Angiotensin-converting enzyme 2 improves liver fibrosis in mice by regulating autophagy of hepatic stellate cells

BACKGROUND Liver fibrosis is the common pathological process associated with the occurrence and development of various chronic liver diseases.At present,there is still a lack of effective prevention and treatment methods in clinical practice.Hepatic stellate cell(HSC)plays a key role in liver fibrogenesis.In recent years,the study of liver fibrosis targeting HSC autophagy has become a hot spot in this research field.Angiotensin-converting enzyme 2(ACE2)is a key negative regulator of reninangiotensin system,and its specific molecular mechanism on autophagy and liver fibrosis needs to be further explored.AIM To investigate the effect of ACE2 on hepatic fibrosis in mice by regulating HSC autophagy through the Adenosine monophosphate activates protein kinases(AMPK)/mammalian target of rapamycin(mTOR)pathway.METHODS Overexpression of ACE2 in a mouse liver fibrosis model was induced by injection of liver-specific recombinant adeno-associated virus ACE2 vector(rAAV2/8-ACE2).The degree of liver fibrosis was assessed by histopathological staining and the biomarkers in mouse serum were measured by Luminex multifactor analysis.The number of apoptotic HSCs was assessed by terminal deoxynucleoitidyl transferase-mediated dUTP nick-end labeling(TUNEL)and immunofluorescence staining.Transmission electron microscopy was used to identify the changes in the number of HSC autophagosomes.The effect of ACE2 overexpression on Wu Y et al.ACE2 improves liver fibrosis through autophagy WJG https://www.wjgnet.com 4976 September 7,2023 Volume 29 Issue 33 autophagy-related proteins was evaluated by multicolor immunofluorescence staining.The expression of autophagy-related indicators and AMPK pathway-related proteins was measured by western blotting.RESULTS A mouse model of liver fibrosis was successfully established after 8 wk of intraperitoneal injection of carbon tetrachloride(CCl4).rAAV2/8-ACE2 administration reduced collagen deposition and alleviated the degree of liver fibrosis in mice.The serum levels of platelet-derived growth factor,angiopoietin-2,vascular endothelial growth factor and angiotensin II were decreased,while the levels of interleukin(IL)-10 and angiotensin-(1-7)were increased in the rAAV2/8-ACE2 group.In addition,the expression of alpha-smooth muscle actin,fibronectin,and CD31 was down-regulated in the rAAV2/8-ACE2 group.TUNEL and immunofluorescence staining showed that rAAV2/8-ACE2 injection increased HSC apoptosis.Moreover,rAAV2/8-ACE2 injection notably decreased the number of autophagosomes and the expression of autophagy-related proteins(LC3I,LC3II,Beclin-1),and affected the expression of AMPK pathway-related proteins(AMPK,p-AMPK,p-mTOR).CONCLUSION ACE2 overexpression can inhibit HSC activation and promote cell apoptosis by regulating HSC autophagy through the AMPK/mTOR pathway,thereby alleviating liver fibrosis and hepatic sinusoidal remodeling.

PD-L1、Ki67与非小细胞肺癌患者临床病理特征及预后的关系

目的探讨PD-L1和细胞核相关抗原Ki67与非小细胞肺癌(NSCLC)患者临床病理特征及预后的关系。方法选取2017年11月—2018年11月浙江省立同德医院收治的90例NSCLC患者。HE染色观察肺组织病变,免疫组织化学法检测肺癌组织中PD-L1、Ki67的表达。采用单因素分析和多因素Logistic回归分析NSCLC患者PD-L1、Ki67表达与临床病理特征的关系,生存曲线分析NSCLC患者PD-L1、Ki67表达与预后的关系。结果PD-L1阳性患者62例(68.89%),阴性患者28例(31.11%)。Ki67阳性患者58例(64.44%),阴性患者32例(35.56%)。鳞癌、低分化、淋巴结转移患者PD-L1阳性率较高(P<0.05)。女性、鳞癌、低分化患者Ki67阳性率较高(P<0.05)。Logistic回归分析显示,病理分型、肿瘤分化程度、淋巴结转移是PD-L1高表达的影响因素(P<0.05);性别、病理分型、肿瘤分化程度是Ki67高表达的影响因素(P<0.05)。PD-L1阳性患者2年生存率(48.39%)低于PD-L1阴性患者(78.57%)(P<0.05),PD-L1阳性患者的中位生存时间为42周,低于PD-L1阴性患者的65周(P<0.05)。Ki67阳性患者2年生存率(48.27%)低于Ki67阴性患者(75.00%)(P<0.05),Ki67阳性患者的中位生存时间为41周,低于Ki67阴性患者的66周(P<0.05)。结论NSCLC患者肺癌组织中存在PD-L1和Ki67高表达。NSCLC患者的病理分型、肿瘤分化程度、淋巴结转移与PD-L1高表达有关;NSCLC患者的性别、病理分型、肿瘤分化程度与Ki67高表达有关。PD-L1、Ki67阳性患者预后较差。

