistic fashion. GLTP appears to be a newly discovered host restrictive factor for HCV replication, Up-regulation of GLTP causes spontaneous restriction of HCV replication.

A proof-of-concept study in HCV-infected Huh7.5 cells for shortening the duration of DAA-based triple treatment regimens.

With the development of more effective direct-acting antivirals (DAAs), dual- or triple-therapy regimens represent the major strategy used to cure chronic hepatitis C virus (HCV) infection. Thus, shorter treatment duration regimens with low burden, few adverse effects and good patient adherence are urgently needed. This study theoretically demonstrates a proof-of-concept approach for shortening therapy duration by examining HCV-infected Huh7.5 cells after treatment with a high or low fixed dose of three DAAs (simeprevir + daclatasvir + sofosbuvir) for 6-15 days. The results demonstrated that HCV-infected Huh7.5 cells achieved an ultrarapid virologic response with undetectable HCV RNA and protein and were cured after treatment with the triple-therapy regimen for 15 days. When the treatment duration was shortened, virologic relapse might occur after treatment with a low fixed dose of the three DAAs for 9 days and did occur after treatment with a low fixed dose for 6 days, although HCV was below detectable levels at the end of treatment. However, virologic relapse could be avoided with treatment of a high fixed dose of the three DAAs for 9 or 6 days. Although a virologic breakthrough occurred after an intermittent treatment regimen at the low fixed dose, the high fixed dose cured HCV-positive Huh7.5 cells with intermittent treatment. In conclusion, HCV is persistently present below detectable levels in HCV-infected Huh7.5 cells for a long time after treatment, and a shortened therapy duration is associated with an increased risk of virologic relapse, but virologic relapse or breakthrough might be avoided by treatment with a combination of more highly effective DAAs.

Clinical practice guideline on traditional Chinese medicine therapy alone or combined with antibiotics for sepsis.

BACKGROUND: To develop the clinical practice guidelines for the treatment of sepsis with traditional Chinese medicine (TCM) therapy alone or TCM combined with antibiotics. METHODS: The methods and process for developing the international clinical practice guidelines were fully consulted between a group of doctors. A total of 25 experts from 14 units were involved in the development of this guideline. The major clinical questions that needed to be solved were raised first, and the best available evidence to solve them was researched. Finally, according to the principle set by the GRADE system, the available evidence was graded with levels ranging from high to low. This formed the recommendation strengths, which included strong recommendation and weak recommendation, or an expert consensus recommendation. RESULTS: The guideline identified the terms and definition for sepsis. For example, it identified its epidemiological characters, the advantages of TCM treatment on sepsis, the diagnosis and its features, the complications, and its rehabilitation and health maintenance. The guideline has put forward 14 recommendations, among which 4 were strong recommendations and 6 were weak recommendations, in addition to 4 expert consensus recommendations. CONCLUSIONS: The methods and processes for developing international clinical practice guidelines were fully consulted under the guide of relevant laws and regulations, and relevant technical documents. Based on the best existing evidence, and combined with the characteristics of TCM and the clinical realities, we developed Clinical practice guidelines for the treatment of sepsis with TCM therapy alone or TCM combined with antibiotics, with full reference to the experts' experience and patients' preferences.

Farnesoid X receptor agonist GW4064 indirectly inhibits HCV entry into cells via down-regulating scavenger receptor class B type I.

Farnesoid X receptor (FXR) agonists play important regulatory roles in bile acid, lipid and glucose metabolism in vitro and in vivo. Thus, FXR agonists exhibit potential therapeutic effects on metabolism-related diseases that are associated with extrahepatic manifestations induced by hepatitis C virus (HCV) infection. This study investigated the effect and mechanism of FXR agonist GW4064 against HCV in vitro to explore the potential application of FXR agonists. Results showed that GW4064 and other FXR agonists have potent antiviral activity against HCV in Huh7.5 cells. GW4064 down-regulated the expression of scavenger receptor class B type I protein via FXR and thereby indirectly inhibited HCV entry into cells, leading to interruption of HCV life cycle. GW4064 also exhibited synergistic anti-HCV effect with known direct-acting antiviral agents (DAAs) used in the clinic and remained sensitive to DAA-resistant HCV mutations. Therefore, FXR agonists are also a kind of antiviral agent, and might be helpful in treatment of HCV-induced hepatic and extrahepatic manifestations.