1.Expression and significance of high molecular weight cytokeratin in breast cancer
Yaxin ZHAO ; Zhiguang ZHAO ; Zhiqiang ZHENG
Chinese Journal of General Surgery 2008;23(6):450-453
Objective To study the expression of high molecular weight cytokeratin (CK34βE12) in breast carcinoma and the relationship between the high molecular weight cytokeratin expression and patient's prognosis. Methods We detected the high molecular weight cytokeratin expression in 100 patient's of breast cancer tissues and paracancerous breast tissue in 30 cases immunohistochemically. The relationship between the expression of high molecular weight cytokeratin in breast cancer tissues and age, menstrual status, estrogen receptor expression, progesterone receptor expression, histology grading, TNM staging, pathology type, and patient survival was analyzed. Results (1) There was no significant correlation between high molecular weight cytokeratin expression of breast -cancer tissues and age, menstrual status, estrogen receptor expression, progesterone receptor expression (all P>0.05). But the correlation was statistically significant between that expression and histology grading, TNM staging, pathology category (all P<0.05). (2) By univariate analysis, the 5-year disease-free survival rate in patients with negative expression was lower than those with positive expression(P<0.05). By multivariate analysis, the expression of high molecular weight cytokeratin was unfavorably related with tumor free survival. Conclusion The expression of high molecular weight cytokeratin in breast carcinoma tissues was correlated with biologic malignancies of breast cancer, the negative high molecular weight cytokeratin expression of breast cancer tissues predicts poor prognosis.
2.Advances in the study of deubiquitinating enzymes in cancer research
Chenxin DAI ; Jian YUAN ; Yaxin ZHENG
China Oncology 2013;(7):547-550
Deubiquitinating enzymes, reversing protein ubiquitination, most of the researches focused on the ifeld of molecular biology. However, it have not yet attracted enough attention in translational medicine research. In fact, target proteins of deubiquitinating enzymes affect the tumor progression through various ways, for example, cell apoptosis and autophagy, the link between inlfammation and cancer, tumor hypoxia, signal transduction, cell cycle regulation and DNA damage. This paper reviewed the research progress on the relations between deubiquitinating enzymes and the correlated factors of tumor.
3.The significance of peripheral blood CK19 mRNA expression in breast cancer patients
Yaxin ZHAO ; Jiefan LIU ; Zhongmin SUN ; Minghua JIANG ; Zhiqiang ZHENG
Chinese Journal of General Surgery 2012;27(8):656-659
ObjectiveTo evaluate the significance of CK19 mRNA monitoring in peripheral blood of breast cancer patients.MethodsPeripheral blood samples were collected from 137 breast cancer patients preoperatively,one day post-operation,7 days,1,3,6,12,18,and 24 months after operation.RTPCR was used to detect CK19 mRNA expression.The relationships were analyzed between CK19 mRNA expression and treatment result, between CK19 mRNA expression and clinicopathological parameters.ResultsThere was no significant difference between the level of CK19 mRNA as tested preoperatively,and 1,7 days postoperatively(P > 0.05 ).From one month and thereafter,CK 19 mRNA decreased significantly when compared with that immediate perioperatives ( P < 0.05 ). Postoperative peripheral CK19 mRNA expression increased in those found with recurrent or metastasized tumors ( P < 0.01 ). CK19 mRNA expression does not correlate with patient's menstrual status,estrogen receptor expression,progesterone receptor expression,HER-2 expression and Ki67 proliferation expression (P > 0.05 ). But there was statistically significant correlation between CK19 mRNA expression and tumour size,histology grading,pathological type,lymph node metastasis ( P < 0.01 ).ConclusionsIn breast cancer patients peripheral blood CK19 mRNA expression is correlated with risk clinicopathological parameters, and increases in patients with postoperative recurrence and tumor metastasis.
