Table of Content

Volume 25 Issue 11
26 November 2020

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Effects of antimalarial drugs on mitochondrial respiration in erythrocytic stages of Plasmodium falciparum 3D7

MA Ji, CUI Zhao, WANG Huajing, LI Shuo, QIN Tingting, LI Canghai, JIANG Tingliang

2020, 25(11):  1201-1213.  doi:10.12092/j.issn.1009-2501.2020.11.001

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AIM: The Seahorse XFe96 analyzer was used to evaluate the effects of thirteen types of international first-line antimalarial drugs in six categories on the mitochondrial electron transport chain (ETC) of Plasmodium falciparum  3D7 (P. falciparum 3D7). METHODS: The antimalarial activity of in vitro drugs acting on P. falciparum 3D7 was evaluated using the three-day inhibition method and SYBR Green I fluorescence analysis method. MACS technology was used to separate and purify P. falciparum 3D7. The mitochondrial oxygen consumption rate (OCR) of Seahorse XF analysis system was used to characterize the bioenergy of P. falciparum 3D7 mitochondria at different times to investigate the effects of antimalarial drugs on mitochondrial aerobic respiration of Plasmodium falciparum. RESULTS: The results of flow cytometry showed that the Plasmodium of trophozoite stages was enriched successfully. The results of in vitro antimalarial activity evaluation showed that, except for the antimalarial drug proguanil (Pro), the other twelve antimalarial drugs were all of the nmol/L level against P. falciparum 3D7. The results of the mitochondrial aerobic respiration showed that the five concentrations of dihydroartemisinin (DHA) and chloroquine (CQ) (0.4, 1, 5, 10, 50×IC50) on P. falciparum 3D7 mitochondria aerobic respiration had no significant effect. At the concentration of 5×IC50 for thirteen antimalarial drugs, Artemisinin (ART), arteether (ARE) and artemether (ARM) can significantly increase mitochondrial maximum respiration, Quinine (QN) and Pro only had a decoupling effect on the oxidative phosphorylation of mitochondrial ETC of P. falciparum 3D7, which did not completely destroy the function of mitochondrial ETC. Atovaquone (Ato) had a significant inhibitory effect on P. falciparum 3D7 mitochondrial aerobic respiration, significantly decrease mitochondrial maximum respiration and proton leak, completely destroying the function of mitochondrial ETC, and the remaining ten antimalarial drugs had no significant effect on P. falciparum 3D7 mitochondrial aerobic respiration. CONCLUSION: The target of most antimalarial drugs in thirteen antimalarial drugs is not the ETC pathway of Plasmodium mitochondrial aerobic respiration.

Inhibition of PI3K/AKT signaling pathway by sesamin and vitamin E improves left ventricular fibrosis in spontaneously hypertensive rats

ZHANG Junxiu, SHEN Yuanyuan, YANG Jieren, ZHENG Shuguo, HAO Wei

2020, 25(11):  1214-1222.  doi:10.12092/j.issn.1009-2501.2020.11.002

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AIM: Sesamin (Ses) is one of the major lignans in sesame seeds with antihypertensive and antifibroticactivities, but its effect is weak. In this experiment, we combined vitamin E (VitE) and Ses to observe their effects on blood pressure and left ventricular fibrosis in spontaneously hypertensive rats (SHRs), and to explore its possible mechanism.  METHODS: Male SHRs were randomly divided into SHRs+Ses+VitE group, SHRs+Ses group, SHRs+VitE group, and SHRs model group. Wistar-Kyoto (WKY) rats were setted as a normal control group. After 10 weeks of administration, systolic blood pressure (SBP) of rats was measured by tail cuff method, and the left ventricle was taken to calculate the organ index. Collagen deposition was observed by Masson staining, and cardiac collagen volume fraction (CVF) was analyzed. Expressions of collagen I, collagen III, transforming growth factor-β1 (TGF-β1), connective tissue growth factor (CTGF), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase inhibitor-1 (TIMP-1), phosphorylated protein kinase B (p-AKT), protein kinase B (AKT) and phosphoinositol 3 kinase (PI3K) were observed by Western blot. RESULTS: SBP, HW/BW, LVW/HW and CVF in SHRs+Ses+VitE group decreased significantly, and the decrease was higher than that in SHRs+Ses and SHRs+VitE group (P<0.01 or P<0.05). The expression of collagen I, collagen III, TGF-1, CTGF, MMP-9, TIMP-1, p-AKT and PI3K in SHRs+Ses+VitE group decreased significantly, and the decrease was higher than that in SHRs+Ses and SHRs+VitE group (P<0.01 or P<0.05). CONCLUSION: Ses combined with VitE can reduce the expression of TGF-1, CTGF, MMP-9 and TIMP-1 by inhibiting the activation of AKT/PI3K signaling pathway, thus exerting the effect of lowering blood pressure and improving left ventricular fibrosis.

