Active Ingredients of Reduning Injection Maintain High Potency against SARS-CoV-2 Variants / 中国结合医学杂志

OBJECTIVE@#To investigate the anti-coronavirus potential and the corresponding mechanisms of the two ingredients of Reduning Injection: quercetin and luteolin.@*METHODS@#A pseudovirus system was designed to test the efficacy of quercetin and luteolin to inhibit SARS-CoV-2 infection and the corresponding cellular toxicity. Luteolin was tested for its activities against the pseudoviruses of SARS-CoV-2 and its variants. Virtual screening was performed to predict the binding sites by Autodock Vina 1.1.230 and PyMol. To validate docking results, surface plasmon resonance (SPR) was used to measure the binding affinity of the compounds with various proteins of the coronaviruses. Quercetin and luteolin were further tested for their inhibitory effects on other coronaviruses by indirect immunofluorescence assay on rhabdomyosarcoma cells infected with HCoV-OC43.@*RESULTS@#The inhibition of SARS-CoV-2 pseudovirus by luteolin and quercetin were strongly dose-dependent, with concentration for 50% of maximal effect (EC50) of 8.817 and 52.98 µmol/L, respectively. Their cytotoxicity to BHK21-hACE2 were 177.6 and 405.1 µmol/L, respectively. In addition, luetolin significantly blocked the entry of 4 pseudoviruses of SARS-CoV-2 variants, with EC50 lower than 7 µmol/L. Virtual screening and SPR confirmed that luteolin binds to the S-proteins and quercetin binds to the active center of the 3CLpro, PLpro, and helicase proteins. Quercetin and luteolin showed over 99% inhibition against HCoV-OC43.@*CONCLUSIONS@#The mechanisms were revealed of quercetin and luteolin inhibiting the infection of SARS-CoV-2 and its variants. Reduning Injection is a promising drug for COVID-19.

Inhibition of abnormal GRK2 transmembrane transfer to reduce maturation of dendritic cells in CIA mice by regulating JAK1-STAT1 signaling pathway / 中国药理学通报

Institute of Clinical Pharmacology of Anhui Medical University, Key Laboratory of And-inflammatory and Immune Medicine, Ministry of Education .Anhui Collaborative Innovation Center of And-Inflammatory and Immune Medicine, Rheumatoid Arthritis Research Center of Anhui Medical University, Jlefei ,230032, China,Aim To reveal the role of the abnormal activation of G protein-coupled receptor kinase 2(GRK2)and abnormal signal transduction of JAK1-STAT1 in the abnormal immune response of rheumatoid arthritis(RA)by exploring the effects of GRK2 on the JAK1-STAT1 signaling pathway in dendritic cells(DCs)of collagen-induced arthritis(CIA)mice and the undelying mechanisms,so as to provide a basis for revealing the new mechanism of RA.Methods The CIA model was established,and the co-stimulatory molecular level of DCs was detected by flow cytometry,the cytokine levels of plasma in mice were detected by ELISA,and the expression of p-JAK1,p-STAT1 and GRK2 in spleen tissues was detected by immunohistochemistry.Bone marrow cells were induced into DCs in vitro and stimulated with IFN-α and PGE2 for 48 h.Flow cytometry was used to detect the level of co-stimulatory molecules and phagocytosis of DCs,and ELISA to detect the level of cytokines in cell supernatant.CO-IP was employed to detect the co-localization of GRK2 and JAK1 in DCs.Western blotting was used to detect the expression of JAK1-STAT1 and the cell membrane expression of GRK2.Imaging flow cytometry was applied to detect the nucleation rate of p-STAT1.Results In vivo the level of co-stimulatory molecules of dendritic cells of CIA mouse increased,and the expression of GRK2 and p-JAK1,p-STAT1 in spleen was positively correlated.The co-localization of GRK2 and JAK1 in spleen of the CIA group decreased significantly.In vitro GRK2 inhibitors reduced the level of costimulatory molecules,cytokines IL-6 and TNF-α,the expression of JAK1 and STAT1,the expression of GRK2 in the cell membrane,and the rate of p-STAT1 nuclear translocation,and increased the Ag uptake capacity of DCs and the co-localization rate of GRK2 and JAK1.Conclusions The abnormal GRK2 transfer to the cell membrane in DCs mediates the maturation of DCs and the activation of the JAK1-STAT1 signaling pathway.Inhibition of GRK2 transfer membrane can restore its control of the JAK1-STAT1 signal transduction of DCs,reduce the maturation of DCs,and play an important role in improving mouse CIA.

