OBJECTIVETo investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).

METHODSThe levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.

RESULTSCompared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.

CONCLUSIONThe siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

Alzheimer Disease ; etiology ; Amyloid beta-Peptides ; metabolism ; Apoptosis ; Cell Line, Tumor ; Gene Silencing ; Humans ; Neuroblastoma ; metabolism ; pathology ; Peptide Fragments ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Receptors, Nicotinic ; genetics ; metabolism ; Signal Transduction ; Transfection ; bcl-2-Associated X Protein ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Aim To investigate the immunomodulatory effects of sea cucumber fucoidan ( SC-FUC) on macro-phage and the signaling pathways. Methods Cell via-bilities in response to different concentrations of SC-FUC were analyzed by MTT, phagocytosis ability was detected by neutral red,and nitric oxide ( NO) produc-tion was examined by Griess reaction kit. The mRNA expression levels of IL-6 , IL-10 , Toll-like receptors (TLRs) and related signal molecules MyD88, TRIF, NF-κB were assayed by real-time PCR. All the experi-ments were based on murine RAW264. 7 cell line. Re-sults SC-FUC could promote RAW264 . 7 cell prolif-eration, phagocytosis as evidenced by uptake of neutral red and release of NO. The effects were significant at the early stage (6 h and 12 h) . SC-FUC could up-reg-ulate the expression of IL-6 , IL-10 , TLR4 , TLR5 , TLR9. Moreover, mRNA expressions of TLRs signaling molecules were increased, as well as MyD88, TRIF, NF-κB. Conclusions SC-FUC could activate macro-phage, and then promote the immune function by pro-moting production or expression of NO, IL-6, IL-10. It is speculated to be relevant to activated cell surface re-ceptors in macrophage, including TLR4, TLR5, TLR9, and NF-κB signaling pathways.

OBJECTIVETo follow up the cohort of a certain factory and analyze the death cause of the employees.

METHODSA dynamic cohort was adopted and the study population consisted of all workers stayed at the factory for more than 1 year. The cohort data was mainly from the personnel ministry in the factory,the death data provided by the personnel ministry,the labour union and the hospital. All cause of death of the all employee and the death condition of radiation group and no-radiation group were analyzed and compared.

RESULTSThe mortality of the workers in the factory was significantly lower than national population, the SMR of all cause of death in all employee, radiation group and no-radiation group were 0.41 (95% CI: 0.37-0.45), 0.24, 0.75 respectively; all cancer death in the three group was 0.59, 0.40 and 0.92. But the death order was different in radiation group and no-radiation group,the order of liver cancer was list first in radiation group, which was unlike that of the nation order and the no-radiation order.

CONCLUSIONThere have no excess death in the factory, but the order of liver cancer is precedence.

Cause of Death ; Cohort Studies ; Female ; Humans ; Male ; Middle Aged ; Neoplasms, Radiation-Induced ; mortality ; Nuclear Reactors ; Occupational Exposure ; Power Plants ; Prospective Studies ; Workplace

It has gradually become a consensus in the industry that the traditional Chinese medicine gypsum should be decocted first, but the understanding of decocting method is not completely unified in the works of doctors since ancient times, and there are occasional disputes about whether it is necessary to decocting first. In this study, the phase determination, physical and chemical characterization, qualitative and quantitative analysis of inorganic and organic components of the decoctions of herbal pairs and the whole prescription Maxingshigan decoction with gypsum as the center, and the pre-decoctions and co-decoctions of them were carried out to explore the scientific connotation of the pre-decoctions of gypsum. Results show that decoction phases were different between the co-decoctions and pre-decoctions of licorice-gypsum (Gancao-Shigao, GC-SG), ephedra-gypsum (Mahuang-Shigao, MH-SG) and almond-gypsum (Xingren-Shigao, XR-SG). The results of the micromorphology, particle size and zeta potential of herbal pairs and prescription (Quanfang, QF) showed that the supramolecular particles in pre-decoctions were smaller, more uniform and more stable than the co-decoctions. The results of organic components analysis showed that different cooking methods did not change the organic composition and content. ICP-OES results showed that the content of inorganic components in pre-decoctions was higher than in co-decoctions for the same boiling time of gypsum. The IR results showed that the pre-decoctions had stronger chemical functional group effect than the co-decoctions. To sum up, compared with the co-decoction, the pre-decoction of gypsum has different phase state and chemical composition interaction, and the difference of inorganic composition is an important material basis affecting the change of phase state compared with the co-decoction. It indicates that the material basis of traditional Chinese medicine decoction is indeed different whether gypsum is decocted first or not, which can provide a basis for the clinical application of decocted gypsum.

