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ObjectiveTo investigate the therapeutic effect of Qingjie Huagong decoction (QJHGD) on a mouse model of severe acute pancreatitis (SAP) and the mechanism of action of QJHGD against inflammatory response. MethodsA total of 36 male C57BL/6J mice were randomly divided into blank group, model group, Western medicine group (ulinastatin), and low-, middle-, and high-dose QJHGD groups, with 6 mice in each group. All mice except those in the blank group were given 5% sodium taurocholate by retrograde pancreaticobiliary injection to establish a model of SAP. After modeling, the mice in the low-, middle-, and high-dose groups were given QJHGD (1, 2, and 4 g/kg, respectively) by gavage, and those in the Western medicine group were given intraperitoneal injection of ulinastatin (5×104 U/kg), for 7 days in total. HE staining was used to observe the histopathological changes of the pancreas; ELISA was used to measure the levels of α-amylase, lipase, interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-18 (IL-18), and tumor necrosis factor-α (TNF-α) in mice; RT-qPCR was used to measure the mRNA expression levels of NOD-like receptor protein3 (NLRP3), Toll-like receptor 4 (TLR4), and nuclear factor-kappa B (NF-κB) in pancreatic tissue; immunohistochemistry was used to measure the positive expression rates of NLRP3, TLR4, and NF-κB in pancreatic tissue; Western blot was used to measure the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6. An analysis of variance was used for comparison of continuous data between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group, the model group had diffuse destruction of pancreatic tissue structure, focal dilatation of pancreatic lobular septum, pancreatic acinar atrophy, and massive inflammatory cell infiltration, as well as significant increases in the content of α-amylase, lipase, IL-1β, IL-6, IL-8, IL-18, and TNF-α (all P<0.05), the mRNA expression levels and positive expression rates of NLRP3, TLR4, and NF-κB (all P<0.05), and the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6 (all P<0.05). Compared with the model group, the low-, middle-, and high-dose QJHGD groups and the Western medicine group had slightly tighter and more intact structure of pancreatic tissue, ordered arrangement of pancreatic acinar cells, a small amount of inflammatory cell infiltration, and hemorrhagic foci of pancreatic lobules, as well as significant reductions in the content of α-amylase, lipase, IL-1β, IL-6, IL-8, IL-18, and TNF-α (all P<0.05), the mRNA expression levels and positive expression rates of NLRP3, TLR4, and NF-κB (all P<0.05), and the protein expression levels of NLRP3, TLR4, NF-κB, IL-1β, and IL-6 (all P<0.05). ConclusionQJHGD may exert a protective effect on the pancreatic tissue of SAP mice by inhibiting the activation of NLRP3/TLR4/NF-κB signaling pathway-related proteins, reducing the release of inflammatory mediators, and preventing the enhancement of inflammatory cascade response.
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Acute pancreatitis (AP) is a common clinical acute abdominal disease, which is characterized by acute onset, rapid development, severe disease, many complications, and high mortality rate. It can progress to severe AP (SAP) if not treated promptly in the early stage. The pathogenesis of AP is complex and involves multiple cellular and molecular levels. It is now clear that oxidative stress and reactive oxygen species (ROS) production are involved in the physiopathological process of AP, which is associated with a low quantity and activity of antioxidant enzymes in pancreatic cells. Nuclear factor E2-related factor 2 (Nrf2) serves as the ''golden key'' to maintain redox homeostasis in tissue cells and constitutes an important signaling pathway for antioxidant response and inflammation in vivo by collaborating with downstream antioxidant enzymes such as heme oxygenase-1 (HO-1). Traditional Chinese medicine has unique efficacy in treating diseases due to its multi-component, multi-target, multi-drug delivery, and multi-formulation characteristics. Based on the concept of synergy between traditional Chinese and Western medicine, traditional Chinese medicine is becoming a new craze in the treatment of AP. The level of oxidative stress and Nrf2/HO-1 signaling pathway in AP pancreatic tissue are in a dynamic change process, and the intervention of traditional Chinese medicine can clean ROS production, affect the inflammatory pathway, and reduce oxidative stress damage, so as to protect against pancreatic injury. This suggests that this pathway plays an important role in AP. This article reviews the recent literature on the regulation of the Nrf2/HO-1 signaling pathway by traditional Chinese medicine for AP and summarizes that the monomers of traditional Chinese medicine targeting this pathway are mainly heat-clearing and detoxifying, blood-activating and blood-stasis-removing, and Qi benefiting and middle warming, and the compounds of traditional Chinese medicine include Yinchenhao Decoction and QingYi Ⅱ, so as to provide a new direction for the prevention and treatment of AP and further drug development.
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The multi-campus mode is an important way to give full play to the advantages of public hospitals and promote the expansion of high-quality medical resources and balanced regional layout. The authors summarized the practical experience of the Second Affiliated Hospital Zhejiang University School of Medicine in promoting multi-campus cultural integration, including vertical dimensional initiatives including raising cultural construction to a strategic level, improving the working mechanism of cultural construction, and building a distinctive cultural identity system; horizontal dimensional initiatives including creating equal status and intergroup cooperation conducive to cultural integration, building a variety of forms of the main cultural communication platform, and building a unified and diverse cross-campus communication bridge. Through cultural integration, the internal cohesion of the hospital was enhanced and the influence of the hospital brand was improved. The authors suggested that cultural integration should always be based on the principle of " seeking common ground while preserving minor differences" , focusing on the construction of systems and standards, and focusing on the construction of communication platforms.
