1.Influence of cerebral hemorrhage stress on the proliferation and senescence of endothelial progenitor cells
Chinese Journal of Behavioral Medicine and Brain Science 2010;19(11):986-988
Objective To explore the influence of cerebral hemorrhage stress on endothelial progenitor cells(EPCs) proliferation and senescence. Methods Mononuclear cells from peripheral blood of patients with cerebral hemorrhage (CH group) and patients with the same underling diseases (control group) as the front were isolated and induced to EPCs. Proliferation, numbers of colonies and cells, senescence of EPCs were checked and compared between the two groups. Results Compared with control group, proliferation (0.270 ± 0. 074 vs 0. 185 ±0.050, t=3.802) ,numbers of colonies( (5. l0 ±0.79) cell/HPF vs ( 1.90 ±0.41 ) cell/HPF, t= 14.407) and cells( (75.15 ± 10.48) cell/HPF vs(57.10 ± 9.28 ) cell/HPF, t = 5. 157 ) of EPCs in CH group on the stress of cerebral hemorrhage were increased significantly (P < 0.01 ), but senescence of EPCs ( (45.54 ± 5.74 ) cells/100 cells vs ( 46.86 ± 5.06 ) cells/100 cells, t = 0. 686 ) in two groups were not different (P > 0.05 ). Conclusion Cerebral hemorrhage stress can increase proliferation of EPCs,but can not change the senescence destiny of EPCs.
2.Study on the Mechanism of Trimethoprim-Sulfamethoxazole Resistance in Stenotrophomonas Maltophilia
Suying ZHAO ; Hongxiang LIU ; Min LI ; Huiling CAO ; Hengfang SHI
Journal of Modern Laboratory Medicine 2014;(6):25-27,31
Objective Class 1 integrase(intI 1),qacE△1-sul1 and sul2 genes were detected in trimethoprim-sulfamethoxazole resistant Stenotrophomonas maltophilia by PCR to assess the relationship between the antibiotic resistance mechanism for trimethoprim-sulfamethoxazole (TMP-SMZ)and these genes distribution.Methods S.maltophilia isolates were collected from patients treated in Affilitated hospital of Nanjing University of TCM during January to May in 2013,DNA was ab-stracted by boiling method and genes were detected by polymerase chain reaction.Results intI 1 genes were observed posi-tive in 25 of 28 strains resistant for TMP-SMZ,qacE△1-sul1 genes were positive in 21 while sul2 genes positive in 15,the positive rates of intI 1,qacE△1-sul1 and sul2 genes were 89.29%,75% and 53.57%;in 18 trimethoprim-sulfamethoxazole sensitive strains,5 were intI 1 positive,4 were qacE△1-sul1 positive while sul2 were none,the positive rates of intI1 and qacE△1-sul1 were 27.78% and 22.22%.Conclusion Most of stenotrophomonas maltophilia resisted trimethoprim-sulfa-methoxazole had intI 1,qacEΔ1-sul 1 and sul2 genes.
3.Effects of atorvastatin on the gene expressions of cyclooxygenase-2 and membrane-associated prostaglandin E2 synthase-1 in carotid atherosclerotic plaques of rabbits
Min ZHANG ; Jianping WANG ; Meimei ZHANG ; Yuanzheng ZHAO ; Wang MIAO ; Zunjing LIU ; Hengfang LIU
Chinese Journal of Geriatrics 2012;31(6):525-528
Objective To explore effects of atorvastatin on the expressions of cyclooxygenase-2(COX-2) and membrane-associated prostaglandin E2 synthase-1 (mPGES-1) in the carotid atherosclerotic plaques of rabbits.Methods Totally 33 male New Zealand white rabbits(≥ 36months of age ) were assigned into normal control group (n=8) and animal model group with carotid atherosclerotic stenosis (n =25).The rabbit models were randomly divided into non-intervention group,celecoxib treatment group (15 mg · kg-1 · d-1,twice daily) and atorvastatin treatment group (5 mg · kg-1 · d-1,once daily) (n=8 each).Four weeks after treatment,the mRNA and protein expressions of COX-2 and mPGES-1 in carotid plaques were determined by RT-PCR and Western blot,respectively.Results The mRNA expressions of COX-2 (0.97±0.09,0.44±0.05,0.60±0.04vs.0.23±0.04,F=66.77,P<0.01) and mPGES-1 (0.92±0.07,0.41±0.04,0.61±0.03 vs.0.17±0.03,F=54.87,P<0.01)in carotid atherosclerotic plaques were significantly higher in non intervention group,celecoxib treatment group and atorvastatin treatment group than in normal control group.The mRNA expressions of COX-2 and mPGES-1 were decreased in celecoxib treatment group and atorvastatin treatment group as compared with non-intervention group ( both P < 0.01 ).The protein expressions of COX-2 (0.89±0.06,0.42±0.07,0.62±0.04 vs.0.18±0.05,F=61.75,P <0.01) and mPGES-1(0.91±0.05,0.44±0.05,0.63±0.05 vs.0.21±0.04,F=86.44,P<0.01)in carotid atherosclerotic plaques in non-intervention group,celecoxib treatment group and atorvastatin treatment group were increased as compared with those in normal control group.The mRNA and protein expressions of COX-2 and mPGES-1 were decreased in celecoxib treatment group and atorvastatin treatment group as compared with non-intervention group(all P<0.01 ).The expressions of COX-2 and mPGES-1 in carotid atherosclerotic plaques were reduced in celecoxib treatment group as compared with atorvastatin treatment group (P < 0.01).Conclusions As COX-2 inhibitor celecoxib,atorvastatin may inhibit the expressions of COX-2 and mPGES-1,and interfere with the inflammatory response which plays key role in the pathological progress of carotid atherosclerotic plaques,and thus slow the progress of carotid atherosclerosis.
