INTRODUCTION:Enhanced angiogenesis and impaired vascular integrity facilitate cancer metastasis. There is accumulating evidence that cancer-derived exosomes take a functional role in these processes. In our previous study, we revealed that Golgi Membrane protein 1 (GOLM1) can promote metastasis of Hepatocellular Carcinoma (HCC), and miRNAs can modulate angiogenesis and vascular permeability in HCC. The objective is to reveal that GOLM1 can promote HCC progression in an exosomal miRNA-dependent way. METHODS:Comprehensive bioinformatics analysis and experiments were conducted to associate GOLM1 expression with angiogenesis in HCC. The effect of hepatoma cell-derived exosomes on Human Umbilical Vein Endothelial Cells (HUVEC) was tested. Exosomal miRNA expression was profiled and validated in GOLM1-knockdown HCC cells. Targets of miR-4449 and miR-3651 were predicted with online tools and validated in vitro. Correlation between miR-4449/miR-3651 and microvascular invasion or recurrence in HCC was assessed. RESULTS:GOLM1 correlated with angiogenesis in HCC. HCC cell-derived exosomes can be transferred to endothelial cells, and GOLM1 can regulate exosome-induced angiogenesis and vascular permeability. In vitro experiments showed that GOLM1 knockdown reduced exosomal abundance of miR-4449 and miR-3651, which target KEAP1 and ZO-1, respectively. Elevated miR-4449 and miR-3651 expression were correlated with microvascular invasion and recurrence in HCC patients. DISCUSSION:We demonstrated that GOLM1 can promote HCC progression independent of its role in modulating EGFR/RTK cell-surface recycling, indicating that patients with high GOLM1 expression may benefit more from anti-angiogenic drugs and highlighting the potential of targeting miR-4449 and miR-3651 to prevent angiogenesis and vascular leakiness in HCC. However, in vivo studies are further needed to validate the effect of miR-4449 and miR-3651 inhibitors in compromising angiogenesis and vascular permeability. Besides, a larger validation cohort is indispensable for establishing the correlation between miR-4449/miR-3651 expression and microvascular invasion and tumor recurrence in HCC. CONCLUSIONS:Our findings suggest that, under the control of GOLM1, HCC cell-derived exosomal miR-4449 and miR-3651 increase angiogenesis and vascular permeability by targeting KEAP1 and ZO-1, highlighting the potential of exosomal miRNAs as promising therapeutic targets for HCC.

ABSTRACT This study aimed to investigate the prognostic value of a novel droplet digital polymerase chain reaction (DDPCR) assay in sepsis patients. In this prospective cohort study, univariable and multivariable Cox regressions were used to assess risk factors for 28-day mortality. We also monitored pathogen load together with clinical indicators in a subgroup of the cohort. A total of 107 sepsis patients with positive baseline DDPCR results were included. Detection of poly-microorganisms [adjusted hazard ratio (HR) = 3.19; 95% confidence interval (CI) = 1.34–7.62; P = 0.009], high Charlson Comorbidity Index (CCI) score (adjusted HR = 1.14; 95% CI = 1.01–1.29; P = 0.041), and Sequential Organ Failure Assessment (SOFA) score (adjusted HR = 1.18; 95% CI = 1.05–1.32; P = 0.005) at baseline were independent risk factors for 28-day mortality while initial pathogen load was not associated (adjusted HR = 1.17; 95% CI = 0.82–1.66; P = 0.385). Among 63 patients with serial DDPCR results, an increase in pathogen load at days 6–8 compared to baseline was a risk factor for 28-day mortality ( P = 0.008). Also, pathogen load kinetics were significantly different between day-28 survivors and nonsurvivors ( P = 0.022), with a decline overtime only in survivors and an increase from days 3 and 4 to days 6–8 in nonsurvivors. Using DDPCR technique, we found that poly-microorganisms detected and increased pathogen load a week after sepsis diagnosis were associated with poor prognosis. IMPORTANCE This prospective study was initiated to explore the prognostic implications of a novel multiplex PCR assay in sepsis. Notably, our study was the largest cohort of sepsis with droplet digital polymerase chain reaction pathogen monitoring to date, allowing for a comprehensive evaluation of the prognostic significance of both pathogen species and load. We found that detection of poly-microorganisms was an independent risk factors for 28-day mortality. Also, pathogen load increase 1 week after sepsis diagnosis was a risk factor for 28-day mortality, and differential pathogen load kinetics were identified between day-28 survivors and nonsurvivors. Overall, this study demonstrated that pathogen species and load were highly correlated with sepsis prognosis. Patients exhibiting conditions mentioned above face a more adverse prognosis, suggesting the potential need for an escalation of antimicrobial therapy. Registered at ClinicalTrials.gov ( NCT05190861 ).

