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OBJECTIVE@#Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro.@*METHODS@#The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining.@*RESULTS@#The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism.@*CONCLUSION@#Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.
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Objective:To compare the differences in dosimetric parameters of target areas between 125I seed implantation with degradable catheters and by hand under the assistance of a real-time intraoperative treatment planning system (TPS). Methods:Forty-two simulated lesions were divided into a degradable catheter group and a free-hand group, with twenty-one lesions in each group. 125I seeds were implanted according to the TPS.The pre-plan and post-implant dosimetric parameters were collected, including the minimum dose ( Dmin), maximum dose ( Dmax), mean dose ( Dmean), conformal index (CI), external index (EI), homogeneity index (HI), minimum prescription dose delivered to 90% of the target volume ( D90), and the mean percentage of volume receiving 90% of the prescription doses ( V90). The Bland-Altman method was employed to analyze the consistency of pre-plan and post-implant dosimetric parameters and the Wilcoxon rank-sum test was used for the comparison of the two groups. Results:According to the Bland-Altman analysis, the dosimetric parameters of the two groups were all in agreement before and after seed implantation except for the Dmin and V90 of the free-hand group.Furthermore, the degradable catheter group had smaller error ranges of Dmax ( Z=-3.824, P<0.005), CI ( Z=-1.962, P<0.005), HI ( Z=-2.352, P<0.005), D90 ( Z=-2.453, P<0.005), and V90 ( Z=-3.159, P<0.005). Conclusions:The dosimetric parameters of 125I seed implantation with degradable catheters under the assistance of real-time TPS have good pre-plan and post-implant consistency and smaller error ranges.
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Objective:To explore the relation between extended resection and prognosis of primary glioblastoma.Methods:A prospective study was performed. The subjects were from the colony of patients with primary glioblastoma who underwent surgical resection in our hospital from January 2014 from January 2017. Tumor volume and tumor residual volume showed by MR imaging T1 enhanced sequences (Group A and Group B), and tumor volume and tumor residual volume showed by fluid-attenuated inversion recovery (FLAIR) sequences (Group A1 and Group A2) were obtained within one week before surgery and 24 h after surgery, respectively; and the degrees of tumor resection were calculated. Cox proportional hazards regression model was used to determine factors influencing overall survival (OS) in glioblastoma patients. Kaplan-Meier method was used to plot the survival curves of these patients. Log-rank test was used to compare the survival rates of patients from Group A and Group B, and patients from Group A1 and Group A2.Results:A total of 128 patients were included; 71(55.5%) were into Group A, and 17 (13.3%) were into Group A1. Multivariate analysis showed that preoperative Karnofsky performance scale (KPS) scores, promoter methylation status of O-6-methylguanine-DNA methyltransferase (MGMT), tumor residual volume by T1 enhanced sequences, tumor residual volume by FLAIR sequences, and tumor resection degree by FLAIR sequences were independent influencing factors for OS ( P<0.05). Survival rate of patients from Group A was significantly higher than that in Group B( P<0.05); and survival rate of patients from Group A1 was significantly higher than that in Group A2 ( P<0.05). Conclusion:Residual of primary glioblastoma showed by FLAIR sequences is an important factor affecting the prognoses, and extended resection is necessary.
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Objective To explore the efficacy of difficult decompression and its influence factors in microvascular decompression for hemifacial spasm.Methods A retrospective analysis of clinical data and follow-up results of 458 patients who underwent microvascular decompression for hemifacial spasm in our hospital from October 2012 to December 2015 was performed.Aecording to whether there was narrow posterior fossa,compression of vertebral artery with perforating branch of brain stem or long vertebral artery,or vessel passing through cranial nerves,all patients were divided into difficult and general decompression groups (n=50,n=408);the effective rate,delayed remission rate and their influencing factors were compared between the two groups.Results All patients were followed up for one to 3 years,averaging 1.6±0.6 years.Among them,46 patients from difficult decompression group were effective (effective rate=92％ [46/50]);and 390 patients from general decompression group were effective (effective rate=95.6％ [390/408]);no statistical difference was noted between the two groups (x2=0.592,P=0.442).But there was statistical difference in delayed remission rate between the two groups (47.8％ vs.25.1％,x2=10.627,P=0.001).Logistic regression analysis was performed among gender,age,course of disease,offending vessels,location,indentation of facial nerve,complex condition of microvacular decompression,efficacy rate and delayed relief rate,and it showed that indentation of facial nerve was related to delayed relief(OR=7.681,P=0.004,95％CI:10.235-31.223).Conclusion The hemifacial spasm patients with difficult decompression are able to achieve as good effect as general decompression if measures are used properly,but the delayed relief rate might be high.
