AIM:To investigate the correlations between selfreported symptoms of irritable bowel syndrome(IBS) and the gastrointestinal(GI) microbiota composition.METHODS:Fecal samples were collected from a total of 44 subjects d...AIM:To investigate the correlations between selfreported symptoms of irritable bowel syndrome(IBS) and the gastrointestinal(GI) microbiota composition.METHODS:Fecal samples were collected from a total of 44 subjects diagnosed with IBS.Their symptoms were monitored with a validated inflammatory bowel disease questionnaire adjusted for IBS patients.Thirteen quantitative real-time polymerase chain reaction assays were applied to evaluate the GI microbiota composition.Eubacteria and GI bacterial genera(Bifidobacterium,Lactobacillus and Veillonella),groups(Clostridium coccoides/Eubacterium rectale,Desulfovibrio desulfuricans) and distinct bacterial phylotypes [closest 16S rDNA sequence resemblance to species Bifidobacterium catenulatum,Clostridium cocleatum,Collinsella aerofaciens(C.aerofaciens),Coprococcus eutactus(C.eutactus),Ruminococcus torques and Streptococcus bovis ] with a suspected association with IBS were quantified.Correlations between quantities or presence/absence data of selected bacterial groups or phylotypes and various IBSrelated symptoms were investigated.RESULTS:Associations were observed between subjects' self-reported symptoms and the presence or quantities of certain GI bacteria.A Ruminococcus torques(R.torques)-like(94% similarity in 16S rRNA gene sequence) phylotype was associated with severity of bowel symptoms.Furthermore,among IBS subjects with R.torques 94% detected,the amounts of C.cocleatum 88%,C.aerofaciens-like and C.eutactus 97% phylotypes were significantly reduced.Interesting observations were also made concerning the effect of a subject's weight on GI microbiota with regard to C.aerofaciens like phylotype,Bifidobacterium spp.and Lactobacillus spp.CONCLUSION:Bacteria seemingly affecting the symptom scores are unlikely to be the underlying cause or cure of IBS,but they may serve as biomarkers of the condition.展开更多
AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domain...AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domains), p16, RUNX3 (runt-related transcription factor 3), E-cadherin, hMLH1 (mutL homolog 1), ABCG2 (ATP-binding cassette, sub-family G, member 2) and BNIP3 (BCL2/adenovirus E1B 19 kDa interacting protein 3) were analyzed in 80 specimens of PF by quantitative methylation-specific polymerase chain reaction (PCR). Eighty patients were divided into 3 groups; Group A (n=35):the depth of cancer invasion was less than the muscularis propria; Group B (n=31): the depth of cancer invasion was beyond the muscularis propria. Both group A and B were diagnosed as no cancer cells in peritoneal cytology and histology; Group C (n=14): disseminated nodule was histologically diagnosed or cancer cells were cytologically defi ned in the peritoneal cavity.RESULTS: The positive rates of methylation in CHFR, E-cadherin and BNIP3 were significantly different among the 3 groups and increased in order of group A, B and C (0%,0% and 21% in CHFR, P<0.05; 20%, 45% and 50% in E-cadherin, P<0.05;26%,35% and 71% in BNIP3, P<0.05). In addition, the multigene methylation rate among CHFR, E-cadherin and BNIP3 was correlated with group A, B and C (9%,19% and 57%, P<0.001). Moreover, the prognosis was analyzed in group B, excluding 3 patients who underwent a non-curative resection. Two of the 5 patients with multigene methylation showed peritoneal recurrence after surgery, while those without or with a single gene methylation did not experience recurrence (P<0.05).CONCLUSION: This study suggested that gene methylation in the PF could detect occult neoplastic cells in the peritoneum and might be a risk factor for peritoneal metastasis.展开更多
AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected ...AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction （MSP） in four esophageal cancer cell lines （KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines）, nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine （5-aza-dc）, a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS： MSP results were as follows： loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation （com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells）; all nine cases of normal esophageal mucosa were unmethylated; and 54/72 （75.00%） samples from patients with esophageal can- cer were methylated, and 25/72 （34.70%） matched adjacent tissues were methylated （75.00% vs 34,70%, z2 = 23.5840, P = 0.000）. mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue （Z2= 9.143, P = 0.002）. XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation （Fisher＇s exact test, P = 0.004）. Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION： XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.展开更多
