1.Identification the rare p phenotype in serological and molecular biological methods
Xiaozhen HONG ; Xianguo XU ; Faming ZHU
Chinese Journal of Blood Transfusion 1988;0(03):-
Objective To identify the p phenotype. Method P blood group system was identified using p phenotype cells,anti PP 1 P k antiserum,and direct DNA sequencing.Result and Conclusion Proband was typed as p, with rare anti PP 1 P k in the serum,family study suggested that inheritance was autosomal recessive.
2.Detection of RhD antigen by flow cytometry and its clinical applications
Xianguo XU ; Ji HE ; Faming ZHU
Chinese Journal of Blood Transfusion 1988;0(04):-
0.05). The absolute counts of RhD(+) cell of 2 patients at 3 different times were 0.124?10 12 /L, 0.245 ?10 12 /L and 0.517?10 12 /L respectively.Conclusion FCM can be used to detect RhD antigen and perform RhD(+) cell counts in patients with RhD(-) who received incompatible blood.
3.Identification of the para-Bombay phenotype AB h m.
Xiaozhen HONG ; Xianguo XU ; Faming ZHU
Chinese Journal of Blood Transfusion 1988;0(01):-
Objective To identify para-Bombay phenotype AB h m. Method ABO and H phenotype were typed. Absorption and elution were performed. Saliva was tested by inhibitory reaction. Direct sequencing was performed and family study was done. Results Proband was typed as rare para-Bombay phenotype AB h mand anti-H was detected in his serum. Family study suggested that the inheritance was autosomal recessive. Conclusion Rare AB h m phenotype was identified and anti-H has been detected in his serum.
4.Research on molecular genetic basis for Jk(a-b-) phenotype
Faming ZHU ; Xianguo XU ; Xiaozhen HONG
Chinese Journal of Blood Transfusion 1988;0(04):-
Objective To investigate the molecular basis for Jk(a b ) phenotype.Methods Routine serologic testing for phenotype.Genomic DNA covering 4~11 exons and partial introns of JK gene was amplified by ploymerase chain reaction.The PCR products were excised and purified from agarose gels with a kit,then fragments were directly sequenced.Results G mutated to A in the 3'acceptor splice site of intron 5;A to G at 78 site from the 3'end of intron 3;C to T at 84 site from the 5'end of intron 8; A to G at 588 site of exons ( exon 7); G to A at 838 site of exons (exon 9).The splice site mutation (G→A) of intron 5 may cause the skipping of exon 6.Conclusion G to A mutation in the 3'acceptor splice site of intron 5 maybe one of the molecular basis for Jk(a-b-) phenotype
5.Application of painless gastrostomy under endoscop jejunal catheterization in enteral nutrition of severe acute pancreatitis in intensive care unit
Chuanjiang WANG ; Faming HE ; Fang XU
Chongqing Medicine 2014;(20):2551-2553
Objective To observe and compare the operation ,short-term therapeutic effect and long-term therapeutic effect be-tween painless gastrostomy under endoscopy and nasal-jejunal catheterization as well as to explore their clinical feasibility and clini-cal application .Methods 81 cases of patients with severe acute pancreatitis in the hospital ICU from May 2012 to September 2013 , were divided into gastrostomy jejunostomy group(43 cases)and nasojejunal feeding group(38 cases) .The operation time ,gastroin-testinal nutrition tube inner curvature ,removal rateetc were observed to calculate the success rate .Contrast ratio ,plugging rate and catheter related infection rate and several indexes were observed to evaluate the therapeutic effect of two groups of short-term infec-tion ofincision .long-term calculation of two groups with tube ,comfort score and nutritional indexes to evaluate the therapeutic effect index .Results The operation time ,the digestive tract inside area rate ,nutrition tube removal ,catheter related pulmonary infection rate ,average indwelling catheter time ,comfort ,nutrition index of the two groups had statistically significant (P<0 .05) ,plugging rate ,wound infection rate had no statistically significant (P>0 .05) .Conclusion With fewer complications and longer tube time , painless gastrostomy under endoscopy is safe ,simple and feasible ,which is easily accepted by patients .
