Objective To detect plasma leval of FⅦa?Fig and LP(a) in patients with coronary heart diseases and their relationship with coronary heart diseases and severity of coronary artery stenosis.Methods The level of FⅦa?Fig and LP(a) were detected in100 patients with coronary heart diseases,and coronary artery tomography were performed. Results The level of FⅦa?Fig and LP(a) were significantly higher in patients with coronary heart diseases than those of control (P

ObjectiveTo establish a novel rapid detection method based on PCR-single-strand-conformational polymorphism (PCR-SSCP) to determine mutation of streptomycin-resistance associated rpsL and rrs genes in isolates of Mycobacterium tuberculosis (MTB).MethodsStreptomycin-resistance of 112 MTB isolates was detected using the routine drug susceptibility test,and a special PCR-SSCP assay was established.The mutations of rpsL and rrs genes in streptomycin-resistant MTB isolates were detected by PCR-SSCP and PCR direct sequencing (PCR-DS) ; the results from two techniques were compared.Results All isolates had both rpsL and rrs genes.Fifty-two isolates (46.4％) were streptomycin susceptible,in which only 1 isolate showed abnormal PCR-SSCP fragments from rrs gene,and the specificity of PCR-SSCP was 98.1％ (51/52).Sixty isolates (53.6％) were streptomycin-resistant,in which 46 (76.6％) and 11 ( 18.3％ ) isolates presented the abnormal PCR-SSCP fragments of rpsL and rrs gene,respectively.One streptomycin-resistant isolate showed abnormal PCR-SSCP fragments from both rpsL and rrs genes.The sensitivity of PCR-SSCP was 93.3％ (56/60).ConclusionThe PCR-SSCP that established in this study is a specific and sensitive method for rapid detection of the streptomycin-resistance associated mutations in rpsL and rrs genes of MTB.

ObjectiveTo investigate the association between catechol-O-methyl transferase (COMT)gene polymorphism and Gilles de la Tourette' s syndrome(GTS).MethodsUsing Amplification Refractory Mutation System(ARMS) PCR genotyping assay method,a polymorphism (val158met) of COMT gene was genotyped in 112 of all GTS patients ( total GTS group) including 54 GTS-alone patients group,48 GTS + ADHD patients group among of them and 71 healthy controls.The correlation between positive association of polymorphism (val158met)of COMT gene in GTS and the age of onset in patients with GTS was also analyzed.ResultsCompared with healthy controls group,genotype of val158met did not differ in total GTS patients group or alone-GTS patients group (χ2 =0.56,P=0.756;χ2 =1.05,P=0.600 respectively).There was also no significant difference (P＞0.05)in allele distribution of val158met in total GTS patients group or alone-GTS patients group compared with controls group respectively (χ2 =0.18,P=0.669;χ2 =0.29,P=0.593 respectively).However,genotype distribution of val158met was significantly different between GTS + ADHD patients group and controls group( χ2 =6.35,P =0.041 ).The frequency of the val allele of this locus was significantly higher in GTS + ADHD patients group than those in controls group ( χ2 =5.49,P =0.019 ).The mean age of onset (6.80 ± 1.54 ) in 36 children within GTS + ADHD patients group with the val/val geantype of COMT gene val158met polymorphism was significantly earlier than the mean age of onset (8.04 ± 1.54)in 12 children in val/val genotype (P =0.016 ).ConclusionPolymorphism (val158met) of COMT gene may be associated with GTS children with comorbid ADHD,which may play an important role to make the age of onset in children with GTS become earlier.

