Objective To investigate the mechanism by which Senegenin(SEN)alleviates microglial inflammatory response through the nuclear factor erythroid 2-related factor 2(Nrf2)/NOD-like receptor protein 3(NLRP3)pathway.Methods BV2 mouse microglia cells were randomly divided into control group,model group,SEN group and MCC950 group.Cells in control group were not treated,and cells in model group were added with 1 μg/ml lipopolysaccharide(LPS);Cells in SEN group were added with 1 μg/ml LPS+4 μmol/L SEN,and cells in MCC950 group were added with 1 μg/ml LPS+10 μmol/L MCC950 for 24 hours.CCK-8 method was used to detect the effect of different concentrations of SEN on the viability of BV2 cells.Griess method was used to determine the release amount of nitric oxide(NO)in the supernatant.Real-time fluorescent quantitative PCR was used to determine the mRNA expression levels of NLRP3,lymphocyte apoptosis-associated spect-like protein containing a CARD(ASC),caspase-1,interleukin(IL)-1β and IL-18 mRNA.Immunofluorescence staining was used to detect the expression levels of ASC,IL-1β,Nrf2 and heme oxygenase-1(HO-1).Western blotting was used to detect the expression levels of NLRP3,caspase-1,ASC,IL-1β,IL-18,Nrf2,HO-1,nuclear factor kappa B(NF-κB)and inducible nitric oxide synthase(iNOS).Results The results of CCK-8 method showed that there was no significant difference in the viability of BV2 cells treated with 2～20 μmol/L SEN compared with control group(P>0.05).Compared with control group,the viability of BV2 cells in model group decreased significantly(P<0.05).Compared with model group,the viability of BV2 cells in 4 μmol/L SEN group was significantly restored(P<0.05).Compared with control group,the results of Griess method showed that the release amount of NO in cells of model group increased significantly(P<0.05);the results of real-time PCR showed that the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA in cells of model group increased significantly(P<0.05);the results of Western blotting showed that the protein expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 proteins in cells of model group increased significantly(P<0.05),and the immunofluorescence staining results showed that the expression levels of iNOS and NF-κB protein in cells of model group increased,and the expression levels of Nrf2 and HO-1 decreased,with statistically significant differences(P<0.05).Compared with model group,the release amount of NO in cells of SEN group and MCC950 group decreased,and the expression levels of NLRP3,ASC,caspase-1,IL-1β and IL-18 mRNA and proteins decreased,with statistically significant differences(P<0.05);in the SEN group,the expression levels of iNOS and NF-κB decreased,and immunofluorescence staining showed that Nrf2 was translocated into the nucleus,and the expression levels of Nrf2 and HO-1 proteins increased significantly,with statistically significant differences(P<0.05).Conclusions SEN could alleviate the inflammatory response of mouse microglia cells induced by LPS and inhibit the activation and expression of NLRP3 inflammasome,with an effect comparable to that of the inflammasome inhibitor MCC950.The mechanism may be related to the regulation of the expression of upstream factors Nrf2 and HO-1.

The growth of three plague phages from Qinghai Plateau in two Yersinia pestis strains(plague vaccine strains EV76 and 614F)and four non-Yersinia pestis strains(Yersinia pseudotuberculosis PTB3,PTB5,Escherichia coli V517,and Yersinia enterocolitica 52302-2)were detected through a micromethod based on the OmniLogTM microbial identification system and by the drop method,to provide a scientific basis for future ecological studies and classification based on the host range.For plague vaccine strains EV76 and 614F,successful phage infection and subsequent phage growth were observed in the host bacte-rium.Diminished bacterial growth and respiration and a concomitant decrease in color were observed with the OmniLogTM mi-crobial identification system at 33 ℃ for 48 h.Yersinia pseudotuberculosis PTB5 was sensitive to Yersinia pestis phage 476,but Yersinia pseudotuberculosis PST5 was insensitive to phage 087 and 072204.Three strains of non-Yersinia pestis(Yersinia pseudotuberculosis PTB3,Escherichia coli V517,and Yersinia enterocolitica 52302-2)were insensitive to Yersinia pestis pha-ges 087,072204,and 476 showed similar growth curves.The growth of phages 476 and 087,as determined with the drop method,in two Yersinia pestis strains(plague vaccine strains EV76 and 614F)and four non-Yersinia pestis strains(Yersinia pseudotuberculosis PTB3,Escherichia coli V517,and Yersin-ia enterocolitica 52302-2)showed the same results at 37 ℃,on the basis of comparisons with the OmniLogTM microbial i-dentification system;in contrast,phages 072204 did not show plaques on solid medium at 37 ℃ with plague vaccine strains EV76 and 614F.Determination based on the OmniLogTM detection system can be used as an alternative to the traditional determination of the host range,thus providing favorable application val-ue for determining the interaction between the phage and host bacteria.