Study on the mechanism of miR-29a/HMGB1 signaling pathway on H9C2 cardiomyocyte fibrosis induced by high glucose and high fat

LIN Xiaoxin, WANG Zhenhua

2020, 25(11):  1223-1232.  doi:10.12092/j.issn.1009-2501.2020.11.003

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AIM: To investigate the role of miR-29a/HMGB1 signaling pathway in fibrosis H9C2 cells induced by HGHL.  METHODS: DMEM medium containing glucose (33 mmol/L) and palmitate (500 μmol/L) was used to intervene in H9C2 cells for 24 h for subsequent experiments. There were 8 experimental groups, namely NC group, HGHL group, miR-NC group, mimics group, inhibitor group, pc-HMGB1 group, si-HMGB1 group, and miR-29a mimics+pc-HMGB1 group. Flow cytometry was used to detect the apoptosis rate of H9C2 cells in each group. The Western blot experiment detected the expression of TGF-β1, CTGF, MMP-9, PPARγ, and HMGB1 in H9C2 cells of each group. RT-qPCR detected the expression levels of miR-29a, TGF-β1, CTGF, MMP-9, PPARγ, HMGB1 mRNA in each group of cells. The scratch test was used to detect the migration ability of H9C2 cells in each group. RESULTS: After HGHL intervention, the apoptosis rate of H9C2 cells was significantly increased (P<0.05), and the cell migration ability was significantly enhanced (P<0.05). The expression level of TGF-β1, CTGF, and MMP-9 mRNA in cells increased significantly (P<0.05), but the expression level of PPARγ mRNA decreased significantly (P<0.05), and the expression of corresponding proteins also changed with the changes in mRNA (P<0.05). Besides, the expression level of miR-29a in H9C2 cells was also significantly reduced (P<0.05). After the transfection of miR-29a mimics, the increase in apoptosis rate of H9C2 cells caused by HGHL intervention was significantly inhibited (P<0.05), and the cell migration ability was also significantly inhibited (P<0.05). Compared with the HGHL group, TGF-β1, CTGF, and MMP-9 protein expression and mRNA expression levels in H9C2 cells were significantly lower (P<0.05), and PPARγ protein expression and mRNA expression levels were significantly increased (P<0.05). Transfection of miR-29a inhibitor promoted the fibrosis process of H9C2 cells induced by HGHL. miR-29a negatively regulated the expression of HMGB1 protein and its mRNA in H9C2 cells. The results of dual-luciferase reporter gene experiments showed that HMGB1 was a downstream target gene of miR-29a. Transfection of si-HMGB1 and miR-29a mimics had similar effects on H9C2 cell fibrosis induced by HGHL. Simultaneous transfection of miR-29a mimics and pc-HMGB1 had no significant effect on H9C2 cardiomyocyte fibrosis induced by HGHL. CONCLUSION: HGHL intervention can significantly increase the apoptosis rate of H9C2 cells, enhance their migration ability, and the process of fibrosis. At the same time, HGHL intervention can significantly down-regulate the expression level of miR-29a in cells, miR-29a negatively regulates the expression of HMGB1 in cells and then affects HGHL-induced H9C2 cell fibrosis.

