Molecular Mechanism of Resveratrol Against NAFLD Based on Network Pharmacology / 中国实验方剂学杂志

ObjectiveTo predict the molecular mechanism of resveratrol against non-alcoholic fatty liver disease （NAFLD） based on network pharmacology and molecular docking and verify the results on the liver cell model induced by PM2.5 exposure. MethodThe targets of resveratrol were screened out from Traditional Chinese Medicine System Pharmacology Database and Analysis Platform （TCMSP）， PubChem， DrugBank， and SwissTargetPrediction， and the potential targets of NAFLD were retrieved from Comparative Toxicogenomics Database （CTD）， DisGeNET， GeneCards， and Online Mendelian Inheritance in Man （OMIM）. Then the common targets were obtained. STRING 11.5 was used to construct the protein-protein interaction （PPI） network of the common targets. Cytoscape 3.8.2 was used to plot the “target-pathway” network， and the core modules and key targets were selected. Metascape was adopted for Gene Ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses of common targets. SYBYL-X 2.0 was used for molecular docking of resveratrol to key targets. Finally，cell apoptosis and the expression of apoptosis-related proteins were detected by flow cytometry and Western blot in the PM2.5-exposed human liver cell line （HepG2）. ResultA total of 115 common targets of resveratrol and NAFLD were obtained. The key targets included tumor necrosis factor （TNF）， B-cell lymphoma-2 （Bcl-2）， and cysteinyl aspartate-specific protease-3（Caspase-3）. As revealed by KEGG enrichment analysis， 174 signaling pathways， represented by the apoptosis and TNF signaling pathways， were obtained. Molecular docking results showed that resveratrol had strong binding activities to Bcl-2 and Caspase-3. Furthermore，the results of flow cytometry and Western blot demonstrated that resveratrol inhibited cell apoptosis of PM2.5-exposed HepG2 cells by regulating the protein expression of Bcl-2 and Caspase-3. ConclusionResveratrol can treat NAFLD in a multi-pathway and multi-target way. It mainly inhibits liver cell apoptosis by affecting the expression of Bcl-2 and Caspase-3， which provides a theoretical basis for the follow-up research on the anti-NAFLD mechanism of resveratrol.

Effect and mechanism of Bidens pilosa decoction on non-alcoholic fatty liver induced by high fat and high glucose in mice / 中国中药杂志

This study aimed to investigate the effect and possible mechanism of Bidens pilosa decoction on non-alcoholic fatty liver disease(NAFLD) induced by high fat and high glucose in mice. Bald/c mice were randomly divided into normal group, model group, metformin(200 mg·kg~(-1)) treatment group, Bidens pilosa decoction(10 g·kg~(-1)) treatment group, metformin and B. pilosa decoction(100 mg·kg~(-1)+5 g·kg~(-1)) treatment group. Except for the normal group, mice in the other four groups were fed with high-fat and high-glucose diet for 8 weeks to establish the non-alcoholic fatty liver model. After 4 weeks of treatment, blood was collected from the eyeballs, the mice were sacrificed, and relevant indicators were detected. The results showed that compared with the model group, blood lipid and blood glucose levels of each treatment group were significantly lower(P<0.05); HE staining results showed that liver pathological damage in each treatment group was significantly improved; oil red O staining results showed fat distribution in each treatment group significantly reduced(P<0.01); immunohistochemical staining showed that glucose regulated the protein expression of protein 78(GRP78) in liver tissues of each treatment group was also significantly reduced(P<0.01); Western blot results showed that endoplasmic reticulum stress signal pathway-related factors GRP78, phosphorylated-protein kinase R-like ER kinase(p-PERK), eukaryotic translation-initiation factor 2α(eIF2α), activating transcription factor 4(ATF4), C/EBP homologous protein(Chop), inositol requiring 1α(IRE1α), and cleaved-cysteinyl aspartate specific proteinase 12(cleaved-caspase-12) were significantly reduced(P<0.01). The results of the combined drug treatment group were better than those of the single drug treatment group. These results showed that B. pilosa decoction had the effect in improving non-alcoholic fatty liver, and its mechanism may be related to the down-regulation of the expression of endoplasmic reticulum stress(ERS)-related factors, and the reduction of the apoptosis of hepatocytes caused by ERS and the down-regulation of blood lipid and blood glucose levels.

