ObjectiveTo investigate the material basis of the pathogenesis of cerebral ischemic injury with blood stasis and toxin syndrome and to explore the protective effects of Huayu Jiedu prescription （HYJDP） on neutrophil extracellular trap-related cell death （NETosis） in cerebral ischemic injury following acute cerebral infarction. MethodsSeventy-two Sprague-Dawley （SD） rats were randomly divided into six groups （n=12 per group）： sham operation （Sham） group， blood stasis and toxin model （Model） group， low-， medium-， and high-dose HYJDP groups （HYJDP-L， HYJDP-M， and HYJDP-H； 9， 18， and 36 g·kg^-1， respectively）， and butylphthalide （NBP） group （0.06 g·kg^-1）. Except for the Sham group， rats in all other groups were subjected to carrageenan/dry yeast combined with a modified intraluminal filament method to establish a focal cerebral ischemia model of the middle cerebral artery with blood stasis and toxin syndrome. Neurological function was evaluated at 24 h after modeling using the Zea-Longa neurological deficit score. Cerebral infarction rate was assessed by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Pathological morphology of brain tissue was observed using hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was used to determine serum levels of interleukin-8 （IL-8）， myeloperoxidase-DNA complexes （MPO-DNA）， and citrullinated histone H3 （CitH3）. Protein expression of phosphorylated phosphatidylinositol 3-kinase （p-PI3K）， protein kinase B （p-Akt）， mammalian target of rapamycin （p-mTOR）， sequestosome 1 （p62）， and CitH3 in brain tissue was detected by Western blot. Immunofluorescence （IF） was used to detect the expression of neutrophil-specific marker Ly6G， CitH3， and neuron-specific nuclear protein （NeuN） in brain tissue. ResultsCompared with the Sham group， neurological deficit scores and cerebral infarction rates in the model group were significantly increased （P<0.01 for both）. HE staining showed varying degrees of neuronal degeneration and necrosis， characterized by blurred neuronal structures， nuclear pyknosis and fragmentation， cytoplasmic dissolution into a vacuolated reticular pattern， and mild glial cell proliferation. ELISA results showed that serum levels of IL-8， MPO-DNA， and CitH3 were significantly increased （P<0.01）. Western blot analysis demonstrated decreased expression of p-PI3K， p-Akt， p-mTOR， and p62， while CitH3 expression was significantly increased （P<0.01）. IF results showed an increased number of NETs⁺ cells and a significant decrease in NeuN⁺ cells （P<0.01）. Compared with the Model group， neurological deficit scores in the HYJDP-H group were significantly decreased （P<0.05）， and cerebral infarction rates in the HYJDP-H and NBP groups were significantly reduced （P<0.01）. HE staining showed that brain tissue damage was markedly alleviated in the HYJDP-H group. ELISA results showed that levels of IL-8， MPO-DNA， and CitH3 were significantly decreased in the HYJDP-M， HYJDP-H， and NBP groups （P<0.01）. Western blot analysis showed that expression of p-PI3K， p-Akt， p-mTOR， and p62 was significantly increased in the HYJDP-H and NBP groups， while CitH3 expression was significantly reduced in all drug intervention groups （P<0.01）. IF results showed that the number of NETs⁺ cells was significantly decreased and the number of NeuN⁺ cells was significantly increased in all drug intervention groups （P<0.01）. ConclusionNETs may be the material basis of the pathogenesis of cerebral ischemic injury characterized by blood stasis and toxin. HYJDP can regulate the PI3K/Akt/mTOR signaling pathway， reduce the release of pro-inflammatory mediators and NETosis-related products， alleviate cerebral ischemic injury caused by autophagy-dependent NETosis， and thereby exert a neuroprotective effect.

