目的研究妇女血清生殖免疫抗体检测在不孕不育治疗中的意义。方法从我院2013年9月-2014年1月时间段内所接收的不孕不育女性患者中选取100名,采用酶联免疫吸附试验(Enzyme linked immunosorbent as say,ELISA),检测患者血清生殖免疫性抗...目的研究妇女血清生殖免疫抗体检测在不孕不育治疗中的意义。方法从我院2013年9月-2014年1月时间段内所接收的不孕不育女性患者中选取100名,采用酶联免疫吸附试验(Enzyme linked immunosorbent as say,ELISA),检测患者血清生殖免疫性抗体,即抗精子抗体(AsAb)、抗子宫内膜抗体(EMAb)、抗卵巢抗体(AOAb)、抗心磷脂抗体(ACA)四项,分析结果并判断检测血清生殖免疫抗体对不孕不育患者临床意义[1]。结果选择的100名患者抗体阳性率为53%,其中AsAb阳性19%,EMAb阳性8%,AOAb阳性10%,ACA阳性16%。结论血清生殖免疫抗体的产生是导致不孕症的重要原因之一,患者年龄不同则含量不同,血清生殖免疫抗体检测在不孕不育治疗中具有重要的意义。展开更多
A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and t...A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and then usedfor immunization of BALB/C mice. In enzyme-linked immunosorbent assay (ELISA) and competitive inhibition ELISA ex-periments, the derived antiserum exhibited high antibody titer (greater than 1: 250 000) as well as varied cross-reactivity(from 97.8% to 161.7%) to three analogs of EF belonging to fluoroquinolones family. But over the concentration rangestudied, no significant cross-reactivity was observed to other group of antibiotics (chloramphenicol, oxytetracycline, sul-phamethoxazole and nysfungin). It was confirmed that the synthesized immunogen was highly antigenic and elicited specificantibody responses in BALB/C mice against EF.展开更多
AIM: To investigate the frequencies of the expression of main protein antigens of Helicobacter pylori (H py/ori) isolates, such as UreB, VacA, CagA1, HpaA, NapA, FlaA and FlaB and the production of specific antibodies...AIM: To investigate the frequencies of the expression of main protein antigens of Helicobacter pylori (H py/ori) isolates, such as UreB, VacA, CagA1, HpaA, NapA, FlaA and FlaB and the production of specific antibodies in sera from H pylori-infected patients, and to understand the correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori. METHODS: H pylori strains in biopsy specimens from 157 patients with chronic gastritis and peptic ulcer were isolated and serum samples from the patients were also collected. The target recombinant proteins rUreB, rVacA, rCagAl, rHpaA, rNapA, rFlaA and rFlaB expressed by the prokaryotic expression systems constructed in our previous studies were collected through Ni-NTA affinity chromatography. Rabbit antisera against rUreB, rVacA, rCagAl, rHpaA, rNapA, rFlaA and rFlaB were prepared by using routine subcutaneous immunization. By using ultrasonic lysates of the isolates as coated antigens, and the self-prepared rabbit antisera as the first antibodies and commercial HRP-labeling sheep anti-rabbit IgG as the second antibody, expression frequencies of the seven antigens in the isolates were detected by ELISA. Another ELISA was established to detect antibodies against the seven antigens in sera of the patients by using the corresponding recombinant proteins as coated antigens, and the sera as the first antibody and HRP-labeling sheep anti-human IgG as the second antibody respectively. Correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori were statistically analysed. RESULTS: In the 125 isolates of H pylori, the positive rates of UreB, VacA, CagAl, HpaA, NapA, FlaA and FlaB were 100%, 65.6%, 92.8%, 100%, 93.6%, 100% and 99.2% respectively. In the 125 serum samples from the H pylori infected patients, the positive rates of antibodies against recombinant UreB, VacA, CagA1, HpaA, NapA, FIaA and FlaB were 100%, 42.4%, 89.6%, 81.6%, 93.6%, 98.4% and 92.8% respectively. H pylori strains were isolated from 79.6% (125/157) of the biopsy specimens, but no close correlations among the H pylori infection frequencies and different types of chronic gastritis and peptic ulcer could be found (P>0.05, x2 = 0.01-0.87). The VacA positive rate (82.40%) in the strains isolated from the specimens of patients with peptic ulcer and the anti-VacA positive rate (54.3%) in the sera from the patients were significantly higher than those (51.5%, 32.3%) from the patients with chronic gastritis (P<0.01, x2= 13.19; P<0.05, x2= 6.13). When analysis was performed in the different types of chronic gastritis, the VacA in the strains isolated from the specimems of patients with active gastritis showed a higher expression frequency (90.0%) than those from superficial (47.9%) and atrophic gastritis (30.0%) (P<0.05, x2 = 5.93; P<0.01,x2 = 7.50). While analysis was carried out in the strains isolated from the specimens with superficial (93.8%) and active gastritis (100%), NapA showed a higher expression frequency compared to that from atrophic gastritis (60.0%) (P<0.01, x2 = 8.88; P<0.05, X2=5.00). CONCLUSION: The types of chronic gastritis and peptic ulcer and their severity are not associated with H pylori infection frequency but closely related to the infection frequency of different virulent H pylori strains. The optimal antigens for developing vaccine and diagnostic kit are UreB, FlaA, HpaA, FlaB, NapA and CagAl, but not VacA.展开更多
AIM:To study the diagnosis of Helicobacter pylori(H pylori) infection through the determination of serum levels of anti- H pylori IgG and IgA antibodies,and the levels of anti-H pylori IgA antibodies in duodenal fluid...AIM:To study the diagnosis of Helicobacter pylori(H pylori) infection through the determination of serum levels of anti- H pylori IgG and IgA antibodies,and the levels of anti-H pylori IgA antibodies in duodenal fluid. METHODS:Data were collected from 93 patients submitted to upper digestive endoscopy due to dyspeptic symptoms. The patients were either negative(group A)or positive (group B)to H pylori by means of both histological detection and urease tests.Before endoscopy,peripheral blood was collected for the investigation of anti-H pylori IgG and IgA antibodies.To perform the urease test,biopsies were obtained from the gastric antrum.For the histological evaluation,biopsies were collected from the gastric antrum (greater and lesser curvatures)and the gastric body. Following this,duodenal fluid was collected from the first and second portions of the duodenum.For the serological assaying of anti-Hpylori IgG and IgA,and anti-Hpylori IgA in duodenal fluids,the ELISA method was utilized. RESULTS:The concentration of serum IgG showed sensitivity of 64.0%,specificity of 83.7%,positive predictive value of 82.0%,negative predictive value of 66.6% and accuracy of 73.1% for the diagnosis of H pylori infection.For the same purpose,serum IgA showed sensitivity of 72.0%, specificity of 65.9%,positive predictive value of 72.0%, negative predictive value of 67.4% and accuracy of 69.8%. If the serological tests were considered together,i.e.when both were positive or negative,the accuracy was 80.0%, sensitivity was 86.6%,specificity was 74.2%,positive predictive value was 74.2% and negative predictive value was 86.6%.When values obtained in the test for detecting IgA in the duodenal fluid were analyzed,no significant difference(P=0.43)was observed between the values obtained from patients with or without H pylori infection. CONCLUSION:The results of serum IgG and IgA tests for H pylori detection when used simultaneously,are more efficient in accuracy,sensitivity and negative predictive value, than those when used alone.The concentration of IgA antibodies in duodenal fluid is not useful in identifying patients with or without H pylori.展开更多