慢性阻塞性肺疾病急性加重期患者血清sTREM-1、suPAR、sICAM-1水平及与病情程度的相关性

目的探讨慢性阻塞性肺疾病急性加重期(AECOPD)患者血清可溶性髓细胞触发受体-1(sTREM-1)、可溶性尿激酶型纤溶酶原激活物受体(suPAR)、可溶性细胞间黏附分子-1(sICAM-1)水平及与病情程度的相关性。方法选取2018年1月-2019年8月浙江省立同德医院收治的AECOPD患者68例作为AECOPD组。另取同期该院42例COPD患者作为COPD组,42例健康体检者作为对照组。检测3组一秒用力呼气容积(FEV1),并计算第一秒用力呼气容积占预计值百分比(FEV1%pred)。采用酶联免疫吸附试验检测3组血清sTREM-1、suPAR、sICAM-1水平。分析AECOPD组血清sTREM-1、suPAR、sICAM-1与FEV1%pred的相关性。结果 3组血清sTREM-1、suPAR、sICAM-1水平比较,经方差分析,差异均有统计学意义(P<0.05);COPD组和AECOPD组高于对照组(P<0.05)。AECOPD组高于COPD组(P<0.05)。Pearson相关性分析显示,AECOPD患者血清sTREM-1、suPAR、sICAM-1与FEV1%pred均呈负相关(r=-0.868、-0.911和-0.908,P<0.05)。结论 AECOPD患者血清sTREM-1、suPAR、sICAM-1水平升高,且其水平越高,FEV1%pred越低。sTREM-1、suPAR、sICAM-1可以作为早期诊断AECOPD病情程度的指标。

Olfactory ensheathing cells promote nerve regeneration and functional recovery after facial nerve defects

Olfactory ensheathing cells from the olfactory bulb and olfactory mucosa have been found to increase axonal sprouting and pathfinding and promote the recovery of vibrissae motor performance in facial nerve transection injured rats. However, it is not yet clear whether olfactory ensheathing cells promote the reparation of facial nerve defects in rats. In this study, a collagen sponge and silicone tube neural conduit was implanted into the 6-mm defect of the buccal branch of the facial nerve in adult rats. Olfactory ensheathing cells isolated from the olfactory bulb of newborn Sprague-Dawley rats were injected into the neural conduits connecting the ends of the broken nerves, the morphology and function of the regenerated nerves were compared between the rats implanted with olfactory ensheathing cells with the rats injected with saline. Facial paralysis was assessed. Nerve electrography was used to measure facial nerve-induced action potentials. Visual inspection, anatomical microscopy and hematoxylin-eosin staining were used to assess the histomorphology around the transplanted neural conduit and the morphology of the regenerated nerve. Using fluorogold retrograde tracing, toluidine blue staining and lead uranyl acetate staining, we also measured the number of neurons in the anterior exterior lateral facial nerve motor nucleus, the number of myelinated nerve fibers, and nerve fiber diameter and myelin sheath thickness, respectively. After surgery, olfactory ensheathing cells decreased facial paralysis and the latency of the facial nerve-induced action potentials. There were no differences in the general morphology of the regenerating nerves between the rats implanted with olfactory ensheathing cells and the rats injected with saline. Between-group results showed that olfactory ensheathing cell treatment increased the number of regenerated neurons, improved nerve fiber morphology, and increased the number of myelinated nerve fibers, nerve fiber diameter, and myelin sheath thickness. In conclusion, implantation of olfactory ensheathing cells can promote regeneration and functional recovery after facial nerve damage in rats.