Objective To investigate the effects of Zuogui Jiangtang Tongmai Prescription on short-chain fatty acids(SCFAs)and G protein-coupled receptor 109A(GPR109A)in diabetic atherosclerotic mice;To explore its mechanism of improving diabetic atherosclerosis.Methods ApoE-/-mice were fed with high sugar and high fat diet and intraperitoneal injection of streptozotocin to establish atherosclerosis model of diabetes.After modeling,the mice were randomly divided into the model group,Western medicine group(metformin hydrochloride+atorvastatin),and Zuogui Jiangtang Tongmai Prescription low-,medium-and high-dosage groups(19,38,76 g/kg);and the other C57BL/6J mice were set as the control group,with 6 mice in each group.Each group was given solution for gavage,once a day for 4 weeks.Blood glucose of the mice were detected,HE staining was used to observe the morphology of the aorta,TG,TC,LDL-C and HDL-C contents were detected by fully automated biochemistry analyzers,ELISA was used to detect the contents of serum HbA1c,fasting insulin(FINS),interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α,gas chromatography was used to detect the content of intestinal SCFAs,and RT qPCR and Western blot were used to detect the mRNA and protein expressions of GPR109A in ileal tissue and nuclear factor(NF)-κB p65 in aortic tissue,respectively.Results Compared with the control group,obvious plaques and inflammatory cells infiltration were seen in the aorta of the model group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents increased(P<0.01),FINS and HDL-C content decreased(P<0.01),intestinal acetate,propionate,butyric acid,isobutyric acid and isovaleric acid contents decreased(P<0.01),GPR109A mRNA and protein expression in ileal tissue decreased,NF-κB p65 mRNA and protein expression in aortic tissue increased(P<0.01).Compared with the model group,the aortic plaque area and inflammatory cells infiltration were significantly improved in each drug intervention group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents decreased(P<0.05,P<0.01);the FINS and HDL-C content increased(P<0.05,P<0.01),intestinal acetic,propionic,butyric acid,isobutyric acid and isovaleric acid contents increased(P<0.05,P<0.01),GPR109A mRNA and protein expression in ileal tissue increased and NF-κB p65 mRNA and protein expression in ileal tissue decreased decreased(P<0.05,P<0.01).Conclusion Zuogui Jiangtang Tongmai Prescription can improve glucolipid metabolism and inflammatory response in diabetic atherosclerotic mice,which may be related to regulating SCFAs/GPR109A pathway.

Objective:A bidirectional Mendelian randomization approach was applied to dissect the causal association between dairy product consumption and osteoarthritis (OA), so to offering novel insights for the prevention, treatment, and prognosis assessment of knee osteoarthritis (KOA).Methods:A two-sample Mendelian randomization (MR) analysis was conducted using public available genome-wide association studies. The primary analysis was performed using the inverse variance weighted (IVW) method. To assess the stability and reliability of the results, we compared IVW and MR-Egger regression using Cochran′s Q test to evaluate SNP heterogeneity. MR-Egger regression was employed to assess pleiotropy, while a leave-one-out approach was used for sensitivity analysis. Results:Based on 207 instrumental variables (IVs), two-sample MR analysis revealed that for each standard deviation increase in genetically predicted cheese intake, the risk of OA was significantly negatively associated [ OR(95% CI)=0.98(0.98, 0.99), P<0.001], so did the risk of KOA [ OR(95% CI)=0.73(0.63, 0.85), P<0.001]. However, no significant association was found between OA risk and the intake of full-fat milk (30 IVs), semi-skimmed milk (15 IVs), skimmed milk (32 IVs), soy milk (25 IVs), or other types of milk (13 IVs), yogurt (11 IVs), or butter (3 IVs). Conclusion:The two-sample MR analysis revealed causally inverse associations between cheese intake and OA as well as KOA.

Chronic liver diseases with common causes including viral infections， alcohol abuse， and autoimmune diseases. Alkaloids， as a class of plant-derived compounds， have shown significant potential in regulating chronic liver diseases. Recent studies have shown that alkaloids are able to exert a therapeutic effect on chronic liver diseases through multiple pathways. These compounds have a regulatory effect on key pathological processes such as liver fibrosis， inflammatory response， oxidative stress， and cell apoptosis， and they also regulate the metabolic homeostasis of hepatocytes by modulating multiple signaling pathways， thereby playing a role in regulating chronic liver diseases. This article reviews the role and mechanism of alkaloids in the treatment of chronic liver diseases， in order to provide new ideas and directions for the treatment of chronic liver diseases.