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BACKGROUND:Vascular endothelial growth factor is found the strongest to promote angiogenesis, which show strong expression in synovial cells in rheumatoid arthritis. OBJECTIVE:To evaluate the relation between the expression of vascular endothelial growth factor and microvessel density in rheumatoid arthritis. METHODS:Col agen II and complete Freund’s adjuvant were used to induce experimental arthritis in Sprague-Dawley rats. Synovium expression of vascular endothelium growth factor and microvessel density were assayed at different time points using immunohistochemistry method, and changes in new vessels, vascular endothelial growth factor expression and arthritis index were observed along the time course. The correlations between vascular endothelial growth factor expression and number of new blood vessels as wel as between arthritis index and vascular endothelial growth factor expression were studied with linear correlation analysis and rank correlation analysis, respectively. RESULTS AND CONCLUSION:With the time lasting, new blood vessels in the synovium proliferated, the synovium was thickened, and the arthritis index rose gradual y as wel as vascular endothelial growth factor expression and number of new microvessels were on the rise. Vascular endothelial growth factor expression was positively correlated with arthritis index (r=0.535, P<0.05) and significantly positively correlated with number of new microvessels (r=0.860, P<0.01). Vascular endothelial growth factor is involved in the course of synovium pannus response, and vascular endothelial growth factor and microvessel density may play an important role in rheumatoid arthritis pathogenesis.
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Objective To explore the association of peroxisome proliferators-activated receptor-γ coactivator-1 (PPARGC1) Gly482Ser with apolipoprotein E (ApoE) variations in longevity (aged above 90 yrs) Hans in Guangxi Yongfu and to explore the potential association between the variations and metabolic traits.Methods Based on our survey in Guangxi Yongfu in 2008-2011,212 elderly cases (aged 90～105 years) were included as longevity group and 207 cases without longevity history were included as control group.By household survey,we collected the longevity related parameters,blood glucose,blood lipid,blood pressure and other related metabolic traits.Peripheral blood was collected to extract DNA,the gene variations of Gly482Ser and ApoE were genotyped,and the database with genome and traits information were set up.By univariate analysis and multivariate genetic statistical analysis,the association between the variations and longevity and metabolic traits was assessed.Results Compared with the control group,the levels of fasting blood glucose,total cholesterol and low density lipoprotein were lower in the longevity group.Gly482Ser was genotyped in all samples and fully fulfilled the Hardy Weinberg equilibrium.After the Bonferroni correction,recessive model failed to find association between GG genotype and longevity.Stratified analyses by ApoEε4 allele revealed that,in the subgroup with no ApoEε4,PPARGC-1 GG genotype was positively associated with longevity in the recessive model,even after Bonferroni correction (OR =1.72,P＜0.05).In addition,longevity group with Gly482Ser GG genotype seemed to have relativelower fasting blood glucose (P ＜ 0.05) and higher high density lipoprotein levels (P ＜ 0.05).Conclusions Longevity Hans in Guangxi Yongfu preserve better metabolic state compared with the control group.GG genotype of Gly482Ser in PPARGC-1 is positively associated with longevity,which depends on not carrying the risk allele of ApoE ε4.
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OBJECTIVE@#To investigate the spatiotemporal expression of neuromedin B receptor (NMBR) in mice myometrium at different pregnant stages, as well as its mechanism and relation with parturition.@*METHODS@#The pregnant mice were divided into no-pregnancy (NP), early pregnancy (EP), mid-pregnancy (MP), late-pregnancy (LP), parturition (PT) and postpartum (PP) groups (12 mice in each group), according to pregnant stage. The mRNA and protein expression of NMBR, HSP70 and IL-6 were detected in myometrium in pregnant mice by semi-quantitative RT-PCR and Western blot, while NF-kappaB-P65 DNA binding activity was determined by NoShift transcription factor assay kits, respectively. Their relation with parturition was analyzed.@*RESULTS@#The mRNA expression level of NMBR in the PT group was significantly higher than that in the NP, EP, LP and PP groups (P0.05). The NMBR protein in PT group was significantly higher than that in the other 5 groups (P<0.01). NF-kappaB-P65 DNA binding activity at PT group was remarkably higher than that in the NP, LP and PP groups (P<0.05). The expression of IL-6 mRNA was significantly higher than that in the NP, LP and PP groups (P<0.05), its protein expression in PT and LP groups was significantly higher than that in the NP and PP groups (P<0.05). The expression of HSP70 mRNA in the PT group was significantly higher than that in the NP and PP groups (P<0.05), and the protein of HSP70 was significantly up-regulated in PT and PP groups compared with in NP and LP groups (P<0.05). The DNA-binding activity of P65 was positively correlated to the mRNA expression of NMBR and IL-6 (r=0.40, P<0.01; r=0.30, P<0.05), so were positively correlated to DNA-binding activity of P65, mRNA expression of HSP70 and NMBR ( r=0.40, P<0.01; r=0.49, P<0.01). DNA-binding activity of P65 did not correlate with the mRNA expression of HSP70.@*CONCLUSION@#The mRNA and protein expressions of NMBR reach a peak at the onset of labor. NMBR may play an important role in the parturition via NF-kappaB P65-IL-6 signal transduction pathway. It may also influence the onset of labor by regulating HSP70, but this role does not rely on P65 pathway.