Effects of telmisartan on intestinal flora and its metabolite TMAO in atherosclerosis

LI Tianxiang, LI Sujuan, HAO Xiangyu, ZHU Zhibo, GUO Jianqiang

2020, 25(11):  1233-1241.  doi:10.12092/j.issn.1009-2501.2020.11.004

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AIM: To investigate the effect of telmisartan on intestinal flora and metabolite TMAO in atherosclerosis. METHODS: Seventeen ApoE-/-mice were randomly divided into two groups: a control group (n=8) and a telmisartan (10 mg/kg, intragastric administration) treatment group (n=9). All mice were fed a high-fat diet. After 12 weeks, the mice were sacrificed. Venous blood was collected from the retro-orbital sinus to detect TMAO using high-performance liquid phase chromatography-tandem mass spectrometry. The severity of atherosclerosis was determined by measuring the area of the plaque at the root of the aorta. The plaque stability was determined by analyzing the expression of interleukin-6 (IL-6), monocyte chemoattractant protein 1 (MCP-1) and macrophage infiltration in plaques and plaque morphology. Colonic fecal 16S- RNA V3-V4 region sequencing was used to analyze intestinal flora. RESULTS: Compared with the control group, the plaque area of the telmisartan treatment group decreased significantly, and the expression of IL-6, MCP-1, and infiltration macrophages also decreased significantly. Plasma TMAO levels were significantly lower in the telmisartan-treated group than in the control group. Meanwhile, the blood pressure and body weight of the mice treated with telmisartan were lower than those of the control group. Intestinal flora analysis showed that telmisartan significantly changed the composition of intestinal flora and reduced six bacteria known to produce TMAO, including Anaeroplasmataceae, Bacteroidaceae, Clostridiaceae, Lachnospiraceae, Ruminococcaceae, and Proteus. CONCLUSION: In addition to blocking AT1R, telmisartan may reduce TMAO plasma content by changing intestinal flora composition.

Effects of Wuzhi capsule on the pharmacokinetics of simvastatin and its metabolite simvastatin acid in rats

SUN Qing, SUN Jianfang, LI Li, CHANG Huichao, ZHOU Quan

2020, 25(11):  1242-1249.  doi:10.12092/j.issn.1009-2501.2020.11.005

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AIM: To develop LC-MS /MS method for simultaneous determination of simvastatin and simvastatin acid concentrations in rat plasma, and investigate the pharmacokinetic effects of Wuzhi capsule (WZC) on simvastatin and simvastatin acid concentrations in rats.  METHODS: The method was based on simple liquid liquid extraction (LLE) with lovastatin as internal standard. Agilent Eclipse-C18 (2.1 mm×150 mm, 3.5 μm) was used as the chromatographic column, while water-acetonitrile (both of containing 0.1% formic acid) (5∶95, V/V) was used as the mobile phase. Detection was performed with multiple reactions monitoring (MRM) using electrospray ionization (ESI).Twelve male rats were divided into control group and experimental group, with 6 rats in each group. The control group was given 40 mg/kg simvastatin by gavage, and the experimental group was given 150 mg/kg Wuzhi capsule (containing deoxyschizandrin 11.25 mg per capsule) by gavage and followed by 40 mg/kg simvastatin by gavage after 15 minutes. About 0.5 mL of blood was collected from the orbital vein plexus of rats in heparin tube at the time points of 0.25, 0.5, 0.75, 1.0, 2.0, 3.0, 4.0, 6.0, 8.0 and 12.0 hours before and after administration, respectively. The relevant pharmacokinetic parameters were calculated by Phoenix software. RESULTS: The calibration curves of simvastatin and its metabolite simvastatin acid showed a good linearity over the concentration range of 5-5 000 ng/mL with the lower limit of quantitation of 5 ng/mL (simvastatin r=0.994 0, simvastatin acid r=0.994 54). The intra-day and inter-day relative standard deviations were both below 10%, and the absolute recovery could reach more than 80%.The major pharmacokinetic parameters were as follows (control group vs. experimental group), Cmax［(97±24) vs. (590±227) ng/mL, (385±137) vs. (1 322±502) ng/mL］, AUC0-t ［(446±70) vs. (1 315±535) ng·h·mL- 1, (1 305±531) vs. (3 393±1 047) ng·h·mL- 1)］, t1/2 ［(2.37±0.18) vs. (2.93±0.78) h, (2.39±0.43) vs. (3.44±0.80) h] increased significantly (P<0.05). CONCLUSION: The LC-MS/MS method established in this study is fast, simple, sensitive and specific, and is suitable for the determination of simvastatin and simvastatin acid in rat plasma. Wuzhi capsule could affect the pharmacokinetic parameters of simvastatin and simvastatin acid in rats. It is suggested that the combination of two drugs in clinical practice will cause significant interaction.