Innuence of portal pressure change on intestinal permeability in patients with portal hypertension

Objective: To investigate intestinal permeability inpatients with portal hypertension and its relationshipwith portal pressure.Methods: Twenty patients with portal hypertensionwere divided into two groups (A, B), 10 patients pergroup. In group A, patients were treated with com-bined transjugular intrahepatic portosystemic shunt(TIPS) and modified Sugiura. In group B, patientswere treated with modified Sugiura only. Intestinalpermeability was assessed before operation, twoweeks after TIPS. and two weeks after modified Sug-iura; 20 healthy control subjects were also assessed.Results: Intestinal permeability was significantlyhigher in the patients than in the control group (P【0. 01). In group A, portal pressure, intestinal per-meability decreased two weeks after TIPS (P【0. 05), and no obvious change was noted two weeksafter modified Sugiura; but they were significantlylower than those before TIPS (P【0. 05). In groupB, intestinal permeability was not different beforeand after operation. Intestinal permeability in groupA was not different from that in group B beforetreatment, but significantly lower after modified Su-giura (P【0. 05). Portal pressure was significantlycorrelated with intestinal permeability (r=0. 627, P【0. 01).Conclusions: This study shows that combined TIPSand modified Sugiura can lower portal pressure andintestinal permeability, and enhance the therapeuticefficacy on portal hypertension.

Correlation between Surface Area Ratio of Medial to Lateral Tibial Plateau and Knee Alignment in Adults

Objective This study aimed to investigate the correlation between the surface area ratio of medial tibial plateau(MTP)to lateral tibial plateau(LTP)and the mechanical tibiofemoral angle(mTFA).Methods Lower limb computed tomography(CT)images were collected at our hospital.Then,the original CT data were analyzed and reconstructed using medical image processing software.The proximal and distal centres of the femur and tibia were marked.The surface areas of MTP and LTP were identified using image processing software.GraphPad Prism 8.0.2 was used to perform the statistical analysis.Results The surface area ratio of MTP to LTP was significantly correlated with the mTFA in all patients(P<0.0001),male group(P<0.0001),female group(P<0.0001),varus group(P<0.0001),and valgus group(P=0.002).Furthermore,the surface area of MTP and LTP was significantly greater in the male group than in the female group(P<0.0001).There was significant difference in the surface area of the MTP between the varus and valgus groups(P<0.0001).Significant difference was also observed in the surface area ratio of MTP to LTP between the varus and valgus groups(P<0.0001).Conclusion The surface area ratio of MTP to LTP was correlated with the mTFA.Within a certain range,the smaller the mTFA,the greater the surface area ratio of MTP to LTP.For patients undergoing total knee arthroplasty,of whom the surface area of the MTP was basically equal to that of the LTP,it is recommended that the osteotomy should be performed in accordance with mechanical alignment standards,and that a symmetrical tibial plateau prosthesis should be used.For patients whose surface area of MTP is significantly greater than that of the LTP,it is recommended that the osteotomy should be performed in accordance with kinematic alignment standards,and that an anatomical tibial plateau prosthesis should be used.

Effects of granulocyte colony-stimulating factor and stem cell factor, alone and in combination, on the biological behaviours of bone marrow mesenchymal stem cells