Sixty-four patients with Graves' diseases were divided into methimazole group ( n =30 ) and propylthiouracil group( n =34 ).20 healthy volunteers were used as the control group.The levels of interleukin (IL) -2,IL-6,and TSH receptor antibody (TRAb) were determined by ELISA after serum samples were eollected before treatment and after treatment for 3 and 6 months.The results showed that the general data of two groups were not significantly different before treatment.IL-2 and IL-6 levels at 6 months after treatment were significantly different from the baseline (P＜ 0.05 ),IL-2 being gradually increased while IL-6 decreased with time.The level of IL-6 in methimazole group was lower than that in propylthiouracil group after treatment for 6 months ( P＜ 0.05 ).There was no significant difference in TRAb levels between two groups before treatment while differences became significant at 3 and 6 months of treatment ( all P＜ 0.01 ),being gradually decreased with time. IL-2/IL-6 ratio became significantly greater at 3 and 6 months of treatment compared with that before treatment in the same treatment group( P＜0.05 ) and the ratio in methimazole group was higher than that in propylthiouracil group by 6 months of treatment ( P＜0.05 ).IL-6level was positively correlated with FT3 and FT4 levels,and IL-2 level was negatively correlated with FT3,FT4,and TRAb levels in GD patients before treatment.The correlations of IL-2 and IL-6 with FT3 and FT4 disappeared by 3 and 6 months of treatment.IL-2 and IL-6 levels were related with TRAb level before and after treatment in methimazole group. These results suggest that the immunosuppressive effect of methimazole is more evident than that of propylthiouracil in patients with Graves' disease.

The data of 6 patients with tertiary hyperparathyroidism were retrospectively analyzed.The first symptom in 3 patients was bone ache,1 with lower extremity weakness,1 with swelling of lower limbs,and 1 with increase serum alkaline phosphatase.5 patients had past history of renal failure,1 case suffered from severe intestinal malabsorption,and 4 of them had renal transplantation.X-ray showed subperiosteal bone resorption,bone demineralization,or cellulose osteitis.5 subjects presented with osteoporosis,and 1 of them had fractures.Left atrioventricular membrane calcification and gluteal calcification were found in each of 2 cases.4 patients suffered from hypercalcaemia,2 patients with normal serum calcium,4 with hypophosphataemia,and 6 with high levels of PTH.Pathological results showed adenoma in parathyroid gland in 4 patients and enlarged parathyroid glands in 2 patients.Serum calcium and PTH levels returned to normal after operation and calcium supplement.

Objective To investigate the role of pathogenic Leptospira interrogans lipopolysaccha-ride (L-LPS) and outer membrane proteins (L-OMP) in the apoptosis of mouse macrophages (J774A.1) and their association with Fas/FasL pathway .Methods Phenol-water extraction and Triton X-114 phase separation were used to extract L-LPS and L-OMP from L.interrogans serogroup icterohaemorrhagiae serovar Lai strain Lai, respectively.Polymyxin B ( PMB) and protease K ( PK) were used to treat L-LPS and L-OMP, respectively.J774A.1 cells were stimulated by L.interrogans strain Lai with or without ultraviolet inactivation.In parallel, the cells were stimulated by extracted L-LPS and L-OMP with or without PMB and PK treatments .The apoptosis and necrosis of J 774 A.1 cells before and after treatment were detected by flow cytometry.The siRNAs were used to silence the expression of Fas or FasL gene in J 774A.1 cells and their inhibitory effects were further validated by using real-time fluorescent quantitative RT-PCR.Flow cytometry was used to detect the effects of L-LPS or L-OMP on the apoptosis of J774A.1 cells with Fas or FasL gene-knockdown .Results L.interrogans strain Lai with or without ultraviolet inactivation could cause similar early apoptosis rates (47.1%and 55.6%) and late apoptosis/necrosis rates (7.6%and 7.9%).The ear-ly apoptosis rates of 1×105 J774A.1 cells were 40.4%and 34.0%after the treatment with 100 ng of L-LPS and 100 μg of L-OMP for 4 h.The late apoptosis/necrosis rates of the cells were 7.5%and 6.9%upon the treatments with L-LPS and L-OMP, respectively.However, the apoptosis or necrosis of the cells was not ob-served when using L-LPS and L-OMP pre-treated by PMB and PK, respectively.Silenced expression of Fas or FasL gene reduced the L-LPS-induced J774A.1 cells apoptosis (P<0.05), while decreased early apopto-sis rate of J774A.1 cells mediated by L-OMP was only observed in Fas gene-knockdown cells (P<0.05). Conclusion Both L-LPS and L-OMP can cause the Fas/FasL-associated apoptosis of macrophages , which is beneficial for L.interrogans to establish the productive infection in hosts .