This study aims to explore the neuroprotective effect of bilobalide(BB) and the mechanisms such as inhibiting inflammatory response in macrophage/microglia, promoting neurotrophic factor secretion, and interfering with the activation and differentiation of peripheral CD4~+ T cells. BB of different concentration(12.5, 25, 50, 100 μg·mL~(-1)) was used to treat the RAW264.7 and BV2 cells for 24 h. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay and cell counting kit-8(CCK-8) were employed to detect the cytotoxicity of BB and appropriate concentration was selected for further experiment. Lipopolysaccharide(LPS) was applied to elicit inflammation in RAW264.7 and BV2 cells, mouse bone marrow-derived macrophages(BMDMs), and primary microglia, respectively. The effect of BB on cell proliferation and secretion of inflammatory cytokines and neurotrophic factors was detected by enzyme-linked immunosorbent assay(ELISA). Spleen monocytes of C57BL/6 female mice(7-8 weeks old) were isolated, and CD4~+ T cells were separated by magnetic beads under sterile conditions. Th17 cells were induced by CD3/CD28 and the conditioned medium for eliciting the inflammation in BMDMs. The content of IL-17 cytokines in the supernatant was detected by ELISA to determine the effect on the activation and differentiation of CD4~+ T cells. In addition, PC12 cells were incubated with the conditioned medium for eliciting inflammation in BMDMs and primary microglia and the count and morphology of cells were observed. The cytoto-xicity was determined by lactate dehydrogenase(LDH) assay. The result showed that BB with the concentration of 12.5-100 μg·mL~(-1) had no toxicity to RAW264.7 and BV2 cells, and had no significant effect on the activity of cell model with low inflammation. The 50 μg·mL~(-1) BB was selected for further experiment, and the results indicated that BB inhibited LPS-induced secretion of inflammatory cytokines. The experiment on CD4~+ T cells showed that the conditioned medium for LPS-induced inflammation in BMDMs promoted the activation and differentiation of CD4~+ T cells, while the conditioned medium of the experimental group with BB intervention reduced the activation and differentiation of CD4~+ T cells. In addition, BB also enhanced the release of neurotrophic factors from BMDMs and primary microglia. The conditioned medium after BB intervention can significantly reduce the death of PC12 neurons, inhibit neuronal damage, and protect neurons. To sum up, BB plays a neuroprotective role by inhibiting macrophage and microglia-mediated inflammatory response and promoting neurotrophic factors.
Female ; Rats ; Mice ; Animals ; Bilobalides/pharmacology* ; Neuroprotection ; Lipopolysaccharides/toxicity* ; Culture Media, Conditioned/pharmacology* ; Mice, Inbred C57BL ; Macrophages/metabolism* ; Microglia ; Cytokines/metabolism* ; Nerve Growth Factors/pharmacology* ; Inflammation/metabolism*

Aim To explore the protective effect of proanthocyanidin B2 (PC-B2) on oxidative damage of PC 12 cells induced by hydrogen peroxide (H