6.Study of the FCRL5 gene single nucleotide p olymorphism in patients with ankylosing spondylitis
Tong LIU ; Shengqian XU ; Faming PAN ; Chen CHEN ; Jianhua XU
Chinese Journal of Rheumatology 2011;15(9):629-633
ObjectiveTo investigate the association between the FCRL5 gene (rs6427384 and rs12036228) single nucleotide polymorphism(SNP) and patients with ankylosing spondylitis (AS). Methods SNP of FCRL5 gene (rs6427384 and rs12036228) was analyzed in 169 patients with AS and 184 healthy controls by ligase detection reaction based on high temperature(LDR)-PCR. The distribution of FCRL5 genotypes and allele frequencies were detected between the two groups. Chi-square test, one-way ANOVA were used for statistical analysis. ResultsSignificant differences were found in the distribution of allele frequencies of FCRL5 gene between AS patients and health controls(P<0.05). The C allele frequencies of FCRL5 gene at positions of rs6427384 and rs12036228 were 17.3%, 92.3% and 25.0%, 87.2% respectively in the AS group and the control group. And the T allele frequencies of rs6427384 and rs12036228 were 82.7%, 7.7% and 75.0%, 12.8% in the AS group and the control group. The percentages of CC, CT and TT genotype(rs6427384) were 3.7%, 27.2%, and 69.1% in the AS group, which were significantly different from those of the control group(3.9%, 42.2% and 53.9% ) (x2=8.7637, P=0.0125 ). Staging of sacroiliitis in X ray were significantly different during AS patients whose genotype represented as CC, CT and TT (rs6427384)(x2=34.159, P=0.0001 ). Incidences of the initial symptoms (low back pain or inflammation of periphery joint)in the AS group were obviously differed among patients with different genotypes (rs6427384) (x2=7.254, P=0.027), so did the mean duration of morning stiffness (F=4.159, P=0.018) and the average scores of BASDAI (F=4.461, P=0.014). Incidences of the initial symptoms in the AS group were also conspicuously different between the AS patients with different genotypes (rs 12036228 ) (x2=6.640, P=0.036 ). ConclusionOur study suggests that the SNP(rs6427384 and rs12036228) of FCRL5 may be a susceptibility factor for AS in Anhui Han population and the genotype may influence the clinical phenotype of AS.
7.Study of the interleukin-1F7 gene single nucleotide polymorphism in patients with ankylosing spondylitis
Chen CHEN ; Shengqian XU ; Faming PAN ; Jianhua XU
Chinese Journal of Rheumatology 2011;15(8):546-549
Objective To investigate the association between interleukin(IL)-1F7 gene (rs3811047)single nucleotide polymorphism (SNP) and ankylosing spondylitis (AS). Methods SNP of IL-1F7 gene (rs3811047) was analyzed in 158 patients with AS and 181 healthy controls by ligase detection reaction based on high temperature ligase (LDR-PCR). The distribution of IL-1 F7 (rs3811047 ) genotypes and allele frequencies were detected between the two groups. Results Significant differences were found in the distribution of IL-1F7(rs3811047 ) genotypes and allele frequencies between AS patients and healthy controls. The frequency of A allele of IL-1F7 gene at positions rs3811047 was 12.03% and 17.68% in AS group and the control group,and the frequency of G allele was 87.97%, 82.32%, respectively (x2=4.2204, P=0.0399). The percentage of AA, AG and GG genotype was 0, 24.05%, 75.95% in AS group, which differed from the controls group (2.76%, 29.83%, 67.41% ). The difference had remarkable statistical significance (x2=6.2675, P=0.043).The positive rate of HLA-B27 in AS patients which represented as AG genotype was 70.27% (26/37),remarkably lower than that in AS patients which represented as GG genotype 94.23% (98/104), the difference was statistically significant (x2=2.168, P=0.030), erythrocyte sedimentation rate and C reactive protein levels were conspicuously lower than that in GG genotype (t=2.971, P=0.013; t=3.300, P=0.001 ). Conclusion Our study suggests that the SNP (rs3811047) of IL-1F7 may be a susceptibile factor for AS in Anhui Han population, the genotype may influence the clinical phenotypes of AS.Patients who carry the A allele may have less inflammation than patients who do not carry the A allele.