4.Application of On-line SPE-HPLC System in Pharmacokinetic Study of Highly Active Anti-Cancer Compound TEB-415
Man WANG ; Yabin WEN ; Kangning LIU ; Ge SI ; Lei LIU ; Zheng YIN ; Yaxin LU
Chinese Journal of Analytical Chemistry 2014;(12):1729-1734
An on-line solid phase extraction-high performance liquid chromatography ( SPE-HPLC ) system was applied in the plasma pharmacokinetic study of highly active anti-cancer compound tyrosine kinase inhibitors (TEB-415) in mouse. The on-line SPE-HPLC method associated with Ultimate3000 system which was applied to the determination of the blood drug level of TEB-415 in mouse plasma. C18 column ( Venusil MP, 150 mm × 4. 6 mm, 5μm) was used as analytical column and the mobile phase consisted of acetonitrile-5 mmol/L monopotassium phosphate buffer ( pH 3 . 5 ) at a flow rate of 1 . 0 mL/min was used as the isocratic elution. An MF Ph-1 column (10 mm×4 mm, 5 μm) was used as on-line SPE column, and water and water-acetonitrile were used as the washing solvent and elution solvent respectively. The detection wavelength was set at 262 nm. The pharmacokinetic parameters were calculated by WinNonlin 5. 2 software. The linear range of the calibration curve was between 100 and 20000 μg/L, and the limit of qualification was 20 μg/L. The extraction recovery was between 90 . 5% and 94 . 6%. The RSD of intra-day and inter-day precision was less than 3. 5%. The accuracy of short-term stability, freeze-thaw stability and long-term stability were between 91. 49% and 101. 96%. After oral medication, the mean peak time (Tmax) of TEB-415 in mice was 5. 29 h, and the mean maximum concentration ( Cmax) was 3403μg/L. The area under the curve ( AUC) of TEB-415 was 24600 μg/L·h. This drug's mean half-life was 3. 84 h, and its mean retention time (MRT) was 6. 56 h. These parameters suggested that TEB-415 had appropriate rate of absorption and elimination with preferable bioavailability.
5.The Relationship between the Expression of COX-2,PD-ECGF and VEGF in Human Gastric Carcinoma and Tumor Angiogenesis
Xihong HU ; Zhiqiang ZHENG ; Zhiguang ZHAO ; Yaxin ZHAO ; Zhanglin NI ; Tao YOU
Journal of Medical Research 2006;0(01):-
Objective To explore the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma tissue,and analyze and identify the relationship between the expression and tumor angiogenesis together with biological behaviour in gastric carcinoma.Methods ElivisionTM immunohistochemical method was performed to detect the expression of COX-2,PD-ECGF,VEGF and CD34,and evaluate the value of MVD in surgically resected gastric carcinoma specimens and biopsy tissues under gastroscope,including normal gastric mucosa tissues, metaplasia tissues and gastric mucosal atypical hyperplasia tissues. Clinicopathologic data of patients were analyzed retrospectively. Results The positive expression of COX-2, PD-ECGF,VEGF and MVD in gastric carcinoma were significantly higher than those in normal gastric mucosa, metaplasia or gastric mucosal atypical hyperplasia,and also their expression and MVD were closely related to lymph node metastasis and depth of invasion. The MVD in the groups of COX-2,PD-ECGF,VEGF positive-expression were significantly higher than those in negative-expression groups respectively. There were positive correlations among the expression of COX-2,PD-ECGF and VEGF in gastric carcinoma. MVD increased significantly when COX-2, PD-ECGF and VEGF coexpressed. Conclusion The expression of COX-2,PD-ECGF,VEGF and tumor angiogenesis participated in the genesis,invasion and metastasis of human gastric carcinoma.COX-2,PD-ECGF and VEGF participated in the tumor angiogenesis of gastric carcinoma. In the process of tumor angiogenesis and expression in gastric carcinoma, COX-2, PD-ECGF and VEGF could strengthen the effectiveness each other.They could strengthen the effective ness of tumor angiogenesis when they coexpressed. They could be selected as targets for tumor anti-angiogenesis treatment.
