Animals ; Calreticulin ; metabolism ; Male ; Muscle, Skeletal ; metabolism ; Oxidative Stress ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley

Acupuncture Therapy ; Animals ; Animals, Newborn ; Brain ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; therapy ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley

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

Humans ; Accidents, Traffic ; Colon, Sigmoid ; Forensic Medicine ; Autopsy ; Abdominal Injuries ; Thoracic Injuries ; Rupture

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.

Animals ; Conditioning, Classical ; Evoked Potentials, Somatosensory ; Fear ; psychology ; Male ; Rats ; Rats, Sprague-Dawley ; Stress, Psychological

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.

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 ; Humans ; Incidence ; Liver ; Liver Cirrhosis ; Lymph Nodes ; Neoplasm Metastasis ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; Survival Rate

OBJECTIVETo apply the Wright-Giemsa stain in micronucleus test and to explore the stain outcomes of Wright-Giemsa dye of various proportions and staining times.

METHODSUse Wright-Giemsa dye, Wright dye (staining time 3 min) and Giemsa dye (staining time 5 min) to stain HepG2 and then observe the staining effect. The Wright-Giemsa dye was applied under 5 different proportions (3:1-1:3) and different staining times (1, 3, 5, 10, 15 min).

RESULTSAfter stained for 3-5 min with the proportion ratio of 3:1 of Wright-Giemsa dye, the HepG2 cells showed much better staining outcomes compared with the single stain of either Wright or Giemsa.

CONCLUSIONSWright-Giemsa stain can be used in cell micronucleus test to obtain good staining outcomes.

Azure Stains ; Coloring Agents ; Hep G2 Cells ; Humans ; Micronucleus Tests ; Staining and Labeling ; methods

OBJECTIVETo analyze the reversal effect of Buzhong Yiqi Decoction (BYD) on multidrug resistance of human adenocarcinoma of lung cell line A549/DDP, and to study its effect on the expression of survivin by using serum pharmacological methods in vitro. Methods Totally 24 SD rats were divided into the high, medium and low dose groups, and the blank control group by randomized controlled method. The high dose BYD containing serum (1. 134 g/mL, 2 mL), the middle dose BYD containing serum (0.576 g/mL, 2 mL), and the low dose BYD containing serum (0.284 g/mL, 2 mL) were prepared. The inhibitory effects of different dose and concentrations BYD on the proliferation of A549 and A549/DDP cells were detected by MTT assay, and the drug resistance reversal fold was calculated. The expression of Survivin in the two cell strains were detected respectively by immunohistochemical assay, Western blot, and immunofluorescence method.

RESULTSBYD containing serum showed obvious inhibitory effect on the growth of A549 and 549/DDP. The inhibition rates of 10% dose groups were higher than those of 5% dose groups. Besides, it gradually increased along with increased concentrations. Compared with 10% blank control group, the inhibition rate increased in 10% middle and low dose groups (P <0.05). After acted with 10% middle dose BYD containing serum, IC50, of A549 and A549/DDP were both reduced (P <0.05), reversal fold (RF) both increased. Its reversal ratio on A549/DDP cells was 2. 46, decreasing the resistance of A549/DDP to DDP. Compared with A549 in the same group, the expression of Survivin was detected to decrease by immunocytochemical assay, Western blot, and immunofluorescence method (P<0.05). Compared with 10% blank control group, the inhibition rate decreased in 10% middle dose group (P <0. 05).

CONCLUSIONS10% middle dose BYD containing serum could significantly inhibit the apoptosis of A549 and A549/DDP. Besides, it could moderately reverse the multidrug resistance of A549/DDP cells to DDP possibly through reducing the intracellular expression of Survivin and enhancing the sensitivity 549/DDP to chemotherapeutics.

Adenocarcinoma ; metabolism ; Animals ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Inhibitor of Apoptosis Proteins ; metabolism ; Lung Neoplasms ; metabolism ; Microtubule-Associated Proteins ; pharmacology ; Rats