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Objective:To explore the application value of ultrasound diaphragmatic function assessment in weaning from invasive mechanical ventilation in patients with respiratory failure.Method:Fifty-three patients with acute respiratory failure who underwent mechanical ventilation and weaning in People’s Hospital Affiliated to Ningbo University from January 2020 to February 2022 were selected for research. After weaning conditions, T-tube was used for spontaneous breathing test (SBT). At SBT-30 min, beside ultrasound was used to collect the right diaphragm movement (DE) and diaphragm thickness, and the shallow rapid breathing index (RSBI), diaphragm shallow rapid breathing index (D-RSBI) and diaphragm thickness change rate (DTF) were calculated. According to the outcome of weaning, the patients were divided into the successfully weaned group and unsuccessfully weaned group. All patients’ clinical data were collected, and the relationship between ultrasound parameters and clinical indexes was compared between the two groups The efficiency of each index for predicting the success rate of weaning was analyzed with the receiver operating characteristic (ROC) curve.Results:Fifty-three patients were enrolled in this study, including 36 patients in the successfully weaned group and 17 patients in the unsuccessfully weaned group. DE and DTF in the successfully weaned group were higher than those in the unsuccessfully weaned group (all P<0.05), RSBI and D-RSBI in the successfully weaned group were significantly lower than those in the unsuccessfully weaned group (all P<0.05). ROC curve analysis showed that when the cut-off value of DE, DTF, RSBI and D-RSBI were >1.21 cm, >27.5%, <48.68 times/(min·L) and <1.31 times/(min·mm), the sensitivity of predicting the success of weaning was 69.0%, 97.2%, 83.0% and 83.0% respectively, and the specificity was 59.0%, 47.1%, 94.1% and 94.3%, respectively. Conclusions:The evaluation of ultrasound diaphragmatic function can effectively guide the clinical mechanical ventilation patients to withdraw the machine, and improve the accuracy of predicting the success rate of weaning. Therefore, t ultrasound diaphragmatic function assessment has a high application value in guiding the mechanical evacuation of patients with mechanical ventilation, and can be widely applied in clinical practice.
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Objective:To investigate the feasibility and effectiveness of incision subcutaneous porous catheter combined with ropivacaine analgesia in enhanced recovery after laparoscopic gastrointestinal surgery.Methods:The clinical data of 140 patients underwent gastrointestinal surgery from August 2021 to April 2022 in Shanghai Fourth People′s Hospital, School of Medicine, Tongji University were retrospectively analyzed. Among them, 70 patients were given routine postoperative analgesia (control group), and 70 patients were given incision subcutaneous porous catheter combined with ropivacaine analgesia on the basis of routine postoperative analgesia (observation group). The visual analogue score (VAS) 4, 24, 32, 48, 56 and 72 h after operation was evaluated; and the complications of subcutaneous catheterization, incision infection, postoperative nausea vomiting, neurological symptoms, time to extubation, patient satisfaction degree, recovery time of intestinal function and hospital stay were recorded.Results:The VAS 4, 24, 32, 48, 56 and 72 h after operation in observation group was significantly lower than that in control group: 1.000 (- 0.250, 2.250) scores vs. 1.000 (- 1.000, 3.000) scores, 2.000 (1.000, 3.000) scores vs. 4.000 (2.000, 6.000) scores, 1.000 (0.000, 2.000) scores vs. 3.000 (1.000, 5.000) scores, 2.000 (1.000, 3.000) scores vs. 3.000 (1.750, 4.250) scores, (1.100 ± 0.934) scores vs. (2.085 ± 0.943) scores and (0.985 ± 0.842) scores vs. (1.814 ± 0.921) scores, and there was statistical difference ( P<0.05 or <0.01). The recovery time of intestinal function and hospital stay in observation group were significantly shorter than that that in control group: (1.743 ± 0.557) d vs. (2.200 ± 0.714) d and (8.043 ± 1.160) d vs. (8.757 ± 1.221) d, and there were statistical difference ( P<0.01); there were no statistical differences in the rate of incision infection, incidence of postoperative nausea vomiting, time to extubation and patient dissatisfaction rate between two groups ( P>0.05); there were no the complications of subcutaneous catheterization and neurological symptoms in two groups. Conclusions:The incision subcutaneous porous catheter combined with ropivacaine analgesia after laparoscopic gastrointestinal surgery is a safe, effective and feasible method. Multimodal analgesia under enhanced recovery after surgery can increase the postoperative recovery after gastrointestinal operations and shorten the postoperative hospital stay.
