1.Rat bone marrow stromal cells induced into dopaminergic neuron by mesencephalic conditional media and cytokines in vitro
Ping NIU ; Huan-Yi CHEN ; Shuai ZHAO ; Ying-Chun DU ;
Chinese Journal of Neurology 2005;0(10):-
Objective To approach a way to induce MSCs to dopaminergic neuron by mesencephalic conditional media and cytokines in vitro,and supply an ideal cells source for the treatment of Parkinson's disease.Methods The rat MSCs were isolated primarily from the femurs and tibias of the Wistar rats.MSCs were cultured,proliferated and purified by passage culture.Cultuered MSCs were divided into the control and the experimental group.In control group,MSCs were cultured without any induction medium.MSCs of experimental group were first cultured at medium containing bFGF for 24 hours.Then media were replaced with induction media which contained the agents as follows,respectively:GM_1,GDNF, GDNF+GM_1,GDNF+GM_1+mesencephalic conditional media.The surface markers of the differentiated neuron,such as NSE and TH were detected by immunocytoehemistry after MSCs were cultured in induction media for 3 and 7 days.Results In control groups,the NSE expression of MSCs was very lower than experimental groups.The percentage of NSE-positive cells of GDNF+GM_1+mesencephalic conditional media group in 7 day((45.257?5.999)/HP)was significantly more than other groups(control group is 2.214?0.779,GM_1 group is 22.014?3.624,GDNF group is 31.345?2.850,GDNF?GM_1 group is 40.314?4.203,P
2.A quantitative analysis of the kinematics of reaching by hemiplegic stroke patients using microsensor motion capture
Chun LUO ; Zhen HUANG ; Bin XIE ; Shuai HUANG ; Jiankang WU
Chinese Journal of Physical Medicine and Rehabilitation 2012;34(2):104-107
Objective To evaluate the feasibility of using a microsensor motion capture unit (MCU) to assess the reaching characteristics of hemiplegic stroke patients. Methods Twenty-three hemiplegic stroke patients with an average age of (61 ± 11) years and ten normal subjects of matching age were asked to sit on a chair and use the affected arm (for the patients) or the right arm (for the normal subjects) to reach for a cup on a table just in front of the shoulder at arm's length away.Four small sensor boxes which could detect movement and its speed and smoothness in three dimensions were fixed to the subject's spine,upper arm,forearm and hand.The test was repeated four times after two trial runs.The average velocity,peak velocity,degree of joint dispersion,entropy of acceleration,joint coordination and active range of motion ( AROM ) of the shoulder were analyzed.The patients and normal subjects were statistically compared.The results were correlated with Fugl-Meyer upper extremity scores using Spearman correlation analysis. Results All of the measured variables showed significant differences between the patients and normal subjects.In the correlation study,the average velocity,peak velocity,AROM of the shoulder and joint coordination showed significant correlation with the Fugl-Meyer scores.There was,however,no significant correlation between degree of dispersion of the shoulder or elbow joints or the entropy of acceleration ( which represented the quality of movement) and Fugl-Meyer upper extremity scores. Conclusions The results suggest that microsensor MCUs can quantitatively assess the kinematics of reaching among hemiplegic stroke patients.They can provide valuable and objective data about the functional quality of multiple joint movements in three dimensions.
