1.Effect and Mechanism of Qixian Tongluo Formula on Contralateral Corticospinal Tract Remodeling and Motor Functional Recovery in Rats with Cerebral Infarction
Shengqiang ZHOU ; Dahua WU ; Bo LI ; Yanjun CHEN ; Jia HUANG ; Qi WANG ; Wen ZENG ; Lingjuan TAN ; Yihui DENG ; Fang LIU
Traditional Chinese Drug Research & Clinical Pharmacology 2024;35(9):1319-1328
		                        		
		                        			
		                        			Objective To observe the effect of Qixian Tongluo Formula on contralateral corticospinal tract(CST)remodeling and motor functional recovery in rats with cerebral infarction,and to explore its potential molecular mechanism from the perspective of regulating factors related to never remodeling.Methods The rat middle cerebral artery occlusion(MCAO)model was established by silk thread ligation.Fifty model rats were randomly divided into model group,citicoline group(0.054 g·kg-1),Qixian Tongluo Formula low-,medium-and high-dose(7.83,15.66,31.32 g·kg-1)groups,and sham operation group,with 10 rats in each group.The intervention administration was started on the 3rd day after operation once a day for 26 consecutive days.On the 3rd,14th and 28th day after operation,the gross motor function was evaluated by Longa score,and the fine motor function was evaluated by beam-walking test(BWT)score.The contralateral motor cortex was injected with the nerve tracer biotin dextran amine(BDA)on the 14 th day after operation to anterogradely trace the CST.On the 28th day after operation,the expression of axonal growth associated protein-43(GAP-43)and BDA positive fibers in the contralateral motor cortex and cervical spinal cord were detected by immunohistochemistry.The co-localization areas of BDA positive fibers and presynaptic marker protein vesicular glutamate transporter 1(VGLUT1)in the cervical spinal cord gray matter were detected by immunofluorescence.The expressions of brain-derived neurotrophic factor(BDNF),glial cell-derived neurotrophic factor(GDNF),nerve growth factor(NGF)and nerve remodeling-associated inhibitory factor[Nogo-A,oligodendrocyte myelin glycoprotein(OMgp)and myelin-associated glycoprotein(MAG)]in the contralateral motor cortex were detected by Western Blot.Pearson correlation analysis was used to analyze the correlation between Longa score or BWT score and BDA/VGLUT1 co-localization area,respectively.Results Compared with the sham operation group,rats in the model group had obvious symptoms of motor function deficits,and the Longa scores were significantly increased(P<0.01)and the BWT scores were significantly decreased(P<0.05,P<0.01)at each time point.The expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was up-regulated(P<0.05),the number of edge-crossing fibers from the posterior funiculus in cervical cord was increased(P<0.05),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was increased(P<0.05),the expressions of BDNF,GDNF and NGF in the contralateral motor cortex were up-regulated(P<0.05),while the expressions of Nogo-A,OMgp and MAG were down-regulated(P<0.05).Compared with the model group,the Longa scores in each administration group on the 14th and 28th day after MCAO operation were significantly decreased(P<0.01),the BWT scores were significantly increased(P<0.01),the expression of GAP-43 in the contralateral motor cortex and cervical spinal cord was significantly up-regulated(P<0.01).The number of edge-crossing fibers from the posterior funiculus in cervical cord was significantly increased(P<0.01),the co-localization area of BDA/VGLUT1 in the gray matter of the cervical spinal cord was significantly increased(P<0.01).The expressions of BDNF,GDNF and NGF in the contralateral motor cortex were significantly up-regulated(P<0.01,P<0.05),while the expressions of Nogo-A,OMgp and MAG were significantly down-regulated(P<0.05),and the most significant effect was observed in the high dose group.The Longa score was negatively correlated with the co-localization area of BDA/VGLUT1(r=-0.89,P<0.01),and the BWT score was positively correlated with the co-localization area of BDA/VGLUT1(r=0.84,P<0.01).Conclusion Qixian Tongluo Formula can improve motor function through promoting contralateral CST remodeling in MCAO rats after cerebral infarction,and the molecular mechanism may be related to the regulation of the expression of nerve remodeling-associated factor in the contralateral motor cortex.
		                        		
