Objective To investigate the expression level of LMO1 in gastric cancer tissues and human gastric cancer cell lines,and explore the invasive and metastatic potential of LMO1 gene silencing by small interfering RNA on the human gastric cancer cell line MKN28.Methods Immunohistochemical technique was applied to detect the expression of LOM1 protein in gastric cancer tissues and tumor adjacent tissues of paraffin specimens in 30 cases.The expression levels of LMO1 in human gastric cancer cell lines AGS,BGC-823,SGC-7901,MKN28 and human gastric mucosal epithelial cells GES were detected by realtime-PCR and Western blot.Using LipofectamineTM 2000,LMO1 siRNA was transfected into MKN28 cellsin vitro (siRNA transfect group).Negative control was established.Real time-PCR and Western blot was used to examine the difference of LMO1 expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of E-cadherin,MMP-9 and VEGF.Results Positive rate of LOM1 protein in gastric cancer tissues(77％)was higher than that in tumor adjacent tissues (17％) (x2 =21.70,P ＜ 0.01).Positive rate of Vavl protein was higher in lymphatic metastasis group than in non-lymphatic metastasis group(x2 =5.83,P =0.02).Compared with GES,the expression level of LMO1 increased significantly in gastric cancer cell lines,especially in MKN28 (P ＜ 0.01).The expression levels of LMO1 mRNA and protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜0.01).The invasive and metastatic potentials of LMO1-siRNA transfected MKN28 cellssignificantly decreased (t =-11.53,P ＜0.01;t =-10.68,P ＜0.01).The expression levels of E-cadherin were higher than the matched negative control cells;and MMP-9,VEGF protein in LMO1-siRNA transfected MKN28 cells were lower than the matched negative control cells (P ＜ 0.01).Conclusions LMO1 has higher expression level in gastric cancer tissues and some gastric cancer cell lines,and down-regulation of LMO1 can inhibit the invasion and metastasis ability of gastric cancer.

Objective To screen the tumor metastasis related differentially expressed genes in hepatocelluar carcinoma (HCC)cells 7402 after stable transfection with FATE/BJ‐HCC‐2 gene .Methods Total RNA was extracted from FATE/BJ–HCC‐2‐transfected HCC(5B4)cells and empty vector control (Mock)cells respectively .Differentially expressed genes were obtained using cDNA microarray .Results Compared with Mock cells ,a total of 1 694 differentially expressed genes were screened out in 5B4 cells ,the 11 gene expressions had obvious differences ,among which the expression amounts in 7 genes were significantly in‐creased ,including MMP‐1 ,PTGS2 ,FN ,CA9 ,IL‐8 ,ILK and Areg .The fold changes were 81 .80 ,49 .86 ,11 .30 ,16 .26 ,3 .48 ,2 .79 and 2 .20 ,respectively .The expression amounts in 4 genes were significantly decreased ,including E‐cadherin ,E‐cadherin , RHOBTB3 ,ALPP and HLA‐DRB4 .The fold changes were -5 .42 ,-2 .23 ,-5 .93 and -8 .03 ,respectively .Conclusion Adopting gene microarray technology can carefully screen the differentially expressed genes of FATE/BJ‐HCC‐2 involved HCC metastasis ,its final goal is to lay a solid theoretical foundation for studying the HCC metastasis mechanism .

Objective:To apply the strategy of proteomics with two dimensional gel electrophoresis(2DE) in esophageal cancer.Methods:the protein of esophageal cancer tissues and its adjacent normal tissue were separated by 2DE, the gels were analyzed with software ImageMaster4.01 to find out the spots different between cancer and control, and the proteins in the spots were identified by matrix associated laser desorption/ionization time of flight(MALDI TOF) mass spectrometry.Results:a new reasonable strategy of study for proteomics on esophageal cancer was raised and candidate cancer marker of two pyruvate kinase isozyme were found under such strategy.Conclusion:Under reasonable strategy, the analysis of proteomics with 2DE on human tissue is a useful method for discovering valuable cancer marker candidates. Pyruvate kinase isozyme M2/M1 which were found in cancer tussues will be very important in diagnosis and treatment of esophageal cancer.

