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Three novel missense germline mutations in different exons of MSH6 gene in Chinese hereditary non-polyposis colorectal cancer families

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World Journal of Gastroenterology 2007年第37期13卷 5021-5024页

作者： Shi-yan yan xiao-yan zhou Xiang Du Tai-Ming Zhang Yong-Ming Lu San-Jun Cai xiao-Li Xu Bao-Hua Yu Heng-Hua zhou Da-Ren Shi Department of Pathology Cancer Hospital/InstituteFudan University Department of Oncology Shanghai MedicalCollege Fudan University Shanghai 200032 China Department of Abdominal Surgery CancerHospital/Institute Fudan University Shanghai 200032 China

AIM: To investigate the germline mutations of MSH6 gene in probands of Chinese hereditary non-polyposis colorectal cancer (HNPCC) families fulfilling different clinical criteria. METHODS: Germline mutations of MSH6 gene were detected by PCR-based DNA sequencing in 39 unrelated HNPCC probands fulfilling different clinical criteria in which MSH2 and MLH1 mutations were excluded. To further investigate the pathological effects of detected missense mutations, we analyzed the above related MSH6 exons using PCR-based sequencing in 137 healthy persons with no family history. The clinicopathological features were collected from the Archive Library of Cancer Hospital, Fudan University and analyzed. RESULTS: Four germline missense mutations distributed in the 4th, 6th and 9th exons were observed. Of them, three were not found in international HNPCC databases and did not occur in 137 healthy controls, indicating that they were novel missense mutations. The remaining mutation which is consistent with the case H14 at c.3488A>T of exon 6 of MSH6 gene was also found in the controls, the rate was approximately 3.65% (5/137) and the type of mutation was not found in the international HNPCC mutational and SNP databases, suggesting that this missense mutation was a new SNP unreported up to date. CONCLUSION: Three novel missense mutations and a new SNP observed in the probands of Chinese HNPCC families, may play an important role in the development of HNPCC.

关键词： Hereditary non-polyposis colorectal cancer MSH6 Missense mutation Colorectal cancer

MLH1 promoter germline-methylation in selected probands of Chinese hereditary non-polyposis colorectal cancer families

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World Journal of Gastroenterology 2008年第48期14卷 7329-7334页

作者： Heng-Hua zhou Shi-yan yan xiao-yan zhou Xiang Du Tai-Ming Zhang Xu Cai Yong-Ming Lu San-Jun Cai Da-Ren Shi Department of Pathology Cancer Hospital/Institute Fudan University Department of Oncology Shanghai Medical College Fudan University Shanghai 209032 China Department of Gastroenterology Xinhua Hospital Shanghai Jiaotong University School of Medicine Shanghai 200092 China Department of Abdominal Surgery Cancer Hospital/Institute Fudan University Shanghai 200032 China

AIM： To detect the MLH1 gene promoter germline- methylation in probands of Chinese hereditary non- polyposis colorectal cancer （HNPCC）, and to evaluate the role of methylation in MLH1 gene promoter and molecular genetics in screening for ***： The promoter germline methylation of MLH1 gene was detected by methylation-specific PCR （MSP） in 18 probands from unrelated HNPCC families with high microsatellite-instability （MSI-H） phenotype but without germline mutations in MSH2, MLH1 and MSH6 genes. At the same time, 6 kindreds were col- lected with microsatellite-stability （MSS） phenotype but without germline mutations in MSH2, MIH1 and MSH6 genes as controls. The results of MSP were confirmed by clone sequencing. To ensure the reliability of the results, family H65 with nonsense germline mutation at c.2228C 〉 A in MSH2 gene was used as the negative control and the cell line sw48 was used as the known positive control along with water as the blank control. Immunochemical staining of MIH1 protein was performed with Envision two-step method in those patients with aberrant methylation to judge whether the status of MLH1 gene methylation affects the expression of MLH1 ***： Five probands with MIH1 gene promoter methylation were detected in 18 Chinese HNPCC families with MSI-H phenotype but without germline mutations in MSH2, MLH1 and MSH6 genes. Two of the five probands from families H10 and H29 displayed exhaustive-methylation, fulfilling the Japanese criteria （JC） and the Amsterdam criteria （AC）, respectively. The other 3 probands presented part-methylation fulfilling the AC. Of the 13 probands with unmethylation phenotype, 8 fulfilled the JC and the Bethesda guidelines （BG）, 5 fulfilled the AC. The rate of aberrant methylation in MLH1 gene in the AC group （22.2%, 4/18） was higher than that in the JC/BG groups （5.6%, 1/18） in all HNPCC families with MSI-H phenotype but without germline mutations in PISH2, PIIH1 and MSH6 genes. However, no proband with m