Effect of butylphthalide on blood-brain barrier after cerebral hemorrhage in SD rats and the mechanisms / 中南大学学报(医学版)

To investigate the alteration in Golgi and blood-brain barrier after cerebral hemorrhage in SD rats, and to evaluate the effect of butylphthalide on blood-brain barrier. 
 Methods: Sprague-Dawley rats were randomly distributed into 4 groups: a control group, a sham group, an intracerebral hemorrhage (ICH) group, and a butylphthalide group. Brain tissue was collected at 48 h after the blood brain barrier permeability was examined. Western blotting and real-time polymerase chain reaction (real-time PCR) were conducted to explore the change of GM130, Cdc42 and tight junction protein and mRNA expression in rat brain after ICH. Immunohistochemistry (IHC) was performed to explore the distribution of ZO-1 and Occludin in the cerebral vascular endothelial cells around the hematoma.
 Results: The Evans blue (EB) extravasation in the ICH group were much greater than that in the sham group (P<0.05). Butylphthalide treatment significantly decreased Evans blue extravasation compared to the ICH group (P<0.05). Results of Western blotting and real-time PCR showed that GM130, Cdc42, ZO-1/Occludin were decreased (P<0.05). The intervention of butylphthalide significantly upregulated the expressions of Cdc42 as well as ZO-1/Occludin (P<0.05), but exerted no effect on GM130 (P<0.05). Immunofluorescent staining showed that GM130 was co-localized with Cdc42 and administration of butylphthalide improved the expression of Cdc42 around the hematoma without affecting the expression of GM130. IHC showed that expressions of occludin and ZO-1 around the hematoma were significantly decreased in the ICH group (P<0.05), whereas butylphthalide treatment elevated the expressions of ZO-1 and occludin around the hematoma compared with the ICH group (P<0.05).
 Conclusion: Morphology of Golgi apparatus is altered and the blood-brain barrier is destroyed after ICH. The application of butylphthalide can alleviate neurological impairment and blood-brain barrier disruption, which is related to the up-regulation of Cdc42, but not GM130.

Application and prospects of butylphthalide for the treatment of neurologic diseases / 中华医学杂志(英文版)

OBJECTIVE@#The 3-N-butylphthalide (NBP) comprises one of the chemical constituents of celery oil. It has a series of pharmacologic mechanisms including reconstructing microcirculation, protecting mitochondrial function, inhibiting oxidative stress, inhibiting neuronal apoptosis, etc. Based on the complex multi-targets of pharmacologic mechanisms of NBP, the clinical application of NBP is increasing and more clinical researches and animal experiments are also focused on NBP. The aim of this review was to comprehensively and systematically summarize the application of NBP on neurologic diseases and briefly summarize its application to non-neurologic diseases. Moreover, recent progress in experimental models of NBP on animals was summarized.@*DATA SOURCES@#Literature was collected from PubMed and Wangfang database until November 2018, using the search terms including "3-N-butylphthalide," "microcirculation," "mitochondria," "ischemic stroke," "Alzheimer disease," "vascular dementia," "Parkinson disease," "brain edema," "CO poisoning," "traumatic central nervous system injury," "autoimmune disease," "amyotrophic lateral sclerosis," "seizures," "diabetes," "diabetic cataract," and "atherosclerosis."@*STUDY SELECTION@#Literature was mainly derived from English articles or articles that could be obtained with English abstracts and partly derived from Chinese articles. Article type was not limited. References were also identified from the bibliographies of identified articles and the authors' files.@*RESULTS@#NBP has become an important adjunct for ischemic stroke. In vascular dementia, the clinical application of NBP to treat severe cognitive dysfunction syndrome caused by the hypoperfusion of brain tissue during cerebrovascular disease is also increasing. Evidence also suggests that NBP has a therapeutic effect for neurodegenerative diseases. Many animal experiments have found that it can also improve symptoms in other neurologic diseases such as epilepsy, cerebral edema, and decreased cognitive function caused by severe acute carbon monoxide poisoning. Moreover, NBP has therapeutic effects for diabetes, diabetes-induced cataracts, and non-neurologic diseases such as atherosclerosis. Mechanistically, NBP mainly improves microcirculation and protects mitochondria. Its broad pharmacologic effects also include inhibiting oxidative stress, nerve cell apoptosis, inflammatory responses, and anti-platelet and anti-thrombotic effects.@*CONCLUSIONS@#The varied pharmacologic mechanisms of NBP involve many complex molecular mechanisms; however, there many unknown pharmacologic effects await further study.