Objective:To investigate the expression of MEK/ERK signaling pathways in renal cell car-cinoma with bone metastasis,and to analyze the differences of expressions of VEGFR-2,MEK,ERK on the primary and metastasis tissue and its mechanism.Methods:The tissue samples were obtained from 7 renal cell carcinoma patients kindly provided by Department of Urology,Peking University People’s Hos-pital from January 1,2009 to January 1,2010.The expression of MEK/ERK signaling pathways was de-tected in the 7 renal cell carcinoma patients`primary and matched metastatic tissues with ICH,The anti-body concentrations were 1 ∶200,1 ∶25,and 1 ∶250,respectively.The mutation of the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the se-cond exon of the MEK1 gene were detected with PCR.Results:The expression intensities of VEGFR-2, MEK,and ERK were measured by H-score [intensity (1,2,3,or 4)multiplied by the distribution (%)].VEGFR-2,MEK,and ERK expressions were divided into 3 groups according to the positive dis-tribution of the tumor cells:1,0 -5%;2,6% -50%;and 3,>50%,To assess intratumor heteroge-neity,three distinct microscopic fields (×200)from each specimen were used to evaluate the expres-sions,Subsequently,the scores were averaged to obtain a single concatenated score for each tissue. VEGFR-2,MEK,and ERK expressions were assessed by 2 independent pathologists who were blinded to the clinicopathological data.The data were expressed as the mean value of the triplicate experiments.The expressions of MEK,and ERK were higher in the metastatic tissues than in the matched RCC tissues (6.10 ±4.10 vs.1.33 ±0.51,P =0.015;9.10 ±2.24 vs.4.43 ±2.84,P =0.021 )while the ex-pression of VEGFR-2 was not different between the primary and metastatic tissues (P =0.901).No mu-tation was detected on the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the second exon of the MEK1 gene.Conclusion:MEK/ERK signa-ling pathways may play an important role in the metastasis and the resistance of sunitinib in RCC patients with bone metastasis.

Objective To investigate the detection of BJ-75A-9 mHNA, WNX and CK19 mRNA in peripheral blood cells of lung cancer patients and evaluate its diagnostic value. Methods The expression of BJ-TSA-9, LUNX and CK19 mRNA in peripheral blood cells was delected from 84 lung cancer patients, 32 benign lung lesions and 20 healthy volunteers by nested reverse transcription polymerase chain re-action (Nested-RT-PCR). Results The positive detection rates of BJ-75A-9,LUNX and CK19 mRNA in the peripheral blood of the pa-tients with lung cancer were 59.5%,40.4%and 22.6% respectively. In the 32 peripheral blood samples of the patients with benign lung le-sions,the positive detection rate of BJ-TSA-9,LUNX and CK19 mRNA were 9.3%, 12.5% and 6.3% respectively. No expression of BJ-TSA-9,LUNX and CK19 was detected in lhe samples of the healthy volunteers. The expression level of BJ-TSA-9, WNX and CKI9 mRNA in the peripheral blood of lung cancer stage IV patients was significantly higher than those of stages II and ID (P< 0.05). Conclusion RT-PCR amplification of BJ-TSA-9, LUNX and CK19 mRNA are an efficient way to detect early haematogenous dissemination of cancer cells for lung cancer patients. The detection of BJ-TSA-9 expression is sensitive and specific for lung cancer,and it is superior to both LUNX and CK19 mRNA. The expression of BJ-7SA-9 mRNA in peripheral blood might predict the hematogenous metastatic spreading of lung cancer cells.

OBJECTIVETo study the relationship between pregnancy-induced hypertension (PIH) and catecholamine levels.

METHODSCatecholamines levels in maternal and fetal blood were determined in 116 patients with PIH and 40 normal control subjects using high performance liquid chromatography. The normal control subjects and PIH cases were selected from patients at term pregnancy receiving elective cesarean section.

