AIM: To explore the treatment methods of severe migraine. METHODS: 51 cases of severe migraine patients were treated with the combination of radix salviae militiorrhizae, diclofenac and sodium aescinate, and another 62 cases of severe migraine patients were treated with diclofenac alone. RESULTS: The efficacy of the combination is better than that of diclofenac alone (P

BACKGROUND:Transformation growth factor beta 1 is mostly used to induce the chondrogenic differentiation of bone marrow mesenchymal stem cel s, but there is a poor induction efficacy.
OBJECTIVE:To explore the chondrogenic differentiation of bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes or induced by transforming growth factor beta 1.
METHODS:Articular chondrocytes and bone marrow mesenchymal stem cel s from SD rats were harvested and divided into 1:2, 2:1, 1:1 concentration groups. Cel s induced by transforming growth factor beta 1 acted as control group. After 20 days of induced culture, MTT was used to detect cel viability, alcian blue colorimetric assay was applied to measure glycosaminoglycan content, and western blot assay was employed to determine the expression of col agen type II.
RESULTS AND CONCLUSION:The absorbance value in the control group was significantly lower than that in the 1:1 and 2:1 groups (P<0.05). Glycosaminoglycan content and protein expression of col agen type II were also lower in the control group than the 1:2, 1:1, 2:1 groups. But there was no difference between 1:1 and 2:1 groups (P>0.05). The results show that bone marrow mesenchymal stem cel s co-cultured with articular chondrocytes can be induced to differentiate into chondrocytes, and meanwhile, there is a saturation phenomenon during the chondrogenic differentiation of bone marrow mesenchymal stem cel s.

Objective To investigate the level of serum free fatty acid (FFA )after improving the life style in patients with coronary heart disease complicated metabolic syndrome and the effect of therapeutic life style on traditional risk factors of coronary artery disease.Methods A total of 395 patients with coronary heart disease complicated metabolic syndrome were recruited.Pa-tients were divided into intervention group (group A,conventional drug therapy+ intensive life style intervention,n=97)and non-intervention group (group B,conventional drug therapy,n=38)according to the scores of life style.Serum free fatty acid (FFA) was determined by ELASA.The scores of life style was obtained bylife style questionnaire.Results (1)The serum FFA of pa-tients with coronary heart disease complicated metabolic syndrome were positively related to waist circumference and waist-high-ra-tio.(2)Waist circumference,BMI and FFA of group A were significantly lower than those in group B after therapeutic life style in-tervention(P <0.05).(3)Compared with the baseline,the constitution index and FFA in group A were significantly lower after 6-months therapeutic life style intervention(P <0.05).Conclusion Therapeutic life style can reduce the level of FFA and constitution index of the patients with coronary heart disease complicated metabolic syndrome.

Objective To develop a method for treating infertile women with polycystic ovary syndrome(PCOS)by ooeyte in vitro maturation(IVM).Methods On day 3～4 of menstrual cycle,infertile women with PCOS were administrated with(100～150IU)FSH/day for 3 days.On day 8～10,when the follicle diameter reached 10～14mm and endometrium＞5mm,the immature oocytes weFe picked up and cultured in vitro for 28～31 hours.Then,ICSI and embryo transfer were performed.Results 3 clinic pregnancies were established among 12 infertile women.the pregnancy rate was 25.0%.Conclusion This research indicates that IVM is a feasible technology for the infertile women with PCOS.

Objective To clone and express the new gene R049 of uropathogenic Escherichia coli 132.and to investigate the immunopmtective effects of the R049 recombinant protein on mice.Methods The pmkaryotic expression system of gene R049 was constructed by directed cloning.Thereafter,the R049 recombinant protein Was expressed and purified by Ni affinity chromatography.Polyclonal antibody was pre-pared by immunizing BALB/c mice with R049 recombinant protein.The R049 recombinant protein and whole bacterial proteins of UPEC132 were analyzed by SDS-PAGE and Western blot.BALB/c mice were im-munized with R049 recombinant protein before challenged by UPECl32 through urinary tract.Then the differences of urine and renal colony counts between immunization group and control group were compared.Results The recombinant strain E coli BL21(DE3)/pET32a-R049 ORF was constructed successfully,and the relative molecular mass of the R049 recombinant protein was 66.9×103 and its purity was up to 95% af-ter purification.The titer of polyclonal antibody wag≥1:102 400 analyzed by indirect ELISA.Both of the R049 recombinant protein and whole bacterial proteins of UPECl 32 were confirmed to show specffic reactions on the antiserunl throughh Western blot.The animal experiments showed the urine and renal colony counts of immunization group were significantly lower than that of the control group(P<0.01,P<0.05).Conclu-sion The new gene R049 of uropathogenic E.coli 132 had immunopmtective effects on mice and the defini-tive mechanism would be needed to further study.

