3.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
5.Study on the Role of ?-synuclein in Mitochondria Dysfunction Caused by Small Dosage Rotenone
Ling-Ling LU ; Yuan LIANG ; Chun-Li DUAN ; Chun-Li ZHAO ; Huan-Ying ZHAO ; Hui YANG ;
China Biotechnology 2006;0(12):-
Mitochondrial dysfunction has been implicated in the aetiology of sporadic Parkinson's disease but its role in the disease mechanism remains unclear.To investigate the effect of synuclein on mitochondrial dysfunction induced by rotenone.The human dopaminergic SH-SY5Y cells were used as a cell model.The cells over-expressed the wild-type ?-synuclein were treated with complex I inhibitor rotenone.The cell viability,complex I activity,Mitochondrial swelling and O2-content were tested at different time point-1w,2w,4w after rotenone treated.CCK-8 test results showed that the cell viability of overexpressed ?-synuclein(SH-SY5Y-Syn)was much lower than the control group(SH-SY5Y-Ctr).After administrating with rotenone about 1w or 2w the cell viability of SH-SY5Y-Syn became higher than that of SH-SY5Y-Ctr.On the 4th week the results were contrary to the first 2 weeks.Similar results were got when test the mitochondrial function.In the first 2 weeks after roteoone administrating,the mitochondrial function of SH-SY5Y-Syn was better than that of SH-SY5Y-Ctr.This suggest that the ?-synuclein could protect the mitochondrial against the injury induced by rotenone in the early stage-1w,2w,while this effect disappeared in the final stage-4w.
7.N-terminal of ?-Synuclein Involved in Regulation of Mitochondrial Function
Li LV ; Tao ZHANG ; Qi LIU ; Chun-Xiang FAN ; Ling ZHANG ; Huan-Ying ZHAO ; Chun-Li ZHAO ; Hui YANG ;
China Biotechnology 2006;0(12):-
Objective: To identify the functional domain of ?-Synuclein in affecting mitochondrial function and how the function to be impaired,especially,the mitochondrial membrane potential and the release of Cytochrome c.Methods: Harvest of ?-Syn-N and ?-Syn-△N by PCR,then subcloned into the pCMV-Myc mammalian expression vector.The recombinant plasmids were transfected into HEK293T cells by Lipofectamine 2000.After detecting the protein expression by Western blot,the functional domain was detected by co-immunoprecipitation.The mitochondrial membrane potential through flow cytometry and immunofluorescence,at the same time,the release of Cytochrome c through flow cytometry to detect.Results: The recombinant plasmids were constructed successfully.CO-IP has proved that N-terminal may be the functional domain of ?-Synuclein in affecting mitochondria.Over-expression of N-terminal could depolarize the mitochondrial membrane potential and induce the Cytochrome c releasing in MN9D cells.Conclusion: N-terminal may be the functional domain of ?-synuclein and over-expression of N-terminal could decrease mitochondrial activity.
8.Clinicopathological Characteristics of Alpha-Fetoprotein-Producing Gastric Cancer.
Journal of Gastric Cancer 2011;11(1):23-30
PURPOSE: alpha-fetoprotein (AFP)-producing gastric cancer is a rare tumor with high rates of liver metastasis and a poor prognosis. Many studies have been performed but there have been no comprehensive investigations of the clinicopathological and prognosis. MATERIALS AND METHODS: Six hundred ninety four patients with gastric cancer who underwent a curative gastric resection in Hanyang University Hospital from February 2001 to December 2008 were evaluated retrospectively after excluding active or chronic hepatits, liver cirrhosis and preoperative distant metastasis. Among them, thirty five patients had an elevated serum level of AFP (>7 ng/ml) preoperatively. The clinicopathological features of AFP-producing gastric cancer were analyzed. RESULTS: There was poorer differentiation, a higher incidence of lymph node metastasis, more marked lymphatic and vascular invasion in the AFP-positive group than in the AFP-negative group. The 5-year survival rate of the AFP-positive group was significantly poorer than that in the AFP-negative group (66% vs. 80%, P=0.002). A significantly higher incidence of liver metastasis was observed in the AFP-positive group than in the AFP-negative group (14.3% vs. 3.6%, P=0.002) with a shorter median time period from the operation to the metachronous liver metastasis (3.7 months vs. 14.1 months, P=0.043). Multivariate survival analysis revealed the depth of invasion, degree of lymph node metastasis and AFP-positivity to be the independent prognostic factors. CONCLUSIONS: AFP-producing gastric cancers have an aggressive behavior with a high metastatic potential to the liver. In addition, their clinicopathological features are quite different from the more common AFP-negative gastric cancer.