Effect of paeoniflorin-6'-O benzene sulfonate on JAK1/STAT3 signaling pathway in rat macrophages with collagen-induced arthritis through regulating G protein coupled receptor kinase 2 / 中国药理学通报

Aim To investigate the inhibitory effect of peonia-6'-o benzene sulfonate (CP-25) on the JAK1/STAT3 signaling pathway in collagen-induced arthritis (CIA) rats macrophages through regulating G protein coupled receptor kinase 2 (GRK2). Methods SD rats were randomly divided into four groups; normal group, model group, CP-25 (50 mg · kg

Protocol of Jidong Women Health Cohort Study: Rationale, Design, and Baseline Characteristics / 生物医学与环境科学（英文）

OBJECTIVE@#The Jidong Women Health Cohort Study is a prospective cohort study on female-specific characteristics and risks of chronic diseases in Chinese women and focuses on the potential association between menopause and risks of cardiovascular disease (CVD).@*METHODS@#The study includes 4,179 female participants with an age of older than 18 years from Caofeidian district, Tangshan city, northern China. Baseline information on female-specific characteristics and potential cardiovascular risk factors was collected and all the participants underwent a physical examination with blood samples collected in 2013. To establish a better risk assessment tool of female CVD, updated information from questionnaire investigation, physical examinations and occurrence of outcome events will be collected through a longitudinal follow-up annually up to the year 2024.@*RESULTS@#At baseline, Mean age of the participants was 42.3 ± 12.8 years. Reproduction occurred in 2,948 participants (70.5%), menopausal transition in 173 (4.3%), and postmenopause in 1,058 (25.3%). The incidence of arterial hypertension, dyslipidemia, and diabetes showed significant difference across different groups stratified by Stage of Reproductive Aging Workshop (STRAW) system (P < 0.05).@*CONCLUSION@#The Jidong Women Health Cohort Study will contribute to the scientific evidence on association between female-specific characteristics and cardiovascular risks, and will also be helpful to provide a new path for early detection and prevention of CVD.

Protective effect of Angelica sinensis polysaccharides on subacute renal damages induced by D-galactose in mice and its mechanism / 中国中药杂志

To explore the protective effect of Angelica sinensis polysaccharides(ASP) on subacute renal damages induced by D-galactose in mice and its mechanism. Male C57BL/6J mice were randomly divided into 3 groups, with 10 mice in each group. The D-galactose model group was subcutaneously injected with D-galactose (120 mg x kg(-1)), qd x 42; the ASP + D-galactose model group was intraperitoneally injected with ASP since the 8th day of the replication of the D-galactose model, qd x 35; and the normal control group was subcutaneously injected with saline at the same dose and time. On the 2nd day of after the injection, the peripheral blood was collected to measure the content of BUN, Crea, UA, Cys-C; paraffin sections were made to observe the renal histomorphology by HE staining; senescence-associated β-g-alactosidase (SA-β-Gal) stain was used to observe the relative optical density (ROD) in renal tissues; transmission electron microscopy was assayed to observe the renal ultrastructure; the renal tissue homogenate was prepared to measure the content of SOD, GSH-PX, MDA; the content of AGEs and 8-OH-dG were measured by ELISA. According to the result, compared with the D-galactose model group, the ASP + D-galactose model group showed obviously decreases in the content of BUN, Crea, UA, Cysc, AGES, 8-OH-dG, the number of hardening renal corpuscle, renal capsular space and renal tubular lumen, ROD of SA-β-Gal staining positive kidney cells, mesangial cells, basement membrane thickness, podocyte secondary processes fusion and MDA and increases in the number of normal renal corpuscle, ribosome and rough endoplasmic reticulum in podocytes, the activity of SOD and GSH-PX. In Conclusion, A. sinensis polysaccharides can antagonize kidney subacute damages induced by D-galactose in mice. Its protective mechanism may be correlated with the inhibition of the oxidative stress injury.

