The aim of this study is to clone the mouse T-bet promoter and enhancer, construct and identify the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS for T-bet transcription regulation study and its function in signaling of multiple sclerosis. The promoter and CNS of T-bet were predicted by bioinformatics assay. The predicted fragment of mouse T-bet promoter plus CNS was amplified by PCR and cloned into pGL4.10. The recombinant plasmid pGL4.10-TBX21pr-CNS was transferred into Escherichia coli DH5α. The positive clone was identified by double digestion with Kpn I and Sfi I and DNA sequencing. Finally, pGL4.10-TBX21pr-CNS was cotransfected with pRL-TK into 293T cells and Jurkat cells, pRL-TK and pGL4.10 as a control. The luciferase activity in 293T cells (P = 0.012 2) and Jurkat cells (P = 0.002 2) was higher than that of the control group. A fragment of 1 028 bp mouse T-bet promoter plus 1 308 bp CNS was successfully cloned and the firefly luciferase reporter gene plasmid pGL4.10-TBX21pr-CNS was constructed. In 293T cells and Jurkat cells, pGL4.10-TBX21pr-CNS has the promoter functions. This work offers a basic material for the research of T-bet transcription.
Animals ; Gene Expression Regulation ; Genes, Reporter ; Genetic Vectors ; Luciferases ; Mice ; Multiple Sclerosis ; Plasmids ; Promoter Regions, Genetic ; T-Box Domain Proteins ; genetics

Objective: To evaluate the reliability and validity of the Chinese version of Frost's Multidimensional Perfectionism Scale(FMPS).Methods: One thousand two hundred and eighty Chinese undergraduate students participated in the study and responded to the FMPS and instruments measuring psychological distresses such as anxiety,depression,and obsession-compulsion.Both exploratory and confirmatory factor analyses were used to examine the factor structure of the Chinese FMPS.The association between FMPS and psychological distresses was analyzed as an investigation for criterion validity.Results: Factor analyses extracted five of the original six factors of FMPS.PC did not emerge as a dimension.The five subscales were shown to have satisfactory internal consistencies with Cronbach's alpha between 0.64 and 0.81 and test-retest reliability 0.63~0.82.Chinese FMPS was associated with anxiety,depression,and obsession-compulsion.Particularly,CM and DA are correlated with these psychological distresses with coefficients between 0.30 and 0.70.Conclusion: Chinese FMPS is satisfactorily valid and reliable.It is suitable to use in Chinese college students.

BACKGROUND: Sinomenine, an alkaloid monomer extracted from Chinese medicinal herb sinomenium acutum,possesses potent anti-inflammatory, analgesic and immunoinhibitory pharmacological activities. Sinomenine has certain therapeutic effect on rheumatoid arthritis and has been widely applied in the clinic. Dendritic cell (DC) is the known antigen presenting cell with the strongest functions in the body.OBJECTIVE: To observe the effect of different doses of sinomenine on CD80 and CD 86 expression and extracellular interleukin-12 secretion in DCs.DESIGN: A controlled repeated measuring study based on the cells.SETTTNG: Department of Pharmacology, Guangdong Medical College.MATERIALS: The experiment was carried out in the Institute of Immunology, Third Military Medical University of Chinese PLA between September 2004 and May 2005. The peripheral blood of healthy donors was supplied by the blood bank of the Southwest Hospital. RPMI1640 medium was purchased from Hyclone Company. Fluorescein isothiocyanate(FITC)-conjugated mouse monoclonal antibodies against CD80 and CD86 and FITC-conjugated mouse IgG1 were purchased from Bioscience Company. Recombinant human interleukin-4 (rhlL-4), recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and tumor necrosis factor-α (TNF-α) were purchased from PharMingen Company. Sinomenine was supplied by Zhengqing Pharmacy Company in Hunan. ELISA kit specific for IL-12 was purchased from Diaclone Company.METHODS: Peripheral blood was collected from healthy volunteers. 1 ×106 U/L recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) and 5×105 U/Lrecombinant human interleukin-4 (rhlL-4)were added to the peripheral blood. After 7 days, DCs were harvested. ①The DCs were counted and cultured at 2×108 L 1 in fresh medium supplemented with TNF-α and different doses of sinomenine(3, 10, 30 mg/L). After 48 hours, 2×105 cells were collected and washed twice with phosphate buffer solution (PBS). Then the expressions of CD80 and CD86 were analyzed using 20 μL FITC-conjugated mouse monoclone antibodies against CD80 or CD86.FITC-conjugated mouse IgG1 was used as control at 4 ℃ away from light for 30 minutes. After staining, cells were washed twice with PBS and fixed in 10 g/L paraformaldehyde. The samples were analyzed on a flow cytometry. ②The DCs were counted and cultured at 2 ×108 L-1 in fresh medium supplemented with TNF-α and different doses of sinomenine (3, 10, 30 mg/L). After 48 hours, the supernatants were collected and the concentration of interleukin-12 (IL-12) was determined in a sandwich ELISA using kit specific for IL-12 according to the manufacture's instruction.MAIN OUTCOME MEASURES: ①Effect of different doses of sinomenine on the expressions of CD80 and CD86 on DCs. ②Effect of different doses of sinomenine on the level of IL-12 secreted by DCs.RESULTS : ① Expressions of CD80 and CD86 on DCs: There were no significant differences in the percentage of cells positive and fluorescence intensity for CD80 and CD86 on DC between different doses of sinomenine groups and control groups (P ＞ 0.05). ② IL-12 level secreted by DCs: From ELISA measurement, we found that IL-12 level of the sinomenine 3 mg/L, 10 mg/L and 30 mg/L groups was (863.1±33.7), (668.8±31.9), (512.6±29.6) ng/L, respectively,which was higher than that of control group [ (922.2±36.6),P ＜ 0.05-0.01]. The level of IL-12 in the supernatants of DCs co-cultured with sinomenine was significantly lower compared with control, which was dose-dependent.CONCLUSTON: Sinomenine can inhibit the level of IL-12 secreted by DCs in dose-dependent manner, but do not influence the expressions of CD80 and CD86 in DCs; The therapeutic effect of sinomenine on rheumatoid arthritis may be related to its inhibition to secretion of IL-12 in DCs.