Aim To explore the mechanism of grape seed proanthocyanidins (GSPs) targeting astrocytes (AS), so as to regulate the phenotype and function of AS and maximize their neuroprotective effect. Methods The effects of GSPs on the phenotype, secretion of pro-inflammatory factors and neurotrophic factors of Al AS induced by TNF-α, IL-1α and Clq were investigated by RT-PCR, Elisa and Western blot in vitro. And JNK phosphorylation was determined using Western blot. Results GSPs significantly reduced the expression of C3d and Clq of Al AS markers and inhibited the phosphorylation of JNK. Moreover, compared with the model group, GSPs could significantly inhibit the release of pro-inflammatory cytokines IL-6, IL-1 α, IL-17 and H

Objective: To understand the spatial characteristics of echinococcosis and associated factors in the pastoral area of Qinghai province, and provide evidence for the effective prevention and control of echinococcosis. Methods: The number of echinococcosis cases in the pastoral areas of Qinghai in 2019 was collected to perform spatial epidemiological analysis. The thematic map of the distribution of echinococcosis cases was generated with software ArcGIS 10.8 for visual analysis and spatial autocorrelation analysis. The spatial autocorrelation and spatial scanning analysis were performed to estimate the clustering of echinococcosis with software SaTScan 9.5. Software GeoDa 1.14 and ArcGIS 10.8 were used to establish spatial lag model and geographical weighted regression model to analyze the related factors of echinococcosis epidemic. Results: In 2019, the echinococcosis surveillance covered 64 741 people in the pastoral area of Qinghai, and 829 echinococcosis cases were found, with a prevalence rate of 1.28%. The distribution of the cases had spatial correlation (Moran's I=0.41, P<0.001). The most possible clustering areas indicated by spatial scanning analysis included Banma, Jiuzhi, Dari and Gande counties of Guoluo Tibetan Autonomous Prefecture (LLR=460.77, RR=9.20, P<0.001). The prevalence of echinococcosis in the pastoral areas was positively associated with the total annual precipitation (β=0.13, P=0.036), and negatively associated with population density (β=-1.36, P=0.019) and doctors/nurse ratio (β=-25.60, P=0.026). Conclusions: The distribution of echinococcosis cases in the pastoral areas of Qinghai in 2019 had spatial correlation, and the prevalence was affected by total annual precipitation, population density, and doctors/nurse ratio.
China/epidemiology* ; Cluster Analysis ; Echinococcosis/epidemiology* ; Epidemics ; Humans ; Prevalence ; Spatial Analysis

The advantages of western medical research were analyzed and the differences between skin nerve stimulation in western medicine and acupuncturemoxibustion in TCM were compared, so as to inspire the scientific researches of acupuncture and moxibustion in China. The related literature was searched and the research basis, content and achievements of the American stimulating peripheral activity to relief condition (SPARC) program were systematically summarized. From the perspectives of theoretical system, stimulation site, stimulation method and mechanism of action, the similarities and differences between skin nerve stimulation in western medicine and acupuncturemoxibustion in TCM were compared. Through comparative analysis, it is found that the systematic construction of SPARC "high-resolution neural circuit map" is essentially the upgraded version of the traditional distribution map of meridians and acupoints in China, which is similar to the research on origin of nervous system and stimulation site of acupuncturemoxibustion of TCM. Under the impact of "localization of traditional Chinese medicine" in other countries, learning from the international advanced research technology, gathering top-level talents, and encouraging openness and innovation will be the necessary pathway to improve the quality of acupuncturemoxibustion research and master the power of knowledge initiative.
Acupuncture ; education ; Acupuncture Points ; Acupuncture Therapy ; China ; Medicine, Chinese Traditional ; Meridians ; Moxibustion