		                        		
		                        		
		                        	
2.Development of the Spleen Deficiency Evidence Scale for County Residentsand Test of Reliability and Validity
Meng ZHU ; Lingjuan JIA ; Fuzhen PAN ; Huiqing CHEN ; Jing XIAO ; Pengfei SHAO ; Yuxuan GONG ; Weifang ZHENG ; Yongsheng ZHANG ; Xiaqiu WU
World Science and Technology-Modernization of Traditional Chinese Medicine 2024;26(7):1939-1945
		                        		
		                        			
		                        			Objective This study was to develop a"Spleen Deficiency Certificate Scale for County Residents"and test its reliability.It was then developed as an objective tool for Chinese medicine evidence and symptoms for the prevention and control of chronic diseases among county residents.Methods The scale was compiled based on the team's previous foundation.The reliability of the scale was evaluated using internal consistency reliability and split-half reliability,while its validity was evaluated using structural validity,content validity,calibration validity,and discriminant validity.Results The study included 213 adults from Lanxi,of whom 155 were tested for intestinal flora.Seven scale entries were identified:Fatigue,fear of cold,bland mouth,loss of appetite,diarrhea,weak bowel movements,and tooth-marked tongue.In the reliability test,Cronbach's alpha coefficient was 0.828 and McDonald's ω coefficient was 0.825.The"stomach pain"and"bloating"entries did not meet the inclusion requirements and were recommended to be deleted.The Spearman-Brown coefficient was 0.839.The exploratory factor analysis of the two common factors explained 61.6%of the cumulative variance.The calibration validity indicated that the ratio of salivary amylase activity before and after acid stimulation was 0.826±0.253 in the group with spleen deficiency.Significant differences(P<0.05)in the genera Dialister,Shigella,Leuconostoc,Photobacterium,Trabulsiella,and Parvimonas between the spleen deficiency group and the non-spleen deficiency group.Conclusion The Spleen Deficiency Scale for County Residents demonstrates good reliability and validity.
		                        		
		                        		
		                        		
		                        	
3.Research progress on the involvement of nuclear receptor in regulating autophagy.
Lingjuan WU ; Xiaoli XU ; Jun LIN ; Weiwen CAI ; Liqun CHEN
Journal of Biomedical Engineering 2018;35(5):822-828
		                        		
		                        			
		                        			Nuclear receptors are transcriptional regulators involved in almost all biological processes such as cell growth, differentiation, apoptosis, substance metabolism and tumor formation, and they can be regulated by small molecules that bind to them. Autophagy is a special way of programmed cell death and it is a highly conserved metabolic process. Once autophagy defects or excessive autophagy occur, the disease will develop. In recent years, numerous studies have shown that nuclear receptors are related to autophagy. Therefore, this paper mainly reviews the research progress on nuclear receptors involved in the regulation of autophagy, and focuses on the mechanism of several nuclear receptors involved in the regulation of autophagy, aiming at understanding the molecular basis of how nuclear receptors participate in regulating autophagy, as well as providing possible ideas and strategies for the treatment of corresponding diseases.
		                        		
		                        		
		                        		
		                        	
4.The temporal expression patterns of the transcription factors and target genes in cardiomyocyte hypertrophy
Man LUO ; Xiaoyun WU ; Min ZHENG ; Lingjuan LIU ; Jie TIAN
Medical Journal of Chinese People's Liberation Army 2017;42(5):394-401
		                        		