关键词： Hereditary non-polyposis colorectal cancer MLH1 Methylation Germline Methylation-specific PCR Microsatellite phenotype

Visceral hypersensitive rats share common dysbiosis features with irritable bowel syndrome patients

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World Journal of Gastroenterology 2016年第22期22卷 5211-5227页

作者： xiao-yan zhou Ming Li Xia Li Xin Long Xiu-Li Zuo xiao-Hua Hou Ying-Zi Cong yan-Qing Li Department of Gastroenterology Shandong University Qilu Hospital Department of Gastroenterology Union Hospital of Tongji Medical College Huazhong University of Science and Technology Department of Microbiology and Immunology The University of Texas Medical Branch

AIM: To evaluate gut microbial dysbiosis in two visceral hypersensitive models in comparison with irritable bowel syndrome(IBS) patients and to explore the extent to which these models capture the dysbiosis of IBS patients. METHODS: Visceral hypersensitivity was developed using the maternal separation(MS) rat model and post-inflammatory rat model. The visceral sensitivity of the model groups and control group was evaluated using the abdominal withdraw reflex score and electromyography in response to graded colorectal distention. The 16 S ribosomal RNA gene from fecal samples was pyrosequenced and analyzed. The correlation between dysbiosis in the microbiota and visceral hypersensitivity was calculated. Positive findings were compared to sequencing data from a published human IBS ***: Dysbiosis triggered by neonatal maternal separation was lasting but not static. Both MS and postinflammatory rat fecal microbiota deviated from that of the control rats to an extent that was larger than the co-housing effect. Two short chain fatty acid producing genera, Fusobacterium and Clostridium XI, were shared by the human IBS cohort and by the maternal separation rats and post-inflammatory rats, respectively, to different extents. Fusobacterium was significantly increased in the MS group, and its abundance positively correlated with the degree of visceral hypersensitivity. Porphyromonadaceae was a protective biomarker for both the rat control group and healthy human controls. CONCLUSION: The dysbiosis MS rat model and the post-inflammatory rat model captured some of the dysbiosis features of IBS patients. Fusobacterium, Clostridium XI and Porphyromonadaceae were identified as targets for future mechanistic research.

关键词： Animal model Irritable bowel syndrome Microbiota Pyrosequencing 16S rRNA gene

Lipotoxic hepatocyte-derived exosomal miR-1297 promotes hepatic stellate cell activation through the PTEN signaling pathway in metabolic-associated fatty liver disease

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World Journal of Gastroenterology 2021年第14期27卷 1419-1434页

作者： Xin Luo Sheng-Zheng Luo Zi-Xin Xu Cui zhou Zheng-Hong Li xiao-yan zhou Ming-Yi Xu Department of Gastroenterology Shanghai General HospitalShanghai 200080China

BACKGROUND Exosomes play an important role in metabolic-associated fatty liver disease(MAFLD),but the mechanism by which exosomes participate in MAFLD still remain *** To figure out the function of lipotoxic exosomal miR-1297 in *** MicroRNA sequencing was used to detect differentially expressed miRNAs(DEmiR)in lipotoxic exosomes derived from primary *** tools were applied to analyze the target genes and pathways regulated by the *** real-time PCR(qPCR)was conducted for the verification of ***,western blot,immunofluorescence staining and ethynyl-20-deoxyuridine assay were used to evaluate the function of lipotoxic exosomal miR-1297 on hepatic stellate cells(LX2 cells).A luciferase reporter experiment was performed to confirm the relationship of miR-1297 and its target gene *** MicroRNA sequencing revealed that there were 61 exosomal DE-miRs(P2 from palmitic acid treated primary hepatocytes compared with the vehicle control ***-1297 was the most highly upregulated according to the microRNA *** tools showed a variety of target genes and pathways regulated by these DE-miRs were related to liver ***-1297 was overexpressed in exosomes derived from lipotoxic hepatocytes by *** promoting genes(α-SMA,PCNA)were altered in LX2 cells after miR-1297 overexpression or miR-1297-rich lipotoxic exosome incubation via qPCR and western blot *** staining and ethynyl-20-deoxyuridine staining demonstrated that the activation and proliferation of LX2 cells were also promoted after the above *** was found to be the target gene of miR-1297 and knocking down PTEN contributed to the activation and proliferation of LX2 cells via modulating the PI3K/AKT signaling *** miR-1297 was overexpressed in exosomes derived from lipotoxic *** lipotoxic hepatocyte-derived exosomal miR-1297 could promot