Objective:To explore the neuroprotective effect and mechanism of Buyang Huanwu Tang (BYHWT) on experimental autoimmune encephalomyelitis (EAE) at different stages. Method:The 36 female C57BL/6 mice were immunized subcutaneously with myelin oligodendrocyte glycoprotein peptides (MOG_35-55),then randomly divided into 9, 17, 28 d EAE control group. Each BYHWT group was orally given drugs on the 3^rd day after immunization (50 g·kg^-1·d^-1), and EAE control group was given the same volume of normal saline in the same way once a day for 9, 17 and 28 d after immunization. The effect of BYHWT on EAE mice was observed with internationally accepted clinical score. Brain and spinal cord specimens were collected at 9, 17 and 28 d after immunization. The neuroprotective effect of BYHWT was observed by hematoxylin-eosin(HE)staining and solid blue staining (LFB). The expressions of BDNF and GAP-43 in spinal cord and brain were detected by Western blot. Result:After treatment, BYHWT can significantly inhibit myelitis cell infiltration and alleviate myelin loss. Compared with EAE group, the expression of Nogo-A in the spinal cord of each BYHWT group was significantly down-regulated (P<0.01), and the expression of BDNF in the spinal cord was significantly up-regulated (P<0.05, P<0.01) in the BYHWT group 17 and 28 d group compared with EAE group 17 and 28 d group. Compared with EAE 9, 17 d group, GAP-43 expression was significantly up-regulated in the spinal cord of BYHWT 9, 17 d group (P<0.01). Conclusion:BYHWT can improve the local nerve growth microenvironment and promote the expression of NTFs, reduce the expressions of neuroinhibitory factors, and play a role in neuroprotection.

Aim To investigate the expression of formyl peptide receptor-2 (FPR2) in lipopolysaccharide (LPS)-induced-BV-2 cells,and detect FPR2's influence on inflammatory response induced by LPS.Methods After 1 mg · L-1 LPS acting on BV-2 cells at 12 h,the extrinsic inflammatory model was established.We used the Western blot assay to test the levels of FPR2 protein.And the expressions of phosphorylated NF-κB,TNF-α and IL-1β were investigated when the LPS-induced-BV-2 was incubated with FPR2's agonist MMK-1 and antagonist Boc-2.Transwell assay was also used to detect the LPS-inducedBV-2 migration induced by MMK-1 and Boc-2.Resuits LPS up-regulated the expression of FPR2,and when its agonist was acted on LPS-induced-BV-2,the levels of phosphorylated NF-κB,TNF-α and IL-1β were significantly higher than those of LPS group.In addition,the chemotaxis of LPS-induced-BV-2 also increased by MMK-1.These effects were abolished by Boc-2.Conclusions LPS can increase the expression of FPR2 on BV-2 cells,and FPR2 enhances the inflammatory response induced by LPS.

Objective To observe the effects of acupuncture on the level of inflammatory cytokines and toll-like receptor 4 in hypothalamic paraventricular nucleus (PVN) of SHR; To investigate the mechanisms of acupuncture in reduction of mean arterial blood pressure (MAP) in SHR. Methods Thirty 10-week old SHR were randomly divided into SHR group, acupuncture group and non-acupoint group, with 10 rats in each group. 10 WKY rats were set as control group. Acupuncture group received bilateral acupuncture in "Taichong" acupoint, and twisting and diarrhea method was used to stimulate; non-acupoint group received acupuncture at the back of feet, and soothing and diarrhea with twisting method was used to stimulate. Materials were taken two weeks later. The mean arterial pressure of rats was detected every day; the expression of TLR4 mRNA in PVN was detected by RT-PCR; The expression of TLR4 protein in PVN was detected by Western blot; The levels of TNF-αa nd IL-6 were detected. Results Compared with control group, the mean arterial pressure of SHR group increased; TLR4 mRNA and protein expression in PVN increased significantly; the level of TNF-α and IL-6 increased significantly (P<0.05). Compared with SHR group, the mean arterial pressure of acupuncture group decreased significantly;TLR4 mRNA and protein expression in PVN decreased significantly; the level of TNF-α and IL-6 decreased significantly (P<0.05). There was no significant change in non-acupoint group. Conclusion Acupuncture in"Taichong" acupoint can attenuate blood pressure of SHR by inhibiting expression of TLR4 in PVN and reducing the levels of TNF-α and IL-6.