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Objective:Based on the Ancient and Modern Medical Record Cloud Platform, we aimed to analyze the rules of TCM compound patents for the treatment of acute pancreatitis.Methods:Compound patents for acute pancreatitis were retrieved from the National Patent Database. After the steps of data screening, data entry, and data specification, a database of compound patents treated for acute pancreatitis was established. The frequency analysis, attribute analysis, association analysis, cluster analysis, and complex network analysis were performed by using the Ancient and modern medical record cloud platform.Results:A total of 87 compound patents were obtained, comprising 213 herbs, of which the core drugs were Rhei radix et rhizoma, Bupleuri radix, Aurantii fructus immaturus, Glycyrrhizae radix et rhizoma, Magnoliae officinalis cortex, Corydalis rhizoma, Scutellariae radix, Aucklandiae radix, Natrii sulfas, Coptidis rhizoma. The drugs were mainly warm, cold and slightly cold, and the drugs taste mostly bitter and spicy, and the drugs mainly belonged to the spleen meridian and liver meridian. The cluster analysis results contained 5 categories. The associations of drugs included Bupleuri radix - Rhei radix et rhizoma, Aurantii fructus immaturus - Rhei radix et rhizoma, Magnoliae officinalis cortex - Rhei radix et rhizoma, for which complex network analysis yielded a core composition of Rhei radix et rhizoma, Bupleuri radix, Glycyrrhizae radix et rhizoma, Natrii sulfas, Aurantii fructus immaturus, Corydalis rhizoma, Scutellariae radix, Magnoliae officinalis cortex. Conclusion:The eliminating stasis by purging for acute pancreatitis is dominated by Rhei radix et rhizoma, channeling Fu Qi method is based on Aurantii fructus immaturus and Bupleuri radix, and eliminating stasis by purging combined with channeling Fu Qi methods can be used with Magnoliae officinalis cortex, Natrii sulfas, etc.
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Objective:To analyze the potential mechanism of Qifang Weitong granules in the treatment of gastric cancer based on network pharmacology and molecular docking method.Methods:TCMSP, TCMID, and Swiss Target Prediction databases were used to screen out the chemical components and related targets of Qifang Weitong Granules. GeneCards and OMIM databases were used to screen out the gastric cancer targets to obtain common targets of this disease and Qifang Weitong Granules and upload them to STRING database to form a PPI network, and obtain the key targets and analyze the correlation between the key targets and gastric cancer in Oncomine tumor database. In addition, the regulatory network of gastric cancer and Qifang Weitong Granules was constructed by using Cytoscape software, and the CluoGO plug-in and R language of Cytoscape software were used to perform GO and KEGG enrichment analysis on the key targets. The possibility of the binding between the molecules of this medicine and targeted molecules is verified by molecular docking.Results:There were 168 medicinal chemical components obtained in Qifang Weitong Granules, 2 803 gastric cancer targets, and 49 common targets. In the regulatory network of gastric cancer and Qifang Weitong Granules, β-sitosterol, formononet, stigmasterol have higher values of chemical composition. The key targets in the PPI network are MAPK8, FOS, AR, etc. The GO enrichment analysis focused on the positive regulation of mitochondrial outer membrane permeability in the apoptosis signaling pathway, while the KEGG enrichment analysis is significantly enriched in apoptosis access. The result of molecular docking showed good binding and stable conformation.Conclusion:Qifang Weitong Granules can induce the expression of genes and proteins related to gastric cancer, show its effect by affecting the level of hormones, cell apoptosis and other biological processes, and activating the apoptosis signal pathway.
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Objective:To investigate the clinical predictive value of combined diaphragmatic and pulmonary ultrasound in acute respiratory failure patients with mechanical ventilation (MV).Methods:From January 2020 to August 2022, patients with acute respiratory failure admitted to People's Hospital Affiliated to Ningbo University who underwent invasive MV and weaning were enrolled. After meeting the weaning standards, spontaneous breathing test (SBT) was performed using T-tube. Right diaphragm excursion (DE), diaphragm thickness and lung ultrasound score (LUS) were collected by bedside ultrasound at 30 minutes of SBT, and rapid shallow respiratory index (RSBI), diaphragmatic-shallow respiratory index (D-RSBI) and diaphragmatic thickening rate (DTF) were calculated. According to the weaning outcome, the patients were divided into successful weaning group and failed weaning group. The clinical data of all patients were collected, and the ultrasound parameters and clinical indicators were compared between the two groups. Receiver operator characteristic curve (ROC curve) was used to evaluate the predictive value of D-RSBI, RSBI, DE combined with LUS score and DTF combined with LUS score for weaning failure patients.Results:A total of 77 patients were enrolled, including 54 cases in the successful weaning group and 23 cases in the failed weaning group. The right DE and DTF of patients in successful weaning group were significantly higher than those in failed weaning group [right DE (cm): 1.28±0.39 vs. 0.88±0.41, DTF: (32.64±18.27)% vs. (26.43±15.23)%, both P < 0.05], LUS score, RSBI and D-RSBI were significantly lower than those in failed weaning group [LUS score: 11.45±2.67 vs. 18.33±3.62, RSBI (times·min -1·L -1): 72.21±19.67 vs. 107.35±21.32, D-RSBI (times·min -1·mm -1): 0.97±0.19 vs. 1.78±0.59, all P < 0.05]. ROC curve analysis showed that when the cut-off value of D-RSBI and RSBI was 1.41 times·min -1·mm -1 and 56.46 times·min -1·L -1, the area under the ROC curve (AUC) for predicting weaning failure was 0.972 and 0.988; and the sensitivity was 95.7% and 87.0%, respectively; the specificity was 81.0% and 100.0%, respectively. The AUC of right DE combined with LUS score and DTF combined with LUS score in predicting weaning failure were 0.974 and 0.985, respectively, with a sensitivity of 91.3% and a specificity of 98.1%. Conclusions:Combined assessment of diaphragmatic and pulmonary ultrasound is a good parameter to effectively predict weaning failure in MV patients, which has high application value in guiding weaning in MV patients, and is worthy of clinical application.