General considerations of model-based meta-analysis

LI Lujin, DING Junjie, LIU Dongyang, WANG Xipei, DENG Chenhui, JI Shangmin, CHEN Wenjun, MA Guangli, WANG Kun, SHENG Yucheng, XU Ling, PEI Qi, CHEN Yuancheng, CHEN Rui, SHI Jun, LI Gailing, WANG Yaning, WANG Yuzhu, XIE Haitang, ZHOU Tianyan, FANG Yi, ZHANG Jing, JIAO Zheng, HU Bei, ZHENG Qingshan

2020, 25(11):  1250-1267.  doi:10.12092/j.issn.1009-2501.2020.11.006

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With the increasing cost of drug development and clinical trials, it is of great value to make full use of all kinds of data to improve the efficiency of drug development and to provide valid information for medication guidelines. Model-based meta-analysis (MBMA) combines mathematical models with meta-analysis to integrate information from multiple sources (preclinical and clinical data, etc.) and multiple dimensions (targets/mechanisms, pharmacokinetics/pharmacodynamics, diseases/indications, populations, regimens, biomarkers/efficacy/safety, etc.), which not only provides decision-making for all key points of drug development, but also provides effective information for rational drug use and cost-effectiveness analysis.  The classical meta-analysis requires high homogeneity of the data, while MBMA can combine and analyze the heterogeneous data of different doses, different time courses, and different populations through modeling, so as to quantify the dose-effect relationship, time-effect relationship, and the relevant impact factors, and thus the efficacy or safety features at the level of dose, time and covariable that have not been involved in previous studies. Although the modeling and simulation methods of MBMA are similar to population pharmacokinetics/pharmacodynamics (Pop PK/PD), compared with Pop PK/PD, the advantage of MBMA is that it can make full use of literature data, which not only improves the strength of evidence, but also can answer the questions that have not been proved or can not be answered by a single study. At present, MBMA has become one of the important methods in the strategy of model-informed drug  development (MIDD). This paper will focus on the application value, data analysis plan, data acquisition and processing, data analysis and reporting of MBMA, in order to provide reference for the application of MBMA in drug development and clinical practice.

Effect of recipient SLCO1B1 rs2291075 polymorphism on tacrolimus dose-corrected trough concentration in early postoperative period of liver transplantation

WU Yi, WANG Rangrang, WANG Zhaowen, FAN Junwei

2020, 25(11):  1268-1275.  doi:10.12092/j.issn.1009-2501.2020.11.007

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AIM: To explore the relationship between SLCO1B1 rs2291075 polymorphism and tacrolimus (FK506) dose-corrected trough concentration (C/D, ng·mL-1·mg-1·kg-1) during early period of liver transplantation.  METHODS: CYP3A5 rs776746 and SLCO1B1 rs2291075 polymorphisms of 210 liver transplantation patients and their corresponding donor livers were assessed by PCR amplification and DNA sequencing. The influence of gene polymorphisms on C/D values of FK506 was analyzed. The early postoperative period after liver transplantation was divided into convalescence phase (1-14 days) and stationary phase (15-28 days) according to the change of liver function and FK506 C/D values. RESULTS: The combined analysis of donor and recipient CYP3A5 rs776746 could distinguish the metabolic phenotype of FK506 into three groups: extensive metabolism (EM), intermediate metabolism (IM) and poor metabolism (PM), which was entitled the E-I-P classification system. Tacrolimus C/D ratios of recipient SLCO1B1 rs2291075 CT and TT carriers were very close and were significantly lower than those of recipient SLCO1B1 rs2291075 CC genotype carriers in convalescence phase (P=0.019 5) and in stationary phase (P=0.015 2). There were no statistically significant differences between tacrolimus C/D ratios of patients carried with SLCO1B1 rs2291075 CT, TT genotype donors and those carried with SLCO1B1 rs2291075 CC genotype donors. A model consisting of FK506 daily dose, total bilirubin, E-I-P classification, and recipient SLCO1B1 rs2291075 could predict FK506 C/D ratios in convalescence phase by multivariate analysis. However, Recipient SLCO1B1 rs2291075 genotype failed to enter forecast model for C/D ratios in stationary phase. Recipient SLCO1B1 rs2291075 genotype had significant effect on FK506 C/D ratios in convalescence phase (P=0.030) and stationary phase (P=0.040) in PM subgroup, which excluded the interference come from donor and recipient CYP3A5 rs776746. CONCLUSION: SLCO1B1 rs2291075 can be a novel genetic locus associated with FK506 metabolism. The combined analysis of donor and recipient CYP3A5 rs776746, recipient SLCO1B1 rs2291075 genotypes helps to guide the personalized administration of FK506 in early period after liver transplantation.

