Objective To analyze the genomic structure of a novel RHD allele. Methods Through polymerase chain reaction (PCR), sequence specific primer-PCR(SSP-PCR) and genomic DNA sequencing,the RHD gene in a D-negative individual was detected. Results In SSP-PCR tests, the sample was tested negative for exons 3~7, 9~10 and intron 2(Din 2), but was positive for the Rh downstream Box3. The RHCE genotype was Ccee. Three PCR methods were used to detect intron 10(Din10) of RHD gene;all gave negative results. Through a RHD full length coding region sequencing method, the sample was found to have sequences identical to normal RHD in exons 1 and 2, while exons 3~10 were negative. Conclusion The sample is D antigen negative, RHD gene positive, with amalgamative allele of RHD-CE(2~10).

Objective:To study the clinical efficacy and safety of BCG-polysaccharide nucleic acid combined with isotretinoin soft capsules in the treatment of plane warts. Methods:Seventy patients with plane warts were randomly divided into the observation group and the control group(n=35). The control group were treated with isotretinoin soft capsules, limp 10mg, bid, 4 weeks to 10mg, qd. The observation group were given BCG-polysaccharide nucleic acid 1ml, im, every otherday, on the basis of the contrl group. After 8 weeks of treatment, T cell populations and immunoglobulin levels were detected, and the efficacy and adverse reactions were recorded. Results:After the treatment, T cell population level, IgG and IgA of the two groups were significantly improved(P<0. 01), and the improvement in the observation group was significantly better than that in the control group (P<0. 05). The onset time of the observa-tion group was shorter than that of the control group ( P<0. 01). The total effective rate of the observation group was higher than, that of the control group(P<0. 05). The occurrence of adverse reactions in the two groups were no significant(P>0. 05). Conclusion:The clinical efficacy of BCG-polysaccharide nucleic acid combined with isotretinoin soft capsules in the treatment of plane warts is sig-nificant with promising security.

OBJECTIVETo assess the efficacy of Yishen Jiejing Decoction (YJD) in treating poststroke shoulder-hand syndrome (SHS) patients of yin deficiency yang hyperactivity with blood stasis stagnation collaterals syndrome.

METHODSTotally 60 SHS patients of yin deficiency yang hyperactivity with blood stasis stagnation collaterals syndrome were randomly assigned to two groups, the treatment group and the control group, 30 cases in each group. Conventional rehabilitation training and therapeutics were applied in all patients. Besides, patients in the treatment group took 50 mL YJD, twice a day. One month without interruption consisted of one course of treatment. The curative effects of each group were evaluated respectively before treatment and at one month after treatment. The neurologic impairment, TCM syndrome, and the improvement of upper limbs movement were assessed by the neurologic impairment integral, scoring for TCM syndrome diagnostics, Fugl-Meyer Assessment (U-FMA). Adverse reactions were observed at the same time.

RESULTSThe effective rate of stroke was 86.67% and the effective rate of SHS was 90.00% in the treatment group, higher than those of the control group (P < 0.05). Both groups got improvement in neurologic impairment, stroke induced blood stasis syndrome, yin deficiency yang hyperactivity syndrome, and the improvement of upper limbs movement after treatment (all P < 0.05). Besides, all the improvement was obviously superior in the treatment group (P < 0. 05). No adverse reaction occurred during the course of treatment.

CONCLUSIONThe curative effect of YJD combined with conventional rehabilitation training was confirmative and superior to the control group.

Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Reflex Sympathetic Dystrophy ; drug therapy ; etiology ; rehabilitation ; Stroke ; complications ; Treatment Outcome ; Yin Deficiency ; drug therapy ; Yin-Yang

Objective To investigate the quantitative abnormality of CD~+_5B lymphocytes in the bone morrow of the patients with autoimmunic hemocytopenia and its clinical significance.Methods The patients were referred to the Institute of Hematology & Blood Diseases Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College.Quantities of CD~+_5B lymphocytes in the bone morrow of 14 patients with autoimmune hemolytic anemia(AIHA)or Evans syndrome,22 immunorelated pancytopenia(IRP)patients and 10 normal controls were assayed by FACS.The correlation between their clinical and laboratory parameters with CD~+_5B lymphocytes was analyzed.Results The qutantity of CD~+_5B lymphocytes were significantly higher in AIHA and Evans and IRP patients than in normal controls;there was no significant difference between AIHA、Evans syndrome and IRP patients.The qutantity of CD~+_5B lymphocytes in the bone morrow of all cytopenic patients showed negative correlation with C_3 complement.In AIHA、Evans syndrome patients,the quantity of CD~+_5 B lymphocytes in their bone morrow showed positive correlation with IBIL.In Evans syndrome patients,the quantity of CD~+_5B lymphocytes in their bone morrow showed positive correlation with PAIgG and PAIgM.The qutantity of CD~+_5B lymphocytes in the bone marrow of all cytopenic patients showed negative correlation with treatment response,but no correlation with the yields of CFU-E、 BFU-E、CFU-F and CFU-GM cultured from their bone marrow mononuclear cells in vitro.Conclusion CD~+_5B lymphocytes in the bone marrow of the patients with autoimmunic hemocytopenia significantly increase and are related to the disease severity and clinical response.It is suggested that CD~+_5B lymphocytes might have important roles in the pathogenesis of autoimmunic cytopenia.

CAS (Chrome azurol S) assay was a universal method for detecting the bacterial siderophores, and Sugar-Asn liquid medium has been applied to the studies on siderophores from Pieudomonas. In this paper, Asp has been substituted for Asn, and MSA-CAS agar plate was developed by integrating the MSA (sugar-Asp) medium and CAS bright blue dye, which has been used in the universal CAS assay. On the aspect of siderophores detection , 8 strains of 7 species from Pseudomonas had been screened on MSA-CAS agar plates and universal CAS assay respectively. The results showed that MSA-CAS agar was higher-sensitive and lower basic fluorescent than universal CAS assay.