4.The effects of magnesium valproate on expression of the kinin B1 and B2 receptors in the hippocampus of the juvenile rats submitted to pilocarpine model of epilepsy
Min ZHANG ; Jianping WANG ; Meimei ZHANG ; Wang MIAO ; Zunjing LIU ; Lei SHI ; Ningning CHEN ; Hengfang LIU
Chinese Journal of Behavioral Medicine and Brain Science 2012;21(4):324-326
ObjectiveTo investigate the mechanisms and the effects of magnesium Valproate on the expressions of the kinin B1 and B2 receptors in the hippocampus of the juvenile rats submitted to pilocarpine model of epilepsy.Methods 35 healthy Wistar juvenile rats were randomly divided into six groups,that is the model groups:Ⅰ group,Ⅱ group,Ⅲ group,and intraperitoneal injection of saline water control groups:Ⅰ a group,Ⅱ a group,Ⅲ a group,after succession of 15 rats to kindle to establish the model of epilepsy by pilocarpine.To collect hippocampus tissue after the rats were to put to death,and to compared the expression levels of kinin B1 and B2 receptor mRNA by RT-PCR and western blot in the hippocampus of rats.ResultsBy treated with magnesium valproate,kinin B1 receptor mRNA (0.38 ± 0.051 ) and protein expressions(0.58 ± 0.057 ) decreased and kinin B2 receptor mRNA (0.48 ±0.056 ) and protein expressions(0.48 ± 0.044 ) increased in Ⅰ group,compared with that (0.76 ±0.068,0.89 ± 0.034;0.28 ± 0.034,0.32 ± 0.039 ) of Ⅰ a group(P < 0.05 ).Compared with control group,there were more significant upregulation of kinin B1 receptor mRNA and protein expressions (P<0.05) in the Ⅰ and the Ⅱ groups and there were no alteration in Ⅲ group.The expressional levels of B2 receptor mRNA and protein were upregulated in the Ⅰ,Ⅱ and Ⅲ groups.ConclusionThe kinin B1 and B2 receptor may play a role in the onset and maintenance of epilepsy.The magnesium valproate increased the expressional levels of kinin B2 receptor,and decreased the expressional levels of kinin B1 receptor.
5.The clinical and electrophysiological characteristics of flail arm syndrome
Fang LIU ; Changdong SONG ; Hengfang LIU ; Min ZHANG ; Shuyan FENG ; Liuyi LI
Chinese Journal of Physical Medicine and Rehabilitation 2021;43(1):21-24
Objective:To explore the clinical and electrophysiological characteristics of flail arm syndrome (FAS).Methods:Clinical and electrophysiological data were collected on 13 FAS patients and 31 persons with upper limb onset amyotropic lateral sclerosis (UL-ALS), including the amplitude of compound muscle action potential (CMAP) related to median nerve, ulnar nerve, and axillary nerve motor conduction. A split-hand index (SI) was calculated by dividing the CMAP amplitude of abductor pollicis brevis by that of the abductor digiti minimi. Clinical features, the CMAP amplitudes and SIs were compared between the FAS and UL-ALS patients.Results:Compared with UL-ALS patients, the age at onset among the FAS patients was older (averaging 60.9 years). The development to the second stage was longer (24±6 months). The upper limb reflexes of 15% of the FAS patients had disappeared and those of 77% were weakened, while the lower limb reflexes of 54% of the FAS patients were active and 38% were weakened, significantly different from the UL-ALS patients. However, there were no significant differences in the CMAP amplitudes of the median and ulnar nerves, nor in SI between the FAS and UL-ALS patients. The SIs of the FAS patients with upper motor neuron signs were significantly lower than those of FAS patients without such signs. Among the FAS patients, the average CMAP amplitude of the ulnar nerve was the highest, followed by those of the median and axillary nerves. Among the UL-ALS patients, however, the average CMAP amplitude of the ulnar nerve was not significantly different from that of the axillary nerve.Conclusions:FAS patients with upper motor neuron signs are more likely to have slip hand. The CMAP amplitude of their axillary nerve tends to be lower than that of their median and ulnar nerves. FAS seems to be a special type of ALS.
