1.Effect of Yiqi Jianpi Fang on Aberrant Crypt Foci Production in Carcinogenesis Wistar Rat Colorectal Cancer Model and Investigation on the Related Mechanism
Fanchen ZHAO ; Yan LI ; Yingbin LUO ; Yiyang ZHOU ; Xiating XU
Progress in Modern Biomedicine 2017;17(26):5043-5047
Objective:To investigate the inhibitory effect and the related mechanism of Yiqi Jianpi Fang on aberrant crypt focus (ACF) in Wister rat colorectal cancer model.Methods:50 Wistar rats were randomly divided into 5 groups:TCM low dose group(TCM solution diluted 3 times,5 mL/kg daily gastric volume),TCM middle dose group (5 mL/kg daily gastric volume),TCM high dose group (15 mL/kg daily gastric volume),model group,normal group.With 10 rats in each group.The colorectal tissues were observed under microscope after methylene blue staining by immunohistochemistry method.Results:Compared with the model group,the number of ACF and large ACF in each TCM group were decreased (P<0.05),and the number of ACF in the TCM middle dose group reduced most obvious,and the difference was statistically significant (P<0.05).The proliferation indexs of PCNA in intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P<0.05).The apoptosis index of intestinal gland cells 24 h and 48 h after modeling in the TCM groups were higher than those in the model group,and the difference was statistically significant(P<0.05).The ectopic expression of β-catenin in TCM groups were lower than that in the model group,and it was highest in the high dose group,than was the low dose group,and the middle group was lowest(P<0.05).The expression of MMP-7 in TCM groups were lower than that in the model group,and it was highest in the low dose group,than was the high dose group,and the middle dose group was lowest (P<0.05).Conclusion:Yiqi Jianpi Fang can significantly reduce the number of ACF in Wister rats,inhibit the activation of Wnt signaling in colorectal cancer,reduce the incidence of colorectal cancer,and have a certain preventive effect.
2.Extracorporeal membrane oxygenation combined with ultrafiltration technique for treatment of kidney injury after severe hemorrhagic shock in rabbits
Geng TIAN ; Liang ZHAO ; Lin LUO ; Juan XIAO ; Mei LIU ; Fuqin TANG ; Yingbin XIAO
Chinese Journal of Trauma 2014;30(8):843-847
Objective To investigate the effect of extracorporeal membrane oxygenation (ECMO) combined with ultrafiltration in treatment of kidney injury induced by serious hemorrhagic shock in rabbits.Methods Models of pressure-controlled hemorrhagic shock was developed in 24 New Zealand white rabbits which were divided into unresuscitation group (n =8),ECMO combined with ultrafiltration group (combined resuscitation group,n =8),and fluid resuscitation group (n =8) according to the random number table.Heart rate was monitored via electrocardiograph and arterial pressure via fermoral artery catheter.Blood samples were collected pre-and post-shock and after resuscitation to measure levels of lactic acid,serum creatinine,IL-6,and TNF-α.Kidney samples were collected for measurement of histopathological changes via HE staining,expression of heat shock protein 70 (HSP70) via immunohistochemical staining.Results Arterial pressure was (53.1 ± 11.4) mmHg in combined resuscitation group,higher than (41.3 ± 11.1) mmHg in fluid resuscitation group and (25.9 ± 10.5) mmHg in unresuscitation group (F =41.425,P < 0.05).Hemorrhagic shock induced significant up-regulation of lactic acid,serum creatinine,IL-6,and TNF-α(P < 0.05),but all were lowered after resuscitation,especially in combined resuscitation group (P < 0.05).HE staining showed the degree of kidney tissue necrosis and inflammatory cytokine infiltration in combined resuscitation group alleviated notably compared with fluid resuscitation group.Median and interquartile values of HSP70 were 17 828.960 0 (15 779.865 0-21 751.980 0) in unresuscitation group,2 714.270 0 (1 339.215 0-7 616.950 0) in fluid resuscitation group,and 262.930 0 (198.820 0-538.195 0) in combine resuscitation group,with statistical differences among groups(P < 0.05).Conclusion ECMO combined with ultrafiltration is superior to conventional fluid resuscitation in improving hypoxia tissue injury and inflammatory reaction after hemorrhagic shock and is beneficial to attenuating kidney injury.