8.In vitro effects of tea polyphenols on the secretion of TNF-α, IFN-γ and IL-2R by peripheral blood CD8+ T lymphocytes from vitiligo patients
Faming XU ; Aie XU ; Jilong WU ; Jie OUYANG
Chinese Journal of Dermatology 2013;46(6):411-414
Objective To detect the level of cytokines and their receptors secreted by peripheral blood CD8 + T lymphocytes from vitiligo patients,and to evaluate the influence of tea polyphenols on their secretion.Methods CD8+ T lymphocytes were isolated from the peripheral blood of 12 patients with progressive vitiligo,12 patients with stable vitiligo and 10 healthy controls,and cultured in vitro for 20 days.Then,some CD8+ T lymphocytes were treated with tea polyphenols of 100 mg/L for two days.Enzyme-linked immunosorbent assay (ELISA) was carried out to determine the levels of tumor necrosis factor (TNF)-α,interferon (IFN)-γ and interleukin-2 receptor (IL-2R) in the culture supernatants of CD8+ T lymphocytes before and after the treatment with tea polyphenols.Analysis of variance and t test were conducted to assess differences in these parameters among these groups and changes between pretreatment and posttreatment,respectively.Results There was a sequential decrease in the level of TNF-α ((191.302 ± 6.077) vs.(175.966 ± 2.467) vs.(173.664 ± 3.600) ng/L,F =4.784,P < 0.05),but a sequential increase in that of IFN-γ ((280.182 ± 36.070) vs.(371.670 ± 24.352) vs.(447.147 ± 8.432) ng/L,F=9.036,P< 0.01) and IL-2R ((8.375 ± 0.161) vs.(8.845 ± 0.161) vs.(9.345 ±0.125) ng/L,F =9.639,P < 0.01) in the culture supernatant of CD8+ T lymphocytes from patients with progressive vitiligo to patients with stable vitiligo and healthy controls.After two days of tea polyphenol treatment,a statistical decrease was observed in the level of TNF-α in the culture supernatant of CD8 + T lymphocytes from patients with progressive vitiligo ((164.797 ± 1.784) ng/L,P < 0.01),patients with stable vitiligo ((166.150 ± 3.576) ng/L,P < 0.05) and healthy controls ((155.028 ± 5.759) ng/L,P < 0.05),but no statistical changes were noted for IFN-γor IL-2R (all P > 0.05) despite a slight elevation in the level of IFN-γin the culture supernatant of CD8+ T lymphocytes from patients with stable vitiligo and in that of IL-2R from all the three groups as well as a mild reduction in that of IFN-γ from patients with progressive vitiligo and healthy controls.Conclusions The changes of cytokines and their receptors secreted by CD8 + T lymphocytes may be associated with the induction and progression of vitiligo.Tea polyphenols may treat vitiligo via suppressing the secretion of TNF-α by CD8+ T lymphocytes.
9.The relationship between single-nucleotide polymorphism in RANKL, osteoprotegerin gene and rheumatoid arthritis
Biwei PEI ; Shengqian XU ; Tong LIU ; Juan DENG ; Xiaomeng SHEN ; Faming PAN ; Jianhua XU
Chinese Journal of Rheumatology 2012;16(11):732-737
Objective To investigate the relationship between single-nucleotide polymorphism (SNP)in receptor activator for nuclear factor-κB ligand (RANKL),osteoprotegerin (OPG) gene and rheumatoid arthritis (RA).Methods In our study,3 SNPs in the genes of OPG (2 SNP:rs2073618,rs3102735) and RANKL (1 SNP:rs2277438) by ligase detection reactions from 200 RA and 201 controls were examined.BMD values of different areas were assessed using dual-energy X-ray absorptiometry.Clinical and laboratory parameters were collected.Analysis of variance,t-test and x2 test were used for statistical analysis.Results No signi-ficant differences in the distribution of the alleles and genotypes were observed between case group and the control group (P>0.05).The haplotype analysis for RANKL and OPG SNPs showed that the rs2073618/rs2277438/rs3102735 GGG haplotype could reduce the risk of RA (1.5% vs 6.0%,P=0.008; OR 0.216;95%CI:0.081 to 0.575) and the GAG haplotype increased the risk of RA (14.5% vs 8.4%,P=0.007; OR 1.862,95%CI:1.179 to 2.943).Patients with RANKL-rs2277438 AA or GG genotypes (n=6) had significantly higher BMD values compared to those with AG genotypes (n=39) at spine lumber 3 (1.05±0.22 vs 0.93±0.26,t=2.314,P=0.023),spine lumber 4 (1.06±0.24 vs 0.94±0.28,t=2.27,P=0.030),spine lumber 2-4 (1.04±0.21 vs 0.89±0.28,t=2.788,P=0.007).The tender joint counts (13±7 vs 10±6),tender joint index (19±11 vs 13±9),and VAS score (5.7±1.9 vs 4.8±1.8) differed significantly between patients with the OPG-rs2073618 CC or GG genotypes (n=60) and GC genotypes (n=40).Conclusion The rs2073618/rs2277438/rs3102735GGG haplotype may be protective against RA,while GAG haplotype may increase the susceptibility to RA.RANKL gene SNP rs2277438 may affect BMD value at spine lumber,and OPG gene SNP rs2073618 may influence the disease activity of RA patients.