[Objective] To analyze the infection status of hepatitis E virus (HEV) among blood donors in Zhengzhou, so as to provide data support for formulating local blood screening strategies. [Methods] Random samples from blood donors from January to December 2022 were tested for HEV RNA using PCR technology. Reactive samples were sequenced for gene analysis, and the donors were followed up. [Results] Among 21 311 samples, 3(0.14‰) were reactive for HEV RNA, all of whom were male. Genetic sequencing results revealed that one strong positive sample was genotype 4, while sequencing failed for the other two due to low viral load. A follow-up of 25 strong positive donors showed that ALT significantly increased on day 7 after donation, anti-HEV IgM and anti-HEV IgG turned positive. On day 21, ALT returned to normal, and on day 35, HEV RNA turned negative. Notably, anti-HEV IgM and anti-HEV IgG persisted until day 482. [Conclusion] There is HEV infection among blood donors in Zhengzhou, and it is necessary to expand the screening scope to comprehensively explore the prevalence and genotype distribution of HEV among blood donors.

ObjectiveTo explore the regulation of hypothalamic-pituitary-gonadal （HPG） axis by modified Erxian decoction in rats with perimenopausal syndrome （PMS） and to further analyze the expression of proteins related to the silent information regulator 1 （SIRT1）/hypothalamic kisspeptin （Kisspeptin）/gonadotropin-releasing hormone （GnRH） signaling pathway in the arcuate nucleus region （ARC） of the hypothalamus， so as to reveal the potential target of action and molecular biological mechanism of modified Erxian decoction for the treatment of perimenopausal syndrome. MethodsAn animal model was established via the incomplete castration method， with successful modeling confirmed by the exfoliated cervical cell smear method. The 48 rats were divided into six groups based on the randomization principle after successful modeling， including a sham operation group， a model group， an estradiol valerate group （0.09 mg∙kg^-1∙d^-1）， high-， medium-， and low-dose modified Erxian decoction groups （7.614， 3.807，1.903 5 g∙kg^-1∙d^-1）， with 8 rats in each group. The estradiol valerate group and the high-， medium- and low-dose modified Erxian decoction groups were continuously administered by gavage for 28 days， and the indicators were detected 24 hours after the last administration. Body weights and uterine indices were measured. The pathological changes of the uterus were observed by hematoxylin-eosin （HE） staining. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the levels of estradiol （E₂）， follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and gonadotropin-releasing hormone （GnRH）. Real-time quantitative polymerase chain reaction （Real-time PCR） and Western blot were used to determine the expression levels of SIRT1， Kisspeptin， kisspeptin receptor （GPR54）， and GnRH in the ARC region of the hypothalamus and gonadotropin-releasing hormone receptor （GnRH-R） in pituitary. ResultsCompared with the sham operation group， rats in the model group had a significantly increased body weight （P0.01）， reduced wet weight and index of uterus （P0.01）， endometrial thinning or atrophy， glandular atrophy， and a decreasing number of glands. Additionally， serum levels of E₂ and the expression of SIRT1 in the ARC region of the hypothalamus significantly decreased （P0.01）. Serum levels of FSH， LH， and GnRH， the expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus， and GnRH-R in pituitary significantly increased （P0.01）. Compared with the model group， the estradiol valerate group and the high-， medium-dose modified Erxian decoction groups had significantly reduced body weight， serum levels of FSH， LH， and GnRH， and expression of Kisspeptin， GPR54， and GnRH in the ARC region of the hypothalamus and GnRH-R in pituitary （P0.05， P0.01） and significantly increased wet weight and index of uterus， serum level of E₂， and expression of SIRT1 in the ARC region of the hypothalamus （P0.05， P0.01）. In addition， they showed thickened endometrium， increased number of endometrial glands， and improved glandular atrophy. ConclusionModified Erxian decoction regulates the function of the HPG axis through multi-targets， and its mechanism of action may be related to the up-regulation of the expression of SIRT1 in the ARC region of the hypothalamus， the inhibition of the over-activation of the Kisspeptin/GnRH signaling pathway， the regulation of the expression of GnRH-R in the pituitary， the restoration of secretion balance of gonadotropins， and the elevation of the estrogen level. This study provides an experimental basis for the interpretation of the scientific connotation of modified Erxian decoction in the treatment of perimenopausal syndrome and a theoretical reference for the development of a novel therapeutic strategy based on the SIRT1/Kisspeptin/GnRH pathway.