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OBJECTIVE To explore the the reg ulation of intestinal flora and effects of Qingjie huagong decoction on intestinal mucosal barrier in severe acute pancreatitis (SAP)mode rats . METHODS SAP rat model was induced by intraperitoneal injection of caerulein and lipopolysaccharide.The survival state of rats in each group were observed.The levels of serum amylase ,interleukin 10(IL-10),IL-18 and IL- 1β in serum were all detected. The pathological changes of pancreatic and small intestinal tissue were observed. The expressions of Occludin,ZO-1 and HMGB1 were detected in small intestinal tissue of rats. The structure and relative abundance of intestinal microflora in rats were detected by 16S rRNA high throughput sequencing. RESULTS After the intervention of Qingjie huagong decoction ,abdominal distension symptoms of SAP model rats were significantly relieved ,and their mental state recovered better ;the levels of serum amylase and IL- 18 in serum were decreased significantly (P<0.05),while the level of IL- 10 was increased significantly (P<0.05). The necrotic area of pancreatic tissue and the infiltration of inflammatory cells were reduced , the degree of intestinal epithelial cell structural disorder was alleviated ,and the shedding of intestinal mucosal epithelium was reduced.The protein expression of HMGB 1 in small intestinal tissue was decreased significantly (P<0.05),and the protein expression of Occludin and ZO- 1 were increased significantly . Results of 16S rRNA high throughput sequencing showed that Qingjie huagong decoction could increased the relative abundance of probiotics such as Bacteroidea and Lactobacillus in rat intestine ,reduced the colonization of harmful bacteria such as Firmicutes. CONCLUSIONS Qingjie huagong decoction can improve the intestinal barrier by up-regulating the expression of Occludin and ZO- 1 in small intestinal tissue and down-regulating the protein expression of HMGB 1. It can also adjust the relative abundances of different flora to protect the intestinal tract.
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Traumatic brain injury (TBI) is characterized by high incidence, high disability rate, and many sequelae. Depression after TBI is the most common sequelae of mental illness, which can affect the prognosis and quality of life of patients. The specific pathogenesis of post-TBI depression is not yet known, and there is no recognized prevention and treatment plan. This review aims to retrospectively summarize the epidemiology, risk factors, clinical manifestations, evaluation, possible mechanisms, and discoveries of treatment of post-TBI depression in the past 5 years, which will help clinicians manage this difficult disease more effectively and provide a reference for future research.
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Objective:To evaluate the clinical efficacy of TCM Qingjie Huagong Decoction combined with routine internal medicine in the treatment of severe acute pancreatitis with cholelithiasis (bile duct stones) in the early stage.Methods:Thirty-two patients with severe acute pancreatitis combined with cholelithiasis in the first affiliated Hospital of GuangXi University of Traditional Chinese Medicine were selected and randomly divided into two groups with 16 in each, both groups were treated for 14 days. Serum amylase (AMS) was detected by iodine-starch colorimetry, GOT and GPT were detected by continuous monitoring method, and CRP, IL-6 and procalcitonin (PCT) were detected by immune transmission turbidimetry. Acute Physiological and Chronic Health Score Ⅱ (APACHE Ⅱ), CT Severity Index Score (CTSI) and Modified Marshall Score were used to evaluate the severity of SAP. The recovery time of body temperature, the relief time of abdominal distension pain, the recovery time of bowel sounds and the total hospital stay were observed and recorded to evaluate the clinical effect.Results:The total effective rate was 93.8% (15/16) in the treatment group and 75.0% (12/16) in the control group. There was significant difference between the two groups ( χ2=8.19, P=0.042). After treatment, the level of AMS, WBC, CRP, PCT, AST, ALT and IL-6 in the treatment group were lower than those in the control group ( t values were 14.3, 7.24, 9.63, 5.48, 7.05, 7.33, 28.34, respectively, all Ps<0.05); After treatment, the time for body temperature to return to normal [(2.91±0.12)d vs. (3.78±0.38)d, t=8.76], the time for relief of abdominal distension pain [(4.77±0.68)d vs. (7.13±1.55)d, t=9.52], the time for recovery of bowel sounds [(3.90±1.80)d vs. (4.89±1.38)d, t=2.98] and the total hospital stay [(22.60±2.80)d vs. (30.37±3.89)d, t=7.88] in the treatment group were all significantly shorter than those in the control group ( P<0.01); APACHE Ⅱ, CTSI and the Modified Marshall Score in the treatment group were lower than those in the control group ( t values were 11.82, 12.72, 7.71, respectively, all Ps<0.01). Conclusion:Qingjie Huagong Decoction combined with ERCP and conventional western medicine therapy can reduce the level of inflammation in patients with cholelithiasis in the early stage of SAP, relieve clinical symptoms and improve clinical efficacy.