Background Liver hepatocellular carcinoma (LIHC) is a solid primary malignancy with poor prognosis. This study discovered key prognostic genes based on T cell exhaustion and used them to develop a prognostic prediction model for LIHC. Methods SingleR's annotations combined with Seurat was used to automatically annotate the single-cell clustering results of the LIHC dataset GSE166635 downloaded from the Gene Expression Omnibus (GEO) database and to identify clusters related to exhausted T cells. Patients were classified using ConsensusClusterPlus package. Next, weighted gene co-expression network analysis (WGCNA) package was employed to distinguish key gene module, based on which least absolute shrinkage and selection operator (Lasso) and multi/univariate cox analysis were performed to construct a RiskScore system. Kaplan-Meier (KM) analysis and receiver operating characteristic curve (ROC) were employed to evaluate the efficacy of the model. To further optimize the risk model, a nomogram capable of predicting immune infiltration and immunotherapy sensitivity in different risk groups was developed. Expressions of genes were measured by quantitative real-time polymerase chain reaction (qRT-PCR), and immunofluorescence and Cell Counting Kit-8 (CCK-8) were performed for analyzing cell functions. Results We obtained 18,413 cells and clustered them into 7 immune and non-immune cell subpopulations. Based on highly variable genes among T cell exhaustion clusters, 3 molecular subtypes (C1, C2 and C3) of LIHC were defined, with C3 subtype showing the highest score of exhausted T cells and a poor prognosis. The Lasso and multivariate cox analysis selected 7 risk genes from the green module, which were closely associated with the C3 subtype. All the patients were divided into low- and high-risk groups based on the medium value of RiskScore, and we found that high-risk patients had higher immune infiltration and immune escape and poorer prognosis. The nomogram exhibited a strong performance for predicting long-term LIHC prognosis. In vitro experiments revealed that the 7 risk genes all had a higher expression in HCC cells, and that both liver HCC cell numbers and cell viability were reduced by knocking down MMP-9. Conclusion We developed a RiskScore model for predicting LIHC prognosis based on the scRNA-seq and RNA-seq data. The RiskScore as an independent prognostic factor could improve the clinical treatment for LIHC patients.

报道1例53岁男性患者，其每日服用15粒辣木籽，共服用20 d余，而后出现肝功能异常，皮肤瘙痒伴有低热，排除其他原因相关的肝病，肝脏病理检查符合药物性肝损伤(DILI)表现，最终诊断为辣木籽相关的DILI。患者入院后经护肝治疗10 d，病情好转出院，随访至今，病情稳定，未再出现肝功能异常。临床医师在面对不明原因肝病患者时应仔细询问用药史，注意个体差异，警惕所有药物都有可能导致DILI。

Background Sepsis is still a major public health concern and a medical emergency due to its high morbidity and mortality. Accurate and timely etiology diagnosis is crucial for sepsis management. As an emerging rapid and sensitive pathogen detection tool, digital droplet PCR (ddPCR) has shown promising potential in rapid identification of pathogens and antimicrobial resistance genes. However, the diagnostic value and clinical impact of ddPCR tests remains to be studied in patients with suspected sepsis. PROGRESS trial is aimed to evaluate the clinical effectiveness of a novel ddPCR assay compared with standard practice. Methods PROGRESS is a multicenter, open-label, pragmatic randomized controlled trial (pRCT) set in ten hospitals, including departments of infectious disease and intensive care units. In this study, a total of 2292 patients with suspected sepsis will be randomly assigned to two arms: the ddPCR group and the control group with a ratio of 3:1. The primary outcome is the diagnostic efficacy, that is, the sensitivity and specificity of the ddPCR assay compared with the synchronous blood culture. Secondary outcomes include the mortality rates and the mean Sequential Organ Failure Assessment (SOFA) score at follow-up time points, the length of stay in the hospital, the time to directed antimicrobial therapy, duration of broad-spectrum antibiotic use, and the EQ-5D-5L score on day 90. Discussion It is the first multicenter pragmatic RCT to explore the diagnostic efficacy and clinical impact of the ddPCR assay in patients with suspected sepsis, taking advantage of both RCT’s ability to establish causality and the feasibility of pragmatic approaches in real-world studies (RWS). This trial will help us to get a comprehensive view of the assay’s capacity for precise diagnosis and treatment of sepsis. It has the potential to monitor the pathogen load change and to guide the antimicrobial therapy, making a beneficial impact on the prognosis of sepsis patients. Trial registration : ClinicalTrial.gov, NCT05190861. Registered January 13, 2022—‘Retrospectively registered’, https://clinicaltrials.gov/ct2/show/NCT05190861 .

