1.DIA Proteomics Reveals Mechanism of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis Extract in Treating α-Syn Transgenic Parkinson's Disease in Mice
Qi ZHENG ; Yi LU ; Donghua YU ; Liangyou ZHAO ; Chunsheng LIN ; Fang LU ; Shumin LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(8):40-50
ObjectiveTo investigate the mechanism of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis extract (ASH) in treating Parkinson's disease (PD) in mice by Data-Independent Acquisition (DIA) proteomics. MethodsThe α-Synuclein (α-Syn) transgenic PD mice were selected as suitable models for PD, and they were randomly assigned into PD, ASH (61.25 mg·kg-1), and Madopar (97.5 mg·kg-1) groups. Male C57BL/6 mice of the same age were selected as the control group, with eight mice in each group. Mice were administrated with corresponding drugs by gavage once a day for 20 days. The pole climbing time and the number of autonomic activities were recorded to evaluate the exercise ability of mice. Hematoxylin-eosin staining was employed to observe neuronal changes in the substantia nigra of PD mice. Immunohistochemistry (IHC) was employed to measure the tyrosine hydroxylase (TH) activity in the substantia nigra and assess the areal density of α-Syn in the striatum. DIA proteomics was used to compare protein expression in the substantia nigra between groups. IHC was utilized to validate key differentially expressed proteins, including Lactotransferrin, Notch2, Ndrg2, and TMEM 166. The cell counting kit-8 (CCK-8) method was used to investigate the effect of ASH on the viability of PD cells with overexpression of α-Syn. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot were employed to determine the protein and mRNA levels of Lactotransferrin, Notch2, Ndrg2, and TMEM 166 in PD cells. ResultsCompared with the control group, the model group showed prolonged pole climbing time, diminished coordination ability, reduced autonomic activities (P<0.01), and reduced swelling neurons. Compared with the model group, ASH and Madopar reduced the climbing time, increased autonomic activities (P<0.01), and ameliorated neuronal damage. Compared with the control group, the model group showed a decrease in TH activity in the substantia nigra and an increase in α-Syn accumulation in the striatum (P<0.01). Compared with the model group, the ASH group showed an increase in TH activity and a reduction in α-Syn accumulation (P<0.05). DIA proteomics revealed a total of 464 differentially expressed proteins in the model group compared with the control group, with 323 proteins being up-regulated and 141 down-regulated. A total of 262 differentially expressed proteins were screened in the ASH group compared with the model group, including 85 proteins being up-regulated and 177 down-regulated. Kyoto encylopedia of genes and genomes (KEGG) pathway analysis indicated that ASH primarily regulated the Notch signaling pathway. The model group showed up-regulation in protein levels of Notch2, Ndrg2, and TMEM 166 and down-regulation in the protein level of Lactotransferrin compared with the control group (P<0.01). Compared with the model group, ASH down-regulated the protein levels of Notch2, Ndrg2, and TMEM 166 (P<0.05) while up-regulating the protein level of Lactotransferrin (P<0.01). The IHC results corroborated the proteomics findings. The cell experiment results showed that compared with the control group, the modeling up-regulated the mRNA and protein levels of Notch2, Ndrg2, and TMEM 166 (P<0.01), while down-regulating the mRNA and protein levels of Lactotransferrin (P<0.01). Compared with the model group, ASH reduced the mRNA and protein levels of Notch2, Ndrg2, and TMEM 166 (P<0.01), while increasing the mRNA and protein levels of Lactotransferrin (P<0.05, P<0.01). ConclusionASH may Synergistically inhibit the Notch signaling pathway and mitigate neuronal damage by down-regulating the expression of Notch2 and Ndrg2. Additionally, by up-regulating the expression of Lactotransferrin and down-regulating the expression of TMEM166, ASH can address brain iron accumulation, intervene in ferroptosis, inhibit mitophagy, and mitigate reactive oxygen species damage, thereby protecting nerve cells and contributing to the treatment of PD.
2.Summary of traditional Chinese medicine diagnosis, treatment and prevention of community-acquired pneumonia
Fang LIU ; Shuo WANG ; Chunsheng LI
Journal of Chinese Physician 2024;26(2):161-165
Community acquired pneumonia (CAP) is one of the common infectious diseases in clinic, with a high mortality. With the aging population, continuous variation of pathogens, and increasingly severe antibiotic resistance, the treatment difficulty of CAP continues to increase. This disease belongs to the categories of traditional Chinese medicine diseases such as " wind warm lung heat disease" and " cough". Traditional Chinese medicine treatment can effectively improve symptoms of CAP, control disease progression, shorten hospital stay, and also have certain advantages in prevention. This article mainly provides an overview of the diagnosis, etiology and pathogenesis, syndrome differentiation and treatment, and prevention of CAP, providing reference for the clinical diagnosis, treatment, and prevention of CAP in traditional Chinese medicine.