Lipoxins, metabolites of arachidonic acid , are a strong“braking signal” towards inflammatory reac-tion.Due to their anti-inflammatory and pro-resolving properties , lipoxins have emerged to be the central targets in the re-search on inflammation .The present article reviews the research advances of synthesis , biological effects and the protective role of lipoxins in the diseases of liver , pancreas , stomach and colorectum , thus providing a novel approach for the treat-ment of digestive diseases .

Glutamate is an essential excitatory neurotransmitter which regulates brain functions.An increase in extracellular glutamate could excessively activate ionotropic glutamate receptors,initiate calcium overload,and lead to cell death after cerebral ischemia.Glutamate transporter-1 (GLT-1) is one of the major glutamate transporters expressed predominantly in astrocytes.Astrocytes also express the enzyme glutamine synthetase (GS) which converts the glutamate to glutamine; the latter is then 'recycled' into neurons.Pretreatment with ceftriaxone (CEF),ischemia and intermittent hypobaric hypoxia could lead to neuroprotection by increasing the expression of GLT-1 and regulating the activity of glutamate transporter in brain.

Wrist-ankle acupuncture is a type of special acupuncture therapy, featured by simple operation, swift efficiency, and few adverse effects. In the recent years, ankle-wrist acupuncture has become more and more popularized in clinic. By retrieving the relevant medical literatures from VIP, CNKI, and Wan Fang databases, this manuscript was to analyze the history of wrist-ankle acupuncture and to review it from both mechanism and clinical reports. Meanwhile, this article also pointed out the existing problems in the research of wrist-ankle acupuncture during the recent years, and expected further standardization and studies.

Neuropathic pain and cancer pain are two difficult types of chronic pain to be controlled by currently available analgesics.Animal models can provide pivotal systems for preclinical study of pain.This article reviews some new models of chronic pain,such as those of cancer pain,central nerve pain,and the pain caused by peripheral nerve injury and disease-induced peripheral neuropathy.

Ten autopsy cases of bronchiole-alveolar carcinoma (BAC) were reported. Grossly, BAC may be divided into 3 types, i. e. finely nodular, coarsely nodular and diffuse type. Histologic pattern falls into 2 categories: ' (1) alveolar type, (2) papillary type. Metastasis is most frequently mediated through lymphatics, spreading to the regional lymph nodes. Next, they are spread by blood stream. Brain (meningeal carcinomatosis),bone and adrenal glands are the frequent metastasized sites. Pulmonary fibrosis and scar formation seem to be the predisposing factors to BAC.

Objective To observe the clinical efficacy of different doses of warm needling in treating knee osteoarthritis (KOA). Method Sixty-seven KOA patients were randomized into group A of 16 cases, group B of 17 cases, group C of 18 cases, and group D of 16 cases. Group A, B, and C were intervened by warm needling, 1 moxa cone for group A, 2 cones for group B, and 3 cones for group C. Group D was intervened by intra-articular injection with Sodium hyaluronate. The Visual Analogue Scale (VAS) and Lysholm Knee Scoring Scale (LKSS) were evaluated before and after intervention.Result The VAS and LKSS scores were significantly changed in the four groups after intervention (P<0.01). After intervention, the VAS and LKSS scores in group B and C were significantly different from that in group A (P<0.05,P<0.01). After intervention, the VAS and LKSS scores in group B and C were significantly different from that in group D (P<0.05,P<0.01).Conclusion Warm needling is an effective method in treating KOA, and warm needling with 2 moxa cones can produce the best effect.

OBJECTIVE:To observe the efficacy and safety of azithromycin sequential therapy in the treatment of mycoplasma pneumonia in children. METHODS:67 children with mycoplasma pneumonia were randomly divided into control group(34 cases) and observation group (33 cases). All children received conventional treatment,such as fever,cough,supplemented by nutrition support;based on it,control group received 10 mg/kg Azithromycin for injection,adding into 5% Glucose injection by intrave-nous drip in 200 ml,once a day;observation group received azithromycin(the same dosage with control group)after intravenous drip 5 d,switched to 10 mg/(kg·d)Azithromycin dry suspension,orally,then stopped 4 d after continuous 3 d. The treatment course was 7-10 d. Clinical efficacy,disappearance time of fever,cough and lung rales,and lung indexes before and after treat-ment,and the incidence of adverse reactions in 2 groups were observed. RESULTS:There were no significant differences in the to-tal effective rate,disappearance time of fever,cough and lung rales in 2 groups(P>0.05). After treatment,the lung function in-dexes were significantly higher than before,the differences were statistically significant(P<0.01),but there was no significant dif-ference between 2 groups (P>0.05). The incidence of adverse reactions in observation group was significantly lower than control group,the difference was statistically significant (P<0.05). CONCLUSIONS:Based on conventional treatment,azithromycin se-quential therapy shows similar efficacy to without sequential therapy in the treatment of mycoplasma pneumonia in children,but with better safety.