6.Clinical analysis of mitochondrial encephalomyopathy with lactic acidosis and stroke -like episodes in children
Shitao WU ; Hengfang LIU ; Fang LIU ; Min ZHANG ; Yuanzheng ZHAO ; Yapei GUO ; Ningning CHEN ; Jianping WANG
Chinese Journal of Applied Clinical Pediatrics 2016;31(24):1884-1888
Objective To investigate the clinical,imaging,pathological and molecular biological features of mitochondrial encephalomyopathy with lactic acidosis and stroke -like episodes(MELAS)in children.Methods The clinical,imaging,pathological and molecular biological features of 1 2 children with MELAS diagnosed through muscle biopsy or gene sequencing in the Fifth Affiliated Hospital of Zhengzhou University from January 201 1 to December 201 5 were retrospectively analyzed.Results (1 )Clinical features:the main manifestations included headache and vomiting in 1 1 cases,epileptic seizures in 9 cases,short stature in 8 cases,hairy in 7 cases,intolerance fatigue in 7 cases,cogni-tive decline in 7 cases,visual disturbance in 6 cases,hearing disturbance in 6 cases,and 5 cases had positive family history.In addition,7 cases had the serum lactic acid level increase in a rest for 1 0 min after exercise.(2)Imaging fea-tures:4 cases showed bilateral basal ganglia calcification symmetry in 8 patients who underwent head CT scan.The most frequently involved parts of the lesion were occipital in 1 0 cases,temporal in 9 cases and parietal lobe in 7 cases in stroke -like episodes.The lesions were lamellar necrosis.The abnormal areas by MRI showed low signal intensity on T1 weighted imaging,high signal intensity on T2 weighted imaging and fluid attenuated inversion recovery,high or equal signal intensity on diffusion weighted imaging,high or low signal intensity on apparent diffusion coefficient;the lactate peak significantly increased on magnetic resonance spectroscopy.The distribution was not in accordance with the control region of the cerebral vessels.Dynamic observation revealed that the lesions were reversible and migratory.(3)Myo-pathological features:muscle biopsy was performed in all children,and ragged -red fibers were found in 1 0 cases by im-proved Gomori staining,strongly succinate dehydrogenase -reactive were found in 9 cases,and the lipid droplets slight-ly increased in 8 cases by oil red O staining.Besides,the crystalline inclusion bodies in mitochondria were arranged in a parking lotpattern in 9 cases by electromicroscope.(4)Molecular biological characteristics:the mitochondrial gene mutations were analyzed in peripheral blood of 9 children and their parents,including 8 cases with A3243G muta-tion and 1 case with G13513A mutation.Five mothers had the same A3243G mutation site in 8 cases.Conclusions Children with MELAS have complex and varied clinical manifestations and certain characteristic of neuroimaging.More-over,muscle pathology and gene sequencing have important diagnostic value.Fully understanding the clinical,muscle pathology,imaging and molecular biological characteristics of children with MELAS can be helpful to the early diagnosis and treatment,also reduce misdiagnosis.