Objective To investigate the effects of Zuogui Jiangtang Tongmai Prescription on short-chain fatty acids(SCFAs)and G protein-coupled receptor 109A(GPR109A)in diabetic atherosclerotic mice;To explore its mechanism of improving diabetic atherosclerosis.Methods ApoE-/-mice were fed with high sugar and high fat diet and intraperitoneal injection of streptozotocin to establish atherosclerosis model of diabetes.After modeling,the mice were randomly divided into the model group,Western medicine group(metformin hydrochloride+atorvastatin),and Zuogui Jiangtang Tongmai Prescription low-,medium-and high-dosage groups(19,38,76 g/kg);and the other C57BL/6J mice were set as the control group,with 6 mice in each group.Each group was given solution for gavage,once a day for 4 weeks.Blood glucose of the mice were detected,HE staining was used to observe the morphology of the aorta,TG,TC,LDL-C and HDL-C contents were detected by fully automated biochemistry analyzers,ELISA was used to detect the contents of serum HbA1c,fasting insulin(FINS),interleukin(IL)-1β,IL-6 and tumor necrosis factor(TNF)-α,gas chromatography was used to detect the content of intestinal SCFAs,and RT qPCR and Western blot were used to detect the mRNA and protein expressions of GPR109A in ileal tissue and nuclear factor(NF)-κB p65 in aortic tissue,respectively.Results Compared with the control group,obvious plaques and inflammatory cells infiltration were seen in the aorta of the model group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents increased(P<0.01),FINS and HDL-C content decreased(P<0.01),intestinal acetate,propionate,butyric acid,isobutyric acid and isovaleric acid contents decreased(P<0.01),GPR109A mRNA and protein expression in ileal tissue decreased,NF-κB p65 mRNA and protein expression in aortic tissue increased(P<0.01).Compared with the model group,the aortic plaque area and inflammatory cells infiltration were significantly improved in each drug intervention group,the blood glucose and serum HbA1c,TG,TC,LDL-C,IL-1β,IL-6 and TNF-α contents decreased(P<0.05,P<0.01);the FINS and HDL-C content increased(P<0.05,P<0.01),intestinal acetic,propionic,butyric acid,isobutyric acid and isovaleric acid contents increased(P<0.05,P<0.01),GPR109A mRNA and protein expression in ileal tissue increased and NF-κB p65 mRNA and protein expression in ileal tissue decreased decreased(P<0.05,P<0.01).Conclusion Zuogui Jiangtang Tongmai Prescription can improve glucolipid metabolism and inflammatory response in diabetic atherosclerotic mice,which may be related to regulating SCFAs/GPR109A pathway.

Objective:A bidirectional Mendelian randomization approach was applied to dissect the causal association between dairy product consumption and osteoarthritis (OA), so to offering novel insights for the prevention, treatment, and prognosis assessment of knee osteoarthritis (KOA).Methods:A two-sample Mendelian randomization (MR) analysis was conducted using public available genome-wide association studies. The primary analysis was performed using the inverse variance weighted (IVW) method. To assess the stability and reliability of the results, we compared IVW and MR-Egger regression using Cochran′s Q test to evaluate SNP heterogeneity. MR-Egger regression was employed to assess pleiotropy, while a leave-one-out approach was used for sensitivity analysis. Results:Based on 207 instrumental variables (IVs), two-sample MR analysis revealed that for each standard deviation increase in genetically predicted cheese intake, the risk of OA was significantly negatively associated [ OR(95% CI)=0.98(0.98, 0.99), P<0.001], so did the risk of KOA [ OR(95% CI)=0.73(0.63, 0.85), P<0.001]. However, no significant association was found between OA risk and the intake of full-fat milk (30 IVs), semi-skimmed milk (15 IVs), skimmed milk (32 IVs), soy milk (25 IVs), or other types of milk (13 IVs), yogurt (11 IVs), or butter (3 IVs). Conclusion:The two-sample MR analysis revealed causally inverse associations between cheese intake and OA as well as KOA.