目的探讨实时聚合酶链反应（polymerase chain reaction，PCR）技术检测妊娠晚期孕妇B族溶血性链球菌（group B Streptococcus，GBS）的准确性。方法本研究为多中心研究。选择2009年3月1日至12月31日在北京大学第一医院妇产科、首都医...目的探讨实时聚合酶链反应（polymerase chain reaction，PCR）技术检测妊娠晚期孕妇B族溶血性链球菌（group B Streptococcus，GBS）的准确性。方法本研究为多中心研究。选择2009年3月1日至12月31日在北京大学第一医院妇产科、首都医科大学附属北京妇产医院产科和北京大学第三医院妇产科产前保健的妊娠35～37周孕妇，取阴道下1/3分泌物及肛周分泌物，采用常规细菌培养法及实时PCR方法进行GBS检测。采用基因测序作为矫正方法，分析实时PCR方法检测GBS的敏感性和特异性。结果（1）3家医院共收集1395份标本，细菌培养法检测GBS阳性40例（2.9%），实时PCR方法检测GBS阳性114例（8.2%）。（2）仅实时PCR方法检测GBS阳性者77例，采用事先设计好的第2对引物扩增后进行测序，检测鉴定为GBS序列的共66例，11例为非GBS。（3）以细菌培养法加测序法校正作为金标准，实时PCR方法检测GBS的敏感性为97.2%（103/106），特异性为99.1%（1278/1289）。常规细菌培养法漏诊率62.3%（66/106）。（4）细菌培养法加测序法校正3家医院孕妇妊娠晚期GBS携带率为7.6%（106/1395）。结论实时PCR方法检测GBS具有较高的敏感性和特异性，有望成为妊娠晚期常规检测GBS的方法。展开更多
基金Supported by The Finnish Funding Agency for Technology and Innovation (Tekes grant No 40039/03)the Finnish Graduate School on Applied Biosciences and the Centre of Excellence on Microbial Food Safety Research,Academy of Finland
文摘AIM:To investigate the correlations between selfreported symptoms of irritable bowel syndrome(IBS) and the gastrointestinal(GI) microbiota composition.METHODS:Fecal samples were collected from a total of 44 subjects diagnosed with IBS.Their symptoms were monitored with a validated inflammatory bowel disease questionnaire adjusted for IBS patients.Thirteen quantitative real-time polymerase chain reaction assays were applied to evaluate the GI microbiota composition.Eubacteria and GI bacterial genera(Bifidobacterium,Lactobacillus and Veillonella),groups(Clostridium coccoides/Eubacterium rectale,Desulfovibrio desulfuricans) and distinct bacterial phylotypes [closest 16S rDNA sequence resemblance to species Bifidobacterium catenulatum,Clostridium cocleatum,Collinsella aerofaciens(C.aerofaciens),Coprococcus eutactus(C.eutactus),Ruminococcus torques and Streptococcus bovis ] with a suspected association with IBS were quantified.Correlations between quantities or presence/absence data of selected bacterial groups or phylotypes and various IBSrelated symptoms were investigated.RESULTS:Associations were observed between subjects' self-reported symptoms and the presence or quantities of certain GI bacteria.A Ruminococcus torques(R.torques)-like(94% similarity in 16S rRNA gene sequence) phylotype was associated with severity of bowel symptoms.Furthermore,among IBS subjects with R.torques 94% detected,the amounts of C.cocleatum 88%,C.aerofaciens-like and C.eutactus 97% phylotypes were significantly reduced.Interesting observations were also made concerning the effect of a subject's weight on GI microbiota with regard to C.aerofaciens like phylotype,Bifidobacterium spp.and Lactobacillus spp.CONCLUSION:Bacteria seemingly affecting the symptom scores are unlikely to be the underlying cause or cure of IBS,but they may serve as biomarkers of the condition.
基金Supported by Fund from the Department of Surgery,Saga University Faculty of Medicine
文摘AIM:To investigate whether gene methylation in the peritoneal fluid (PF) predicts peritoneal recurrence in gastric cancer patients.METHODS: The gene methylation of CHFR (checkpoint with forkhead and ring finger domains), p16, RUNX3 (runt-related transcription factor 3), E-cadherin, hMLH1 (mutL homolog 1), ABCG2 (ATP-binding cassette, sub-family G, member 2) and BNIP3 (BCL2/adenovirus E1B 19 kDa interacting protein 3) were analyzed in 80 specimens of PF by quantitative methylation-specific polymerase chain reaction (PCR). Eighty patients were divided into 3 groups; Group A (n=35):the depth of cancer invasion was less than the muscularis propria; Group B (n=31): the depth of cancer invasion was beyond the muscularis propria. Both group A and B were diagnosed as no cancer cells in peritoneal cytology and histology; Group C (n=14): disseminated nodule was histologically diagnosed or cancer cells were cytologically defi ned in the peritoneal cavity.RESULTS: The positive rates of methylation in CHFR, E-cadherin and BNIP3 were significantly different among the 3 groups and increased in order of group A, B and C (0%,0% and 21% in CHFR, P<0.05; 20%, 45% and 50% in E-cadherin, P<0.05;26%,35% and 71% in BNIP3, P<0.05). In addition, the multigene methylation rate among CHFR, E-cadherin and BNIP3 was correlated with group A, B and C (9%,19% and 57%, P<0.001). Moreover, the prognosis was analyzed in group B, excluding 3 patients who underwent a non-curative resection. Two of the 5 patients with multigene methylation showed peritoneal recurrence after surgery, while those without or with a single gene methylation did not experience recurrence (P<0.05).CONCLUSION: This study suggested that gene methylation in the PF could detect occult neoplastic cells in the peritoneum and might be a risk factor for peritoneal metastasis.