4.Study on Anti-tumor Activities of Physalins B in Vitro
Journal of Guangzhou University of Traditional Chinese Medicine 2015;(4):652-655,660
Objective To study the in-vitro anti-tumor activities of physalin B (PhB) extracted from Physalis angulata L.. Methods Th e effect of PhB on the cell proliferation and the morphology changes of tumor cell strains HepG2 and SGC7901 was observed by methyl thiazolyl tetrazolium (MTT) assay and by phase contrast microscopy respectively, and the results were compared to positive control anti-cancer drug hydroxycamptothecine (HCTP) . Apoptosis of HepG2 and SGC7901 cells induced by PhB was determined by 4’ , 6-diamidino-2-phenylindole ( DAPI) staining method. Results MTT results showed that the growth of HepG2 and SGC7901 was significantly inhibited by PhB in dose-and time-dependent manner, and the inhibitory effect of high concentration of PhB was better than the positive control medicine HCTP. Phase contrast microscopy results showed that the cellular morphology was improved by PhB, which was accorded with the results showed by MTT assay. DAPI staining results showed that the apoptosis of HepG2 and SGC7901 was enhanced by PhB. Conclusion PhB extracted from Physalis pubescens L. exerts certain inhibitory effect on the proliferation of HepG2 and SGC7901 cells.
5.Regulating the EMT of human gastric cancer cell line in vitro through miRNA sponge
Suli LI ; Fang ZHOU ; Qingyu ZHANG ; Wenliang JIA ; Anling ZHANG ; Lei HAN ; Chunsheng KANG
Chinese Journal of Clinical Oncology 2014;45(11):684-688
Objective:To explore the effect and mechanism of miRNA sponge on the epithelial-mesenchymal transition (EMT) of gastric carcinoma cell lines SGC7901. Methods:Synthetic ZEB2 3'UTR plasmid and siRNA targeting ZEB2 were transfected into the SGC7901 cell line by Lipofectamine 2000. Real-time quantitative polymerase chain reaction was performed to evaluate the expres-sion levels of miR-200a/b/c. Finally, the migratory, invasive, and proliferative activities of the gastric carcinoma cells in vitro were ana-lyzed by the scratch test, the Transwell cell invasion, and the cell cloning assay. The expression of the target protein was detected by Western blot. Results:Compared with the control group, the expressions of miR-200a/b/c significantly decreased, and their migration, invasion, and proliferation capabilities were considerably higher after they were transfected with ZEB2 3'UTR. Although the expres-sions of miR-200a/b/c significantly increased, the migratory, invasive, and proliferative activities of SGC7901 cells also degraded after they were transfected with siRNA targeting ZEB2. The expression of ZEB2 increased, and that of E-cadherin decreased at the protein level after they were transfected with ZEB2 3'UTR. The protein expression of Vimentin in SGC7901 cells significantly increased. The indicators show the opposite trend when cells were transfected with siZEB2, and the differences between the control and mutation groups were insignificant. Conclusion:ZEB2 3'UTR can regulate EMT course by regulating the miR-200a/b/c expression in gastric car-cinoma, consequently regulating the invasion and migration of carcinoma cells.
6.Effects of ZEB2 3′UTR Transfection on Proliferation, Invasion and Migration in Human Gastric Epithelial Cell GES-1
Suli LI ; Fang ZHOU ; Qingyu ZHANG ; Wenliang JIA ; Anling ZHANG ; Lei HAN ; Chunsheng KANG
Tianjin Medical Journal 2014;(5):401-405
Objective To explore the effects of zinc finger E-box binding protein (ZEB)2 3′UTR gene transfection on proliferation, invasion and migration in human gastric epithelial cell line GES-1. Methods The synthetic ZEB2 3′UTR and miR-200b micmics were transfected into GES-1 cell line by lipofectamine 2000. We set up control grop, the mutation group and ZEB2 3′UTR group. Real-time quantitative PCR was performed to evaluate the expression levels of miR-200a/b/c and ZEB1/ZEB2 mRNAs after transfection.And then we set up control group, ZEB2 3′UTR group, ZEB2 3′UTR+negative control group and ZEB2 3′UTR+miR-200b micmics group. The protein expression levels of ZEB1, ZEB2, matrix metallopro-teinases (MMP) 2/9 and proliferating cell nuclear antigen (PCNA) were detected by Western blot assay. The invasion and mi-gration capability were analyzed by transwell assay and wound healing test. MTT assay was used to detect the proliferation ability. Results Compared with control group and mutation group, the expressions of miR-200a/b/c were significantly de-creased, especially for miR-200b. And the expressions of ZEB1/ZEB2 were significantly increased at both mRNA and pro-tein levels after transfected with the ZEB2 3′UTR, enhancing the capability of migration,invasion,and proliferation (P <0.05). Compared with ZEB2 3′UTR group, the capabilities of proliferation,invasion and migration were significantly lower in combined group. Conclusion ZEB2 3′UTR can increase the ability of cell proliferation, invasion and metastasis through regulating the levels of miR-200a/b/c, and then influence the regulation of transcription of the target gene, which could lead to malignant transformation of GES-1 cells.