A novel stable blue agar plate which is convenient to preparing and more effective than the universal chrome azurol sulfonate (CAS) assay established by Schwyn and Neilands was designed by replacing MM9 growth medium and pipes with certain concentrate of phosphorus buffer solution which pH could be stabled at 6.8. It is more suitable for screening over- siderophores production bacteria. Since OD_ 630 of the sample is usually out of the range of spectrophotometer with CAS assay solution when quantifying the siderophores and the outcome is not steady,the measuring wavelength had been changed to 680 nm corresponding to the middle of max absorbance and the correlation between siderophores concentrations and OD was unchanged. But the detecting sensitivity is elevated by enlarged the absorbance differences among samples with different productivity of siderophores at 680 nm .

A secretive expression vector of Pichia pastoris system which can be used for the direct cloning of PCR products was constructed,and was verified through the expression of recombinant cellobiohydrolase II in Pichia pastoris.A randomly selected fragment was amplified with properly designed primers by PCR.The XhoI and Eam1105Ⅰ restriction sites were included in the 5'end of the fragment,and the Eam1105Ⅰ and XbaI restriction sites were included in its 3'end.The PCR amplified product was inserted into the P.pastoris expression plasmid pPICZ?A through XhoI and XbaI restriction sites and the resultant plasmid was digested with Eam1105Ⅰ,and lastly the big fragment was recovered,generating the P.pastoris expressive Tvector pPICZ?T.Then the cellobiohydrolase II of T.reesei was successfully expressed in P.pastoris with this expressive Tvector.Such results indicated that the constructed expression Tvector was convenient for PCR product cloning,and was effective for heterologous protein expression in P.pastoris.On the other hand,the application of the expression Tvector avoided the introduction of additional amino acids at the Nterminus of the expressed protein,which generally occurred when normal expression vectors were used in secretive expression system.

Adolescent ; Adult ; Female ; Humans ; Male ; Mercury Poisoning ; diagnosis ; therapy ; Middle Aged ; Occupational Diseases ; diagnosis ; therapy ; Retrospective Studies ; Young Adult

Objective To study the correlation between single nucleotide polymorphisms of -238,-308 G/A in promoter region of tumor necrosis factor -?(TNF-?) gene and the type of juvenile idiopathic arthritis (JIA).Methods Clinical data and blood preparation of 127 children with JIA and 106 healthy children were evaluated.Subgroups of JIA were defined according to the Edmonton criteria.The -238 G/A and -308 G/A polymorphisms in DNA analysis in this study were extracted from the whole blood.The restricted fragment length polymorphisms were determined in the cases of all JIA children and control group.Results 1.The TNF-?-238 G/A allele frequencies of JIA group and control group:allele frequency of JIA group was 92.9% and 7.1%,and the control group was 95.3% and 4.7%.The distribution of allele frequencies was no significantly different between JIA group and control group(?2=1.149 P=0.284).But there were significant difference between polyarticular JIA (RF negative) and control group(?2=7.621 P=0.006).2.The TNF-?-308 G/A allele frequencies of JIA group and control group:allele frequency of JIA group was 94.1% and 5.9%,the control group was 95.3% and 4.7%.The distributions of allele frequencies was no significantly different between JIA group and control group(?2=0.322 P=0.571).There were significantly difference between polyarticular JIA (RF negative) and control group (?2=7.621 P=0.006).Conclusions The TNF-?-238,-308 polymorphisms of A in the-238 and-308 TNF-? gene are important to the joint destruction of JIA.The study will be beneficial to provide indirect support to the application of anti-TNF drugs to the treatment of JIA.

Objective To investigate the correlation between the single nucleotide polymorphism(SNP) of macrophage migration inhibitory factor(MIF) gene -173G/C and the susceptibility in patients with juvenile idiopathic arthritis(JIA).Methods Study group consisted of 97 children of JIA.All patients included in this study met the International League of Associations for Rheumatology criteria for JIA.Control group consisted of 102 healthy individuals.Germline DNA was extracted from peripheral blood by AxyPrep blood genomic DNA miniprep kit.Polymerase chain reaction-restrictivon fragment length polymorphism(PCR-RFLP) was used for genotyping the -173G/C polymorphism of MIF.Genotype distribution and allele frequencies were obtained by direct counting.Statistical analysis was performed by using SPSS Bosoftware.Allele and genotype distributions were compared using the chi-square test.The relative risk of alleles was described by odds ratios (OR) and 95% confidence intervals (95%CI).Hardy-weinberg equilibrium was confirmed with the chi-square test.Results We detected 3 kinds of genotypes at the MIF-173 locus.The frequency of each genotype was 54.6%(GG),42.3%(GC),3.1%(CC) in JIA group,and 79.4%(GG),20.6%(GC),0(CC) in control group.The C allele frequencies in the JIA and control group were 24.7% and 10.3%,respectively.There was significant difference was observed between the JIA and control group in the frequencies of mutant genotype(GC and CC) of MIF-173G/C polymorphism(?2=13.872 P=0).Individuals possessing a MIF-173C allele did have an increased risk of JIA(OR=2.79,95% CI 1.62-4.81 P=0).When the genotype and allele distributions of the MIF-173 gene in the subtypes of JIA and contronl group were compared,a significant difference wad found in the systemic JIA and control group (P0.05).Conclusions MIF-173G/C SNP may be associated with the sensitivity of JIA.MIF-173 C allele may increase susceptibility to JIA.