AIM. To evaluate the serologic IgG response to Hpylori and CagA across age groups and in healthy children and adults.METHODS： Totally, 386 children aged 1-15 years and 200 adults aged 20-60 years, were enrolled to st...AIM. To evaluate the serologic IgG response to Hpylori and CagA across age groups and in healthy children and adults.METHODS： Totally, 386 children aged 1-15 years and 200 adults aged 20-60 years, were enrolled to study. The serum samples of participant were tested for presence of antJ-Hpylori and anti-CagA IgG by using ELISA method.RESULTS： The seroprevalence of Hpylori in adults was significantly higher than that observed in children （67.5% vs 46.6%; P 〈 0.000003）. In children, the seropositivity rate in males （51.9%） was significantly （P 〈 0.05） higher than that observed in females （41.7%）. The prevalence of serum anti-CagA antibody was 72.8% and 67.4% in infected children and adults, respectively. The mean titer of serum anti-CagA antibodies was significantly higher among children in comparison to adults （64.1 Uarb/mL 30.7; P 〈 0.03）. In infected children and adults the prevalence of serum anti-CagA antibody was higher in males compared to females （78.4% vs 66.3%; P = 0.07 and 75.6% vs 54.71%; P 〈 0.04, respectively）. The age-specific prevalence of anti-H py/or/and anti-CagA antibody （in infected subjects） was 37.6% and 59.57% at age 1-5 years, 46.9% and 75% at age 6-10 years, 54.9% and 79.45% at age 11-15, 59.01% and 83.33% at age 20-30 years, 66.6% and 60.52% at age 31-40 years, 73.46% and 63.88% at age 41-50 years and 75.75% and 60% at age 51-60 years with mean titer of anti-CagA antibody of 75.94, 63.32, 57.11, 52.06, 23.62, 21.52 and 21.80 Uarb/mL, respectively. There was significant difference between mean serum anti-CagA antibody in age subgroups （P 〈 0.001）.CONCLUSION： These results showed that anti-Hpylori and anti-CagA antibodies were common in the children and adults. The Hpylori-specific antibodies influenced by age and sex of subjects. Moreover, it seems that males are more susceptible to infection with CagA^＋ strains compared to females. The seroprevalence of anti-CagA antibody was increased with age, up to 30 years and then decreased. It was also found that the magnitude of the IgG response to CagA decreased with advanced age.展开更多
文摘目的研究妇女血清生殖免疫抗体检测在不孕不育治疗中的意义。方法从我院2013年9月-2014年1月时间段内所接收的不孕不育女性患者中选取100名,采用酶联免疫吸附试验(Enzyme linked immunosorbent as say,ELISA),检测患者血清生殖免疫性抗体,即抗精子抗体(AsAb)、抗子宫内膜抗体(EMAb)、抗卵巢抗体(AOAb)、抗心磷脂抗体(ACA)四项,分析结果并判断检测血清生殖免疫抗体对不孕不育患者临床意义[1]。结果选择的100名患者抗体阳性率为53%,其中AsAb阳性19%,EMAb阳性8%,AOAb阳性10%,ACA阳性16%。结论血清生殖免疫抗体的产生是导致不孕症的重要原因之一,患者年龄不同则含量不同,血清生殖免疫抗体检测在不孕不育治疗中具有重要的意义。
基金supported by the National Natural Science Fundation of China(No.30400336)Science and Technology Project of Qingdao(03-1-NSH-2-2).
文摘A two-step zero-length cross-linking procedure using active esters was successfully adopted for conjugating en-rofloxacin (EF) to human serum albumin (HSA). The derived conjugate was characterized by UV spectrum and then usedfor immunization of BALB/C mice. In enzyme-linked immunosorbent assay (ELISA) and competitive inhibition ELISA ex-periments, the derived antiserum exhibited high antibody titer (greater than 1: 250 000) as well as varied cross-reactivity(from 97.8% to 161.7%) to three analogs of EF belonging to fluoroquinolones family. But over the concentration rangestudied, no significant cross-reactivity was observed to other group of antibiotics (chloramphenicol, oxytetracycline, sul-phamethoxazole and nysfungin). It was confirmed that the synthesized immunogen was highly antigenic and elicited specificantibody responses in BALB/C mice against EF.