3.Effect of apolipoprotein E genetic polymorphism in repair of blood brain barrier breakdown following traumatic brain injury
Shuang TANG ; Yong JIANG ; Shuai ZHOU ; Chun ZENG ; Xuehua XIONG ; Xiaochuan SUN
Chinese Journal of Trauma 2014;30(10):1040-1045
Objective To investigate the effect and underlying mechanism of apolipoprotein E (APOE) genetic polymorphism in treating blood brain barrier (BBB) breakdown after traumatic brain injury (TBI).Methods Human APOE knock-in mice (ε3,ε4),APOE knockout mice,and APOE wild-type mice with each numbering 80 were respectively divided into TBI group (n =50),sham-operation group (n =15) and normal control group (n =15) according to the random number table.TBI group was subdivided at 1 day (n=15),3 days (n=15),and7 days (n=20).TBI was induced with a pneumatically operated injury device.BBB permeability to large or small molecules was evaluated by measuring Evans blue (EB) and fluorescein sodium (NaFI) extravasation into the damage area at 1,3,and 7 days postinjury.Brain water content was determined using the dry-wet method.Western blotting and qRT-PCR for tight junction-associated proteins Occludin and Claudin-5 were performed at 7 days postinjury.Results With respect to normal control group,BBB permeability to EB and NaFI was significantly higher in ε4 and APOE knockout mice than in ε3 and APOE wild-type mice.There appeared significant increase in BBB permeability to EB and NaFI in TBI group,with insignificant differences among rats of each genotype at 1 and 3 days postinjury (P > 0.05).Whereas at 7 days postinjury,BBB permeability to EB in APOE wild-type and e3 mice returned to the normal level (P > 0.05),but it re mained at a high level in APOE knockout and ε4 mice (P < 0.01).Meanwhile,BBB permeability toNaFI was significantly higher in ε4 and APOE knockout mice than in ε3 and APOE wild-type mice (P < 0.01).Brain water content was equivalent among rats of each genotype at 1,3 and 7 days postinjury (P >0.05).Western blotting and qRT-PCR demonstrated Occludin and Claudin-5 in ε4 and APOE knockout mice were significantly lower than those in ε3 and APOE wide-type mice (P < 0.05).Conclusion APOE plays an important role in restoration of BBB function after TBI,but ε4 may impede the recovery of BBB breakdown after TBI through its effect on tight junction.
4.Neonatal dengue fever:four cases report
Junping WANG ; Yong ZHANG ; Chuan NIE ; Weiwei GAO ; Chun SHUAI ; Qianqing LIN ; Xiaorui HUANG ; Jie YANG
Journal of Clinical Pediatrics 2016;34(9):661-663
Objective To explore the clinical characteristics of neonatal dengue fever. Methods The clinical data from 4 neonates with dengue fever who were admitted and treated in 2014 were retrospectively analyzed and the related literatures had been reviewed. Results Four cases of neonatal dengue fever were all males. Three cases were mother to child transmission, the age at onset was 1 to 7 days after birth, and their mothers suffered with prenatal fever and were diagnozed of dengue fever during perinatal period. One case was community acquired, the age at onset was day 21 after birth and the neonate was bit by mosquito the day before. All four neonates had fever, two cases had rash, and one case had hemorrhagic spot. None of them had jaundice or cough. All of them had thrombocytopenia ( 30-125 )× 109/L, prolonged activated partial thromboplastin time ( 44 . 0-89 . 8 s), and increased aspartate aminotransferase (AST) ( 46-71 U/L). Three cases had declined ifbrinogen ( 1 . 36-2 . 53 g/L). Two cases had increased CK-MB ( 29-86 U/L). Two cases had increased CRP ( 3 . 00-46 . 05 mg/L). After the treatment of anti-infection and intravenous immunoglobulin, all of them were cured and discharged. The duration of hospital stay was 4-17 days. Conclusion The clinical manifestations of neonatal dengue fever were mainly fever and blood coagulation dysfunction, clinical symptoms are mild and lack of speciifcity, and prognosis are good. Mother to child transmission is one of the ways of dengue virus infection.
5.5-Aza-2 ’-deoxycytidine and 4-phenylbutyric acid exert a cooperative effect on the expression of miR-196 b in chronic myeloid leukemia cells
Yue LIU ; Chun SHUAI ; Jiesheng LI ; Hong YIN ; Yanbin SONG ; Wenli MA
Acta Anatomica Sinica 2014;(4):521-524
Objective To study if 5-Aza-2’-deoxycytidine along or together with 4-phenylbutyric acid could affect miR-196b expression levels in chronic myeloid leukemia cells .Methods K562 cells were treated with DNA methylation inhibitor 5-Aza-2’-deoxycytidine, histone deacetylase inhibitors 4-phenylbutyric acid separately and the combined treatment with both of them, then expression levels of miR-196b were detected using Real-time PCR.Results The half inhibition concentration of 4-phenylbutyric acid was 1.58mmol/L.Comparing with the expression level of miR-196b in normal human bone marrow cells, the expression levels of miR-196b were significantly lower in Aza group , PBA group and negative control cells and nearly consistent among three groups , and as high as normal cells in combined treatment group . Conclusion The expression level of miR-196b in K562 cells could not return to normal treated with 5-Aza-2 ’-deoxycytidine or 4-phenylbutyric acid separately , while could restore normal when treated with both agents , indicating that miR-196b expression level in K562 cells is related with both DNA methylation and histone acetylation .