Preliminary study on the application value of trazodone combined with cognitive behavioral therapy in community detoxification management

GUI Donghui, CAI Dandan, ZHANG Jianbin, ZHU Huaqiang, ZHOU Wenhua

2020, 25(11):  1276-1282.  doi:10.12092/j.issn.1009-2501.2020.11.008

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AIM: To investigate the clinical application value of trazodone combined with cognitive behavioral therapy on heroin addicts during the period of community detoxification.  METHODS: A total of 140 heroin addictive patients treated by community detoxification management were randomly divided into four groups, 35 patients were given trazodone combined with cognitive behavioral therapy (T+P group), 35 patients were given a single trazodone therapy (T group), 35 patients were given a single cognitive behavioral therapy (P group), and 35 patients were only given community detoxification management (S group), the course of the treatment lasted for 12 weeks. At the time-point of baseline and the weekends 4, 8 and 12 after treatment, four groups were scored by Hamilton anxiety rating scale (HAMA), Hamilton depression rating scale (HAMD), Pittsburgh sleep quality index scale (PSQI), psychological craving rating scale (PCS), Chinese perceived stress scale (CPSS), and urine morphine positive rate. The safety was evaluated with Treatment Emergent Side-effect Scale (TESS) and laboratory test. RESULTS: A total of 125 patients completed the 12 weeks of treatment. HAMA, HAMD and PSQI scores of T+P group and T group on weekends 4, 8 and 12 were significantly reduced when compared with baseline, respectively (P<0.05). HAMA and HAMD scores of P group on weekends 4, 8 and 12 were significantly decreased when compared with baseline, respectively (P<0.05). PSQI scores of P group on weekends 8 and 12 were significantly decreased when compared with baseline, respectively (P<0.05). HAMA, HAMD and PSQI scores of S group on weekend 12 were significantly reduced when compared with baseline, respectively (P<0.05). There were significant differences on the PCS and CPSS scores between T+P group, T group, P group and S group on weekends 4, 8 and 12, respectively (P<0.05). There also showed statistically significant differences on the urine morphine positive rates between T+P group and S group on weekends 4, 8 and 12, respectively (P<0.05). There was no significant difference on the TESS score between T+P group and T group (P>0.05). CONCLUSION: Trazolone combined with cognitive behavior therapy can significantly improve the negative emotions, sleep quality and other related mental symptom of heroin addiction patients in the community detoxification period, and play a positive role in improving the quality of community detoxification management.

Side effects of lopinavir/ritonavir for COVID-19

SHENG Haoyu, QUAN Bin, LIANG Manman, WU Qiongle, YANG Jianghua

2020, 25(11):  1283-1287.  doi:10.12092/j.issn.1009-2501.2020.11.009

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AIM: To study the regularity and characteristics of side effects of lopinavir/ritonavir for COVID-19.  METHODS: The type of side effects, general information, medical history and prognosis in 61 confirmed and suspected COVID-19 patients with lopinavir/ritonavir were analyzed. RESULTS: Among the 61 patients, 41(67.21%) had lopinavir/ritonavir related side effects, mainly manifested as gastrointestinal reactions (82.93%) and liver function damage (53.66%). Old age, long course of disease and chronic gastrointestinal disease are independent risk factors for side effects. CONCLUSION: Lopinavir/ritonavir has a high incidence of side effects and can be used in COVID-19 patients under the condition of close observation of the patient's symptoms and test results. Special population should improve pharmaceutical care to ensure the safety of drug use.