10.Association of tumor necrosis factor-α receptor gene single nucleotide polymorphism in patients with ankylosing spondylitis
Chen CHEN ; Biwei PEI ; Shengqian XU ; Juan DENG ; Tong LIU ; Faming PAN ; Jianhua XU
Chinese Journal of Rheumatology 2013;(1):31-36
Objective To investigate the value of tumor necrosis factor (TNF)-α receptor gene,TNFRSF1A+36A/G(rs767455) and-383A/C(rs2234649),TNFRSF1B+196T/G(rs1061622) single nucleotide polymorphism (SNP) for the susceptibility to ankylosing spondylitis (AS) and the relationship between SNP and AS.T test,Chi-square test,and ANOVA were used for statististical analysis.Methods Two hundred and fifteen patients who had definite diagnosis of AS and 216 healthy blood donors were involved in this study.SNPs of TNF-α receptor gene:TNFRSF1A +36A/G(rs767455),-383A/C(rs2234649) and TNFRSF1B+196T/G (rs1061622) were detected with the ligase detection reaction (LDR-PCR) method.Results ① Distribution frequencies of A alleles(86.8%,91.5%) and G alleles (13.2%,8.5%) of TNFRSF1A(rs767455) in AS and controls were significantly different with each other (x2=4.627,P=0.0315),while the distribution frequency in group of homozygotes (AA or GG genotype) in AS and controls were 74.6%(150/201) and 83.9%(177/211),the frequencies in group of heterozygotes (AG) were 25.4% (51/201) and 16.1%(34/211)(x2=5.390,P=0.020).Frequency of alleles and the genotypes of TNFRSF1A (rs2234649) and TNFRSF1B (rs1061622) between AS and control group were similar(P>0.05).It also demonstrated that TNF-αreceptor gene haplotype (rs1061622T-rs2234649A-rs767455G) carriers apparently increased the susceptibility to AS (11.5% vs 6.9%)(OR:1.753,95%CI:1.078~2.852,P=0.022).② Analysis of variance found that the duration of morning stiffness (F=3.168,P=0.044) and peripheral joint tenderness counts (F=4.598,P=0.011) among the three genotype groups of TNFRSF1B (rs1061622) in patient with AS were evidently differed with each other.Bath AS functional index (BASFI) among different genotype groups of TNFRSF1A (rs2234649) in AS had remarkable diversity (F=5.783,P=0.004).None of above indicators among groups of different genotypes of TNFRSF1A (rs767455) in AS were uniform (P>0.05).③ Forty-four patients were treated with TNF-α antagonist (entanercept),25 mg,subcutaneous injection,twice weekly for 3 months,then followed with Sulfaslazine (SASP) 2.0 g/d and Celecoxib 0.4 g/d for another 9 months.ASAS20 was the primary endpoint for the evaluation of therapeutic effect at the visit of 3 month and 12 month.No associations were found between SNP and short or long term outcome of treatment with TNF-α antagonist in AS (P>0.05).Conclusion TNFRSF1A (rs767455) SNP correlates with susceptibility to AS in Anhui Han local patients.Carriers of TNF-α receptor gene haplotype (rs1061622T-rs2234649A-rs767455G) may increase the susceptibility to AS.SNP of TNFRSF1B (rs1061622) is associated with disease activity in AS,while SNP of TNFRSF1A(rs2234649)relates to functional index of the disease.There is no association between SNP of TNFRSF1A / TNFRSF1B and short or long term outcome of treatment with TNF-α antagonist in AS.