7.Expression of Vascular endothelial growth factor mRNA and protein in endothelial progenitor cells in patients during acute cerebral hemorrhage associated with hypertension
Wang MIAO ; Qidong YANG ; Weixin GUO ; Xiaoyun XIE ; Hengfang LIU ; Hongbo LUO ; Juan BAO
Chinese Journal of Neurology 2011;44(2):101-104
Objective To explore the expression of vascular endothelial growth factor (VEGF)mRNA and protein in endothelia progenitor cells (EPCs) and VEGF in the culture medium and serum of patients during acute period of cerebral hemorrhage associated with hypertension (APCHH). Methods Mononuclear cells from peripheral blood of patients with APCHH ( 16 patients) and hypertension ( 16 patients) were isolated and induced to EPCs. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis was performed to assay VEGF mRNA. VEGF protein was assessed by Western blotting. The VEGF protein level in patient's serum and culture medium ( at day 7 ) were assayed using VEGF ELISA Kit and compared between APCHH group and hypertension group. Results Compared with hypertension group, VEGF mRNA (0. 186 ±0. 035 versus 0.090 ±0.031, t =8.318, P <0.0l ) and protein (0. 223 ± 0. 028 versus 0.169 ± 0. 022, t = 3. 744, P < 0. 01 ) expression of EPCs, the concentration of VEGF protein in the supernatant (414 ±37 versus 316 ±29, t =8. 270, P <0. 01 ) and in serum (408 ±49versus 222 ±34, t = 12.406, P <0. 01 ) were all significantly increased in APCHH group. Conclusion The VEGF protein levels in serum of patients and in the culture medium, VEGF mRNA and protein expression in EPCs were all significantly increased during acute periods of cerebral hemorrhage.
8.Effects of thyroid hormone on NF-κB and TNF-α in cerebra cortex of rats with bran ischemic reperfusion injury
Zheng MA ; Min ZHANG ; Hengfang LIU ; Shitao WU ; Yapei GUO ; Chao DOU ; Qiongzhe ZHANG
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(1):8-12
Objective To investigate the effect of thyroid hormone on the expression of NF-κB and TNF-oα in the ischemic cortex of rats after focal cerebral ischemia-reperfusion.Methods 96 male SD rats were randomly divided into sham-operation group,sham-operation+T3 group,IR group and IR+T3 group.Using suture legal method to establish a rat model of middle cerebral artery occlusion for 2 h followed by reperfusion.In sham-operation+T3 group and IR+T3 group,T3 was given 3 days before ischemia and 1 hour after ischemia,respectively,intraperitoneal injection T3 10 μg/100 g for rats.Rats in other groups were given the same volume normal saline at the same time.The infarct size was determined by TTC staining at 24 h after reperfusion.HE staining was used to observe the morphological and structural changes of brain tissue.Using Real-time PCR method and immunohistochemical staining method to detect the expression of NF-κB mRNA,TNF-α mRNA and protein in ischemic cortex of rats.Results Compared with sham-operation group and sham-operation+T3 group,the pathological damage of brain tissue in IR group was obvious,while the pathologic damage of IR +T3 group was less than that in IR group.Immunohistochemistry assay showed that the expression of NF-κB was(49.19±5.55)in sham-operation group,(45.75±2.12) in sham-operation+T3 group,(56.88±2.23)in IR group and(50.25±1.67)in IR +T3 group,the expression of TNF-α was (22.50±3.07) in sham-operation group,(24.13±2.03) in sham-operation+T3 group,(37.25±2.82) in IR group and (30.25±1.67) in IR +T3 group,and the NF-κB,TNF-α in IR group were obviously higher than that in sham-operation group and sham-operation+T3 group(P<0.05),while IR+T3 group were lower than that in IR group(P<0.05).Real-time PCR showed that NF-κB mRNA,TNF-α mRNA level in IR group was the highest,which was higher than that of sham-operation group and sham-operation+T3 group(P<0.05),and the NF-κB mRNA,TNF-oα mRNA expression in IR+T3 group were significantly decreased compared with that in IR group(P<0.05).Conclusion Thyroid hormone has a protective effect on cerebral ischenia reperfusion injury,which may be achieved by reducing the expression of inflammatory factor NF-κB and TNF-oα.