关键词： Metabolic-associated fatty liver disease miRNA-1297 Exosome Hepatic stellate cell PTEN Liver fibrosis

Human epidermal growth factor receptor type 2 protein expression in Chinese metastatic prostate cancer patients correlates with cancer specific survival and increases after exposure to hormonal therapy

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Asian Journal of Andrology 2008年第5期10卷 701-709页

作者： Bo Dai Yun-Yi Kong Ding-Wei Ye Chun-Guang Ma xiao-yan zhou Xu-Dong Yao Department of Urology Cancer Hospital Fudan University Shanghai 200032 China Department of Pathology Cancer Hospital Fudan University Shanghai 200032 China Department of Oncology Shanghai Medical College Fudan University Shanghai 200032 China

Aim： To investigate human epidermal growth factor receptor type 2 （HER2） protein expression and gene amplification in Chinese metastatic prostate cancer patients and their potential value as prognostic factors. Methods： Immunohistochemistry （IHC） was performed to investigate HER2 protein expression in prostate biopsy specimens from 104 Chinese metastatic prostate cancer patients. After 3-11 months of hormonal therapy, 12 patients underwent transurethral resection of the prostate （TURP）. HER2 protein expression of TURP specimens was compared with that of the original biopsy specimens. Of these, 10 biopsy and 4 TURP specimens with HER2 IHC staining scores ≥ 2＋ were investigated for HER2 gene amplification status by fluorescent in situ hybridization （FISH）. Results： Of the 104 prostate biopsy specimens, HER2 protein expression was 0, 1＋, 2＋ and 3＋ in 49 （47.1%）, 45 （43.3%）, 8 （7.7%） and 2 （1.9%） cases, respectively. There was a significant association between HER2 expression and Gleason score （P = 0.026）. HER2 protein expression of prostate cancer tissues increased in 33.3% of patients after hormonal therapy. None of the 14 specimens with HER2 IHC scores 〉 2＋ showed HER2 gene amplification. Patients with HER2 scores 〉 2＋ had a significantly higher chance of dying from prostate cancer than those with HER2 scores of 0 （P = 0.004） and 1＋（P = 0.034）. Multivariate Cox regression analysis showed that HER2 protein expression intensity was an independent predictor of cancer-related death （P = 0.039）. Conclusion： An HER2 IHC score 〉 2＋ should be defined as HER2 protein overexpression in prostate cancer. Overexpression of HER2 protein in cancer tissue might suggest an increased risk of dying from prostate cancer. HER2 protein expression increases in some individual patients after hormonal therapy.

关键词： prostatic neoplasms human epidermal growth factor receptor type 2 immunohistochemistry gene amplification prostate cancer prognosis

Two novel germline mutations of MLH1 and investigation of their pathobiology in hereditary non-polyposis colorectal cancer families in China

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World Journal of Gastroenterology 2007年第46期13卷 6254-6258页

作者： Chao-Fu Wang xiao-yan zhou Tai-Ming Zhang Ye Xu San-Jun Cai Da-Ren Shi Department of Pathology Cancer Hospital of Fudan UniversityShanghai 200032 Department of Oncology Shanghai MedicalCollege Fudan University Shanghai 200032 China Department of Abdominal Surgery CancerHospital of Fudan University Shanghai 200032 China