		                        			
		                        			Objective To investigate the temporal expression patterns of the related transcription factors and target genes in cardiomyocyte hypertrophy,and provide valuable clues for further researches on cardiomyocyte hypertrophy.Methods Isoproterenol (ISO) was used to induce ventricular myocytes hypertrophy in neonatal mice.The survival rate of cardiomyocyte was detected by CCK-8,and the average diameters and surface areas of cells were measured by computer photograph analysis system.The mRNA and protein expression levels of related genes were respectively measured by RT-PCR and Western blotting.Results The model of cardiomyocyte hypertrophy stimulated by ISO was constructed successfully.The expression levels of GATA4,MEF2C,GATA5,BNP and ANP increased 24 hours after ISO treatment,the expression levels of P300,α-MHC and TBX5 increased 12 hours after ISO treatment,and of SRF and β-MHC mRNA increased 6 hours after ISO treatment (P<0.05).The expression levels of GATA4,α-MHC,β-MHC,SRF and P300 mRNA increased firstly,and then decreased in cardiomyocytes induced by ISO.The expression levels of GATA4,SRF,α-MHC,β-MHC and P300 mRNA were still higher than normal (P<0.05),but of MEF2C decreased to normal (P>0.0S) 72 hours after ISO treatment.The expression levels continuously elevated of GATA5,TBX5,ANP and BNP mRNA than that of controls (P<0.05),while no fluctuation was found in NKX2.5 mRNA expression (P>0.05).The expression of GATA4 protein increased,while of HEY2 protein decreased 48 hours after ISO treatment (P<0.05).Conclusions In hypertrophic cardiomyocytes,the expression pattern of MEF2C is similar to,but the patterns of GATA5,GATA4,TBX5 and SRF are different with that in the development of heart,implying these genes are important during the process from compensatory stage to decompensation stage.The expression patterns of GATA4,MEF2C and SRF are similar to that of acetylase P300,implying the temporal expressions could be regulated by P300 in cardiomyocyte hypertrophy.
		                        		
		                        		
		                        		
		                        	
5.Analysis of clinical characteristics of different degrees of myocardial noncompaction in children
Huiming ZHOU ; Lingjuan LIU ; Yao WU ; Tiewei LYU ; Jie TIAN
Journal of Clinical Pediatrics 2017;35(10):751-754
		                        		
		                        			
		                        			Objectives To explore the clinical characteristics and prognosis of different degrees of noncompaction of ventricular myocardium (NVM) in children and to investigate the clinical significance of the ratio of the thickness of the non-compact myocardium to the dense myocardium (N/C). Methods The clinical data of the children with NVM were analyzed retrospectively. The differences between children with N/C > 2.0 and children with N/C 1.4~2.0 were compared. Results There were 26 children in each of the two groups. There was no significant difference in age, sex, the ratios of low birth weight and malnutrition, heart disease, and complication (P all > 0.05). When discharged, 8 cases were improved, 15 cases has no obvious improvement, and 3 cases died in N/C>2.0 group; while 18 cases were improved, 8 cases has no obvious improvement, and none died in N/C 1.4~2.0 group, and the effective rate of treatment between two groups was statistically difference (χ2=7.69,P=0.006). In 8 years of follow-up, the mortality rate in N/C > 2.0 group was higher than that in N/C 1.4~2.0 group, and the difference was statistically different (χ2=6.993, P=0.008). Conclusions The treatment efficiency was lower and the prognosis was worse when the N/C value was higher in children with NVM.
		                        		
		                        		
		                        		
		                        	
6.Rhabdomyosarcoma of breast resembles acute leukemia:a case report and review of the literature
Jingjing ZHU ; Zhengyang LI ; Haifei CHEN ; Jie LI ; Yuhao ZHANG ; Tianqin WU ; Hongshi SHEN ; Jieqing TANG ; Jing WANG ; Longmei QIN ; Lingjuan JIN
Cancer Research and Clinic 2015;(7):469-472
		                        		