Objective To detect the expression of Zbtb7 gene in Phorbol esters(PMA) induced differentiation of leukemia cell lines.Methods 50 nmol/L PMA was used to induce the differentiation of U937 and K562 cell lines.The expression of Zbtb7 was detected by real-time PCR in the cells after induction on day 0,1,3 and 5,respectively. Results The expression of Zbtb7 in leukemia cell lines was markedly enhanced after treated by PMA(P

OBJECTIVETo construct a lipL32/1-lipL21-OmpL1/2 fusion gene and its prokaryotic expression system, and to establish an enzyme-linked immunosorbent assay (ELISA) using the rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for sensitive and specific detection of IgM in the serum of patients with leptospirosis.

METHODSlipL32/1-lipL21-OmpL1/2 fusion genes were constructed using a primer-linking PCR. The target recombinant protein antigens, rLipL32/1, rLipL21, rOmpL1/2 and rLipL32/1-LipL21-OmpL1/2, were expressed and the purified antigens were then immobilized to the surface of microplate wells for ELISA-based detection of IgM in the sera of leptospirosis patients.

RESULTSOf 493 acute leptospirosis patients, 95.7% and 97.8% were positive by rLipL32/1-LipL21- OmpL1/2-IgM-ELISA using different serum dilutions, which was higher than the rLipL32/1-IgM-ELISA (93.1% and 90.3%), rLipL21-IgM-ELISA (90.3% and 87.0%), and rOmpL1-IgM-ELISA (85.6% and 81.1%) (P<0.01). All IgM-ELISAs tested negative against 56 non-leptospirosis patients with typhoid fever, hemorrhagic fever or dengue fever.

CONCLUSIONTrigeminal fusion antigen increases ELISA sensitivity and the rLipL32/1-LipL21-OmpL1/2- IgM-ELISA is a sensitive and specific serological diagnostic method for clinical leptospirosis.

Antigens, Bacterial ; genetics ; metabolism ; Bacterial Outer Membrane Proteins ; genetics ; metabolism ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Immunoglobulin M ; immunology ; Leptospirosis ; diagnosis ; metabolism ; Lipoproteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

Recombinant protein SMB(PRG4) containing two Somatomedin B domains and a small amount of glycosylation of repetitive sequences of proteoglycan 4 was cloned according to PGR4 gene polymorphism. Mature purification process was established and recombinant protein SMB(PRG4), with high-level expression was purified. By using size-exclusion chromatogaraphy and dynamic light scattering, we found that the recombinant protein self-aggregate to dimeric form. Structure prediction and non-reducing electrophoresis revealed that SMB(PRG4), was a non-covalently bonded dimer.
Glycosylation ; Protein Multimerization ; Proteoglycans ; chemistry ; Recombinant Proteins ; chemistry ; Somatomedins ; chemistry

ObjectiveTo evaluate the clinical efficacy and toxicity of gefitinib combined with Zhenqi Fuzheng capsule in treatment of senile non-small-cell lung cancer (NSCLC) at the middle and late stages. Methods88 patients with NSCLC at Ⅲ b-Ⅳ were randomly divided into combined drugs group ( gefitinib combined Zhenqi Fuzheng capsule,n =46 cases) and single drug group (gefitinib,n=42 cases).After treatment for 60 d,the short-term efficacy,side effects and quality of life were observed and evaluated. The objective response and survival rate were assessed after following up for 2 years. Results The short-term efficacy rate in the combined drugs group (19.6 ％ ) was higher than single drug group (11.9 ％ ),but there was no significant difference between the two groups (x2=0.096,P＞0.05).The rate of Karnofsky score in improvement and stableness was 71.7％ in combined drugs group and 50.0％ in single drug group,and quality of life improved after combined drugs compared with single drug treatment (x2 =4.376,P＜0.05).The adverse effects in combined drugs group was some lower than in single drug group with no statistical significance.There was no differences in the survival rates of 2 years between the two groups(x2 =0.556,P＞0.05). ConclusionsThe gefitinib combined Zhenqi Fuzheng capsule in treatment of middle-and late-stage NSCLC is worthy to be popularized due to positive efficacy,less side effects and higher quality of life.