The pathogenesis of migraine has not been clarified yet.The cnrrent dominate theories on the pathogenesis of migraine included:vasogenic theory,neurogenic theory,biochemical factors,genetic factors,and infection factors.Study progress migraine in recent years was summarized in the paper.

BACKGROUND: As a main gene engineering vector, adeno-associated virus (AAV) is characterized by its extensive host cells, lasting and stable expression and less immune response to hosts, and is applied widely. But AAV is a kind of defective virus, and need incasing cells to supply E1 protein. As important and special AAV incasing cells, AAV-293 cells can produce E1 in trans. But AAV-293 cells are delicated and cultivated difficultly, and the biological character is easy to be changed. Therefore, it is necessary to establish a culture method of AAV-293 cells to meet the need of gene engineering.OBJECTIVE: To establish a culture method of AAV-293 cells in vitro.DESIGN: An opening study.SETTING: Department of Neurology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: AAV-293 cells line was provided by Stratagene Corporation; high-carbohydrate OMEM (H-DMEM) powder by Gibco Company; there plasmids in AAV Helper-Free by Stratagene Company.METHODS: This experiment was carried out in the neurology laboratory of Tongji Hospital in Wuhan during the period from October 2006 to April 2007. AAV-293 cells were resuscitated and cultivated with H-DMEM growth medium in vitro, and were passaged and stored in liquid nitrogen when the cells monolayer confluence reached 50%. At the same time, their growing state was observed by inverted microscope, and their growth curve was noted. According to whether AAV-293 cells could give out green fluorescence or not (observed by fluorescence inverted microscope) after they were cotransfected with the there AAV system plasmids and infected with AAV supernatant, their biological character of packing AAV was assessed.MAIN OUTCOME MEASURES: ① Morphological observation of AAV-293 cells; ② the growth curve; ③ the package of AAV.RESULTS: ① AAV-293 cells observed by fluorescence inverted microscope were growing adhesively well with irregular polygons, light endochylemas and ambiguous nuclei appearances. ②The growth curve showed that the growing adaptive phase was the first day after AAV-293 cells were passaged, the actively growing phase was from the second day to the fifth day, and the growing platform phase was after the sixth day. ③ AAV-293 cells with green fluorescence observed by fluorescence inverted microscope, and cotransfection of the there AAV system plasmids was successful. AAV-293 cells gave out green fluorescence after infected with AAV supernatant, and AAV package succeeded.CONCLUSION: The culture method established by the authors in the experiment is simple and useful, and the cultured AAV-293 cells remain a good function of AAV package.

Objective To investigate the clinical significance of the changes of electrocardiogram (ECG) and cardiac markers for myocardial damage after on-pump coronary artery bypass grafting (CABG) and off-pump coronary artery bypass grafting (OPCABG). Method Monitoring 25 patients of OPCABG (OPCABG group) and 25 patients of CABG (CABG group) R wave amplitude of V4 and V5 on antorior electrocardiographic lead and simultaneously determining cardiac markers for myocardial damage creatine phosphokinase isoenzyme MB (CPK-MB), tropenin I (cTnI) and heat-shock protein 70 (HSPT0) on different lime. Results R wave amplitude of V4 and V5 on anterior electrocardiographic lead had no significant changes on 0, 6, 18 and 24 hours after OPCABG. On the contrary, R wave amplitude of V4 and V5 on anterior electrocardiographic lead decreased significantly on 0, 6 and 18 hours after CABG (P<0.01), and came back to preoperative values 24 hours after operation. The levels of CPK-MB and cTnI reached its peak and higher significantly for CABG than those for OPCABG on 24 hours after operation, 29.29 μg/L vs 5.98 μg/L and 6.74 μg/L vs 1.91 μg/L respectively. HSP70 increased significantly on 6 hours after operation in two groups, but median of HSP70 was higher significantly in CABG group (11044.5 pmol/L vs1702.0 pmol/L). In the first day after operation the HSP70 peak was correlated significantly with the level of CPK-MB(r=0.370, P<0.01) and cTnI (r=0.458,P<0.01). Conclusions Myocardial damage is significantly alleviated for patients of OPCABG comparing with those of CABG. The HSF70 in circulation may indicate the degree of myocardial damage.

Objective To observe the therapeutic effect on thymus radiotherapy in the patients with myasthenia gravis (MG).Methods The clinical data of 68 patients with MG treated by thymus radiotherapy (X ray radiotherapy in the deep for 22 cases, 60 Co treatment for 46 cases)in our hospital from 1957 to 1999 were analysed retrospectively.The patients were followed up long term.Results Total remission rate of patients with MG treated by radiotherapy was 90%.Clinical results were significant,the life quality of patients with MG improved greatly.The side effect with the skin injury for 10 cases and the alimentary canal symptom for 9 cases were found mostly.Conclusion The following patients can obtain obvious therapeutic effect with radiotherapy,they are those with poor efficiency to drugs,those with invasive thymoma during the operation,those with instable symptoms after the operation.