基金Supported by the Excellent Young Teacher Fund of Chinese Education Ministry and the General Science Technology Research Program of Zhejiang Province, No. 001110438
文摘AIM: To investigate the frequencies of the expression of main protein antigens of Helicobacter pylori (H py/ori) isolates, such as UreB, VacA, CagA1, HpaA, NapA, FlaA and FlaB and the production of specific antibodies in sera from H pylori-infected patients, and to understand the correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori. METHODS: H pylori strains in biopsy specimens from 157 patients with chronic gastritis and peptic ulcer were isolated and serum samples from the patients were also collected. The target recombinant proteins rUreB, rVacA, rCagAl, rHpaA, rNapA, rFlaA and rFlaB expressed by the prokaryotic expression systems constructed in our previous studies were collected through Ni-NTA affinity chromatography. Rabbit antisera against rUreB, rVacA, rCagAl, rHpaA, rNapA, rFlaA and rFlaB were prepared by using routine subcutaneous immunization. By using ultrasonic lysates of the isolates as coated antigens, and the self-prepared rabbit antisera as the first antibodies and commercial HRP-labeling sheep anti-rabbit IgG as the second antibody, expression frequencies of the seven antigens in the isolates were detected by ELISA. Another ELISA was established to detect antibodies against the seven antigens in sera of the patients by using the corresponding recombinant proteins as coated antigens, and the sera as the first antibody and HRP-labeling sheep anti-human IgG as the second antibody respectively. Correlations among the different clinical types of chronic gastritis and peptic ulcer and the infection and virulence of H pylori were statistically analysed. RESULTS: In the 125 isolates of H pylori, the positive rates of UreB, VacA, CagAl, HpaA, NapA, FlaA and FlaB were 100%, 65.6%, 92.8%, 100%, 93.6%, 100% and 99.2% respectively. In the 125 serum samples from the H pylori infected patients, the positive rates of antibodies against recombinant UreB, VacA, CagA1, HpaA, NapA, FIaA and FlaB were 100%, 42.4%, 89.6%, 81.6%, 93.6%, 98.4% and 92.8% respectively. H pylori strains were isolated from 79.6% (125/157) of the biopsy specimens, but no close correlations among the H pylori infection frequencies and different types of chronic gastritis and peptic ulcer could be found (P>0.05, x2 = 0.01-0.87). The VacA positive rate (82.40%) in the strains isolated from the specimens of patients with peptic ulcer and the anti-VacA positive rate (54.3%) in the sera from the patients were significantly higher than those (51.5%, 32.3%) from the patients with chronic gastritis (P<0.01, x2= 13.19; P<0.05, x2= 6.13). When analysis was performed in the different types of chronic gastritis, the VacA in the strains isolated from the specimems of patients with active gastritis showed a higher expression frequency (90.0%) than those from superficial (47.9%) and atrophic gastritis (30.0%) (P<0.05, x2 = 5.93; P<0.01,x2 = 7.50). While analysis was carried out in the strains isolated from the specimens with superficial (93.8%) and active gastritis (100%), NapA showed a higher expression frequency compared to that from atrophic gastritis (60.0%) (P<0.01, x2 = 8.88; P<0.05, X2=5.00). CONCLUSION: The types of chronic gastritis and peptic ulcer and their severity are not associated with H pylori infection frequency but closely related to the infection frequency of different virulent H pylori strains. The optimal antigens for developing vaccine and diagnostic kit are UreB, FlaA, HpaA, FlaB, NapA and CagAl, but not VacA.