6.Effect of Xiangshahewei Pill on secretion in anorectic rat models induced by Fenfluramine
Jing WANG ; Ying HONG ; Yaxin YANG ; Sheng ZHAO ; Jianfeng XING ; Xiankun WANG ; Xiaohe ZHENG
Chinese Traditional Patent Medicine 1992;0(08):-
AIM:To explore the effect of Xiangshahewei Pill(Radix Aucklandiae,Rhizoma Cyperi,Fructus Amomi,etc.)on secretion in anorectic rat model induced by Fenfluramine.METHODS:Anorectic rat model was induced by Fenfluramine in dose of 15 mg/kg for 8 days.Xiangshahewei Pill was administered orally to the rats also for 8 days.The volume of gastric juice,total gastric acid,mucus,acidity of gastric acid and activity of pepsin of rats were measured.RESULTS:Xiangshahewei Pill could improve the turbulence of gastric secretion in anorectic rat induced by Fenfluramine.Xiangshahewei Pill in dose of 0.75 g/kg(P
7.CRMP-2 is involved in hypoxic preconditioning-induced neuroprotection against cerebral ischemic injuries of mice
Caiyan ZHANG ; Sujuan FENG ; Xu LIU ; Xiangning BU ; Nan ZHANG ; Yaxin ZHENG ; Xiaowen YUAN ; Xiaoguang LI ; Junfa LI
Basic & Clinical Medicine 2009;29(11):1133-1138
Objective To investigate whether conventional protein kinase C (cPKC ) βⅡ-interacting collapsin response mediator protein-2 (CRMP-2) provides neuroprotection against cerebral ischemic (I) injuries. Methods Male BALB/c mice were randomly divided into normoxic control (Nor) , HPC, Nor + Sham, HPC + Sham, Nor + I and HPC + I groups (n = 6 per group). Using our HPC and MCAO mouse models, we applied immunoprecipita-tion, two-dimensional electrophoresis and mass spectrometry to characterize cPKCβⅡ-interacting proteins and combined with SDS-PAGE and Western blot to quantitatively analyze CRMP-2 phosphorylation and degradation levels in the brain of mice after HPC and MCAO. Results The expression level of 10 cPKCβⅡ-interacting proteins changed obviously in cerebral cortex of HPC mice when compared with Nor group. One of these proteins, CRMP-2 protein level increased in particulate fraction and decreased in cytosolic fraction of cerebral cortex of HPC mice. CRMP-2 phosphorylation level in ischemic core (Ic) of cerebral cortex decreased significantly ( P < 0. 05 , n = 6) as compared with that of Nor + sham group, but CRMP-2 phosphorylation level in HPC +I group increased significantly as compared with that of Nor +I group ( P < 0. 05, n = 6). In ischemic cortex, CRMP-2 degradation (proteolysis) was observed as the appearance of 55 ku breakdown products (BDP). However, the CRMP-2 degradation level, BDPs products decreased significantly in penumbra ( P) of ischemic cortex from HPC +I group when we compared with that of Nor +I group (P < 0. 05, n = 6 ). Conclusion CRMP-2 is involved in attenuating the decrease of CRMP-2 phosphorylation in ischemic core and in inhibiting its degradation in penumbra of cerebral cortex of mice thereby to lessen the ischemic injuries.
8.The inhibitory effects of sorafenib on gastric cancer cells MGC80-3
Yi CHEN ; Shuo CHEN ; Juanhong ZHENG ; Yaxin ZHAO ; Jie JIA
China Modern Doctor 2015;(21):8-10
Objective To explore the inhibitory effective and mechanism on the proliferation and apoptosis of sorafenib in vitro gastric cancer MGC80-3 cells. Methods The inhibitory effective of sorafenib at different concentrations for gastric cancer cell MGC80-3 were analyzed by MTT assay. Apoptosis rate was observed by AnnexinV/PI staining. Re-sults The proliferation inhibition of sorafenib on gastric cancer MGC80-3 cell was time-dose dependent manner, the difference was statistically significant (P<0.05). Conclusion The apoptosis of gastric cancer MGC80-3 cells is different and the apoptotic rate is increased with the increase of sorafenib concentration.
9.miR-9 on the proliferation of breast cancer cells by targeting hexokinase 2
Yuanbin WANG ; Yaxin ZHAO ; Lijun YANG ; Jinying ZHENG ; Jundong TANG ; Ruiming MA
Chinese Journal of Endocrine Surgery 2020;14(1):52-55
Objective:To study the regulatory effect of miR-9 on the proliferation of breast cancer cells by targeting hexokinase 2 (HK2) .Methods:Breast cancer tissues and paracancer tissues were collected and the expression levels of miR-9 and HK2 were detected. MCF-7 cells were cultured and divided into blank control group, NC group, miR-9 group, NC-siRNA group and HK2-siRNA group. The cell viability, expression levels of HK2 and cleaved caspase-3 were detected, the targeted binding of miR-9 to HK2 was verifed.Results:the expression level of miR-9 in breast cancer was lower than that in paracancer tissue (0.52±0.08 vs 1.05±0.25, t=16.685, P<0.000) , and the expression level of HK2 was higher than that in paracancer tissue (0.73±0.14 vs 0.34±0.08, t=17.587, P<0.000) , and the expression level of miR-9 was negatively correlated with HK2; the OD490 level, the expression level of HK2 and the fluorescence activity of double luciferase reporter gene containing HK2 mRNA 3 'UTR in miR-9 group were lower than those in NC group (0.58±0.09 vs 1.04±0.21, 0.51±0.08 vs 1.18±0.24, 41.11±9.28 vs 148.28±29.59, t/P=4.027/0.007, 5.297/0.002, 6.912/0.001) , and the expression level of cleaved caspase-3 was higher than that in NC group (1.08±0.26 vs 0.42±0.09, t/P=4.797/0.003) . The OD490 level and the expression level of HK in HK-siRNA group were higher than that in NC-siRNA group, and the expression level of cleaved caspase-3 was higher than that of NC-siRNA group. Conclusion:miR-9 can inhibit the proliferation of breast cancer cells, and its mechanism may be related to the targeted inhibition of HK2 expression and the increase of downstream cleaved caspase-3 expression.