OBJECTIVETo analyze the differences between the bacterial diversities in the saliva of caries-free and caries-susceptible adolescents through polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE).

METHODSTwenty adolescent subjects aged 12-18 years were recruited and subdivided into two groups: caries-free adolescents (n = 10) and caries-susceptible adolescents (n = 10). Saliva samples were collected. Total DNA was isolated directly from each sample. A portion of the 16S rRNA gene locus was PCR-amplified by using universal primers. Microbial diversity was analyzed through PCR-DGGE.

RESULTSAnalyzing the DGGE profile, we found that the composition of the saliva microbiome exhibited great intra-individual differences; the average band numbers of the caries-free adolescent group and the caries-susceptible adolescent group were 32.5 ± 3.7 and 27.3 ± 3.4, respectively. The differences between the groups were statistically significant (P = 0.008). Shannon-Wiener's indexes of the caries-susceptible adolescent group and the caries-free adolescent group were 2.5 ± 0.2 and 2.6 ± 0.2, respectively, but the differences between the groups were not significant (P = 0.405). Clustering analysis results suggested that most of the samples in the same group clustered together; this observation showed a high community structure similarity.

CONCLUSIONThe microbial diversity and complexity of bacteria in saliva are significantly higher in caries-free adolescents than in caries-susceptible adolescents. During caries development, bacterial diversity in the saliva likely decreases.

Adolescent ; Bacteria ; Child ; DNA, Bacterial ; analysis ; Denaturing Gradient Gel Electrophoresis ; Dental Caries ; microbiology ; Dental Caries Susceptibility ; Humans ; Microbiota ; Mouth ; microbiology ; Polymerase Chain Reaction ; RNA, Ribosomal, 16S ; Saliva ; microbiology

AIM: To further investigate the immunologic mechanism of myasthenia gravis(MG). METHODS: Acetylcholine receptor antibodies(AchRab) and presynaptic membrane antibodies (PsMab) was determined by ELISA in 285 cases of MG patients. Tumor necrosis factor- alpha(TNF-?) and RBC-C3b receptor rosette rate(RBC-C3bRR)and RBC-IC rosette rate (RBC-ICR)was determined in 202 patients with MG. Furthermore, the peripheral blood lymphocyte subsets was examined in 104 cases MG patients. RESULTS: The positive rates of the AchRab and PsMab in MG were very significantly higher than that of control.The positive rate of PsMab in patients with negative AchRab was very significantly higher than that of patients with the positive AchRab.In 202 MG patients, the TNF-? was significantly higher, but the RBC-C3bRR was significantly lower than those of control, while the RBC-ICR has no significant change. CONCLUSION: MG is an autoimmune disease with damaged postsynaptic membrane by the AchRab, and some patients with both the AchRab and PsMab could manifest disfunction in the postsynaptic and presynaptic membrane. While the patients who just has the PsMab maybe damaged only in the presynaptic membrane. Other body fluid immune or/and cell immune mechanism may be involved in MG patients without AchRab and PsMab. [

Objective To observe the effect of local mild hypothermia and Naloxone in the treatment of acute intracerebral hemorrhage. Methods Forty-five patients with acute intracerebral hemorrhage were randomly divided into 4 groups:a control group(12 patients),a hypothermia group(11 patients),a Naloxone group(11 patients)and a hypothemrmia plus Naloxone group(11 patients).The patients in the control group were managed with conventional interventions including the administration of 6-aminocaproic acid within 24 hours and dehydrant when intracranial pressure was high.Those in the hypothermia and Naloxone groups were treated with local hypothermia at 33～34 ℃ for 3 days or intravenous transfusion of Naloxone at 4 mg/d in addition to the conventional intervention.Those in the combination group were treated with local hypothermia and intravenous Naloxone in addition to the conventional intervention.Immediately after admission and 2 weeks after treatment,head CT scans were conducted to observe the volume of cerebral hematoma and edema.The patients' neurological function was scored according to the European Stroke Standards(ESS)before and after treatment. Results There was no significant difference among the 4 groups in terms of the volume of hematoma and edema or in their ESS scores before treatment.After treatment,any differences among the 4 groups with regard to hematoma volume were not significant.The volume of edema in the hypothermia group was similar to that in the combination group and significantly lower than that in the Naloxone andcontrol groups.Hematoma volume in the Naloxone group was significantly lower than that in the control group.After treatment,the ESS scores were significantly higher in the combination group than that in hypothermia group,and scores in the hypothermia group were significantly higher than in the Naloxone group.ESS scores in the Naloxone group were significantly higher that in the control group. Conclusion Local mild hypothermia and Naloxone treatment can inhibit cerebral edema and enhance recovery of neurological function in patients with intracerebral hemorrhage.Local mild hypothermia has advantages over Naloxone in inhibiting the development of cerebral edema and in promoting recovery of neurological function.Local mild hypothermia in combination with Naloxone further inhibits edema,and it can enhance neurological function to a greater extent.