6.Application of Cell Counting Kit-8 in detecting the growth inhibiting effect of 5-Aza-2 ’-deoxycytidine on chronic myeloid leukemia cells
Yue LIU ; Chun SHUAI ; Jiesheng LI ; Hong YIN ; Yanbin SONG ; Wenli MA
Acta Anatomica Sinica 2014;(4):582-584
Objective To study the application of Cell Counting Kit-8(CCK-8) in detecting the growth inhibiting effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cell .Methods The proliferation of K562 cells was detected by CCK-8 with different concentrations of 5-Aza-2 ’-deoxycytidine and the cell cycle and apoptosis of K 562 cells were detected after K562 treated by 50% inhibitory concentration of 5-Aza-2 ’-deoxycytidine .Results The 50% inhibitory concentration of 5-Aza-2’-deoxycytidine was 15.55nmol/L, after treated with this concentration , K562 cells showed that G2 phase arrest occurred , proliferation inhibited and apoptosis peaks appeared .Conclusion Inhibition of proliferation of K562 cells with different concentrations of 5-Aza-2’-deoxycytidine varied in a dose-dependent relationship , and 5-Aza-2’-deoxycytidine could promote apoptosis of K 562 cells.CCK-8 can be used in detecting the effect of 5-Aza-2’-deoxycytidine on chronic myeloid leukemia cells .
7.Effect of Schisandra chinensis lignans on neuronal apoptosis and p-AKT expression of rats in cerebral ischemia injury model.
En-Ping JIANG ; Shuai-Qun WANG ; Zhuo WANG ; Chun-Rong YU ; Jian-Guang CHEN ; Chun-Yan YU
China Journal of Chinese Materia Medica 2014;39(9):1680-1684
OBJECTIVETo observe the effect of Schisandra chinensis lignans (SCL) on neuronal apoptosis and PI3K/AKT signaling pathway of rats in the cerebral ischemia injury model, and study its possible mechanism.
METHODRats were orally administered SCL high, middle and low dose groups (100, 50, 25 mg x kg(-1)) for 14 days. The cerebral ischemia injury model was established by using the suture-occluded method to rate the neurological functions. The cerebral infarction area was observed by TTC staining. The pathological changes in brain tissues were determined by HE staining. Bcl-2 and Bax expressions were detected by immunohistochemical assay. The protein expressions of p-AKT and AKT were assayed by Western blotting.
RESULTCompared with the model group, SCL high, middle and low dose groups showed reduction in the cerebral infarction area to varying degrees, improve the pathological changes in brain tissues, promote the expression of apoptin Bcl-2 and p-AKT, and inhibit the expression of apoptin Bax.
CONCLUSIONSCL shows a protective effect on rats with cerebral ischemia injury. Its mechanism may be related to the increase in p-AKT ability and antiischemic brain injury capacity and the inhibition of nerve cells.
Administration, Oral ; Animals ; Apoptosis ; drug effects ; Blotting, Western ; Brain Ischemia ; metabolism ; pathology ; prevention & control ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Immunohistochemistry ; Lignans ; administration & dosage ; pharmacology ; Male ; Neurons ; drug effects ; metabolism ; pathology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Phytotherapy ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rats ; Rats, Sprague-Dawley ; Schisandra ; chemistry ; Signal Transduction ; drug effects ; bcl-2-Associated X Protein ; metabolism
8.Effects of guizhi fuling capsule on sex hormone levels and breast issue morphology of mammary gland hyperplasia model rats.
Xiu-juan CHANG ; Jun ZHOU ; Shuai ZHANG ; Jian CHEN ; Chun-miao CHEN ; Zhen-zhong WANG ; Wei XIAO
China Journal of Chinese Materia Medica 2014;39(21):4139-4142
OBJECTIVETo research the effects of Guizhi Fuling capsule on sex hormones levels in blood serum and breast issue morphology of hyperplasia of mammary glands model rats.