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Objective:To investigate the expression and diagnostic value of microRNA-335-5p (miR-335-5p) and Human Vascular endothelial growth factor receptor 1 (VEGFR-1, FLT1) in serum exosomal bodies of patients with triple negative breast cancer (TNBC) .Methods:Gene Expression Omnibus database (GEO) analysis was performed and differentially expressed miRNA in breast cancer tissues was screened. From Jan. 2016 to Nov. 2017, the peripheral blood samples of 56 TNBC patients in People’s Hospital Affiliated to Ningbo University were collected, and the exosomes were isolated and identified. FLT1 was selected as the target gene of miR-335-5p by using bioinformatics analysis. Expression levels of miR-335-5p and FLT1 in serum exosome were detected by qRT-PCR. The relationship between miR-335-5p and clinicopathological parameters of TNBC patients was analyzed. The diagnostic value of miR-335-5p was evaluated by receiver operating characteristic (ROC) , and the survival prognosis of miR-335-5p and FLT1 was analyzed by Kaplan Meier survival curve.Results:The expression of miR-335-5p in the serum exosome of TNBC patients was lower than that of the control group, while the expression of FLT1 in the serum exosome was higher than that of the control group ( P<0.05) . The expression of miR-335-5p was related to tissue grade ( χ2=22.02, P<0.000 1) , degree of differentiation ( χ2=20.67, P<0.000 1) and lymph node metastasis ( χ2=4.667, P=0.030 8) in patients with TNBC ( P<0.05) . The area under ROC diagnosed by miR-335-5p was 0.809 8 (95% CI: 0.726 3-0.893 2, P<0.000 1) . Kaplan-Meier survival analysis showed that the overall survival time of patients with low expression of miR-335-5p was significantly shorter than that of patients with high expression of miR-335-5p ( P=0.004 5) . The high expression of its target gene FLT1 was associated with low survival rate ( P=0.048 0) . Conclusion:Serum exosomal miR-335-5p can be used as an important index to predict the prognosis of TNBF and is expected to play a role in diagnosis and treatment of TNBC.
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Background@#Arctic-like (AL) lineages of rabies viruses (RABVs) remains endemic in some Arctic and Asia countries. However, their evolutionary dynamics are largely unappreciated. @*Objectives@#We attempted to estimate the evolutionary history, geographic origin and spread of the Arctic-related RABVs. @*Methods@#Full length or partial sequences of the N and G genes were used to infer the evolutionary aspects of AL RABVs by Bayesian evolutionary analysis. @*Results@#The most recent common ancestor (tMRCA) of the current Arctic and AL RABVs emerged in the 1830s and evolved independently after diversification. Population demographic analysis indicated that the viruses experienced gradual growth followed by a sudden decrease in its population size from the mid-1980s to approximately 2000.Genetic flow patterns among the regions reveal a high geographic correlation in AL RABVs transmission. Discrete phylogeography suggests that the geographic origin of the AL RABVs was in east Russia in approximately the 1830s. The ancestral AL RABV then diversified and immigrated to the countries in Northeast Asia, while the viruses in South Asia were dispersed to the neighboring regions from India. The N and G genes of RABVs in both clades sustained high levels of purifying selection, and the positive selection sites were mainly found on the C-terminus of the G gene. @*Conclusions@#The current AL RABVs circulating in South and North Asia evolved and dispersed independently.
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Real time fluorescent quantitative PCR (RT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. However, due to the low viral load in patient throats and the limitations of RT-PCR, significant numbers of false negative reports are inevitable, which results in failure to timely diagnose, early treat, cut off transmission, and assess discharge criteria. To improve this situation, an optimized droplet digital PCR (ddPCR) was used for detection of SARS-CoV-2, which showed that the limit of detection of ddPCR is significantly lower than that of RT-PCR. We further explored the feasibility of ddPCR to detect SARS-CoV-2 nucleic acid from 77 clinical throat swab samples, including 63 suspected outpatients with fever and 14 supposed convalescents who were about to discharge after treatment, and compared with RT-PCR in terms of the diagnostic accuracy. In this double-blind study, we tested, surveyed subsequently and statistically analyzed 77 clinical samples. According to our study, 26 samples from COVID-19 patients with RT-PCR negative were detected as positive by ddPCR. No FPRs of RT-PCR and ddPCR were observed. The sensitivity, specificity, PPV, NPV, NLR and accuracy were improved from 40% (95% CI: 27-55%), 100% (95% CI: 54-100%), 100%, 16% (95% CI: 13-19%), 0.6 (95% CI: 0.48-0.75) and 47% (95% CI: 33-60%) for RT-PCR to 94% (95% CI: 83-99%), 100% (95% CI: 48-100%), 100%, 63% (95% CI: 36-83%), 0.06 (95% CI: 0.02-0.18) and 95% (95% CI: 84-99%) for ddPCR, respectively. Moreover, 14 (42.9 %) convalescents still carry detectable SARS-CoV-2 after discharge. Overall, ddPCR shows superiority for clinical diagnosis of SARS-CoV-2 to reduce the false negative reports, which could be a powerful complement to the current standard RT-PCR. It also suggests that the current clinical practice that the convalescent after discharge continues to be quarantined for at least 2 weeks is completely necessary which can prevent potential viral transmission.