Objective:To establish a noninvasive prediction model for liver fibrosis in chronic hepatitis B (CHB) virus infection patients with alanine aminotransferase (ALT) less than upper limit of normal level.Methods:A total of 183 CHB patients with ALT <40 U/L admitted in the First Affiliated of Wenzhou Medical University from January 2014 to September 2017 were enrolled. There were 149 cases of non-marked liver fibrosis (F0-F2) and 34 cases of marked liver fibrosis (F3-F6) according to the Ishak scoring system. The clinical data, liver stiffness measurement (LSM), liver ultrasound imaging, serum biochemical features and hepatitis B virus related indexes of patients were retrospectively analyzed. The independent predictors of liver fibrosis were screened and a prediction model was constructed based on the results of multivariate logistic regression analysis. The receiver operating characteristic (ROC) curve was applied to evaluate the established model and other indicators in predicting liver fibrosis.Results:Multivariate logistic regression analysis showed that LSM ( OR=1.327, 95% CI 1.149-1.531), liver ultrasound scores ( OR=6.610, 95% CI 2.704-16.156) were independent predictors of liver fibrosis. The area under ROC curve(AUC)of the established model (LU) in predicting liver fibrosis was 0.873(95% CI 0.799-0.947), which was significantly greater than that of the ultrasound score, LSM, FIB-4, APRI and GPR (AUC=0.790, 0.804, 0.654, 0.673 and 0.770; Z=3.394, 1.982, 2.077, 3.168 and 2.165, all P<0.05 or <0.01). With the cut-off value of -1.787, the sensitivity and specificity of LU model were 85.3% and 77.2%, respectively. Conclusion:The established model (LU) can effectively predict the liver fibrosis in CHB patients with ALT less than upper limit of normal.

BACKGROUND:Coronavirus disease 2019 (COVID-19) is an emerging viral disease. Here, we report the clinical features, management, and short-term outcomes of COVID-19 patients in Wenzhou, China, an area outside Wuhan.METHODS:Patients admitted to the Infectious Diseases Department of Ruian People's Hospital in Wenzhou, from January 21 to February 7, 2020, were recruited. Medical data on epidemiological history, demographics, clinical characteristics, laboratory tests, chest computerized tomography (CT) examination, treatment, and short-term outcomes were retrospectively reviewed. Blood biochemistry and routine tests were examined using standard methods and automatic machines. CT examination was performed several times during hospitalization as necessary.RESULTS:A total of 67 confirmed COVID-19 cases were diagnosed; 64 (95.4%) were common cases and three (4.5%) were severe cases. The most common symptoms at admission were fever (86.6%), cough (77.6%), productive cough (52.2%), chest distress (17.9%), and sore throat (11.9%), followed by diarrhea (7.4%), headache (7.4%), shortness of breath (6.0%), dizziness (4.5%), muscular soreness (4.5%), and running nose (4.5%). Thirty patients (47.8%) had increased C-reactive protein levels. The CT radiographs at admission showed abnormal findings in 54 (80.6%) patients. The patients were treated mainly by oxygen therapy and antiviral drugs. By March 3, 2020, all 67 patients completely recovered and had negative nucleic acid tests. The patients were discharged from the hospital and transferred to a medical observation isolation center for further observation.CONCLUSION:Cases of COVID-19 in Wenzhou are milder and have a better prognosis, compared to those in Wuhan. Timely and appropriate screening, diagnosis, and treatment are the key to achieve good outcomes.

目的 探讨胆维他片对肝纤维化大鼠肝中抗氧化体系的影响.方法 选取30只大鼠按照随机数字表法分为肝纤维化模型组、胆维他片组和正常对照组,每组10只.除正常对照组外,其余各组均要求制备大鼠肝纤维化模型.正常对照组和肝纤维化模型组不给药,胆维他片组于造模开始给予灌胃给药,其余组给予蒸馏水灌胃,于第12周测定肝中丙二醛(MDA)、抗氧化体系的过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、谷胱甘肽(GSH),同时采用Masson's染色肝组织胶原行纤维半定量评估纤维化的程度.结果 纤维化模型组和胆维他片组CAT、SOD、GSH低于正常对照组,MDA高于正常对照组,差异有统计学意义(<0.05);胆维他片组CAT、SOD、GSH均高于纤维化模型组,MDA低于纤维化模型组,差异有统计学意义(<0.05).纤维化模型组和胆维他片组肝纤维化分期高于正常对照组,差异有统计学意义(<0.05);胆维他片组肝纤维化分期低于纤维化模型组,差异有统计学意义(<0.05).结论 胆维他片能有效降低肝纤维化肝内的脂质过氧化,促进肝内氧化及抗氧化体系的恢复,具有减轻肝纤维化的作用.