Humans ; Interleukin-17 ; Liver Diseases ; immunology ; Th17 Cells ; immunology

OBJECTIVETo investigate class I integron of Shigella flexneri, its prevalence in children, and its relation to bacterial resistance to antimicrobial agents.

METHODSTotally 51 strains of Shigella flexneri were isolated from fecal samples of children suffering from bacterial diarrhea seen between June 2004 and November 2004 at Children's Hospital of Fudan University. Polymerase chain reaction (PCR) was employed to amplify various integron markers, including intI1, gene cassette region and 3' conserved region of class I intrgron; susceptibility of Shigella flexneri strains to 7 antimicrobial agents was determined by K-B (Kriby-Bauer) method.

RESULTSForty-six strains of Shigella flexneri had intI gene with a positive rate of 90.2% (46/51); 24 strains of Shigella flexneri were positive for qacEDelta1-sul1, the positive rate was 47.1% (24/51); proportion of the isolates positive for all the three regions of class I integron was 43.1% (22/51); 46 strains of intI positive Shigella flexneri were all positive for ant (3'')-I. Among 46 strains of intI positive isolates, proportions of the isolates positive and negative for qacEDelta1-sul1 were 47.8% (22/46) and 52.2% (24/46), respectively. In the class I integron positive Shigella flexneri, the resistance rates of ampicillin (chi(2) = 10.13, P < 0.01) and chloramphenicol (chi(2) = 19.97, P < 0.01) were significantly higher than those in the class I integron-negative group.

CONCLUSIONSClass I integron was detected in 90.2% of Shigella flexneri in children; carriage of class I integron is related to antimicrobial resistance of Shigella flexneri.

Anti-Bacterial Agents ; pharmacology ; Child ; DNA, Bacterial ; genetics ; Diarrhea ; microbiology ; Drug Resistance, Bacterial ; genetics ; Dysentery, Bacillary ; drug therapy ; microbiology ; Feces ; microbiology ; Humans ; Integrons ; drug effects ; genetics ; Polymerase Chain Reaction ; Retrospective Studies ; Shigella flexneri ; drug effects ; genetics ; isolation & purification

OBJECTIVETo evaluate the feasibility and efficacy of surgical treatment for patients with primary hyperparathyroidism (pHPT).

METHODSThe studies were analyzed for 55 patients with pHPT who underwent parathyroidectomy in our hospital from 1985 to 2002.

RESULTSEight patients were asymptomatic. The skeletal manifestations were found in 24 cases, urinary stones in 10 cases, and both skeletal manifestations and urinary stones in 13 cases. Pathological bone fractures occurred in 16 cases. Hypercalcemia was discovered in all patients with the average value of (3.1 +/- 0.4) mmol/L, ranging from 2.7 to 3.9 mmol/L and. Fifty patients showed elevated parathyroid hormone (PTH) with the average value of (489.2 +/- 69.2) pg/ml, ranging from 102 to 2,000 pg/ml. Preoperatively all patients underwent sonography, CT and/or scintigraphy. The overall preoperative image-directed localization rate was 90.9%. Follow-up was done from 6 months to 2 years after surgery. The symptoms and signs of all patients relieved postoperatively with the improving of osteoporosis and healing of bone fracture. Of all cases, 39 presented with temporary hypocalcemia, 37 showed circumoral paresthesia in whom 10 showed tetany, 15 showed eucalcemia and one had mild hypercalcemia after operation. The serum calcium was normal in all cases with hypocalcemia by Rocaltrol and calcium supplementation for 1 - 3 weeks. PTH level decreased to normal fro 2 weeks to 2 months in 47 cases and was still mildly higher than normal in 3 cases.

