1.Anti-inflammatory mixture alleviates acute lung injury induced by sepsis in rats by modulating Beclin-1-mediated autophagy
Weilan LU ; Guoliang YAN ; Yifan SHEN ; Haitao LI ; Sai WU ; Tongrui WENG ; Rui ZHANG ; Yanwen HUO
Chinese Critical Care Medicine 2024;36(7):717-722
Objective:To investigate the protective effects of an anti-inflammatory mixture on acute lung injury (ALI) induced by sepsis in rats, as well as its possible mechanisms.Methods:A total of 40 Sprague-Dawley (SD) rats were randomly divided into the sham group, septic ALI model group (model group), 3-methyladenine (3-MA) control group, and anti-inflammatory mixture pretreatment group, with 10 rats in each group. Cecal ligation and perforation (CLP) was performed to reproduce a septic ALI model. The rats in the sham group only underwent opening and closing the abdomen without perforation and ligation. Both groups were given saline gavage and intraperitoneal injection for 3 consecutive days before surgery. The 3-MA control group was given intraperitoneal injection of saline and autophagy inhibitor 3-MA 15 mg/kg for 3 consecutive days before modeling. The anti-inflammatory mixture pretreatment group was given 8.8 mL/kg of anti-inflammatory mixture by gavage [the composition of anti-inflammatory mixture: rhubarb 15 g (after the next), coptis chinensis 15 g, baical skullcap root 12 g, magnoliae cortex 12 g, dahurian patrinia herb 30 g] and saline intraperitoneal injection for 3 consecutive days before modeling. The rats in each group were anesthetized 24 hours after surgery and died due to abdominal aortic blood collection. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum inflammatory cytokines interleukins (IL-1β and IL-6). Lung tissue was taken and then the bronchoalveolar lavage fluid (BALF) was collected, and the levels of IL-1β and IL-6 were detected by ELISA. Lung wet/dry weight (W/D) ratio was measured. After hematoxylin-eosin (HE) staining, the histopathological changes of the lungs were observed under light microscopy. Western blotting was used to detect the expression of autophagy markers microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ (LC3-Ⅱ/Ⅰ) and Beclin-1 protein in lung tissue. Autophagosomes in lung tissue were observed with transmission electron microscopy.Results:Compared with the sham group, the rats in the model group exhibited severe destruction of lung tissue structure, with significant infiltration of inflammatory cells, the lung W/D ratio and the levels of IL-1β and IL-6 in serum and BALF were significantly increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein were down-regulated, the autophagosomes were more. The rats in the 3-MA control group exhibited more severe lung tissue injury as compared with the model group, the lung W/D ratio and the levels of inflammatory cytokines in serum and BALF were further increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein still showed a decrease tendency as compared with the sham group, and the autophagosomes were less than that in the model group. Compared with the model group, the anti-inflammatory mixture pretreatment group showed milder lung tissue injury with a minimal amount of inflammatory cell infiltration, the lung W/D ratio was significantly reduced (7.07±1.02 vs. 11.33±1.85, P < 0.05), the levels of IL-1β and IL-6 in both serum and BALF were significantly decreased [IL-1β (ng/L): 26.04±3.86 vs. 40.83±5.46 in serum, 17.75±2.02 vs. 26.86±4.32 in BALF; IL-6 (ng/L): 91.28±10.15 vs. 129.44±13.05 in serum, 76.06±7.51 vs. 120.91±7.47 in BALF, all P < 0.05], and the ratio of LC3-Ⅱ/Ⅰ and Beclin-1 protein expression were significantly increased [LC3-Ⅱ/Ⅰ ratio: 1.23±0.02 vs. 0.60±0.02, Beclin-1 protein (Beclin-1/GAPDH): 2.37±0.33 vs. 0.62±0.05, both P < 0.05]. Furthermore, an increase in the number of autophagosomes was observed. Conclusion:The anti-inflammatory mixture improves lung injury in rats with sepsis induced by CLP and reduce inflammation levels, potentially through upregulation of Beclin-1-mediated autophagy.