الموضوعات
Animals , Female , Male , Mice , Pregnancy , Gestational Age , HSP70 Heat-Shock Proteins , Metabolism , Interleukin-6 , Metabolism , Myometrium , Metabolism , Parturition , Metabolism , RNA, Messenger , Genetics , Metabolism , Random Allocation , Receptors, Bombesin , Genetics , Metabolism , Signal Transduction , Transcription Factor RelA , Metabolismالملخص
OBJECTIVE: To investigate the effects of Xiehuo Bushen Decoction (XHBSD), a compound Chinese herbal medicine, on the survival and differentiation of transplanted neural stem cells (NSCs) in brains of rats with intracerebral hemorrhage, and to explore the mechanism of Xiehuo Bushen formula in promoting the survival of transplanted NSCs. METHODS: NSCs separated from hippocampuses of neonatal SD rats were cultured. Sixty-five panel reactive antibody (PRA) positive SD rats were selected by lymphocytotoxicity methods. The PRA positive rats were made into intracerebral hemorrhagic model and divided into three groups: cerebral hemorrhage group (n=15), NSCs transplanted group (n=25) and XHBSD group (n=25). XHBSD was orally administered after 5-bromodeoxyuridine (BrdU)-marked NSCs were transplanted in brains of rats with intracerebral hemorrhage in the XHBSD group. Rats in the other two groups were administered distilled water. The expressions of interferon gamma (IFN-gamma) and interleukin-4 (IL-4) mRNAs were measured by reverse transcription polymerase chain reaction (RT-PCR); the numbers of BrdU and 200 kD neurofilament (NF200) positive cells were detected by double-labeling immunofluorescence method. RESULTS: The expression of IFN-gamma mRNA was down-regulated significantly in the XHBSD group, but the expression of IL-4 mRNA was up-regulated significantly (P<0.05). The numbers of BrdU and NF200 positive cells were also increased remarkably in the XHBSD group. CONCLUSION: XHBSD can promote the survival and differentiation of transplanted NSCs, which may be related to inducing the expression of IL-4 mRNA and inhibiting the expression of IFN-gamma mRNA.
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Objective To analyze and identify the proteins relevant to Ganyang Huafeng Syndrome(GYHF) and Yinxu Shengfeng Syndrome(YXSF) of cerebral infarction,and investigate the essence of GYHF and YXSF.Methods 3 groups,GYHF of cerebral infarction,YXSF of cerebral infarction and healthy people were designed with 15 cases of each group.Total protein were extracted from lymphocyte of peripheral venous blood,and two dimensional electrophoresis was used to separate total proteins from norma land model ratsynovium.Having been stained,proteins were analyzed by PDquest software.Some selected protein spots were identified by peptide mass fingerprint(PMF) based on Matrix assisted laser desorption/ ionizationtime of flight mass spectrometry(MALDI-TOF-MS) and database.Results 23 different protein spots and 8 identical peotein spots were found out from GYHF and YXSF group,22 proteins were identified from the different protein spots including 15 kinds of proteins and 6 protein spots including 5 kinds of proteins.GYHF of cerebral infarction related with proteins such as metabolic enzymes,signal transmission proteins,cytoskeletal protein,antioxydation proteins and so forth.Conclusion GYHF and YXSF of cerebral infarction have different proteins and identical proteins,which indicates that the two syndromes have the same matter substructure and different essential connotation.