METHODThe unpregnancy SD rat models of hyperplasia of mammary glands were established by injecting 0.5 mg x kg(-1) benzoate estradiol. After five weeks doses,the effects of Guizhi Fuling capsule 2.0, 1.0, 0.5 g x kg(-1) and Rupixiao tablet 0.5 g x kg(-1) on the changes of papilla diameter, height and breast issue morphology of the naimal models were explored, and sex hormones levels in blood serum were measured.
RESULTGuizhi Fuling capsule can inhibitnipple swell, improve breast tissue morphology pathological profiles of the animal models, and decrease oestradiol (E2) level and increase progesterone (P) level in blood serum.
CONCLUSIONThese results suggested that Guizhi Fuling capsule could, improve mammary gland pathological profiles. Regulating sex hormone levels may be its important mechanism for treatment of hyperplasia of mammary glands.
Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadal Steroid Hormones ; blood ; Hyperplasia ; Mammary Glands, Animal ; drug effects ; pathology ; Rats
9.Observation of clinical curative effect in the patients with internal endometriosis by interventional therapy
Peng WANG ; Xin-yan NG ZHA ; Shuai WANG ; Shun-ji SUN ; Xiu-chun WANG ; Zhao-cheng JIAN ; Ye-quna SUN
Chinese Journal of Postgraduates of Medicine 2012;35(3):8-10
ObjectiveTo explore the curative effect in the patients with internal endometriosis by interventional therapy.MethodsUsing Seldinger technique,34 cases with internal endometriosis wereperformed bilateral uterine artery embolization.Observed postoperative menstrual quantity,dysmenrrhea degree,anemia and the change of the volume of uterine lesions.ResultsAll the patients were followed up for 1-3 years,menstrual quantity average decreasd of 59.1%P < 0.05 ),the symptoms of dysmenorrhea was significantly eased in 28 cases (82.4%,28/34).All the patients of anemia haemoglobin were back to normal,volume of uterus average reduced 43.8%P < 0.05 ),lesion was obviously smaller or disappear.Ultrasonography showed myometrium and blood flow signal of lesion was was obviously reduced.Conclusion Internalendometriosis by interventional therapy can get good results,symptoms improve significantly.
10.Determination of oleanic acid and paeoniflorin in Paeonia lactiflora by ultrasound-assisted ionic liquid-reversed phase liquid chromatography.
Wei LIU ; Dong-dong LI ; Hong-shuai YANG ; Yuan-yuan CHEN ; Jin-feng WEI ; Wen-yi KANG ; Xiu-chun GUO
China Journal of Chinese Materia Medica 2015;40(3):443-449
Four kinds of ionic liquids [BMIM] Br, [BMIM] BF4, [BMIM] PF6, [HMIM] PF6 were used to analyze the content of oleanic acid and paeoniflorin in Paeonia lactiflora with ultrasonic-assisted extraction coupled with HPLC. The chromatographic column, Purospher star RP-C18 (4.6 mm x 250 mm, 5 μm), was used. Acetonitrile and water (90:10) as mobile phase was used to determine the content of oleanic acid with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 210 nm, chromatographic column temperature at room temperature. Paeoniflorin content was determined using acetonitrile and water (18:82) as mobile phase with a gradient elution and flow rate at 1.00 mL · min(-1), detection wavelength at 250 nm, the chromatographic column temperature at room temperature. The result show that oleanic acid has the highest extraction yield when the conditions are solid-liquid ratio of 1:80 (g · mL(-1)), and the [BMIM] Br methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, the content of oleanic acid from 0.24 to 3.76 μg showed a good linearity (r = 0.9999), the average recovery was 97.20%. Paeoniflorin has the highest extraction yield when the conditions are solid-liquid ratio of 1:130 (g · mL(-1)), and the [C4 MIM] PF6 methanol solution concentration of 0.6 mol · L(-1). Under the optimal extraction conditions, paeoniflorin content from 0.42 to 4.20 μg showed a good lin- earity (r = 1.000), the average recovery was 98.84%. This method is simple and reliable, its repeatability is also very good. It has important significance in the study P. lactiflora of ionic liquid microextraction.
Chromatography, Reverse-Phase
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methods
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Glucosides
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analysis
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Ionic Liquids
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chemistry
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Monoterpenes
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analysis
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Oleanolic Acid
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analysis
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Paeonia
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chemistry
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Ultrasonics