7.Validation of 5 prediction models for acute kidney injury and its outcome after cardiac surgery procedures in Chinese patients
Wuhua JIANG ; Xiaoqiang DING ; Yi FANG ; Lan LIU ; Chunsheng WANG ; Jie TENG
Chinese Journal of Nephrology 2013;29(6):413-418
Objective To assess the clinical usefulness and value of the 5 models for the prediction of acute kidney injury (AKI),severe AKI which renal replacement treatment was needed (RRT-AKI) and death after cardiac surgery procedures in Chinese patients.Methods One thousand and sixty-seven patients who underwent cardiac surgery procedures in the department of cardiac surgery in the Zhongshan Hospital,Fudan University between May 2010 and January 2011 were involved in this research.The predicting value for AKI (AKICS),RRT-AKI (Cleveland,SRI and Mehta score) and death (EURO score) after cardiac surgery procedures was evaluated by Hosmer-Lemeshow goodness-of-fit test for the calibration and area under receiver operation characteristic curve (AUROC)for the discrimination.Results The incidence of AKI was 20.34%(217/1067),and 63.13% of their renal function recovered completely.The incidence of RRT-AKI was 3.56%(38/1067) and the mortality of AKI and RRT-AKI was 9.68% (21/217) and 44.73% (17/38) respectively.The total mortality was 3.28% (35/1067).The discrimination and calibration for the prediction ofAKI of AKICS were low.For the prediction ofRRT-AKI,the discrimination and calibration of Cleveland score were high enough,but the predicated value was lower than the real value (1.70% vs 3.86%).The discrimination of Mehta score and the calibration of SRI were low.The discrimination and calibration for the prediction of death of EURO score was low.Conclusion According to the 2012 KDIGO AKI definition,none of the 5 models above is good at predicting AKI after cardiac surgery procedures.Cleveland score has been validated to have a proper impact on predicting RRT-AKI after cardiac surgery procedures,but the predicting value is still in doubt.EURO score has been validated to have an inaccurate predicting value for death after cardiac surgery procedures.
8.Toxicity assessment of hydroxyapatite-coated mechanical heart valve embedded with collagen to vascular endothelial cells
Guangcun CHENG ; Chunsheng LI ; Yu YAN ; Lan WANG ; Zhongya YAN ; Le LUO ; Xiaodong FANG ; Ruhua TAO
Chinese Journal of Tissue Engineering Research 2013;(51):8894-8900
BACKGROUND:In early experiments, we prepared hydroxyapatite-coated mechanical heart valve embedded with col agen using impulse laser sediment method.
OBJECTIVE:To further analyze the histocompatibility and toxicity of hydroxyapatite-coated mechanical heart valve embedded with col agen.
METHODS:After passage, canine vascular endothelial cellsuspension was inoculated onto the hydroxyapatite-coated mechanical heart valve embedded with col agen. One group was inoculated in 5%CO2, 37 ℃ incubator for 3 weeks static culture, and the other group was inoculated in 5%CO 2 , 37 ℃ incubator for 3 weeks spinner culture. Scanning electron microscope was used to observe cellattachment on the material. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to the proliferative capacity of vascular endothelial cells cultured with the hydroxyapatite-coated mechanical heart valve embedded with col agen.
RESULTS AND CONCLUSION:During the spinner culture, adherent cells were found on the surface of mechanical heart valve, and the cells distributed evenly and confluent at 21 days to cover the surface of the material. The number of adherent cells in the spinner culture was higher than that in the static culture. The cells during the static culture were aggregated and distributed irregularly. The mechanical heart valve exhibited no effects on the proliferation of canine vascular endothelial cells which grew wel . These findings indicate that the hydroxyapatite-coated mechanical heart valve embedded with col agen exert no effect on proliferation of vascular endothelial cells, has no toxicity and has good biocompatibility.