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Objective To investigate the comparative proteomes difference of PBMC between health adults and Parkinson Disease(PD) patients with Ganyang Huafeng syndrome and Xuexu Shengfeng syndrome.So to approach the essence of the two TCM syndromes from the protein expression level.Methods A series of methods,including immobilolized pH gradient two-dimensional polyacrylamide gel electrophpresis(2-DE),Coomassie Brilliant Blue staining,PDQuest 2-DE analysis software,peptide massfingerprint based on matrix-assisted laser desorption/ionization time of flying mass spectrometry(MALDI-TOF-MS) and SWISS-PROT database searching,were used to separate and indentify the proteome of PBMC between health adults and PD patients with Ganyang Huafeng syndrome and Xuexu Shengfeng syndrome.Results The good 2-DE pattern including resolution and reproducibility was obtained.After Coomassie Brilliant Blue staining,15 spots were incised and analyzed by MALDI-TOF-MS.The typic peptide masses were searched in the SWISS-PROT database by Mascot software.All proteins were preliminarily identified,which ralated to cytoskeleton,anti-oxidative stress,protein degradation,signal transduction and cell cycle,etc.Conclusions The well-resolved,reproducible 2-DE patterns of Ganyang Huafeng syndrome and Xuexu Shengfeng syndrome PD were established and revealed the protein expression difference of PBMC between the two syndromes of PD patients.This research is helpful to provide substance evidence in investigation of the essence of Ganyang Huafeng syndrome and Xuexu Shengfeng syndrome on protein level.
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BACKGROUND: Pathological change of synovium in rheumatoid arthritis (RA) has the characteristic of tumor-like growth, it appears thickening of the synovium tissue and the formatiom of pannus, which generate periarticular erosion and destruction. Multiplicate cell factors and growth factors participate in the development course of tumor-like lesion of synovium, and the tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF)play important roles in the development of RA and the formation of pannus.OBJECTIVE: To observe the contents of plasma TNF-α of collagen-induced arthritis and the expression change of VEGF of synovium at different time point, and investigate the effect and correlation of TNF-α and VEGF in the pathogenesis of RA.DESIGN: Randomized grouping experiment taking animals as subjects.SETTING: Institute integrated traditional and western medicine of Xiangya Hospital of Central South University.MATERIALS: The experiment was finished from July to November 2003 in the laboratory of institute integrated traditional and western medicine.Forty SD rats aged 45-50 days were selected, the rats were randomly divided into the normal control group (n=10) and collagen-induced arthritis model group(n=30).METHODS: 10 mg Cattle collagen type Ⅱ and 5 ml full Freund adjuvant were grinded together, 100 μL of which were intradermally injected at the root of tails of rats, collagen-induced arthritis model of rats were re-immunized with the above-mentioned methòd and dosage after 21 days. The accumulated points of arthritis index was evaluated based on degree and extent of flare and the condition of tumefaction and deformation, the higher the accumulated points of arthritis index were, the more serious the arthritis symptom was. The blood was obtained by decapitation after 25 days in normal control group, and in the collagen-induced arthritis model group the blood was obtained by decapitation 25,30,35,40,45 days after immunization (model establishing), the contents of plasma TNF-α of collagen-induced arthritis rats at different time point was detected by radio-immune assay, the level of expression of VEGF in synovium tissue were detected with immunohistochemical method simultaneously, the correlation of invasion time and the neovascularization of synovium. The accumulated points of arthritis index . TNF-α and VEGF was observed. The correlation of TNF-α and VEGF was analyzed with the linear correlation analysis, the correlation of the accumulated points of arthritis index and TNF-α. VEGF was analyzed with the rank correlation analysis.MAIN OUTCOME MEASURES: The correlation of invasion time of collagen-induced arthritis with the accumulated points of arthritis index, with the plasma content of TNF-α and the expression of VEGF, the correlation analysis of the plasma content of TNF-α of collagen- induced arthritis rats with the expression of VEGF in synovium.RESULTS: Forty rats attended the experiment, all of them entered the final analysis. the neovascularization of synovium was increased, synovium was thickening, the accumulated points of arthritis index was gradually increased, and the contents of TNF-α and level of VEGF were increased accordingly with the process of the invasion time of the collagen-induced arthritis rats; Its accumulated points of arthritis index had the positive correlation with the level of expression of VEGF (r=0.535 ,P ＜ 0.05)and had the correlated increasing tendency with the contents of TNF-α, but there was no significant difference(r=0.371 ,P ＞ 0.05 ). the plasma contents of TNF-α had the positive correlation with the level of expressions of VEGF (r=0.893,P ＜ 0.01 ).CONCLUSION: The TNF-α and VEGF have an important effect on the inflammatory reaction of RA and Cytokine network of neovascularization of synovium, they are possibly mutually influenced and promoted and have the effect of mediating the malignant network circulation; They are the key factors among multiplicate ones which mediate the generation and development of RA, bone erosion and Mutilation.