目的 探讨聚乙二醇干扰素联合利巴韦林治疗尿毒症合并丙型肝炎病毒(HCV)感染患者的疗效及安全性.方法 收集2005年1月至2015年9月在温州医科大学附属第三医院接受治疗的4例尿毒症合并HCV感染患者,初始接受聚乙二醇干扰素α-2a 135μg治疗,每周1次,皮下注射,并联合利巴韦林200 mg,每天1次.根据血红蛋白(Hb)水平调整利巴韦林剂量.观察治疗前,治疗后4、12、48、72、127、158、167周HCV-RNA,Hb,血小板计数(Plt),肝、肾功能及电解质情况,观察治疗疗效及其安全性.结果 4例患者应用聚乙二醇干扰素联合利巴韦林治疗后,1例4周时、3例12周时均未检测到HCV-RNA,2例随访48周时、1例随访20周时仍检测不到HCV-RNA,1例12周时检测不到HCV-RNA,未进一步随访.4例患者均因出现Hb下降而调整利巴韦林剂量.3例患者出现Plt减少,但可维持耐受.结论 尿毒症合并HCV感染可以应用聚乙二醇干扰素联合利巴韦林抗病毒治疗,疗效较好,需严密监测不良反应.

AIM To investigate the role of Gadd45a in hepatic fibrosis and the transforming growth factor (TGF)-β/Smad signaling pathway. METHODS Wild-type male BALB/c mice were treated with CCl4 to induce a model of chronic liver injury. Hepatic stellate cells (HSCs) were isolated from the liver of BALB/c mice and were treated with small interfering RNAs (siRNAs) targeting Gadd45a or the pcDNA3.1-Gadd45a recombinant plasmid. Cellular α-smooth muscle actin (α-SMA), β-actin, type I collagen, phospho-Smad2, phospho-Smad3, Smad2, Smad3, and Smad4 were detected by Western blots. The mRNA levels of α-SMA, β-actin, and type I collagen were determined by quantitative real-time (qRT)-PCR analyses. Reactive oxygen species production was monitored by flow cytometry using 2,7-dichlorodihydrofluorescein diacetate. Gadd45a, Gadd45b, anti-Gadd45g, type I collagen, and SMA local expression in liver tissue were measured by histologic and immunohistochemical analyses. RESULTS Significant downregulation of Gadd45a, but not Gadd45b or Gadd45g, accompanied by activation of the TGF-β/Smad signaling pathways was detected in fibrotic liver tissues of mice and isolated HSCs with chronic liver injury induced by CCl4 treatment. Overexpression of Gadd45a reduced the expression of extracellular matrix proteins and α-SMA in HSCs, whereas transient knockdown of Gadd45a with siRNA reversed this process. Gadd45a inhibited the activity of a plasminogen activator inhibitor-1 promoter construct and (CAGA)9 MLP-Luc, an artificial Smad3/4-specific reporter, as well as reduced the phosphorylation and nuclear translocation of Smad3. Gadd45a showed protective effects by scavenging reactive oxygen species and upregulating antioxidant enzymes. CONCLUSION Gadd45a may counteract hepatic fibrosis by regulating the activation of HSCs via the inhibition of TGF-β/Smad signaling.

目的探讨复方甘草酸苷注射液（SNMC）治疗妊娠晚期慢性乙肝患者对婴幼儿生长发育的影响。方法选择妊娠晚期慢性乙肝患者160例，分为SNMC组和思美泰注射液（SAM）对照组，每组80例。随访测量婴幼儿出生后6、12和24个月时的身高、体重、头围和胸围；同时采用贝利婴幼儿发展量表的智力发展指数（MDI）和运动发展指数（PDI）对婴幼儿的智力和运动状况进行评估。结果两组婴幼儿在6、12和24个月时的身高、体重、头围和胸围均数比较以及MDI和PDI均数比较，均无统计学差异（均P＞0.05）。结论复方甘草酸苷注射液治疗妊娠晚期慢性乙肝患者，对婴幼儿生长发育无明显影响。