Mechanism and advance of COVID-19 on immune related pathogenesis and immunotherapy

LI Hongpeng, ZHANG Liu, LI Qingyun

2020, 25(11):  1288-1294.  doi:10.12092/j.issn.1009-2501.2020.11.010

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The novel coronavirus pneumonia (COVID-19) is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and is pandemic globally. The progress and evolution of the disease is closely related to the immune defense function. The host immune promotes virus clearance. The dysfunction of immune compromises virus clearance, and incurs uncontrolled inflammatory response which causes immune pathological injury and makes the disease worse. Therefore, the application of immunotherapy for immune disorders caused by SARS-CoV-2 infection is expected to be an effective strategy for the treatment of COVID-19.

Application and progress of pharmacokinetics study in bioequivalence evaluation of orally inhaled drug products

ZHANG Jisheng, HUANG Kai, CHU Nannan, HE Qing

2020, 25(11):  1295-1308.  doi:10.12092/j.issn.1009-2501.2020.11.011

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Orally inhaled drug products (OIDPs) have a pivotal position and great clinical demand in the treatment of asthma and chronic obstructive pulmonary disease. The development of local generic drugs which are bioequivalent to branded drugs in efficacy and safety while with less price will not only help to solve the problem of drug accessibility, but also greatly reduce the public health burden.OIDPs are complex combinations of formulation and device, and have special drug delivery route and characteristic of local release. Thus, generic drugs of OIDPs are difficult to develop and get registration. Until now, Food and Drug Administration (FDA), European Medicines Association (EMA), Health Canada (HC) and National Medical Products Administration (NMPA) all consider that pharmacokinetics (PK) method can be used to evaluate systemic exposure (safety) in human bioequivalence (BE) study of OIDPs, but there is no consensus on its role in the evaluation of pulmonary deposition (efficacy). The possible reason lies in that the efficacy of OIDPs is determined by both the amount and the region of drug pulmonary deposition. Nevertheless, PK study is still more sensitive and economical in assessing potential differences among products, when compared with pharmacodynamics or clinical endpoint study. Here we mainly compared the domestic and international guidelines and evaluation methods of OIDPs BE study, and introduced the experimental design of PK study and its application and progress in lung deposition study. 

Research progress of melatonin and pancreatic diseases#br#

ZHANG Chenhan, NIE Jingyun, XIN Pengfei, CHAI Chen, LI Bo

2020, 25(11):  1309-1314.  doi:10.12092/j.issn.1009-2501.2020.11.012

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Melatonin is an indoleamine secreted by the pineal gland and plays a key role in a variety of physiological activities, including the regulation of circadian rhythm, immune response, oxidative process and apoptosis. In recent years, the anti-tumor properties of melatonin have attracted more and more attention. A large number of studies have found that melatonin plays a protective role in the early process of acute pancreatitis and can inhibit the production of pancreatic adenocarcinoma cells through apoptosis; moreover, it can regulate the synthesis and secretion of pancreatic endocrine hormones through autophagy and affect the development of pancreatic endocrine tumors. Therefore, melatonin may affect the development of pancreatic disease, but the mechanism of its action on pancreatic diseases has not been fully understood. This paper reviews the research progress of melatonin and pancreatic diseases in recent years, analyzes the role of melatonin in pancreatic diseases and the therapeutic effect of melatonin combined with chemotherapy drugs on pancreatic cancer. 

Research progress of autophagy in viral pneumonia

WANG Weilu, MAO Yicheng, ZHU Yizhun

2020, 25(11):  1315-1320.  doi:10.12092/j.issn.1009-2501.2020.11.013

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Autophagy, as a highly conserved process of recovery and degradation of eukaryotic components, plays an important role in cell hunger response, maintenance of normal cell activity and specific functions in lung tissue. When cells are damaged due to physiological, pathological or chemical factors, autophagosomes and lysosomes fuse to produce a large number of autophagosomes, thus inducing the initiation of autophagy process, which is beneficial to protect cells from injury. Recent studies have found that autophagy related signaling pathways are highly correlated with the occurrence and development of a variety of viral pneumonia, such as swine influenza A (H1N1) virus, highly pathogenic avian influenza virus (H5N1), severe acute respiratory syndrome (SARS), middle east respiratory syndrome (MERS) and new coronavirus pneumonia (COVID-19). This paper will introduce the five viral pneumonia that have a great impact on China and even the whole world, and elaborate the mechanism of autophagy in these disease. Hopefully it could provide theoretical basis and effective methods and means for targeted autophagy treatment of viral pneumonia.