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BACKGROUND: Cytochrome C oxidase(CCO) is the terminal enzyme in respiration chain of mitochondrion, and it plays a key role in aerobic metabolism and energy production during the process of oxidative phosphorylation. Recently, it is found that energy production of mitochondrion is closely related to the cellular apoptosis, and the changes of CCO activity is closely related to the neuronal impairment after cerebral ischemia and anoxia.OBJECTIVE: To investigate the protective mechanisn of compound pinggan xifeng decoction on the neuronal impairment in cerebral hemorrhage (CH) according to the mitochondrial energy metabolism and cellular apoptosis in neurons.DESIGN: A randomized and controlled trial based on experimental animals.SETTING: Institute of Integrated Traditional Chinese Medicine and Western Medicine, Xiangya Hospital, and Center of Telemedicine.MATERIALS: The experiment was completed in the animal laboratory(key laboratory of province) of Institute of Combination of Traditional Chinese Medicine and Western Medicine from November 2 to 9 in 2003. A total of 80 healthy male SD rats were selected from Experimental Animal Center of Xiangya School of Medicine, Public Health Ministry.METHODS: CH rat models were induced with collagenase Ⅶ, CCO activity was assayed with histochemistry combined with semi-quantification of gray scale, and the cellular apoptosis was evaluated with Tunel method.MAIN OUTCOME MEASURES: CCO activity of CH rats in lateral hippocampal CA1;lateral cellular apoptosis of CH rats.RESULTS: After 12-hour model establishment, CCO activity in CH group was decreased dramatically compared with that in sham operation group (P＜ 0.01), which was 52.12 ±3.75 and 26.98 ±6.32 respectively in lateral hippocampal CA1. And the cellular apoptosis in CH group was increased notably compared with that in sham operation group(P ＜ 0.01),which was(13.56 ± 1.72)/sight and(4. 32 ± 1.04)/sight respectively.Then the two had deteriorated afterwards. After the treatment with pinggan xifeng decoction, CCO activity can be maintained, and the cellular apoptosis was reduced.CONCLUSION: Neuronal injury was closely related to the decrease of CCO activity and the cellular apoptosis in CH. Pinggan xifeng decoction could maintain CO activity of mitochondrion, improve the cellular aerobic netabolism, and reduce the cellular apoptosis, which might be one of the protective mechanisms for secondary neuronal injury in CH.
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Objective:With proteomic approach,to find the differentially expressed proteins spots in the peripheral blood mononuclear cells(PBMCs) of healthy people and intracerebral hemorrhage patients with liver yang forming wind syndrome(LYFWS) treated by Zhenganxifeng decoction(ZGXFD) to investigate the possible proteins of LYFWS and the cure mechanism of ZGXFD.Methods: 10 intracerebral hemorrhage patients with LYFWS were treated by ZGXFD for 7-10 days.10 healthy people were selected taken as healthy controls.10ml venous blood was drawed from every subjects and PBMCs were isolated from blood by using lymphocytes separation medium.The total proteins was extracted from PBMCs.The total proteins from either intracerebral hemorrhage patients or healthy controls were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis.After Coomassie brilliant blue G250 staining,gel-image analysis was performed by PDQuest.The differentially expressed proteins spots were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALD I-TOF-MS).Results: Gel-image analysis revealed that there were 22 proteins spots expressed differentially.20 proteins spots were expressed differentially in the intracerebral hemorrhage patients with LYFWS as compared with healthy control,of which 8 proteins were down-regulated,7 proteins were up-regulated and 5 proteins were lost.After treated by ZGXFD,4 proteins were down-regulated,5 proteins were up-regulated and 2 proteins were lost,9 proteins weren’t expressed di erentially as compared with healthy control.8 of 20 di erentially expressed proteins spots in the intracerebral hemorrhage patients with LYFWS identi ed by MALDI-TOF-MS as compared with healthy control.These proteins are related to cell metabolism,signal transduction and so forth,of which 6 proteins were down-regulated,1 proteins were up-regulated and 1 proteins were lost.After treated by ZGXFD,2 proteins were down-regulated,1 proteins were lost,5 proteins weren’t expressed di erentially as compared with healthy control.Conclusion: 2-DE pro les of intracerebral hemorrhage patients with LYFWS in PBMCs had been established.The proteins of intracerebral hemorrhage patients with LYFWS had been identi ed,which were related to cell metabolism,signal transduction and so forth.This research presented that ZGXFD could adjust multiple di erentially expressed proteins of intracerebral hemorrhage patients with LYFWS.It was also indicated that the cure mechanism of ZGXFD was to regulate multitarget proteins.