CONCLUSIONSParathyroidectomy is an effective approach to patients with pHPT. With preoperative image-directed localization techniques, minimally invasive parathyroidectomy is a valid surgical strategy.

Adolescent ; Adult ; Aged ; Female ; Humans ; Hyperparathyroidism ; diagnosis ; surgery ; Male ; Middle Aged ; Parathyroidectomy ; methods ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo construct an inducible eukaryotic vector carrying red fluorescent protein (DsRed) and evaluate the regulation of DsRed gene expression in vitro.

METHODSThe vector pRS17-RUDsRed containing DsRed gene, promoter and RU486-inducible system was constructed using molecular biological methods. To minimize potential interference, the two transcriptional elements were spaced with a 1.6 kb insulator. Fluorescence microscopy and flow cytometry were used to observe the activation of this regulatable vector after transfection in MFC cells.

RESULTSThe vector was identified by digestion with different restriction enzymes, sequencing and PCR. In the absence of RU486, the cells transfected with the vector exhibited very low DsRed protein expression, and the addition of RU486 induced efficient DsRed expression in the cells.

CONCLUSIONThe RU486-inducible eukaryotic vector carrying DsRed protein allows effective regulation of the target gene expression in vitro, which provides a useful tool for gene regulation and gene therapy studies.

Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Mifepristone ; pharmacology ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Tumor Cells, Cultured

OBJECTIVETo compare the safety and efficacy of myocardial contrast enhancement (MCE)-guided and angio-pressure (AP)-guided transcoronary ablation of septal hypertrophy (TASH) for patients with hypertrophic obstructive cardiomyopathy (HOCM).

METHODSTASH was performed under MCE-guide (n = 47, group I) or AP-guide (n = 25, group II) for drug-refractory patients with HOCM. Myocardial perfusion imaging (MPI) data as well as other clinical data were compared.

RESULTSTASH both under MCE-guide or AP-guide resulted in similar and significant reduction of left ventricular outflow tract gradient (PG) and associated with significant symptom improvement (all P < 0.001). Dosage of ethanol use, peak-level of CK-MB and ablated myocardial area and incidence of arrhythmia were also similar between the two groups.Similar left ventricular/atrial dimension changes post TASH were observed in the 2 groups during follow-up. However, the first selected septal vessels were changed under MCE in 6 patients.

CONCLUSIONSOur data demonstrated that the MCE-guided TASH was not superior to AP-guided TASH in safety and efficacy. However, MCE-guided TASH can avoid the misplace of ethanol to avoid innocent myocardial ablation.

Adult ; Cardiac Catheterization ; methods ; Cardiomyopathy, Hypertrophic ; diagnostic imaging ; therapy ; Catheter Ablation ; methods ; Female ; Humans ; Male ; Middle Aged ; Myocardial Perfusion Imaging ; Ultrasonography

OBJECTIVE: To explore the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus after ovariectomy in mice. METHODS: BDNF levels were detected by immunohistochemistry combined image analysis in hippocampal CA regions and dentate gyrus of ovariectomized mice. RESULTS: The expression of BDNF in hippocampus of mice decreased significantly after the ovariectomy after 4 days. The recovery BDNF expression started 14 days after the ovariectomy and after 28 days, the expression of BDNF in hippocampus recovered to the normal level. CONCLUSION The decrease of estrogen in ovariectomized mice can weaken the expression of BDNF in hippocampus during the early stage.
Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Dentate Gyrus ; metabolism ; Estrogens ; blood ; Female ; Hippocampus ; metabolism ; Mice ; Ovariectomy ; RNA, Messenger ; biosynthesis ; genetics