10.Effects of short-chain fatty acids on gut microbiota and hippocampal TLR4/MyD88/NF-κB pathway proteins in depression model mice
Wenjuan HAN ; Yaxin ZHENG ; Lan WANG ; Fengya ZHEN ; Yan ZHANG ; Cuixia AN
Chinese Journal of Behavioral Medicine and Brain Science 2024;33(7):583-589
Objective:To investigate whether short chain fatty acid(SCFAs) intervention has an antidepressant effect by improving gut microbiota dysregulation and regulating the TLR4/MyD88/NF-κB inflammatory pathway in depression model mice.Methods:Totally 60 SPF grade male C57BL/6 J mice aged 6-8 weeks were randomly divided into three groups: control group, depression model group, and SCFAs group, with 20 mice in each group.The mice in depression model group and SCFAs group were given the chronic unpredictable mild stress (CUMS) stimulations for 8 weeks to establish the depression model.From the 6th week, SCFAs group mice were given a mixed solution of short chain fatty acid salts for drinking, until modeling was completed, meanwhile mice in the model group were given 0.78% NaCl solution for drinking.The depression-like behavior was assessed using the sucrose preference test (SPT) and forced swimming test (FST) following modeling, and the open field test (OFT) was employed to evaluate the anxiety-like behavior of mice.16S rRNA gene sequence was used to analyze the gut microbiota of mice.The activation of astrocytes and TLR4/MyD88/NF-κB inflammatory pathway in hippocampus was determined by immunofluorescence staining and Western blot.SPSS 26.0 was used for statistical analysis, one-way ANOVA was used for comparison among the three groups, and LSD- t test was used for further pairwise comparisons. Results:There were statistically significant differences in the sugar water preference rate, the immobility time in FST, and the percentage of activity time in OFT among the three groups ( F=10.554, 10.912, 12.599, all P<0.05).The the sugar water preference rate and the percentage of activity time in OFT of the depression model group were both lower than those of the control group (both P<0.05), and the immobility time in FST was higher than that of the control group ( P<0.05).The sugar water preference rate in SCFAs group((84.7±3.5)%, (75.3±6.0)%)and the percentage of activity time in OFT((7.4±1.4)%, (3.2±0.9)%) were both higher than those in the depression model group(both P<0.05 ), while the immobility time in FST was shorter than that in the depression model group((110.5±21.5) s, (148.0±20.1) s, P<0.05).There was a statistical difference in the β diversity of gut microbiota among three groups ( P=0.001).At the family level, compared with the depression model group, the relative abundance of Rikenellaceaee and Bacteroidaceae increased in the SCFAs group, while the relative abundance of Clostridia_UCG-014 decreased.At the genus level, the relative abundance of Clostridia_UCG-014 and Prevotella decreased, while the relative abundance of Alistipes increased (all P<0.05).The immunofluorescence results showed that there was a statistically significant difference in GFAP expression levels among the three groups of mice ( F=16.565, P=0.004).The GFAP expression in the depression model group was higher than that in the control group and SCFAs group (both P<0.05).The Western blot results showed that there were statistically significant differences in the expression levels of TLR4, MYD88, and NF-κB ptoteins in the hippocampal tissue of the three groups ( F=70.59, 174.39, 14.40, all P<0.05).The protein levels of TLR4, MyD88, and NF-κB in the depression model group were all higher than those in the control group and SCFAs group (all P<0.05). Conclusion:SCFAs can ameliorate the depressive-like behavior in depression model mice and reduce the activation of astrocytes in the hippocampus, which may be associated with the improvement of dysregulated gut microbiota and down-regulation of the TLR4/MyD88/NF-κB pathway protein.