Biological mechanisms of human-derived leukemia stem cells senescence regulated by Angelica sinensis polysaccharide / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.</p><p><b>METHOD</b>Acute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.</p><p><b>RESULT</b>The purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.</p><p><b>CONCLUSION</b>ASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.</p>

Effect of combined administration of Angelica polysaccharide and cytarabine on liver of human leukemia NOD/SCID mouse model / 中国中药杂志

Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.

Research of anti-aging mechanism of ginsenoside Rg1 on brain / 中国中药杂志

Neurodegenerative disease is common and frequently occurs in elderly patients. Previous studies have shown that ginsenoside Rg1 was able to inhibit senescent of brain, but the mechanism on the brain during the treatment remains elucidated. To study the mechanism of ginsenoside Rg1 in the process of anti-aging of brain, forty male SD rats were randomly divided into normal group, Rg1 normal group, brain aging model group and Rg1 brain aging model group, each group with 10 rats (brain aging model group: subcutaneous injection of D-galactose (120 mg kg(-1)), qd for 42 consecutive days; Rg1 brain aging model group: while copying the same test as that of brain aging model group, begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Rg1 normal group: subcutaneous injection of the same amount of saline; begin intraperitoneal injection of ginsenosides Rg1 (20 mg x kg(-1)) qd for 27 d from 16 d. Normal: injected with an equal volume of saline within the same time. Perform the related experiment on the second day after finishing copying the model or the completion of the first two days of drug injections). Learning and memory abilities were measured by Morris water maze. The number of senescent cells was detected by SA-beta-Gal staining while the level of IL-1 and IL-6 proinflammatory cytokines in hippocampus were detected by ELISA. The activities of SOD, contents of GSH in hippo- campus were quantified by chromatometry. The change of telomerase activities and telomerase length were performed by TRAP-PCR and southern blotting assay, respectively. It is pointed that, in brain aging model group, the spatial learning and memory capacities were weaken, SA-beta-Gal positive granules increased in section of brain tissue, the activity of antioxidant enzyme SOD and the contents of GSH decreased in hippocampus, the level of IL-1 and IL-6 increased in hippocampus, while the length of telomere and the activity of telomerase decreased in hippocampus. Rats of Rg1 brain aging group had their spatial learning and memory capacities enhanced, SA-beta-Gal positive granules in section of brain tissue decreased, the activity of antioxidant enzyme SOD and the contents of GSH increased in hippocampus, the level of IL-1 and IL-6 in hippocampus decreased, the length contraction of telomere suppressed while the change of telomerase activity increased in hippocampus. Compared with that of normal group, the spatial learning and memory capacities were enhanced in Rg1 normal group, SA-beta-Gal positive granules in section of brain tissue decreased in Rg1 normal group, the level of IL-1 and IL-6 in hippocampus decreased in Rg1 normal group. The results indicated that improvement of antioxidant ability, regulating the level of proinflammatory cytokines and regulation of telomerase system may be the underlying anti-aging mechanism of Ginsenoside Rg1.

Reversion effects of curcumin on multidrug resistance of MNNG/HOS human osteosarcoma cells in vitro and in vivo through regulation of P-glycoprotein / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>P-glycoprotein (P-gp) encoded by ATP-binding cassette sub-family B member 1 (ABCB1) gene is a kind of ATP-dependent drug transporter, which plays important roles in multidrug resistance (MDR) of human cancers, such as osteosarcoma. Curcumin is a natural phenolic coloring compound originating from the rhizomes of Curcuma longa, which is proved to possess antitumor biological activities including reversion of MDR. However, the effect and molecular mechanisms of curcumin to osteosarcoma MDR remain unclear.</p><p><b>METHODS</b>We established a human osteosarcoma drug-resistant cell line MNNG/HOS/MTX by pulse exposure to methotrexate (MTX) and verified that the new cell lines were cross-resistant to other anticancer agents. Then, according to the cytotoxicity assay, we reversed MDR of MNNG/HOS/MTX by 30 µmol/L curcumin, and detected the mechanisms of curcumin reversing MDR through Real-time PCR, Western blotting assay, and Rhodamine123 (Rh123) transport test. Finally, we evaluated the effect of curcumin reversing MDR in vivo by MNNG/HOS/MTX cells xenograft-nude mice model.</p><p><b>RESULTS</b>MNNG/HOS/MTX was proved to be a human osteosarcoma MDR cell line. MTT tumor chemosensitivity test indicates that 30 µmol/L curcumin attenuates the half maximal inhibitory concentration (IC50) and resistance index (RI) to MTX, diamminedichloroplatinum (DDP), adriamycin (ADM), ifosfamide (IFO), and epirubicin (EPI) in MNNG/HOS/MTX cells (P < 0.05). Real-time PCR and Western blotting assays demonstrated that curcumin down-regulated P-gp expression of MNNG/HOS/MTX cells. Rh123 transport test showed that curcumin inhibited the transport function of P-gp in vitro. In vivo studies showed that curcumin displayed the features of sensitizing antitumor drugs and inhibiting the proliferation, invasion, and metastasis of osteosarcoma MDR cells.</p><p><b>CONCLUSION</b>Down-regulation of P-gp and inhibition of the function of P-gp efflux pump may contribute to MDR reversion induced by curcumin in vitro and in vivo.</p>