In order to investigate the neuroprotective effects of cyclin-dependent kinase-5 (cdk-5) inhibition in mice with Niemann-Pick disease type C (NPC) (npc(-/-)), recombinant adeno-associated virus (rAAV) carrying the small interfering RNA (siRNA) specific for cdk-5 gene was injected into 3-day-old npc(-/-) mice intracerebroventricularly. The rAAV-GFP-injected age-matched npc(-/-) mice and non-surgery age-matched npc(-/-) mice were employed as controls (n=6-10/group). From the 4th to 8th week after the treatment, mice were weighed, and evaluated for limb motor activity by using the coat hanger test once a week. Eight-week-old npc(-/-) mice were sacrificed by decapitation, and brains were quickly dissected and halved sagittally. Immunohistochemistry, Western blotting, and HE staining were used to evaluate the neuropathology in npc(-/-) mice. The results showed that rAAV-cdk-5-siRNA-GFP significantly reduced the number of axonal spheroids, delayed the death of Purkinje neurons, ameliorated motor defects in npc(-/-) mice, and significantly attenuated the hyperphosphorylation of tau proteins. These data suggested that inhibition of cdk-5 activity has neuroprotective effect on neurons in NPC mice.

Objective To study the influence of inhibited steroidogenic factor-1 on human adrenocortical H295R cells, and explore its role in the pathogenesis of adrenal tumors. Methods The plasmids pGenesil1-SF-1-shRNA which containing U6 promoter and SF-1-specific short hairpin RNA (shRNA) and pGenesil1-negative-shRNA containing unspecific shRNA were transfected into H295R cell. The expression of SF-1 was measured by Western blot and real-time polymerase chain reaction(RT-PCR). Cell proliferation was analyzed by WST-1 assay and cell count. Ki-67 expression was detected by immunohistochemistry and cell apoptosis was examined by TUNEL assay. Results Compared with those in control cells, the protein and mRNA level of SF-1- transfected cells were reduced by 69.7% and 71.2% (P＜0. 01). WST-1 and cell count method showed that SF-1 gene silencing obviously inhibited cell proliferation(P＜0. 01). By contrast, there was a 3. 7-fold increase in the percentage of apoptotic H295R cells in SF-1-inhibited group than that of control group (P＜0. 01). Immunohistochemistry showed that Ki-67 positive cells in SF-1-inhibited cells were lower than the negative control cells (16.90±2.17) % and (33. 48±3.16)%,(P＜0. 01). Conclusion SF-1 gene silencing can inhibit the proliferation of adrenocortical cells, and it is expected to become a key protein in understanding pathogenesis of adrenal tumors or treating them.

Objective To investigate the clinical characteristics of Takayasu's arteritis (TA) with pulmonary hypertension (PAH) in order to improve the diagnosis and treatment earlier. Methods Twelve out of 191 patients with TA registered in Peking Union Medical College Hospital from 1987 to 2007 were diagnosed as PAH, the clinical data of 12 patients were analyzed. Results Ten patients were females. The range of age were from 14 to 47 years old, the average age was (27±10) years old. Eleven patients had the clinical manifestations or/and signs of pulmonary artery involvement. Seven patients presented with short breath after exercise or hemoptysis as the first manifestation, four patients with fatigue, four patients with intermittent claudication or pain or numbness of extremities, three patients with dizziness. Seven patients belonged to type Ⅰ+Ⅳ, one patient to type Ⅱ+Ⅳ, three patients to type Ⅲ+Ⅳ, one patient to type Ⅴ. Elevated ESR/CRP was found in ten patients. All patients took the glucocorticoid and DMARDs, stent implantation in pulmonary artery was done in one patient, Bentall was operated in another patient.The symptoms of all patients improved except one patient died for low cardiac output after operation. Conclusion PAH is one of the severe complications in late stage of TA, and other arteries are usually involved too. Because it is difficult to observe PAH in TA patients in early stage, CTA or pulmonary angiography and UCG should be taken in early stage. The stent implantation or dilating the artery should be considered aa a treatment, but at on the same time, glucocorticoid and DMARDs should be taken to avoid the relapse.