基金Supported by Grants from the National Basic Research Program(973 Program), No. 2012CB934002, 2010CB912802National Key Scientific Instrument Special Programme of China, No.2011YQ03013405National Science Foundation of China,No. 81121004, 81071953 and 81161120432
文摘AIM： To explore epigenetic changes in the gene encod- ing X chromosome-linked inhibitor of apoptosis-associ- ated factor 1 （XAF1） during esophageal carcinogenesis. METHODS： Methylation status of XAF1 was detected by methylation-specific polymerase chain reaction （MSP） in four esophageal cancer cell lines （KYSE30, KYSE70, BICl and partially methylated in TE3 cell lines）, nine cases of normal mucosa, 72 cases of pri- mary esophageal cancer and matched adjacent tissue. XAF1 expression was examined by semi-quantitative reverse transcriptional polymerase chain reaction and Western blotting before and after treatment with 5-aza- deoxycytidine （5-aza-dc）, a demethylating agent. To investigate the correlation of XAF1 expression and methylation status in primary esophageal cancer, immu- nohistochemistry for XAF1 expression was performed in 32 cases of esophageal cancer and matched adjacent tissue. The association of methylation status and clini-copathological data was analyzed by logistic regression. RESULTS： MSP results were as follows： loss of XAF1 expression was found in three of four esophageal cell lines with promoter region hypermethylation （com- pletely methylated in KYSE30, KYSE70 and BIC1 cell lines and partially in TE3 cells）; all nine cases of normal esophageal mucosa were unmethylated; and 54/72 （75.00%） samples from patients with esophageal can- cer were methylated, and 25/72 （34.70%） matched adjacent tissues were methylated （75.00% vs 34,70%, z2 = 23.5840, P = 0.000）. mRNA level of XAF1 mea- sured with semi-quantitative reverse transcription poly- merase chain reaction was detectable only in TE3 cells, and no expression was detected in KYSE30, KYSE70 or BIC1 cells. Protein expression was not observed in KYSE30 cells by Western blotting before treatment with 5-aza-dc. After treatment, mRNA level of XAF1 was detectable in KYSE30, KYSE70 and BIC1 cells. Protein expression was detected in KYSE30 after treatment with 5-aza-dc. Immunohistochemistry was performed on 32 cases of esophageal cancer and adjacent tissue, and demonstrated XAF1 in the nucleus and cytoplasm. XAF1 staining was found in 20/32 samples of adjacent normal tissue but was present in only 8/32 samples of esophageal cancer tissue （Z2= 9.143, P = 0.002）. XAF1 expression was decreased in cancer samples compared with adjacent tissues. In 32 cases of esophageal can- cer, 24/32 samples were methylated, and 8/32 esopha- geal cancer tissues were unmethylated. XAF1 staining was found in 6/8 samples of unmethylated esophageal cancer and 2/24 samples of methylated esophageal cancer tissue. XAF1 staining was inversely correlated with XAF1 promoter region methylation （Fisher＇s exact test, P = 0.004）. Regarding methylation status and clinicopathological data, no significant differences were found in sex, age, tumor size, tumor stage, or metas- tasis with respect to methylation of XAF1 for the 72 tis- sue samples from patients with esophageal cancer. CONCLUSION： XAF1 is frequently methylated in eso- phageal cancer, and XAF1 expression is regulated by promoter region hypermethylation.
文摘目的探讨实时聚合酶链反应（polymerase chain reaction，PCR）技术检测妊娠晚期孕妇B族溶血性链球菌（group B Streptococcus，GBS）的准确性。方法本研究为多中心研究。选择2009年3月1日至12月31日在北京大学第一医院妇产科、首都医科大学附属北京妇产医院产科和北京大学第三医院妇产科产前保健的妊娠35～37周孕妇，取阴道下1/3分泌物及肛周分泌物，采用常规细菌培养法及实时PCR方法进行GBS检测。采用基因测序作为矫正方法，分析实时PCR方法检测GBS的敏感性和特异性。结果（1）3家医院共收集1395份标本，细菌培养法检测GBS阳性40例（2.9%），实时PCR方法检测GBS阳性114例（8.2%）。（2）仅实时PCR方法检测GBS阳性者77例，采用事先设计好的第2对引物扩增后进行测序，检测鉴定为GBS序列的共66例，11例为非GBS。（3）以细菌培养法加测序法校正作为金标准，实时PCR方法检测GBS的敏感性为97.2%（103/106），特异性为99.1%（1278/1289）。常规细菌培养法漏诊率62.3%（66/106）。（4）细菌培养法加测序法校正3家医院孕妇妊娠晚期GBS携带率为7.6%（106/1395）。结论实时PCR方法检测GBS具有较高的敏感性和特异性，有望成为妊娠晚期常规检测GBS的方法。