Application and prospects of butylphthalide for the treatment of neurologic diseases / 中华医学杂志(英文版)

Objective@#The 3-N-butylphthalide (NBP) comprises one of the chemical constituents of celery oil. It has a series of pharmacologic mechanisms including reconstructing microcirculation, protecting mitochondrial function, inhibiting oxidative stress, inhibiting neuronal apoptosis, etc. Based on the complex multi-targets of pharmacologic mechanisms of NBP, the clinical application of NBP is increasing and more clinical researches and animal experiments are also focused on NBP. The aim of this review was to comprehensively and systematically summarize the application of NBP on neurologic diseases and briefly summarize its application to non-neurologic diseases. Moreover, recent progress in experimental models of NBP on animals was summarized.@*Data sources@#Literature was collected from PubMed and Wangfang database until November 2018, using the search terms including "3-N-butylphthalide," "microcirculation," "mitochondria," "ischemic stroke," "Alzheimer disease," "vascular dementia," "Parkinson disease," "brain edema," "CO poisoning," "traumatic central nervous system injury," "autoimmune disease," "amyotrophic lateral sclerosis," "seizures," "diabetes," "diabetic cataract," and "atherosclerosis."@*Study selection@#Literature was mainly derived from English articles or articles that could be obtained with English abstracts and partly derived from Chinese articles. Article type was not limited. References were also identified from the bibliographies of identified articles and the authors’ files.@*Results@#NBP has become an important adjunct for ischemic stroke. In vascular dementia, the clinical application of NBP to treat severe cognitive dysfunction syndrome caused by the hypoperfusion of brain tissue during cerebrovascular disease is also increasing. Evidence also suggests that NBP has a therapeutic effect for neurodegenerative diseases. Many animal experiments have found that it can also improve symptoms in other neurologic diseases such as epilepsy, cerebral edema, and decreased cognitive function caused by severe acute carbon monoxide poisoning. Moreover, NBP has therapeutic effects for diabetes, diabetes-induced cataracts, and non-neurologic diseases such as atherosclerosis. Mechanistically, NBP mainly improves microcirculation and protects mitochondria. Its broad pharmacologic effects also include inhibiting oxidative stress, nerve cell apoptosis, inflammatory responses, and anti-platelet and anti-thrombotic effects.@*Conclusions@#The varied pharmacologic mechanisms of NBP involve many complex molecular mechanisms; however, there many unknown pharmacologic effects await further study.

Protective effect of earthworm active ingredients against endoplasmic reticulum stress induced L-02 hepatocyte apoptosis / 中国中药杂志

This study focused on the protective effect of earthworm active ingredients (EWAs) on hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS) in L-02 cells. The L-02 cells were cultured in vitro. The cell viability was measured with CCK-8, the apoptosis of L-02 cells was detected by flow cytometry, and the relevant protein and mRNA expressions were detected by Western blot and qPCR. According to the findings, tunicamycin (TM) could obviously reduce the survival rate of L-02 cells in a time-dependent and dose-dependent manner. Compared with normal group, the apoptosis rate in model group was significantly increased (<0.05 or <0.01). The protein and mRNA expressions of ERS-related signal molecules, such as GRP78, PERK, eLF2α, ATF4, CHOP and Bax, were significantly up-regulated (<0.05 or <0.01), while Bcl-2 was significantly down-regulated (<0.05 or <0.01). After the administration with different concentrations of EWAs, compared with model group, EWAs could significantly increase the survival rate ofL-02 hepatocyte and decrease the cell apoptosis rates. It could also reduce the protein and mRNA expressions of ERS-related signal molecules, such as GRP78, PERK, eLF2α, ATF4, CHOP and Bax, in a dose-dependent manner (<0.05 or <0.01) and increased the protein and mRNA expressions of Bcl-2(<0.05 or <0.01). These results showed that EWAs had a significantly protective effect on hepatocyte apoptosis induced by ERS in L-02 cells. Its mechanism may be related to the down-regulation of mRNA and protein expressions of GRP78, PERK, ATF4, eLF2α, CHOP and Bax, and the up-regulation, the relief of ERS and the promotion of the proliferation of impaired L-02 cells.