2.Functional characteristics of YAP-positive hepatocytes expression in an early stage of NASH with transcriptome sequence analysis
Weilan ZENG ; Jiaen LIANG ; Yaxue LIU ; Yan WANG
Chinese Journal of Hepatology 2022;30(6):649-655
Objective:To analyze and compare the differentially expressed genes (DEGs) of Yes-associated protein (YAP)-positive and negative hepatocytes and further understand the preliminary functional characteristics of YAP-positive hepatocytes in an early mouse model of nonalcoholic steatohepatitis (NASH) with transcriptome sequence (RNA-Seq).Methods:C57BL/6 mice were fed with methionine-choline deficiency (MCD) diet for 2 weeks to establish an early NASH model, and the control group was fed with normal diet. Liver tissue was stained with hematoxylin-eosin (HE) and Sirius red, and the pathological score was recorded. The expression of YAP and P-YAP were determined by immunohistochemistry (IHC) in liver tissues. Primary hepatocytes with viability greater than 90% were isolated and purified by collagenase perfusion combined with Percoll density gradient centrifugation. YAP-positive and negative hepatocytes were assessed by YAP antibody, flow cytometry and RNA-Seq analyses. Sequencing results were screened by GO, KEGG and interaction network analysis methods. RT-PCR was used to verify the expression levels of YAP and some DEGs in liver tissue model group. Two samples mean was compared by independent samples t-test. Results:Compared with the control group, the HE-stained liver tissue of MCD-induced mice at 2 weeks showed steatosis (pathological score 1.07±0.21), accompanied by lobular inflammation (pathological score 1.13±0.32) and ballooned hepatocyte (pathological score 0.80) ±0.20). Sirius red staining showed non-significant liver fibrosis (pathological score 0.40±0.40). IHC showed partial YAP-positive hepatocytes expression in an early stage of NASH. RNA-Seq analysis showed that clean reads of YAP-positive and negative hepatocytes were 49 310 604 and 5 4820 036, respectively. Compared with YAP-negative hepatocytes, YAP-positive hepatocytes had differential expression of 5 565 genes, including 1 662 up-regulated genes and 3 903 down-regulated genes. GO analysis of up-regulated genes showed that the metabolic processes related to mitochondrial functions, such as purine nucleoside triphosphate and nucleoside triphosphate were significantly enriched in biological processes (BP), while down-regulated gene analysis showed that olfactory-related receptor were significantly enriched in BP. KEGG analysis showed that DEGs were enriched in 292 pathways, and oxidative phosphorylation (OXPHOS) pathway was significantly enriched in signaling pathway. RT-PCR validated that inflammatory factors (interleukin-1β, interleukin-6), YAP and its target genes (Cyr61, Ankrd1), and Cox5b and Sdhc genes were significantly up-regulated in the OXPHOS pathway, which was consistent with the sequencing results. In addition, eight key genes with interaction network analysis were predicted.Conclusion:Changes in hepatocyte metabolic levels may be associated with increased YAP activity in an early stage of NASH.
3. Emodin regulates microRNA expressions of ileum in sepsis mice model
Zhi YONG ; Zhijun JIANG ; Meiyuan ZHANG ; Zhiwen FAN ; Weilan SUN ; Yan TANG
Chinese Journal of Emergency Medicine 2019;28(11):1366-1372
Objective:
To investigate the regulation of emodin on microRNA expressions in mouse model with sepsis by GeneChip microRNA array.
Methods:
Forty two c57 mice were randomly (random number) divided into 3 groups: sham operation group (sham group,
4. Application of three semi-quantitative risk assessment methods in electroplating enterprises
Weilan YAN ; Junfeng LIU ; Kun CHEN ; Jian WANG ; Shibiao SU ; Rongzong LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(6):420-425
Objective:
To apply the semi
5.Progress in treatment of elderly patients with acute myeloid leukemia
Liangming MA ; Weilan YAN ; Duoping LIU
Journal of Leukemia & Lymphoma 2017;26(4):193-198
The incidence of elderly patients with acute myeloid leukemia (AML) is increased year by year, and the median onset age is 67 years old. As old patients often have the viscera dysfunction, it is still lack of unified treatment clinically. This article summarizes the latest research progress of elderly patients with AML in the 58th American Society of Hematology Annual Meeting.