AIM： To detect germline mutations of MLH1, and investigate microsatellite instability and expression of MLH1 in tumor tissues of hereditary non-polyposis colorectal cancer （HNPCC） with two novel germline mutations, and further investigate the pathobiology of the two novel mutations of MLH1. METHODS： RNA was extracted from the peripheral blood of 12 patients from 12 different families that fulfilled the Amsterdam 11 Criteria for HNPCC. Germline mutations of MLH1 were determined by RT-PCR, followed by cDNA sequencing analysis. PCR-GeneScan analysis was used to investigate microsatellite instability with a panel of five microsatellite markers （BAT26, BAT25, D5S346, D2S123 and mfd15）, along with immunohistochemical staining to detect the expression of MLH1 protein in two patients＇ tumor tissues with novel mutations. RESULTS： Three germline mutations were found in four patients, one of the mutations has previously been reported, but the other two, CGC→TGC at codon 217 of exon 8 and CCG→CTG at codon 581 of exon 16, have not been reported. The two patients＇ tumor tissues with novel mutations had high-frequency microsatellite instability that showed more than two unstable loci, and both tumors lost their MLH1 protein expression. CONCLUSION： The two novel germline mutations of MLH1 in HNPCC families i.e. CGC→TGC at codon 217 of exon 8 and CCG→CTG at codon 581 of exon 16, are very likely to have pathological significance.

关键词： Colorectal cancer Hereditary non-polyposiscolorectal cancer MLH1 gene Germline mutation Microsatellite instability Gene sequencing

Heterochronic triple primary malignancies with Epstein-Barr virus infection and tumor protein 53 gene mutation:A case report and review of literature

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World Journal of Clinical Cases 2021年第5期9卷 1184-1195页

作者： Wen-Xia Peng Xin Liu Qi-Feng Wang xiao-yan zhou Zhi-Guo Luo Xi-Chun Hu Department of Medical Oncology Fudan University Shanghai Cancer CenterShanghai 200032China Department of Pathology Fudan University Shanghai Cancer CenterShanghai 200032China

BACKGROUND The diagnosis and etiology of multiple primary malignant neoplasms(MPMNs)are difficult to ***,we report a case of heterochronic triple primary malignancies with gastric cancer,nasopharyngeal squamous cell cancer,and then rectal *** SUMMARY The patient was first diagnosed with gastric cancer at the age of 33 in 2014 and underwent distal gastrectomy and gastrojejunostomy and six cycles of adjuvant *** years later,he was diagnosed with nasopharyngeal cancer and treated with radical chemoradiotherapy in ***,a mass in the middle of the rectum was resected and reported as ulcerative,moderately to poorly differentiated *** on the etiology of MPMNs showed that Epstein-Barr virus(EBV)infection may be the cause of gastric cancer and nasopharyngeal squamous cell cancer since these two primary lesions were positive for transcripts of EBV-encoded ribonucleic acid using an in situ hybridization EBV-encoded ribonucleic acid probe in formalin-fixed,paraffinembedded *** cause of rectal cancer may be due to a somatic mutation of tumor protein 53 gene in exon 8(c.844C>T,***282Trp)through highthroughput sequencing for the rectal *** standard therapy for each primary cancer was administered,and the patient has no evidence of cancer disease to *** To our knowledge,this is the first report on heterochronic triple primary malignancies whose cause may be associated with EBV infection and tumor protein 53 genetic *** etiological research may not only elucidate the cause of MPMN but also has implications in clinical management.

关键词： Multiple primary malignant neoplasms Epstein-Barr virus infection Epstein-Barr virus-encoded RNA TP53 mutation Etiology Case report

xiaojianzhong decoction prevents gastric precancerous lesions in rats by inhibiting autophagy and glycolysis in gastric mucosal cells

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World Journal of Gastrointestinal Oncology 2023年第3期15卷 464-489页

作者： Jia-Xiang Zhang Sheng-Chuan Bao Juan Chen Ting Chen Hai-Liang Wei xiao-yan zhou Jing-Tao Li Shu-Guang yan College of Basic Medicine Shaanxi University of Chinese MedicineXianyang 712046Shaanxi ProvinceChina Key Laboratory of Gastrointestinal Diseases and Prescriptions in Shaanxi Province Shaanxi University of Chinese MedicineXianyang 712046Shaanxi ProvinceChina Department of General Surgery The Affiliated Hospital of Shaanxi University of Chinese MedicineXianyang 712000Shaanxi ProvinceChina Department of Gastroenterology The Affiliated Hospital of Shaanxi University of Chinese MedicineXianyang 712000Shaanxi ProvinceChina Departments of Infectious Disease The Affiliated Hospital of Shaanxi University of Chinese MedicineXianyang 712000Shaanxi ProvinceChina