		                        			
		                        			Objective To investigate the clinical pathological characteristics, diagnosis and treatment of breast rhabdomyosarcoma, and to enhance the awareness of malignancy infiltration to bone marrow (BM). Methods The data of one case of Rhabdomyosarcoma of breast were analyzed retrospectively. BM aspirate and biopsy, morphology, immunology, cytogenetics, molecular biology (MICM) in different parts of BM, peripheral blood smear, fine puncture of breast mass, final biopsy of breast mass by Mammotome System and whole body PET-CT were performed. The immunochemistry stain of specimen of breast mass was used. Results The peripheral blood smear of this patient showed immature erythrocytes, leucocytes and classification of unknown cells which were consistent with BM morphology. The results of BM aspirate and biopsy depicted a hypercellular specimen with disseminated unknown cells infiltration. Unknown cells were positive for CD56 and negative for any hematopoietic markers by flow cytometry. The whole body PET-CT showed that uptake of 18F-FDG of bilateral breast and whole BM was increased, whereas the mass of breast was not presented by CT. PET-CT suggested a probable malignant hematologic disease. The enough specimen of breast mass got from Mammotome System showed embryonal rhabdomyosarcoma, and the tumor cells were positive for MyoD1, Vimentin and Desmin. Conclusions It is a challenge for early diagnosis of solid sarcoma with unknown origin which diffusely infiltrating into BM. Negative expression of hematopoietic markers by flow cytometry plays a role on differential diagnosis in this setting, whereas PET-CT only provides a valuable reference. Enough specimen and immunohistochemical staining could provide solid evidences of diagnosis.
		                        		
		                        		
		                        		
		                        	
7.Shenqi Fuzheng Injection (SFI) inhibits irradiation-induced brain injury
Jian ZHANG ; Ruiguang ZHANG ; Qian CAI ; Lingjuan CHEN ; Jihua DONG ; Gang WU ; Xiaorong DONG
Chinese Journal of Radiological Medicine and Protection 2015;35(6):419-422
		                        		
		                        			
		                        			Objective To explore the effect of SFI in radiation-induced mice brain injury after 20 Gy cranial radiation.Methods The mice were divided into three groups:(1) control group,(2) RT-only group:the whole brain was irradiated with a dose of 20 Gy,(3) RT and SFI group:SFI at 20 ml/kg/d from 4 weeks after 20 Gy cranial radiation theraty(CRT).Results Morris water maze test showed that the latency of the irradiated group was longer than control group and SFI improved the cognitive function of mice (t =6.34,6.70,P <0.05).The expression of TNF-α reached to the highest level at 3 h after irradiation,and then it decreased but got the second higher level again at 4 weeks after irradiation.The expression of IL-1 β reached to the highest level at 72 h after irradiation and decreased until 4 weeks after irradiation.SFI decreased both expressions of TNF-α (t =11.34,9.70,6.07,P < 0.05) and IL-1 β (t =12.27,5.70,7.52,P < 0.05).Immune florescence staining showed that SFI reduced the number of activated microglia (t =12.35,8.64,7.82,P < 0.05)and inhibited the translocation of p65 of microglia after irradiation.Conclusions Findings suggest that SFI may decrease microglial activation and suppress the expression of TNF-α and IL-1β by inhibiting the translocation of NF-κB p65 and then attenuate irradiation-induced brain injury.
		                        		
		                        		
		                        		
		                        	
8.oxidized low-density-lipoprotein inhabits the proliferation and Oct-4 express of rat bone marrow mesenchymal stem cells in vitro
Tiewei LV ; Huichao SUN ; Lei ZHANG ; Lingjuan LIU ; Xiaoyun WU ; Xiaoyan LIU ; Jing ZHU ; Zhenguo LIU ; Jie TIAN
Chongqing Medicine 2014;(14):1737-1740
		                        		
		                        			
		                        			Objective To detect the effect of ox‐LDL on self‐renewal and Oct‐4 express of MSCs in vitro .Methods MSCs cul‐tured in vitro were divided into 4 groups :blank control(no reagents in culture system ) ,ox‐LDL (1 ,5 ,10 ,20 μg/mL ox‐LDL were added into culture system) ,ox‐LDL+NAC(corresponding ox‐LDL were added into culture system after NAC treatment ) ,negative control(corresponding nLDL were added into culture system ) .Growth curve were drawn at different time ,Oct‐4 ,a stem cell special marker ,were detected by real‐time PCR ,the production of ROS (reactive oxygen species ,ROS) in culture system were measured with electron paramagnetic resonance spectroscopy .Results proliferation of MSCs was inhibited by ox‐LDL ,when concentration of ox‐LDL was more than 5μg/mL ;apoptosis of MSCs appeared as well as attenuated expression of Bcl‐2 ,ox‐LDL generated a signifi‐cant amount of ROS in the culture system ,which was completely prevented by NAC .Conclusion The proliferation and Oct‐4 ex‐pression of MSCs were inhibited by ox‐LDL ,which may be related to increase of ROS in culture system .
		                        		