Objective To investigate the effects of dexmedetomidine on inflammatory responses in brain tissues of the patients undergoing carotid endarterectomy.Methods A total of 40 ASA physical status Ⅱ or Ⅲ patients,aged 65-80 yr,scheduled for elective unilateral carotid endarterectomy under general anesthesia,were randomly divided into 2 groups (n =20 each) using a random number table:dexmedetomidine group (group Dex) and control group (group C).In group Dex,dexmedetomidine 0.03 μg · kg-1 · min-1 was infused over 10 min before induction of anesthesia,and after tracheal intubation dexmedetomidine was then infused at a rate of 0.3 μg · kg-1 · min-1 until 30 min before the end of operation.The equal volume of normal saline was given in group C.At 20 min before induction of anesthesia (T0),10 min after induction of anesthesia (T1),15 min after carotid artery clamping (T2),15 min after carotid artery unclamping (T3),and at 6 and 24 h after operation (T4,5),blood samples were drawn from the ispilateral jugular bulb for determination of serum concentrations of malondialdehyde (MDA) (by TBA) and S100B,tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) (by ELISA).Results Compared with group C,the serum S100B concentrations were significantly decreased at T3-5,the serum TNF-α and IL-6 concentrations were decreased at T2.5,and the serum MDA concentration was decreased at T3 in group Dex.Conclusion Dexmedetomidine can reduce the brain damage through mitigating inflammatory responses in brain tissues of the patients undergoing carotid endarterectomy.

AIM:To evaluate the efficacy and safety of gabapentin in the treatment of migraine.METHODS:A randomized,double-blind,placebo-controlled study was taken.104 patients(aged 18 to 60 year)with migraine were randomly assigned to receive placebo(control group,49 patients)or gabapentin(experimental group,55 patients)for 4 weeks.The efficacy,side effects,TCD and EEG were assessed at the beginning of trials,and 4th,8th weeks.RESULTS:Comparing with the placebo group,the frequency,duration and intensity of migraine were markedly decreased in gabapentin group,there was significant difference between groups(P0.05).CONCLUSION:Gabapentin is an effective and safe agent from migraine.

Objective To construct the secretory expression vector of recombinant human C-reactive protein(rhCRP) for its se-cretory expression in Pichia pastoris ,rhCRP was expressed as a secretory protein and purified ,and the immunity reactivity of the purified protein was identified .Methods The DNA fragment of rhCRP which was designed and synthesized was cloned into pPICZαA vector .Recombinant plasmid pPICZαA/rhCRP was linearized by SacⅠand transformed into Pichia pastoris X-33 by elec-trotransformation .The rhCRP was secreted into the medium under the methanol induction .RhCRP was purified by Histamine affin-ity chromatography .The purified rhCRP was identified by SDS-PAGE and Western blotting ,and its immunity reactivity and stabili-ty was identified by indirect ELISA .Results The pPICZαA/rhCRP expression vector was successfully constructed .The rhCRP of 23 × 103 was inducted and successfully expressed as a secretory protein by the recombinant Pichia pastoris strains .The rhCRP was purified by one step up to 90 .42% purity ,and it was showed good immunity and stability by indirect ELISA .Conclusion The rh-CRP with higher purity and immunoreactivity was successfully obtained by using the Pichia pastoris expression system ,which pro-vided an important experimental basis for producing anti-human CRP antibodies and developing testing CRP reagent .

Fertilization is a crucial step for origin of life.During Assisted Reproductive Technologies (ART),total fertilization failure is complex and unpredictable.Total fertilization failure may related to some abnormal cellular mechanistic events,such as:any stage of sperm and cumulus-oocyte-complexes penetration,sperm-zona pellucida binding / penetration,sperm-oocyte membrane binding,oocyte activation,sperm discondensation or pronuclear formation.Most of total fertilization failure could be solved by intracytoplasmic sperm injection.But oocytes of some patient still can't fertilize successfully,even though assisted oocyte activation be used.As for total fertilization failure patients in ART,combining the mature of oocyte,sperm quality and some trail to improve clinical protocol in later cycle may prevent failure to happen again.

AIM: To investigate the CdTe quantum dots coated with MPA and explore its biocompatibility with living cells. METHODS: CdTe quantum dots coated with MPA were prepared in aqueous phase and MPA CdTe QDs were Characterized with TEM, fluorospectrophotometer and ultraviolet spectrophotometer. QDs were Modified with with avidin, purified and prepared as flurescent probe. LSCM was used to observe the expression of MHC Ⅱ antigen on PMφ cells, which was labeled by QDs. Cell culture and MTT assays were used to determine the biocompatibility of MPA coated CdTe quantum dots with the B-16 cells as target cells. RESULTS: The particle diameter of CdTe quantum dots prepared in aqueous phase was well distributed. They had good photological performance and greater stability after coated with MPA. MHC Ⅱ antigen on PMφ was labeled with the QDs-Avidin fluorescent probe showed great fluorescence intensity, which was easy to be detected by fluorescence microscope and LSCM. MPA CdTe QDs showed cytotoxicity when its density was very high, but they showed little cytotoxicity during the normal use of influence label density limit. CONCLUSION: MPA CdTe QDs can be used as new fluorescent lable as they are of even size, not easy to bleach or quench, have good photological performance and stability and good biocompatibility.