BACKGROUND Gastric precancerous lesions(GPL)precede the development of gastric cancer(GC).They are characterized by gastric mucosal intestinal metaplasia and dysplasia caused by various factors such as inflammation,bacterial infection,and *** in autophagy and glycolysis affect GPL progression,and their effective regulation can aid in GPL treatment and GC *** decoction(XJZ)is a classic compound for the treatment of digestive system diseases in ancient China which can inhibit the progression of ***,its specific mechanism of action is still *** To investigate the therapeutic effects of XJZ decoction on a rat GPL model and the mechanisms underlying its effects on autophagy and glycolysis regulation in *** Wistar rats were randomly divided into six groups of five rats each and all groups except the control group were subjected to GPL model construction for 18 *** rats’body weight was monitored every 2 wk starting from the beginning of *** histopathology was examined using hematoxylin-eosin staining and Alcian blue-periodic acid-Schiff *** was observed using transmission electron *** expressions of autophagy,hypoxia,and glycolysis related proteins in gastric mucosa were detected using immunohistochemistry and *** expressions of the following proteins in gastric tissues:B cell lymphoma/Leukemia-2 and adenovirus E1B19000 interacting protein 3(Bnip-3),microtubule associated protein 1 light chain 3(LC-3),moesin-like BCL2-interacting protein 1(Beclin-1),phosphatidylinositol 3-kimase(PI3K),protein kinase B(AKT),mammalian target of rapamycin(mTOR),p53,AMP-activated protein kinase(AMPK),and Unc-51 like kinase 1(ULK1)were detected using western *** relative expressions of autophagy,hypoxia,and glycolysis related mRNA in gastric tissues was detected using reverse transcription-polymerase chain *** Treatment with XJZ increased the rats’body we

关键词： xiaojianzhong decoction Gastric precancerous lesions Autophagy Glycolysis Gastric mucosal cells Herb

A Confocal Technique Applicable to Studies of Cellular pH-related Signaling in Plants

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Journal of Integrative Plant Biology 2008年第6期50卷 682-690页

作者： Bing-Bing Lit Zhi-Hui Gao xiao-yan zhou Hui-Bo Ren Min Xie Yi-Juan Fan Jian-Fang Hu Wen-Suo Jia College of Agronomy and Biotechnology China Agricultural University Beijing 100094 China

pH may act as a crucial signal in both animal and plant cells. It is very difficult to monitor pH signals and this has largely hindered progress in the investigation of pH signaling, particulady systematic pH signaling. Here, we report the development of a confocal technique to monitor leaf apoplastic pH in intact plants, which is particularly suitable for the studies on root to shoot signaling. A variety of different pH indicators and plant species were tested. It was found that different pH indicators, for example, 2＇,7＇-Bis-（2-carboxyethyl）-5-（and-6）-carboxyfluoresce （BCECF）, SNARF-4F 5-（and-6）- carboxylic acid （SNARF） and DM-NERF （NERF）, were of different properties, and to successfully monitor pH at a sub-cellular level, the comparability between the pH indicator and plant species must be involved according to their suitable pH range and loading characteristics. The loading characteristics of different pH indicators differ with different plant species, cell types and their developing stages. No matter what methods were adopted, BCECF and SNARF could not be loaded specifically in the leaf apoplast in sunflower, tomato, and Comelina communis L. In contrast, regardless of the methods adopted, NERF could be loaded efficiently and specifically in the leaf apoplast in C. communis, but not in other plants. In C. communis, the determination coefficient for in vitro and in situ calibration of NERF was very high, which was respectively 0.9951 and 0.991 6, and therefore, the adoption of NERF together with C. communis could construct an ideal experimental system that is suitable for the investigation of pH systematic signaling. Ratio image analysis demonstrated that the leaf apoplastic pH was about 5.5 in non-stressed conditions, and water deficit could trigger an increase in pH by about half a pH unit, which is the first evidence to directly indicate that pH is able to act as a systematic signal under water deficit conditions.

关键词： confocal microscope pH signaling water deficit.