alpha-Fetoproteins
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9.Effect of Melatonin on CD4~+CD_(25)~+ Regulatory T Cell and Airway Inflammation in Asthmatic Rat
min, WANG ; jun-lan, YAO ; an-xiu, XIONG ; guang-huan, ZHANG ; qun-xing, WANG ; chun-hua, LUO
Journal of Applied Clinical Pediatrics 1994;0(04):-
Objective To explore the effect of Melatonin(MT) on CD4+CD25+ regulatory T cell (CD4+CD25+Tr)and airway inflammation in asthmatic rat.Methods Thirty-two SD rats were randomly divided into 4 groups,8 rats in each group.Asthmatic group:rats were immunized on day 1 and 7 by intraperitoneal inject of mixture of ovalbumin(OVA) and aluminumhydroxide.From day 14,the animals were allenged with aerosolized OVA for 20 min per day for 7 consecutive days.MT group:OVA-sensitized rats were injected intraperitoneally with 0.1 mg/kg MT 30 min before each OVA challenge.Dexamethasone group:OVA-sensitized rats were injected intraperitoneally with 0.5 mg/kg Dexamethasone 30 min before each OVA challenge.Control group:OVA for inhalation and MT for intraperitoneal injection was replaced with saline.After the last challenge,peripheral blood was stained to count the percentage of eosinophil(EOS).Then the rats were lavaged and total leukocytes counts in bronchoalveolar lavage fluid(BALF) were performed after staining with Wright-Giemsa staining.The EOS counts around the airway was counted after the histological section of lung staining with hematoxylin and eosin staining.The serum level of immunoglobulin E(IgE) was detected by immunoenhancement.The change of CD4+CD25+Tr was assessed with flow cytometry.SPSS 10.0 software was applied to analyze data. Results In asthmatic rats,the CD4+CD25+ Tr/ CD4+T cells ratio had significant negative relationship with the EOS counts around the airway and the total leukocytes counts in BALF (r=-0.73 P0.05).There was a significant decrease in the percentage of the eosinophils in peripheral blood,the eosinophil counts around the airway,the total leukocytes counts in BALF and the serum level of IgE in MT group compared with asthmatic group (Pa
10.Changes of Serum Zinc Level of Rotaviral Enteritis in Children and Therapeutic Effect of Zinc
chun-hua, KANG ; xiao-shan, QIU ; li, HUA ; huan-kui, HUANG
Journal of Applied Clinical Pediatrics 2003;0(10):-
Objective To explore the change of serum zinc level of children with rotavirus(RV) enteritis and efficacy of zinc supplementation on children with rotaviral enteritis.Methods One thousand one hundred and eighty-three healthy children were enrolled in this study,who were divided into different groups,and the zinc levels of children in every different age group were detected.These levels were used as normal levels of children in Dongguan district.One hundred and forty-three diarrhea patients in this 2 years were enrolled.Confirmed dia-gnosis was depended on the RV test in patients′ stool.The children with diarrhea were randomly divided into zinc treatment group (n=72) and conventional therapy group (n=71).The serum zinc levels in RV enteritis was analyzed.The patients in conventional therapy group were received diarrhea conventional therapy inchuded smectite and fluid infusion and so on;in addition to diarrhea conventional therapy,zinc gluconate 20 mg/d (10 mg/d below 6 months of age) for successive 14 days were given the patientsin zinc treatment group.The different efficacy of the 2 groups and the changes of serum zinc in treatment group were analyzed.Results Compared with normal children,the ones suffering from RV enteritis had lower serum zinc level(P0.05).Conclusions The ones suffering from RV enteritis have lower serum zinc level.In addition to diarrhea conventional therapy,giving zinc treatment has better effect on enhancing curative effect of RV enteritis.On the other hand,the suggestion of zinc supplement made by WHO is also suitable for Chinese children.