Intervention effectiveness of intensive training in medical interns’occupa-tional exposure protection / 中国感染控制杂志

Objective To study the intervention effectiveness of intensive training in medical interns’occupational exposure protection.Methods Medical interns in seven colleges and universities in Wuhan between 2011 and 2015 were chosen and divided into control group (675 medical interns in 2011 - 2013,comprehensive training about healthcare-associated infection control was conducted for once before internship )and intervention group (682 medical interns in 2014 - 2015,hospital-department-tutor three steps of intensive training was conducted),the occurrence of occupational exposure and exposure knowledge awareness between two groups were compared. Results The awareness rate of knowledge about occupational exposure protection and the implementation rate of protective measures in intervention group were both higher than control group (all P <0.05).Incidence of occupa-tional exposure in intervention group was lower than control group (21 .70%[n=148]vs 65.33%[n=441 ],χ2 =262.91 ,P <0.01);percentage of interns who conducted active serological virus detection and intensified vaccination of hepatitis B virus were both higher than control group (14.66%[n=100]vs 2.96%[n=20];11 .73%[n=80]vs 2.67%[n=18],respectively).Occupational exposure before and after intervention occurred most frequently when interns were preparing medicine,occupational exposure reporting rate in intervention group was higher than control group (72.97%[108/148]vs 50.11 % [221/441 ],χ2 =52.78,P <0.01 ).Conclusion The intensive training of occupational exposure protection among medical interns can improve the awareness and skills of occupational protec-tion,and reduce the occurrence of occupational exposure.

Effect of kaolin combined with propranolol on paraquat concentration in lungs of poisoned mice / 上海预防医学

[ Objective ] To observe the effect of kaolin and propranolol on paraquat ( PQ ) concentration in the lungs of poisoned mice. [ Methods] A group of 144 ICR mice were randomly divided into 3 groups:PQ, treatment, and control.Then 100 mg/kg PQ were intragastrically administrated ( ig) in PQ group and treatment group, while only the same volume normal saline was given in control group.And then 48 g/kg kaolin combined with 3.2 mg/kg propranolol were administered in treatment group immediately after poisoning while only the same volume of normal saline was given in the other two groups. Pathological examination was done and PQ concentration in lungs of the mice detected 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h after poisoning. [ Results] In the lungs of the mice in PQ group occurred alveolar capillary expansion, endothelial cell swelling, small or large sheet-shaped inflammation cell infiltration and mainly neutrophils while in treatment group the above lesions were apparently alleviated.In PQ and treatment groups, PQ concentration in lungs of both groups rose significantly 0.5 h after poisoning and up to peak at 4 h.But PQ concentration in lungs decreased significantly in treatment group from 4 h to 24 h after poisoning ( P<0.05) , as compared with that in PQ group. [ Conclusion] PQ concentration in lungs of the poisoned mice was decreased and the injury alleviated when they were treated with kaolin combined with propranolol.It is held that further research is worth doing in clinical practice.

Factors related to Glycophorin A mutation frequency of workers exposed to benzene / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze related factors which affect GPA mutation frequency of workers exposed to benzene, with the Glycophorin A (GPA) mutation assay and explore the possibility of GPA mutation frequency as an index of predicting the risk of benzene poisoning.</p><p><b>METHODS</b>The erythrocytes were bound with fluorescent-labeled monoclonal antibody after isolated and fixed from the peripheral blood, and then the GPA mutation assay was performed using the flow cytometry (FCM). The related factors of GPA mutation frequency were analyzed by statistical methods.</p><p><b>RESULTS</b>The GPA mutation frequency of chronic benzene poisonings was significantly higher than that of their controls (P < 0.05). Significant direct correlation was found between age, length of service, accumulative exposure score and the GPA mutation frequency of workers exposed to benzene (P < 0.01). However, there was significantly inverse correlation between the 3AB index and the GPA mutation frequency (GPAN0: r(s) = -0.589, P < 0.01, GPANN: r(s) = -0.615, P < 0.01). In the multiple factor regression analysis on GPA mutation frequency, benzene exposure and individual susceptibility both entered model of multiple factors analysis, the coefficient of determination of benzene-exposed workers was 0.819.</p><p><b>CONCLUSION</b>Exposure to benzene and individual susceptibility are the most important factors that affect GPA mutation frequency. GPA mutation frequency increases with the benzene exposure and individual susceptibility.</p>