A case-control study on the duration of sleeping and cerebral infarction / 中华流行病学杂志

Objective To explore the relationship between duration of sleeping and cerebral infarction.Methods A case-control study involved 1037 cerebral infarction patients admitted by the Second Affiliated Hospital of Harbin Medical University,December 2011-December 2012 as cases.Another 1205 adults free from cerebro-vascular diseases who had undergone physical examination in the hospital at the same period,were served as controls.All the subjects were interviewed with unified questionnaire.Chi-square test,u-test and multivariate logistic regression analysis were performed.Results After adjustment for potential confounding factors including age,sex,body mass index,wrist-hip ratio,smoking,alcohol intake,hypertension,diabetes mellitus,coronary artery disease and lipid parameters,data from the multivariate logistic regression analysis showed that the risk of cerebral infarction was greater in people who slept less than 6 hours per night than those who slept between 6 hours and 8 hours per night,with an odds ratio (95％ CI) as 2.81 (95％ CI:1.68-4.70).There was no significant association between factor as ‘sleeping longer than 8 hours/pre day' and cerebral infarction.Through the subgroup analysis,data showed that the association between ‘ shorter than 6 hour sleep/night' and cerebral infarction consistently exsited,across the categories of sex,and the degree of association was greater in women than in men,with the odds ratio as 5.58 (95％CI:1.78-17.52) and 2.00(95％CI:1.10-3.64) respectively.Conclusion Short sleeping duration might increase the risk of developing cerebral infarction.

Expression of bone morphogenetic protein-4 mRNA in the cervical posterior longitudinal ligament after decompression / 中国组织工程研究

BACKGROUND: It has been thought that posterior cervical decompression increases the risk of cervical instability and the stress on posterior longitudinal ligament may change growth factor expression in vivo. However, the association between BMP-4 expression and heterotopic bone formation has not been confirmed. Bone morphogenetic protein 4 (BMP-4) is one of factors that can lead to heterotopic bone formation alone.OBJECTIVE: To observe the effects of posterior cervical decompression on BMP-4 mRNA expression in the posterior longitudinal ligament.DESIGN, TIME AND SETTING: A randomized controlled animal experiment was performed in Taishan Medical College between January 2005 and December 2006.MATERIALS: A total of 48 adult Sprague Dawley rats of either gender and of clean grade, weighing 270-350 g, were included in this study.METHODS: All rats were randomly and evenly divided into 3 groups: experimental, sham-operated, and blank control. In the experimental group, C36 laminectomy for decompression was performed.In the sham-operated group: only skin incision was made. Rats in the blank control group received no any treatment. Four rats were allocated for each time point (1, 2, 4, and 8 weeks post-surgery) for harvesting posterior longitudinal ligament.MAIN OUTCOME MEASURES: Detection of BMP-4 mRNA expression in the cervical posterior longitudinal ligament by semi-quantitative reverse transcription-polymerase chain reaction.RESULTS: In the experimental group, obvious BMP-4 mRNA expression was found at 1-8 weeks post-surgery. While no BMP-4 mRNA expression was found at all times in the sham-operated and blank control groups.CONCLUSION: Posterior cervical decompression can increase BMP-4 mRNA expression in the posterior longitudinal ligament.Endogenous BMP-4 may contribute to ligamentous ossification and development post-surgery.