9.Effects of bone marrow mononuclear cells transplantation on ERK1/2 signal pathway after cerebral infarction in rats
Ningning CHEN ; Jianping WANG ; Xiaojie FU ; Hengfang LIU ; Min ZHANG ; Yuanzheng ZHAO
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(8):678-683
Objective To explore the neuroprotection and mechanisms of bone marrow mononuclear cells (BMMNCs),and evaluate whether ERK1/2 signaling pathway was involved in it.Methods384 healthy male SD rats,which were 6-8 week old,weighting 250-280 g,were selected.The middle cerebral artery occlusion (MCAO) model was established in SD rats using the suture method.The rats were randomly divided into sham operation group,model group,BMMNCs group and ERK1/2 inhibitor group,with 96 rats in each group.At the time of 24 h after the successful modeling,200 μl PBS solution was injected into the caudal vein of the rats in the model group,200 μl PBS solution containing 5×106 BMMNCs was injected into the rats in the BMMNCs group and the ERK1/2 inhibitor group.meanwhile,5 μl PD98059 was injected into the lateral ventricle of the brain of rats in the ERK1/2 inhibitor group.At the time points of 3 d,7 d and 14 d,the modified neurological severity scores (mNSS) was used to evaluate the neurological function,the volume of cerebral infarction was assessed by TTC staining,the pERK1/2,Bax,Bcl-2 and caspase-3 levels were detected by Western blot,and the effect of BMMNCs on activation of microglia was detected by immunofluorescence assay.Results(1)At each time point,the mNSS and the volume of cerebral infarction of the model group were significantly higher than those of the sham operation group (P<0.05),while the mNSS and the volume of cerebral infarction of the BMMNCs group were lower than those of the model group,higher than those of the sham operation group,and it was gradually decreased with the treatment time extension (P<0.05).There was no difference in comparison between the ERK1/2 inhibitor group and the model group (P>0.05).(2)At each time point,the pERK1/2,Bcl-2,Bax and caspase-3 protein levels of the model group were significantly higher than those of the sham operation group (P<0.05).The pERK1/2 and Bcl-2 protein levels of the BMMNCs group(pERK1/2:(0.38±0.16),(0.39±0.15),(0.40±0.20),Bcl-2:(0.38±0.14),(0.39±0.15),(0.37±0.13)) were higher than those of the model group(pERK1/2:(0.17±0.05),(0.14±0.04),(0.13±0.03),Bcl-2:(0.23±0.11),(0.24±0.12),(0.27±0.14),Bax:(0.39±0.13),(0.40±0.14),(0.45±0.23),caspase-3:(0.52±0.26),(0.56±0.27),(0.58±0.28)),while Bax and caspase-3 protein levels(Bax:(0.25±0.13),(0.19±0.06),(0.21±0.08),caspase-3:(0.35±0.13),(0.34±0.16),(0.29±0.09)) were decreased (P<0.05).The pERK1/2 protein level of ERK1/2 inhibitor group was lower than other groups,There was no difference in the level of Bcl-2,Bax and caspase-3 between the ERK1/2 group and the model group.(P>0.05).(3) At each time point,microglia (Iba1 positive) in ischemic penumbra of the BMMNCs group was significantly more than those of the model group,and it was increased with the time extension (P<0.05).There was no difference in comparison between the ERK1/2 inhibitor group and the model group (P>0.05).ConclusionBMMNCs can reduce the apoptosis through ERK1/2 signaling pathway,thus improving the neurological function and reducing the infarct scope.
10.Effect of thyroid hormones on expressions of CytC and AIF after cerebral ischemia-reperfusion injury in rats
Qiongzhe ZHANG ; Hengfang LIU ; Min ZHANG ; Ming CUI ; Yapei GUO ; Shitao WU
The Journal of Practical Medicine 2017;33(1):67-71
Objective To investigate the effect of thyroid hormone(T3)on the expressions of cytochrome c (CytC) and apoptosis?inducing factor(AIF) after cerebral ischemia reperfusion injury in rats and its mechanism. Methods SD male rats were randomly divided into four groups: sham operation group(sham1),sham operation group + T3(sham2) ,ischemia?reperfusion group (IR) ,and thyroid hormone treatment group(T3). A rat model of cerebral ischemia?reperfusion injury was established by right middle cerebral artery occlusion for 2 h,followed by reperfusion for 24 h. Thyroid hormones (10μg/100 g) or normal saline were given at 1 h after onset of ischemia and 6 h after reperfusionby intraperitoneal injection. Neurological deficit scores were evaluated 24 h after reperfusion. Cerebral infarction volume was evaluated by TTC staining. Histological changes was observed by HE staining. Expressions and mRNA levels of CytC and AIF in ischemic brain tissue were evaluated by immunohistochemistry and real?time fluorescent quantitative RT?PCR. Results Ascompared with those in sham operation groups ,the expressions and mRNA levels of CytC and AIFincreased significantlyin IR groups. As compared with those in IR groups ,the indexes were remarkably decreasedin T3 groups (P < 0.01). Nerve function was markedly improved andinfarction area narrowed.Conclusions Thyroid hormone plays a certain role in protection of cerebral ischemia?reperfusion injury in rats,whose mechanism may be associated with inhibition of the expression of apoptosis factors?CytC and AIF.