Aim: The effects of granulocyte colony- stimu-lating factor (G-CSF) and stem cell factor (SCF) on the proliferation and osteogenic differentia-tion capacity of bone marrow mesenchymal stem cells (MSCs) were studied in the experi-ment. Methods: Bone marrow MSCs were col-lected from rabbits successfully, and treated with various concentrations of G-CSF, SCF or a combination of the two. Flow cytometric ana-lyse, MTT test, CFU-F assay, and alkaline phosphatase (ALP) activity measurement were employed. Results: The results of flow cytome-try showed that immunophenotype of the cells were CD29+/CD45-, CD105+/ CD34–, CD90+/ HLADR–. MSCs were shown to constitutively express low levels of c-kit which could be en-hanced by SCF. G-CSF and SCF had an obvious facilitative effect on the proliferation of MSCs in a dose-dependent fashion. In addition, G-CSF and SCF would be effective in reversibly pre-venting their differentiation, as showed by the decrease of ALP activity, leading to self-renewal rather than differentiative cell divisions. The effects of G-CSF were superior to SCF. And cells in the group treated with combination of G-CSF and SCF showed more powerful effects than the groups treated with G-CS, SCF, or none of the two. Conclusion: On the whole, these studies demonstrated that MSCs responsed to G-CSF, SCF, and to G-CSF plus SCF in a manner that suppressed differentiation, and promotes proliferation and self-renewal, and support the view that these factors could act synergistically.

Integrated multi-omics analysis reveals liver metabolic reprogramming by fish iridovirus and antiviral function of alpha-linolenic acid

Iridovirus poses a substantial threat to global aquaculture due to its high mortality rate;however,the molecular mechanisms underpinning its pathogenesis are not well elucidated.Here,a multi-omics approach was applied to groupers infected with Singapore grouper iridovirus(SGIV),focusing on the roles of key metabolites.Results showed that SGIV induced obvious histopathological damage and changes in metabolic enzymes within the liver.Furthermore,SGIV significantly reduced the contents of lipid droplets,triglycerides,cholesterol,and lipoproteins.Metabolomic analysis indicated that the altered metabolites were enriched in 19 pathways,with a notable down-regulation of lipid metabolites such as glycerophosphates and alpha-linolenic acid(ALA),consistent with disturbed lipid homeostasis in the liver.Integration of transcriptomic and metabolomic data revealed that the top enriched pathways were related to cell growth and death and nucleotide,carbohydrate,amino acid,and lipid metabolism,supporting the conclusion that SGIV infection induced liver metabolic reprogramming.Further integrative transcriptomic and proteomic analysis indicated that SGIV infection activated crucial molecular events in a phagosome-immune depression-metabolism dysregulation-necrosis signaling cascade.Of note,integrative multi-omics analysis demonstrated the consumption of ALA and linoleic acid(LA)metabolites,and the accumulation of L-glutamic acid(GA),accompanied by alterations in immune,inflammation,and cell death-related genes.Further experimental data showed that ALA,but not GA,suppressed SGIV replication by activating antioxidant and anti-inflammatory responses in the host.Collectively,these findings provide a comprehensive resource for understanding host response dynamics during fish iridovirus infection and highlight the antiviral potential of ALA in the prevention and treatment of iridoviral diseases.

Neuroendoscopy Combined with Lateral Ventricle Drainage and Urokinase Infusion in the Treatment of 20 Patients with Hypertensive Intraventricular Hemorrhage

Objective:To explore the clinical efficacy of neuroendoscopy combined with lateral ventricle drainage and urokinase infusion for the treatment of hypertensive intraventricular hemorrhage.Methods:A retrospective analysis of 20 patients diagnosed with intraventricular hemorrhage between March 2017 and August 2018 at the First Affiliated Hospital of Medical College,Shihezi University(Shiohezi,China)was performed.While actively treating primary lesions,20 patients with intraventricular hemorrhage were selected and treated using minimally invasive neuroendoscopic removal of intraventricular hematoma combined with lateral ventricular drainage and urokinase perfusion according to each patient’s condition.Results:One day after surgery,head computed tomography examination revealed that>50% of the hematoma in the lateral ventricle was removed in 14 cases,and 20%-50% of the hematoma in the other 6 cases.After follow-up,13 patients exhibited satisfactory recovery,5 died(3 due to lung infection,1 due to intracranial infection,and 1 due to multiple organ failure),and 2 were in a vegetative state.Conclusion:Neuroendoscopy should be used as much as possible in treating intraventricular hemorrhage given its advantages of minimal injury,complete removal of hematoma,high safety,and wide clinical applicability.