9.The risk factors and prognosis of acute kidney injury after cardiac surgery: a prospective cohort study of 4007 cases
Jiarui XU ; Jie TENG ; Yi FANG ; Bo SHEN ; Zhonghua LIU ; Shaowei XU ; Jianzhou ZOU ; Lan LIU ; Chunsheng WANG ; Xiaoqiang DING
Chinese Journal of Internal Medicine 2012;(12):943-947
Objective To investigate the risk factors and prognosis influential factors of acute kidney injury (AKI) after cardiac surgery.Methods The clinical data of patients who were hospitalized and underwent cardiac surgery from April 2009 to May 2011 were collected prospectively.Demographic characteristics,types of surgeries,preoperative renal function,pre-and intra-operative conditions and clinical outcomes,etc were recorded.Results A total of 4007 patients underwent cardiac surgery were recruited.The overall incidence of AKI was 31.2% (1250/4007).The incidence of AKI requiring renal replacement treatment (AKI-RRT) was 2.6% (104/4007).The overall hospital mortality was 1.9% (77/4007),and was significantly higher in AKI group than in non-AKI group (5.4% vs 0.3%,P <0.01).The hospital mortality of AKI-RRT group was 36.5% (38/104).Grouped by type of surgery,cardiac transplantation had the highest AKI incidence (73.0%) and highest in-hospital mortality (18.9%),followed by coronary artery bypass grafting (CABG) combined with valve surgery (AKI incidence 57.8%,in-hospital mortality 6.1%) and aneurysm surgery (AKI incidence 52.0%,in-hospital mortality 5.5%).Multivariate logistic regression analysis showed that man,age,BMI,hypertension,chronic heart failure,pre-operative serum creatinine (SCr) > 106.0 μmol/L,intra-operative cardiopulmonary bypass time,intra-operative hypotension and aneurysm surgery were the risk factors of AKI after cardiac surgery.Multivariate logistic regression analysis showed that pre-operative SCr > 106.0 μmol/L and intra-operative hypotension were independent risk factors of renal recovery after cardiac surgery while recovery of urine output was the favorable factor.Conclusions Cardiac surgery usually induces high AKI incidence and poor prognosis,which closely associated with many risk factors in peri-operative stage.The incidence of AKI is related to a number of perioperative risk factors.Heart transplantation,aneurysm surgery,CABG combined valve surgery are high risk surgeries.
10.Prognostic investigation of AKIN criteria combined with APACHE Ⅱ and SOFA scoring system in acute kidney injury after cardiac surgery
Wei JIN ; Jie TENG ; Yi FANG ; Zhonghua LIU ; Bo SHEN ; Yan XU ; Yanyan HENG ; Zhaohua YANG ; Chunsheng WANG ; Xiaoqiang DING
Chinese Journal of Nephrology 2011;27(3):170-175
Objective To explore the prognostic value of Acute Kidney Injury Network (AKIN)criteria combined with Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ)and Sequential Organ Failure Assessment(SOFA)scoring system in acute kidney injury(AKI)after cardiac surgery. Methods Clinical data of patients who underwent open-heart surgery in Zhongshan Hospital,Fudan University from April 2009 to August 2009 were prospectively collected.AKI after cardiac surgery was classified by AKIN staging system.APACHE Ⅱ and SOFA scores were evaluated according to the worst value of physiologic variables in the 1st 24 h after surgery.Discrimination and calibration of these three models were assessed by receiver operating characteristic(ROC) curve and Hosmer-Lemeshow goodness-of-fit test.Besides,their effects on inhospital mortality were evaluated by multivariate Logistic regression analysis. Results Of the 993 admissions,309 patients developed AKI and the incidence was 31.1%.The median time that developed postoperative AKI and reached the Scr peak were 1 d and 2 d respectively.Either APACHE Ⅱ or SOFA scores,which was positively correlted with the severity of AKI(APACHEⅡr=0.37,P<0.01;SOFA r=0.42,P<0.01)was higher in AKI patients compared with that in nonAKI patients(P<0.01).The mortality rose corresponding to the severity of kidney injury.However,the predicted death rate-adjusted(PDR-A)calculated by APACHE Ⅱ scoreS Was higher than the actual value in non-AKI patients and AKIN stage 1(P<0.01),while it was lower in AKIN stage 3 (P<0.01).The areas under the ROC curve of APACHEⅡ,SOFA and AKIN criteria were all above 0.8 and the results of Hosmer-Lemeshow goodness-of-fit test indicated good calibration of three models.Multivariate analysis showed that APACHE Ⅱ≥19(OR=4.26)and AKIN stage 3(OR=76.151 were independent predictors of in-hospital mortality. Conclusions AKI can be classified by AKIN criteria in the early stage after cardiac surgery and the AKIN staging system may serve the prediction of prognosis.The APACHE Ⅱ and SOFA scores just evaluated in the 1st 24 h after operation can discern the severity of patients'illness.Three models all present good discrimination and calibration in predicting patients'outcome.APACHE Ⅱ≥19 along with AKIN stage 3 are found to be the independent predictors of in-hospital mortality.It should be noticed that the deviation between PDR-A and the actual mortality in subgroups,dynamic evaluation may raise the accuracy of scoring system.

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