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Background@#High concentrations of particulate matter less than 2.5 μm in diameter (PM2.5) in poultry houses is an important cause of respiratory disease in animals and humans. Pseudomonas aeruginosa is an opportunistic pathogen that can induce severe respiratory disease in animals under stress or with abnormal immune functions. When excessively high concentrations of PM2.5 in poultry houses damage the respiratory system and impair host immunity, secondary infections with P. aeruginosa can occur and produce a more intense inflammatory response, resulting in more severe lung injury. @*Objectives@#In this study, we focused on the synergistic induction of inflammatory injury in the respiratory system and the related molecular mechanisms induced by PM2.5 and P. aeruginosa in poultry houses. @*Methods@#High-throughput 16S rDNA sequence analysis was used for characterizing the bacterial diversity and relative abundance of the PM2.5 samples, and the effects of PM2.5 and P. aeruginosa stimulation on inflammation were detected by in vitro and in vivo. @*Results@#Sequencing results indicated that the PM2.5 in poultry houses contained a high abundance of potentially pathogenic genera, such as Pseudomonas (2.94%). The lung tissues of mice had more significant pathological damage when co-stimulated by PM2.5 and P. aeruginosa, and it can increase the expression levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-α through nuclear factor (NF)-κB pathway in vivo and in vitro. @*Conclusions@#The results confirmed that poultry house PM2.5 in combination with P. aeruginosa could aggravate the inflammatory response and cause more severe respiratory system injuries through a process closely related to the activation of the NF-κB pathway.
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Objective:To evaluate the value of lung ultrasound (LUS) in the early assessment of patients with acute respiratory distress syndrome (ARDS).Methods:A prospective double-blind cohort study was conducted. Patients with ARDS conformed to the Berlin diagnosis criteria admitted to the Intensive Care Unit (ICU) of Ningbo Yinzhou People’s Hospital from July 2016 to January 2020. According to the oxygenation index (OI), the patients were divided into the mild to moderate group (100 mmHg<OI≤300 mmHg) and the severe group (OI≤100 mmHg); Patients underwent LUS and transpulmonary thermodilution technique (TPTD) monitoring and chest CT on the first day after diagnosis. Acute physiology and chronic health status (APACHEⅡ) score, lung injury score (LIS), and PaO 2/FiO 2 (OI) were recorded at admission. LUS score, EVLWI, OI, APACHEⅡ score and LIS were compared. The correlation between LUS and EVLWI, OI, APACHEⅡ score and LIS score was measured by Pearson linear correlation anaysis. Receiver operating characteristic (ROC) curve were used to evaluate LUS score in predicting ARDS. Results:A total of 52 patients with ARDS were enrolled, 34 patients in the mild to moderate group and 18 patients in the severe group. Compared with chest CT scans, the accuracy of LUS diagnosis was 93.12%, the sensitivity was 91.33%, the specificity was 95.31%, the positive predictive value was 95.95%, and the negative predictive value was 90.03%. Pearson linear correlation analysis showed that LUS was positively correlatied with EVLWI ( r=0.756, P<0.01), LIS score ( r=0.817, P<0.01), and APACHEⅡ score ( r=0.655, P<0.01), while was negatively correlated with OI ( r=-0.823, P<0.01). The areas under the receiver operating characteristic curves of LUS and EVLWI measured by TPTD were 0.922 ( P<0.01) and 0.972 ( P<0.01), LUS score threshold value of 19.5 had the sensitivity of 0.833 and specificity of 0.791 for prediction of severe ARDS. Conclusions:LUS is convenient and easy to perform, and LUS score has a high value, which is a better prognostic indicator for early assessment of ARDS patients.
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Objective:To evaluate the value of lung ultrasonography score (LUS) on assessing extravascular lung water (EVLW) and prognosis in patients with acute respiratory distress syndrome (ARDS).Methods:The clinical data of 46 patients meeting ARDS Berlin definition admitted to intensive care unit (ICU) of Ningbo Yinzhou People's Hospital from July 2016 to December 2019 were retrospectively collected. The general data, vital signs, blood lactic acid (Lac), oxygenation index (OI), LUS, extravascular lung water index (EVLWI), sequential organ failure assessment (SOFA) score, clinical pulmonary infection score (CPIS) and the length of ICU stay were collected. According to the prognosis of patients during ICU treatment, the patients were divided into survival group and non-survival group, and the clinical characteristics between the two groups were compared. The correlation between LUS and OI, EVLWI, SOFA, and CPIS were analyzed by Pearson correlation analysis. Receiver operator characteristic (ROC) curve was plotted to determine the prognostic value of LUS for ARDS patients during ICU treatment.Results:Forty-six patients were enrolled in the analysis, of whom 32 patients survived (69.6%), and 14 patients died (30.4%) during ICU treatment. There was no significant difference in gender, age, left ventricular ejection fraction (LVEF) or heart rate (HR) between the two groups. Compared with the survival group, the mean arterial pressure (MAP) and OI in the non-survival group were significantly lowered [MAP (mmHg, 1 mmHg = 0.133 kPa): 57.48±33.34 vs. 85.45±19.56, OI (mmHg): 74.50±18.40 vs. 233.06±28.28, both P < 0.05], while Lac, LUS, EVLWI, SOFA and CPIS were significantly increased [Lac (mmol/L): 6.78±2.56 vs. 2.21±1.42, LUS score: 23.57±2.03 vs. 15.58±2.24, EVLWI (mL/kg): 22.93±2.56 vs. 12.96±2.18, SOFA score: 20.21±3.35 vs. 12.43±2.97, CPIS score: 8.07±1.38 vs. 4.59±1.04, all P < 0.01], and the length of ICU stay was significantly shortened (days: 9.33±3.28 vs. 16.89±4.12, P < 0.05]. Pearson correlation analysis showed that a significant negative linear correlation was found between LUS and OI ( r = -0.823, P < 0.01), and positive linear correlations were found between LUS and EVLWI, SOFA, CPIS ( r values were 0.745, 0.614, 0.757, respectively, all P < 0.01). ROC curve analysis showed that both LUS and EVLWI could predict the prognosis of ARDS patients during ICU treatment, and the areas under ROC curve (AUC) of LUS and EVLWI were 0.936 and 0.991, respectively. When the cut-off of LUS score was 20.5, the sensitivity and specificity were 85.7% and 81.2% respectively. Conclusions:LUS score has a good correlation with EVLWI monitored by pulse index continuous cardiac output (PiCCO), which can reflect lung water content. LUS score can be used as an early prognostic indicator for ARDS patients.