OBJECTIVE To evaluate the efficacy and safety of stronger neo-minophagen C injection in treatment of chronic hepatitis B(CHB) patients with pregnancy and the newborns and its effect on immune response of hepatitis B vaccine(HBVac) and hepatitis B immune globulin(HBIG) in the newborns.METHODS A total of 100 chronic hepatitis B patients with in late pregnancy were collected and randomly divided into two groups: the group A and the group B.The cases in group A were treated with stronger neo-minophagen C injection(the experimental group),while the cases in group B were treated with ademetionine injection(the control group).A course of treatment was four weeks,and the alanine aminatransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(Tbil)were tested regularly.The neonatal weight and length were measured,and the Apgar score was calculated.HBVac 20μg,10μg,10μg were injected into the neonatal hips anterolateral muscle after the birth of 12 hours,a month and six months correspondently,while the HBIG 200 IU were respectively injected into the neonatal hips anterolateral muscle after the birth of 12 hours and a month.At the age of one,the influence of combined immunization in the newborns were assessed by hepatitis B surface antibody assay.RESULTS There was no statistical difference in the maternal age,gestational weeks,and the neonatal weight,length or apgar score between the two groups.The nomalization of ALT was 82.0% in the experimental group versus 60.0% in the control group(P=0.027),the experimental group was better than the control group,and so was the nomalization of AST(88.0% vs 68.0%,P=0.028).The difference in the pre-treatment level of TBil between the two groups was not statistically significant,and so was the level of TBil after the treatment.The positive rate of the hepatitis B surface antibody(HBsAb) was 92.0% of the group A and 94.0% of the group B,the difference was not statically significant.The average anti-HBs titer was(553.9±72.35)mIU/ml in the group A and(537.6±53.54)mIU/ml in the group B,the difference was not statistically significant;the combined immunization of the newborns in the two groups was similar.CONCLUSION The stronger neo-minophagen C injection can improve the liver function of the chronic hepatitis B patients with late pregnancy,without the combined immunization of the newborns being affected.

BACKGROUND:The aim of this study was to compare the effect of combination lamivudine (LAM) and adefovir dipivoxil (ADV) versus entecavir (ETV) monotherapy for naïve HBeAg-positive chronic hepatitis B (CHB) patients.MATERIAL/METHODS:Fifty enrolled patients with CHB were evenly divided into 2 groups: a group treated with of lamivudine (LAM) (100 mg/day) plus adefovir (ADV) (10 mg/day) combination, and a group treated with entecavir (ETV) (0.5 mg/day). Serum levels of ALT, AST, creatinine, bilirubin, HBsAg, HBeAg and HBV viral load, and genotypic resistance were analyzed at 0, 12, 24, 52, and 104 weeks. HBV DNA levels were determined by real-time PCR and HBsAg and HBeAg by chemiluminescence. Serum levels of ALT, AST, creatinine, and bilirubin were measured by an automatic biochemical analyzer. Data analysis was performed with SPSS 12.0 software.RESULTS:There were no significant differences in the virological response (VR) rates between LAM+ADV and ETV cohorts at 24, 52, and 104 weeks (P>0.05). The HBeAg seroconversion rates were 28% and 20%, and the biochemical response (BR) rates were 88% and 84% at week 104 in the LAM+ADV and ETV groups, respectively. The rates of undetectable HBV DNA, HBeAg seroconversion, and ALT normalization rates were similar in both cohorts. No virological breakthrough or serious adverse effects were noted for any patient during the study period.CONCLUSIONS:Both LAM+ADV combination therapy and ETV monotherapy were effective and safe in the treatment of -naïve HBeAg-positive CHB patients. However, further studies are needed to obtain long-term results.

OBJECTIVE To analyze the distribution and drug resistance of the pathogens causing intravascular catheter-related bloodstream infections,so as to direct the treatment of intravascular catheter-related infections.METHODS A total of 35 patients diagnosed as intravascular catheter-related infections were retrospectively analyzed,the distribution and drug resistance of the pathogens were statistically significant.RESULTS A total of 41 strains of pathogens were isolated from 35 patients,gram-positive bacteria accounted for 56.10%,43.90% of which were Staphylococcus,gram-negative bacteria accounted for 39.02%,12.20%of which were Acinetobacter and Klebsiella,fungi accounted for 4.88%;the drug resistance rates of gram-positive bacteria to teicoplanin and vancomycin were 0,the resistance rates to rifampicin and tetracycline were 18.18% and 13.64%;the drug resistance rates of gram-negative bacteria to amikacin,cefoperazone/sulbactam,meropenem and imipenem were 25.00%;no strains of fungi resistant to fluconazole,itraconazole,amphotericin B and 5-fluorocytosine.CONCLUSION It's very important for the treatment of intravascular catheter-related bloodstream infections to learn about the distribution and drug resistance of the pathogens causing intravascular catheter-related bloodstream infections.