		                        		
		                        		
		                        	
9.Impact of low dietary glycemic index on decreasing of the risk of coronary heart disease: a systematic review
Meiqin XUE ; Liping SUN ; Wenwen WU ; Shahshan LIU ; Lingjuan ZHANG
Chinese Journal of Practical Nursing 2013;(7):46-49
		                        		
		                        			
		                        			Objective To evaluate the impact of low dietary glycemic index on the risk of coronaryheart disease (CHD).Methods We searched the Cochrane Database of Systematic Reviews,JBI Database of Systematic Reviews,MEDLINE,EMbase,CBMdisk,and CNKI to collect cohort studies from 2000 to 2011.The quality of cohort studies was critically appraised and data were extracted by 4 reviewers independently.Meta-analysis were conducted for the eligible cohort studies using RevMan 5.0.Results Five cohort studies were included,and 128 911 samples were researched.The result of meta analysis indicated that low dietary glycemic index was a protective factor of CHD,RR value was 1.25,95% CI was (1.13,2.15).Further more,the result was only effective to female,RR value was 1.35,95% CI was (1.16,1.57),but not to male,RR value was 0.96,95%CI was (0.76,1.31).Conclusions Low dietary glycemic index can decrease the risk of CHD,but it is still to be proved whether it is suitable to male.
		                        		
		                        		
		                        		
		                        	
10.Vibrio vulnificus induces acute necrosis of dendritic cells via toll-like receptor 2 and 4 pathways
Zhigang WANG ; Zhan WU ; Shuiling XU ; Ge CUI ; Lingjuan RUAN
Chinese Journal of Infectious Diseases 2013;(5):263-268
		                        		
		                        			
		                        			Objective To explore how Vibrio vulnificus (Vv) invades dendritic cells (DC) and induces acute necrosis of DC via toll-like receptor (TLR) 2 and 4 pathways.Methods Vv 1.1758 strain and DC 2.4 mixed culture model was established,observed the infection rates of DC with optical microscope,the location of Vv and structural changes of DC by transmission electron microscope.The expression levels of TLR2 and TLR4 mRNA were determined by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and tumor necrosis factor-α (TNFα) protein titers were measured by enzyme-linked immunosorbent assay (ELISA).DNA ladder qualitative test was used to detect cell apoptosis,while flow cytometry was used to quantify cell apoptosis and necrosis rates.Statistical analysis was done by chi-square test and one-way ANOVE.Results The infection rates of DC after 0.5,1,2,4 and 6 h of mixed culture were (7.8±0.8) %,(13.9± 1.1) %,(34.6±4.9) %,(77.8± 10.2)% and (95.8 ± 13.1)%,respectively.Vv was generally located in the internal cell membrane of DC 2.4.After 2 h co-culture,nuclear chromatins of DC became active and intranuclear apoptosis bodies appeared.After 4 h,cytoplasmic vacuoles appeared,chromatin gathered,and cell membranes were seriously damaged.After 6 h,mitochondria was highly swelled and distorted,and cell apoptosis and necrosis occurred.TLR2 and TLR4 mRNA levels reached peak values after co-culture for 0.5 h; TNF-α level began to increase at 1 h (P<0.05) and reached peak values at 2 h.DNA Ladder electrophoresis presented scouring necrosis after 2 h culture and apoptotic bands appeared between 720 bp and 900 bp after 4 to 5 h culture.Early apoptosis rates of DC after 2,4 and 6 h culture were (3.1±3.8)%,(7.8±4.7)% and (12.7±8.2)%,and necrosis rates of DC were (16.7±12.5)%,(41.6±25.9)% and (75.5±33.6)%,higher than that of control group (all P<0.05).Conclusions Vv infects DC and induce DNA degradation through up-regulated expression of TLR2 and TLR4 and increasing of TNF-α inflammatory mediators.During cell degradation,apoptosis and necrosis coexist,while necrosis is predominant.
		                        		
		                        		
		                        		
		                        	
            
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