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Objective:To evaluate the role of O-linked-β-N-acetylglucosamine (O-GlcNAc) transferase (OGT) in methane-induced alleviation of oxygen-glucose deprivation and restoration (OGD/R) injury to rat spinal cord neurons and the relationship with Nrf2 expression.Methods:The primarily cultured spinal cord neurons of rats were seeded in 6-well plates at a density of 1×10 5 cells/ml and divided into 6 groups ( n=60 each) using a random number table method: control group (group C), group OGD/R, methane group (group M), and methane plus OGT inhibitor alloxan group (group MA). The medium was replaced with glucose- and serum-free Earle′s salt solution, and the neurons were exposed to 37 ℃ and 5%CO 2-95%N 2 in an incubator for 2 h followed by routine culture to establish the model of OGD/R.In group M, 200 μl methane-saturated saline (final concentration of methane 1.8 mmol/L) was added at oxygen-glucose restoration.Alloxan 8 mmol/L was added at 10 min after oxygen-glucose restoration to inhibit OGT in MA group.At 12 h of oxygen-glucose restoration, the neuronal survival rate, leakage rate of lactic dehydrogenase (LDH) and apoptotic rate of neurons were measured.The expression of OGT and H3K4me3 in Nrf2 promoter was measured by chromatin immunoprecipitation and fluorescent quantitative real-time polymerase chain reaction.Nucleoprotein was extracted for determination of O-GlcNAc glycosylation of RBBP5 (a H3K4 methyltransferase subunit) by immunoprecipitation and Western blot.The spatial proximity of MLL1 subunit of MLL complex to histone H3 was detected by proximity ligation assay.The expression of Nrf2 protein and mRNA was detected by Western blot and quantitative real-time polymerase chain reaction, respectively.The activities of superoxide dismutase (SOD) and catalase (CAT) and malonaldehyde (MDA) concentration were measured by enzyme-linked immunosorbent assay. Results:Compared with group C, the survival rate of neurons was significantly decreased, and the leakage rate of LDH, apoptotic rate and MDA content in supernatant were increased in OGD/R and M groups ( P<0.01). Compared with OGD/R, the survival rate of neurons was significantly increased, the leakage rate of LDH, apoptotic rate and MDA content in supernatant were decreased, the expression of OGT and H3K4me3 in Nrf2 promoter was up-regulated, O-GlcNAc glycosylation and spatial proximity level of MLL1 to histone H3 were increased, the expression of Nrf2 protein and mRNA was up-regulated, and the activities of SOD and CAT were increased in group M ( P<0.01). Compared with group M, the survival rate of neurons was significantly decreased, the leakage rate of LDH, apoptotic rate and MDA concentration were increased, the expression of OGT and H3K4me3 in Nrf2 promoter was down-regulated, O-GlcNAc glycosylation and spatial proximity level of MLL1 to histone H3 were decreased, the expression of Nrf2 protein and mRNA was down-regulated, and activities of SOD and CAT were decreased in group MA ( P<0.05 or 0.01). Conclusion:OGT is involved in methane-induced alleviation of OGD/R injury to rat spinal cord neurons, which is related to up-regulating Nrf2 expression.