文摘AIM:To study the diagnosis of Helicobacter pylori(H pylori) infection through the determination of serum levels of anti- H pylori IgG and IgA antibodies,and the levels of anti-H pylori IgA antibodies in duodenal fluid. METHODS:Data were collected from 93 patients submitted to upper digestive endoscopy due to dyspeptic symptoms. The patients were either negative(group A)or positive (group B)to H pylori by means of both histological detection and urease tests.Before endoscopy,peripheral blood was collected for the investigation of anti-H pylori IgG and IgA antibodies.To perform the urease test,biopsies were obtained from the gastric antrum.For the histological evaluation,biopsies were collected from the gastric antrum (greater and lesser curvatures)and the gastric body. Following this,duodenal fluid was collected from the first and second portions of the duodenum.For the serological assaying of anti-Hpylori IgG and IgA,and anti-Hpylori IgA in duodenal fluids,the ELISA method was utilized. RESULTS:The concentration of serum IgG showed sensitivity of 64.0%,specificity of 83.7%,positive predictive value of 82.0%,negative predictive value of 66.6% and accuracy of 73.1% for the diagnosis of H pylori infection.For the same purpose,serum IgA showed sensitivity of 72.0%, specificity of 65.9%,positive predictive value of 72.0%, negative predictive value of 67.4% and accuracy of 69.8%. If the serological tests were considered together,i.e.when both were positive or negative,the accuracy was 80.0%, sensitivity was 86.6%,specificity was 74.2%,positive predictive value was 74.2% and negative predictive value was 86.6%.When values obtained in the test for detecting IgA in the duodenal fluid were analyzed,no significant difference(P=0.43)was observed between the values obtained from patients with or without H pylori infection. CONCLUSION:The results of serum IgG and IgA tests for H pylori detection when used simultaneously,are more efficient in accuracy,sensitivity and negative predictive value, than those when used alone.The concentration of IgA antibodies in duodenal fluid is not useful in identifying patients with or without H pylori.
文摘AIM. To evaluate the serologic IgG response to Hpylori and CagA across age groups and in healthy children and adults.METHODS： Totally, 386 children aged 1-15 years and 200 adults aged 20-60 years, were enrolled to study. The serum samples of participant were tested for presence of antJ-Hpylori and anti-CagA IgG by using ELISA method.RESULTS： The seroprevalence of Hpylori in adults was significantly higher than that observed in children （67.5% vs 46.6%; P 〈 0.000003）. In children, the seropositivity rate in males （51.9%） was significantly （P 〈 0.05） higher than that observed in females （41.7%）. The prevalence of serum anti-CagA antibody was 72.8% and 67.4% in infected children and adults, respectively. The mean titer of serum anti-CagA antibodies was significantly higher among children in comparison to adults （64.1 Uarb/mL 30.7; P 〈 0.03）. In infected children and adults the prevalence of serum anti-CagA antibody was higher in males compared to females （78.4% vs 66.3%; P = 0.07 and 75.6% vs 54.71%; P 〈 0.04, respectively）. The age-specific prevalence of anti-H py/or/and anti-CagA antibody （in infected subjects） was 37.6% and 59.57% at age 1-5 years, 46.9% and 75% at age 6-10 years, 54.9% and 79.45% at age 11-15, 59.01% and 83.33% at age 20-30 years, 66.6% and 60.52% at age 31-40 years, 73.46% and 63.88% at age 41-50 years and 75.75% and 60% at age 51-60 years with mean titer of anti-CagA antibody of 75.94, 63.32, 57.11, 52.06, 23.62, 21.52 and 21.80 Uarb/mL, respectively. There was significant difference between mean serum anti-CagA antibody in age subgroups （P 〈 0.001）.CONCLUSION： These results showed that anti-Hpylori and anti-CagA antibodies were common in the children and adults. The Hpylori-specific antibodies influenced by age and sex of subjects. Moreover, it seems that males are more susceptible to infection with CagA^＋ strains compared to females. The seroprevalence of anti-CagA antibody was increased with age, up to 30 years and then decreased. It was also found that the magnitude of the IgG response to CagA decreased with advanced age.