3.Chemiluminescent immunoassay for high-sensitivity C-reactive protein.
Pingguo LIU ; Guoqiang LI ; Yixin CHEN ; Haifeng LUO ; Dedang HUANG ; Yingbin WANG ; Shengxiang GE ; Jun ZHANG ; Ningshao XIA
Chinese Journal of Biotechnology 2010;26(8):1150-1156
We developed a high-sensitivity C-reactive protein quantifiable chemiluminescent immunoassay (hs-CRP CLIA). The high-purity native CRP was purified from hepatic cirrhosis patient ascetic fluid by affinity and ion exchange chromatography and used as an immunogen to develop the monoclonal antibodies (mAbs) against CRP. Twenty-two mAbs were identified reactive with CRP in ELISA and 13 of them were reactive in the phosphorycholine ligand capture ELISA. The mAbs 10C5 and 10C11 were selected to develop the hs-CRP CLIA. The linearity and performance of the hs-CRP CLIA was characterized. It was showed not reactive when testing against other serum materials (IgG, hemoglobin and triglyceride). The reliable correlation (R2 > 0.993) was obtained between testing value (RLU/S) and the concentration of human serum CRP calibrator. The linearity fell in the range of 0.04-20.38 mg/L. The assay has good accuracy and reproducibility, the mean recovery was 99% and the precision of the intra- and inter assay was CVs (4.2%-5.8%) and (9.0%-11.5%), respectively. In testing of 90 human sera, this assay performed well and correlated comparably with a commercial hs-CRP ELISA kit. Thus, hs-CRP CLIA is an accurate, reliable, quantifiable assay for detection of high-sensitive C-reactive protein in serum, it may be useful to improve the risk assessment of cardiovascular disease and the prognosis of inflammatory bowel disease.
C-Reactive Protein
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analysis
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chemistry
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Humans
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Immunoassay
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methods
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Luminescent Measurements
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methods
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Sensitivity and Specificity
4.Nodal induces epithelial-mesenchymal transition in gallbladder carcinoma via the Smad2/4 pathway
Yang WANG ; Yingbin HUANG ; Guangyu CHEN ; Rui HE ; Jun DU ; Canqiao LUO ; Xingyuan JIAO
Chinese Journal of Hepatobiliary Surgery 2018;24(9):594-599
Objective To study the expressions of Nodal in normal gallbladder,and gallbladders with cholelithiasis,cholecystitis and carcinoma;and to study the impact of inhibiting or promoting Nodal expressions in gallbladder carcinoma on the Smad2/4 pathway and epithelial-mesenchymal transition.Methods Immunohistochemistry was used to detect the expressions and distributions of Nodal protein in 30 normal gallbladders,96 simple cholecystitis/calculous cholecystitis specimens and 42 gallbladder carcinoma specimens.The mRNA and protein expressions of Nodal in normal and malignant gallbladdcr mucosal epithelium cells and breast cancer cells were detected by RT-PCR,western blotting and wound healing tests.The impact of activating agents and inhibitors on the expression levels of Nodal and its signaling pathway Smad2/4 and EMT-related proteins were analyzed.Results Immunohistochemistry showed that the positive rates of Nodal in the gallbladder cancer group was significantly higher than that in the gallbladder stone group and the normal gallbladder group.The results were 83.3% (35/42),44.8% (43/96),6.7% (2/30) respectively (P< 0.01).RT-PCR and Western blotting showed the expressions of Nodal in gallbladder carcinoma cells were higher than normal gallbladder cells (P<0.05).After using rhNodal to up regulate the Nodal expression,the Smad2 protein phosphorylation was promoted and the EMT associated proteins were up-regulated.After using the inhibitor SB431542 to suppress the Nodal expression,the Smad2 protein phosphorylation decreased and the EMT associated proteins were down-regulated.Conclusions The expression of Nodal was closely related to cell proliferation and metastasis in gallbladder carcinoma.Tumor progression was promoted via the smad2/4 pathway through epithelial-mesenchymal transition.
5.Detection of free DNA septin 9 gene methylation in plasma.