Objective To analyze the research trends and frontier of TCM regulation of NF-κB;To provide reference for related research.Methods Relevant literature about TCM regulation of NF-κB was retrieved from CNKI,VIP,Wanfang Data,CBM and Web of Science from January 1,2013 to December 31,2022.VOSviewer 1.6.19 and CiteSpace 6.2.R4 software were used for visualization analysis on authors,institutions and keywords.Results Totally 3 728 articles in Chinese and 995 in English were included,and the number of articles was on the rise.The Chinese and English articles involved 487 and 237 core authors,respectively,forming research teams represented by Yan Guanghai,Liu Jian,Wang Li,and Li Wei,Zhang Li,Zhang Yu,etc.There were 7 and 8 effective clusters in Chinese and English articles respectively.Keyword analysis showed that this research field mainly focused on diseases(inflammatory diseases,tumors,cardiovascular and cerebrovascular diseases,etc.),research methods(in vivo experiment,in vitro experiment)and intervention methods(acupuncture,TCM monomer,TCM compound,etc.).Conclusion TCM regulation of NF-κB mainly focuses on related diseases and intervention methods,and it is a research trend to find drug action targets and conduct experimental verification through network pharmacology and molecular docking technology.

Objective:To evaluate the effectiveness and safety of mesenchymal stem cells (MSCs) in the treatment of knee osteoarthritis (KOA).Methods:The databases PubMed, OVID, Web of Science, CNKI, Wanfang, were systematically searched from inception to May 2023 to collect randomized control trials(RCTs) of MSCs in the treatment of KOA. The literature was selected according to the inclusion and exclusion criteria, and relevant data was extracted. Meta-analysis was conducted using RevMan 5.4 software. Heterogeneity was assessed using the I2 statistic, with the fixed effects model applied when I2 was less than 50%, and the random effects model utilized otherwise. The combined effect size and 95% confidence interval ( CI) were calculated using the inverse-variance method. Sensitivity analysis and assessment of publication bias were performed using Stata 14.0 software. Results:A total of 29 RCTs involving 1 402 participants were included. The outcomes showed that at the 12 month follow-up, MSCs reduced pain [WOMAC pain: MD(95% CI)=-4.38(-7.22, -1.55), P<0.001; VAS: MD(95% CI)=-2.00(-2.67, -1.33), P<0.001 ]. And WOMAC stiffness[WOMAC stiffness:MD(95% CI)=-1.21(-2.32, -0.10), P<0.001]; moreover, MSCs reduced KOA severity and Restored joint function, [WOMAC: MD(95% CI)=-9.40(-15.87,-2.93), P<0.001; Lequesne: MD(95% CI)=-10.57(-15.89, -5.24), P<0.001 ].And the effect lasted for at least 4 years. MRI analysis showed no significant difference between the MSC group and the control group [MD(95% CI)=-6.68 (-18.45, 5.09), P=0.270]. Subgroup analysis showed that there was no significant difference in the effects of the tissue source and dosage of MSC on osteoarthritis pain and joint function. Using WOMRS for subgroup analysis, we found that adipose tissue was the best MSCs source for cartilage repair in osteoarthritis [MD (95% CI) =-30.94(-43.87, -18.01), P<0.001]. For the occurrence of adverse events, no study had reported the occurrence of serious adverse events. Most of the adverse events were self-limited pain and discomfort, such as transient joint swelling and back pain. Conclusion:Based on current evidence, MSCs may be a safety therapy that have a good effect for OA, and the time to maintain the curative effect is no less than 4 years. There is currently no reliable evidence to address the issue of which tissue source and injection cell dosage yield the best therapeutic effect. High-quality clinical trials are needed.