6.Effect of proliferation of triptolide on MV4-11 cells and its mechanism research associated with RAS/ERK/MAPK
Duoping LIU ; Liangming MA ; Yujin LU ; Tao WANG ; Fengli ZHENG ; Weilan YAN
Journal of Chinese Physician 2017;19(6):844-847
Objective To explore the effect of proliferation and apoptosis,and research its mechanism associated with RAS/extracellular signal regulated kinase/mitogen-activated protein kinase (RAS/ ERK/MAPK) in Fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutation acute lymphocytic leukemia cell line MV4-11 cells treated by triptolide (TP).Methods MV4-11 cells were respectively treated by triptolide with diverse concentrations and different times.The proliferation inhibition rate was measured by methyl thiazolyl tetrazolium,the apoptosis rate was detected by flow cytometry,the mRNA expressions of FLT3,RAS,ERK,forkhead box protein M1 (FOXM1),and c-Myc were analyzed by realtime fluorescent quantitative polymerase chain reaction (PCR).Results The proliferation inhibition rate markedly increased in a dose-time dependent manner after MV4-11 cells were treated by TP.After cells were treated with 10,and 20 nmol/L TP,respectively,the apoptosis rates at 48 h were (17.30 ± 0.56) %,(35.77 ±0.55)%,and those at 72 h were (49.83 ±0.45)%,(68.90 ±0.75)% correspondingly.PCR data showed that the mRNA level of FLT3 mRNA decreased obviously,and that of RAS,ERK,FOXM1,and c-Myc also decreased.Conclusions Triptolide could significantly inhibit the MV4-11 cells proliferation and induce cell apoptosis,and its mechanism might be through inhibiting the expression of related genes on RAS/ERK/MAPK signaling pathway.
7.Effects of triptolide on proliferation and apoptosis of acute myeloid leukemia cell line MV411 with FMS-like tyrosine kinase 3 internal tandem duplication and their mechanisms
Weilan YAN ; Liangming MA ; Tao WANG ; Yujin LU
Journal of Leukemia & Lymphoma 2017;26(6):321-324,330
Objective To investigate the effects of triptolide (TP) on proliferation and apoptosis of acute myeloid leukemia cell line MV411 with FMS-like tyrosine kinase 3 internal tandem duplication (FLT3-ITD), and its effect on PI3K-Akt-mTOR pathway. Methods MTT assay was used to detect the proliferation inhibition of MV411 cell proliferation treated by different concentrations of TP in 24, 48 and 72 h. Flow cytometry was used to measure the cell apoptosis rate at 48 and 72 h. Real-time fluorescent quantitative PCR was used to detect the expression of FLT3, PTEN, PI3K, Akt, mTOR mRNA on PI3K-Akt-mTOR pathway. Results After treated by 0, 5, 10, 20, 40, and 80 nmol/L TP in 24, 48 and 72 h, the viability of MV411 cells was inhibited in a time-dose dependence manner. MV411 cells was treated by 0, 10, and 20 nmol/L TP after 48 and 72 h, the cell apoptosis rates were rising with the increasing of drug concentration, there were statistical significances [48 h:(3.30±0.20) %, (17.10±0.36) %, (35.67±0.61) %, 72 h: (7.37±0.32) %, (49.33± 0.40)%, (68.92±0.11)%, all P=0.000]. The expressions of FLT3, PI3K, Akt, and mTOR mRNA were down-regulated and PTEN mRNA expression was up-regulated by TP. Conclusion TP may inhibit the proliferation and induce apoptosis of MV411 cells by affecting the expression of PI3K-Akt-mTOR pathway related genes.