Zhi LUO ; Yingbin HU ; Xiaoyun PU
Journal of Central South University(Medical Sciences) 2021;46(2):127-134
OBJECTIVES:
To explore the correlation between cytosine-phosphoric-guanylic (CpG) site of Septin 9 gene and colorectal cancer, and to develop a real-time PCR detection system in plasma in patients with colorectal cancer.
METHODS:
The methylation of training samples was detected by high-throughput sequencing technology, and the sites highly consistent with the clinical information of colorectal cancer were identified. Then the detection system of real-time PCR was designed to analyze the consistency of plasma and tissue based on methylationa sensitive enzyme digestion. Finally, 100 clinical trials were conducted to evaluate the performance of the detection system with the methylation sensitive enzyme digestion-real-time PCR.
RESULTS:
The highly consistent sites, which were selected by high-throughput sequencing from 71 training set samples, was the 38th CpG. Based on the detection region, the screened methylation sensitive enzymes were
CONCLUSIONS
The 38th CpG site of Septin 9 detected by the detection system of methylation sensitive enzyme digestion-real-time PCR can highly predict the occurrence of colorectal cancer with great clinical application value.
Colorectal Neoplasms/genetics*
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CpG Islands/genetics*
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DNA
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DNA Methylation
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Humans
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Plasma/metabolism*
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Septins/metabolism*
6.Overexpression of the long non-coding RNA ADAMTS9-AS2 suppresses colorectal cancer proliferation and metastasis.
Xiaoyun BU ; Ang QIN ; Zhi LUO ; Yingbin HU
Journal of Central South University(Medical Sciences) 2019;44(7):741-748
To investigate the expression, clinical significance, and biological function of the long non-coding RNA (lncRNA) ADAMTS9-AS2 in colorectal cancer (CRC).
Methods: Gene microarray analysis was performed to explore the expression of ADAMTS9-AS2 in CRC. Real-time PCR was used to verify its expression in 20-paired CRC tissues and adjacent non-tumor tissues. We further explored the relationship between ADAMTS9-AS2 expression and clinicopathological features, and its prognostic role in relapse-free survival (RFS) among early stage CRC patients using Kaplan-Meier and Cox regression analyses. In vitro assays, cell counting kit-8 assay, colony formation assay, and Transwell assay were used to evaluate the biological function of ADAMTS9-AS2 in CRC.
Results: ADAMTS9-AS2 was down-regulated in CRC patients according to the gene microarray analysis, which was confirmed in CRC tissues and cells. High expression of ADAMTS9-AS2 was associated with a higher 5-year RFS rate (83.8% vs 73.5%, P=0.041) and it was an independent prognostic factor for RFS [hazard ratio (HR)=0.528; 95% CI 0.299 to 0.932; P=0.028] at the early stage of CRC. ADAMTS9-AS2 overexpression in CRC cells inhibited cell proliferation, migration, and invasion, while suppression of ADAMTS9-AS2 showed opposite effects.
Conclusion: ADAMTS9-AS2 is a valuable prognostic factor for CRC and may function as a tumor suppressor in CRC via inhibiting cell proliferation and metastasis.
ADAMTS9 Protein
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genetics
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Cell Movement
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Cell Proliferation
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Colorectal Neoplasms
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genetics
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Gene Expression Regulation, Neoplastic
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Humans
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Neoplasm Recurrence, Local
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RNA, Long Noncoding
7.Expert consensus on the use of human serum albumin in adult cardiac surgery.
Fei XIANG ; Fuhua HUANG ; Jiapeng HUANG ; Xin LI ; Nianguo DONG ; Yingbin XIAO ; Qiang ZHAO ; Liqiong XIAO ; Haitao ZHANG ; Cui ZHANG ; Zhaoyun CHENG ; Liangwan CHEN ; Jimei CHEN ; Huishan WANG ; Yingqiang GUO ; Nan LIU ; Zhe LUO ; Xiaotong HOU ; Bingyang JI ; Rong ZHAO ; Zhenxiao JIN ; Robert SAVAGE ; Yang ZHAO ; Zhe ZHENG ; Xin CHEN
Chinese Medical Journal 2023;136(10):1135-1143