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Objective:To evaluate the role of reactive oxygen species (ROS) /nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway in desflurane preconditioning-induced reduction of lipopolysaccharide (LPS)-induced acute cerebral inflammation in rats.Methods:One hundred and twenty clean-grade healthy male Sprague-Dawley rats, aged 8-10 weeks, weighing 200-250 g, were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), LPS-induced acute cerebral inflammation group (group L), desflurane preconditioning group (group D), and N-acetylcysteine plus desflurane preconditioning group (group ND). LPS 1 mg/kg was injected via the tail vein to establish the model of acute cerebral inflammation in L, D and ND groups.In group D, rats inhaled 8.2% desflurane for 1 h once a day for 5 consecutive days, and LPS was intravenously injected at 24 h after the end of the 5th inhalation.In group ND, ROS scavenger N-acetylcysteine 150 mg/kg was intraperitonealy injected at 30 min before each inhalation of desflurane, and the other treatments were similar to those previously described in group D. The expression of Nrf2 and kelch-like ECH-associated protein 1 (Keap1) in microglia and astrocytes in the hippocampal CA1 region was determined by immunofluorescence double staining before LPS injection (at 24 h after the last desflurane preconditioning). Morris water maze test was used to measure the escape latency, space exploration time spent in the original platform quadrant, and frequency of crossing the original platform at 6, 12 and 24 h after injecting LPS.The expression of NOD-like receptor family pyrin domain containing 3 (NLRP3), apoptosis-associated speck-like protein containing CARD (ASC), caspase-1, interleukin-18 (IL-18) and interleukin-1beta (IL-1β) in microglia and astrocytes in the hippocampal CA1 region was determined by immunofluorescence double staining at 6, 12 and 24 h after injecting LPS.Nissl staining and immunofluorescence were used to count normal neurons and activated microglia and astrocytes in the hippocampus at 24 h after LPS injection. Results:Compared with group C, no significant change was found in the expression of Nrf2 and Keap1 in microglia and astrocytes before LPS injection ( P>0.05), and the escape latency was significantly prolonged, the space exploration time spent in the original platform quadrant was shortened, the frequency of crossing the original platform was decreased, the number of normal neurons was decreased, the number of activated microglia and astrocytes was increased, and the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was up-regulated after injecting LPS in group L ( P<0.05). Compared with group L, the expression of Nrf2 was significantly up-regulated, and the expression of Keap1 was down-regulated before injecting LPS ( P<0.05), and the escape latency was shortened, the space exploration time spent in the original platform quadrant was prolonged, the frequency of crossing the original platform was increased, the number of normal neurons was increased, the number of activated microglia and astrocytes was reduced, and the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was down-regulated after injecting LPS in group D( P<0.05). Compared with group D, the expression of Nrf2 was significantly down-regulated, the expression of Keap1 was up-regulated before injecting LPS ( P<0.05), and the escape latency was significantly prolonged, the space exploration time spent in the original platform quadrant was shortened, the frequency of crossing the original platform was decreased, the expression of NLRP3, ASC, caspase-1, IL-18 and IL-1β was up-regulated, the number of normal neurons was reduced, and the number of activated microglia and astrocytes was increased after LPS injection in group ND ( P<0.05). Conclusion:ROS/Nrf2 signaling pathway is involved in desflurane preconditioning-induced reduction of LPS-induced acute cerebral inflammation in rats.
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Objective To evaluate the role of TET3-induced DNA demethylation in methane-in-duced up-regulation of nuclear factor-erythroid 2-related factor 2 ( Nrf2) expression in rat spinal cord neu-rons subjected to oxygen-glucose deprivation and restoration ( OGD∕R) injury. Methods The primarily cultured spinal cord neurons of rats were seeded in 6-well plates at a density of 1×105 cells∕ml and divided into 5 groups ( n=48 each) using a random number table method: control group ( group C) , group OGD∕R, methane group (group M), methane plus TET3-siRNA group (group M+siTET3) and methane plus negative siRNA group (group M+siCon). The medium was replaced with glucose- and serum-free Earle's salt solution, and the neurons were exposed to 5% CO2-95%N2 in an incubator for 2 h followed by routine culture to establish the model of OGD∕R. In group M, 200μl methane-saturated saline ( final concentration of methane 1. 8 mmol∕L) was added at oxygen-glucose restoration. TET3-siRNA 100 pmol∕L and negative siRNA 100 pmol∕L were added at 24 h before oxygen-glucose restoration to perform transfection in M+siTET3 and M+siCon groups, respectively. At 12 h of oxygen-glucose restoration, the neuronal survival rate, release rate of lactic dehydrogenase ( LDH) and apoptotic rate of neurons were measured, and the ex-pression of TET3 and Nrf2 protein and mRNA was detected by Western blot and fluorescent quantitative re-al-time polymerase chain reaction, respectively, and contents of superoxide dismutase (SOD), catalase ( CAT) and malonaldehyde ( MDA) were measured by enzyme-linked immunosorbent assay. Neuronal DNA was extracted for determination of methylation and hydroxymethylation rates of DNA ( by enzyme-linked im-munosorbent assay) and methylation of CpG island in Nrf2 gene promoter ( by fluorescent real-time methyla-tion specific polymerase chain reaction). Results Compared with group C, the survival rate of neurons was significantly decreased, the release rate of LDH and apoptotic rate were increased in group OGD∕R ( P<0. 01) . Compared with OGD∕R, the survival rate of neurons was significantly increased, the release rate of LDH and apoptotic rate were decreased, the expression of TET3 and Nrf2 protein and mRNA was up-regula-ted, DNA hydroxymethylation rate and contents of SOD and CAT were increased, and the DNA and Nrf2 promoter methylation rates and MDA content were decreased in group M ( P<0. 05 or 0. 01) . Compared with group M, the neuronal survival rate was significantly decreased, the release rate of LDH and apoptotic rate were increased, the expression of TET3 and Nrf2 protein and mRNA was down-regulated, the DNA hydroxymethylation rate and contents of SOD and CAT were decreased, and the DNA and Nrf2 promoter methylation rates and MDA content were increased in group M+siTET3 ( P<0. 05 or 0. 01) , and no signifi-cant change was found in the parameters mentioned above in group M+siCon ( P>0. 05) . Conclusion The mechanism by which methane up-regulates Nrf2 expression in rat spinal cord neurons subjected to OGD∕R injury is related to activating TET3 and promoting DNA demethylation in Nrf2 promoter.