8.Application of Diffusion Tensor Image in Diagnosis of Vascular Parkinsonism
Xiaolin TAN ; Yan REN ; Songbai LI ; Zhiqiang ZHANG ; Weilan ZHANG ; Xiaohong LI ; Na LIU ; Fan LOU
Journal of China Medical University 2016;45(4):361-364,370
Objective To study the pathogenesis of lower limb motor disorder in vascular parkinsonism(VP)using diffusion tensor image(DTI). Methods A case?control study was performed in a cohort of 16 VP patients and 32 PD patients. Patients were all recruited from the first affiliated hospital of China Medical University from Dec. 2011 to Dec. 2012. The lower limb motor function of those patients was assessed using measurement scales. All patients received magnetic resonance image(MRI)and diffusion tensor image(DTI). Results Patients in VP group showed more seri?ous movement disorders such as freezing of gait(gait ignition failure)than these in PD group,and the movement disorders of their lower limbs prog?ress more rapidly in VP group than the PD group. Movement disorders of their lower limbs in VP group were related to fibers in bilateral frontal lobes (P=0.008). Movement disorder of their lower limbs in PD group were related to fibers in substantia nigra(P=0.030). There are statistically signifi?cant differences in both the FA values of substantia nigra,bilateral frontal lobes and the ADC values of frontal lobes,and the pars compacta and pars reticulata of substantia nigra between these two groups(all P<0.05). Conclusion Movement disorders of bilateral lower limbs are more seriously manifested in VP patients than in PD patients. The onset movement dysfunction of bilateral lower limbs is closely related to fibers in the frontal lobe.
9.Analysis of 410 Cases of Clinical Pharmacist Consultations and Discussion of Working Mode
Man ZHU ; Yan SUN ; Daihong GUO ; Fei PEI ; Dongxiao WANG ; Weilan WANG ; Zhihui TANG ; Chao FAN ; Zhao REN
China Pharmacist 2014;(11):1912-1914
Objective:To understand the mode and effect of the consultations involved clinical pharmacists, and discuss the work-ing process and mode of the consultations. Methods:Based on the practical work and team discussion, the flow chart and consultation registration form for the clinical pharmacist consultations were established and applied. Totally 410 cases of clinica1 pharmacist consul-tations were retrospectively investigated from March 2009 to December 2012. Results:The consultation flow chart was strictly followed by the clinical pharmacists in our hospital, and the obvious effect was shown. The number of consultation was increased from 59 cases in 2009 to 122 cases in 2013. In all the consultation cases, 21 departments were involved, including the medical departments and the surgery departments. Among the cases,the adjustment in antibacterial drug treatment was the main requirements. Meanwhile, the a-doption rate of the clinical pharmacist suggestions was 95. 12%. Conclusion:Clinical pharmacists are playing an important role in clin-ical treatment. The consultation work can be regulated by the consultation flow chart and registration form with the improved adoption rate and consultation quality.
10.Research on problem design of problem-based learning in pediatric teaching
Haihong XUE ; Jihong QIAN ; Lianwen WANG ; Xiaojun YUAN ; Yi CHEN ; Weilan WU ; Yan CHEN ; Kun SUN
Chinese Journal of Medical Education Research 2012;11(6):582-584
Objective To apply 3C3R model in problem design of PBL for pediatrics teaching.Methods The 3C3R model comprises two classes of components:core components and processing components.Core components of the model are 3C,which are content,context and connection while processing components are 3R,which are researching,reasoning and reflecting.3C3R model was used in the problem design for the PBL case of ‘ Why the mouth of Baobao became purple when he was crying?' Totally 76 eight-year program medical students and 7 tutors were enrolled as teaching object.The anonymous questionnaires from the students were collected for assessment of PBL teaching.Results The percentage of students with scores ≥4 for content in PLB problem design was 90.8%,for context was 80.3%,for connection was 64.5%,for researching was 81.6%,for reasoning was 69.7% and for reflecting was 40.8%.The percentage of tutors with scores ≥4 for content in PLB problem design was 100%,for context was 71.4%,for connection was 28.6%,for researching was 71.4%,for reasoning was100%,for reflecting was57.1%.Both students and tutors held a positive attitude towards the component of content,context,researching and reasoning in problem design model.But the components of connection and reflection needed to be improved.Conclusion The 3C3R model is helpful for problem design of PBL in pediatric teaching.

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