RESULTSPlasma norepinephrine (NE) levels were significantly higher in patients with severe PIH than those in control subjects (P < 0.05). Both patients and control subjects had higher NE levels in the umbilical artery blood than in the umbilical vein blood (P < 0.05). NE levels in the umbilical artery blood were five times higher than those in the maternal blood.

CONCLUSIONThe pathogenesis of PIH may relate to catecholamine concentrations in fetus.

Catecholamines ; blood ; Female ; Fetus ; physiopathology ; Humans ; Hypertension ; blood ; Pregnancy ; Pregnancy Complications, Cardiovascular ; blood

Objective:Through determination of urinary arsenic metabolites in high water arsenic exposed areas of Jilin and Shanxi provinces, to explore the mode and possible influencing factors of arsenic metabolism in different populations.Methods:From October 2018 to August 2019, a cluster sampling was carried out in villages (arsenic in drinking water ≥0.05 mg/L) of some townships (towns) in Lyuliang City, Shanxi Province and Baicheng City, Jilin Province for epidemiological investigation and general health examination. The residents over 35 years old drinking water from local centralized water supply and small well water sources were selected as arsenic exposure group, and people (nearby low-arsenic water source areas) with the same diet and living habits and similar economic conditions were selected as control group. Urine samples were collected. Liquid chromatography-atomic fluorescence spectrometry(LC-AFS) technology was used to separate and detect 4 species of arsenic compounds, including trivalent inorganic arsenic (iAs Ⅲ), pentavalent inorganic arsenic (iAs Ⅴ), methylated arsine (MMA), and dimethylated arsine (DMA). Total arsenic (tAs), inorganic arsenic percentage (iAs%), MMA percentage (MMA%), DMA percentage (DMA%), primary methylation index (PMI) and the secondary methylation index (SMI) were calculated. The influencing factors of arsenic metabolism were analyzed by multiple linear regression. Results:A total of 1 415 villagers were investigated, including 1 256 in arsenic exposure group and 159 in control group. Compared with the control group, there were no significant differences in age, gender ratio and occupation distribution between arsenic exposure group and control group ( P > 0.05), but there were significant differences in smoking, drinking, body mass index (BMI) and education level distribution ( P < 0.05). The median of urinary tAs, iAs%, MMA%, DMA%, PMI and SMI in control group and arsenic exposure group were 12.86 μg/L, 15.03, 5.23, 76.35, 84.97, 93.68 and 69.68 μg/L, 10.24, 8.37, 79.31, 89.76, 90.65, respectively, the levels of urinary tAs, DMA% and PMI in arsenic exposed group were higher than those in control group, while iAs% and SMI were lower than those in control group, the differences were statistically significant ( U=- 13.87, - 4.30, - 6.64, - 6.64, - 1.99, P < 0.05). After analysis of the factors influencing urinary arsenic metabolism in the population, we found that age and BMI had an impact on iAs% ( β=- 0.08, - 0.08, P < 0.05); gender, drinking, BMI and education level were influencing factors of MMA% ( β =- 0.11, - 0.09, - 0.07, 0.08, P < 0.05); DMA% was mainly affected by age, gender, BMI and education level ( β = 0.06, 0.09, 0.10, - 0.09, P < 0.05); PMI was mainly affected by age and BMI ( β = 0.08, 0.08, P < 0.05); while SMI was affected by gender, drinking, BMI and education level ( β=0.09, 0.08, 0.08, - 0.09, P < 0.05). Conclusions:The urinary arsenic metabolism models of different arsenic exposed groups are different. Age, gender, smoking, drinking, BMI and education level may be influencing factors of different arsenic metabolism models.

Objective:To search for the key genes influencing the resistance of rectal cancer to chemoradiotherapy based on the weighted gene co-expression network analysis (WGCNA).Methods:The data were collected from gene expression omnibus. The whole genome expression data GSE119409 of patients receiving radiotherapy and chemotherapy were obtained by gene expression ominibus. The weighted gene co-expression networks of pathological complete response group and non-pathological complete response group were constructed respectively. NetRep conservative evaluation method was used to comprehensively analyze the three key network attributes of gene connectivity, gene significance and module membership of each node in the network module, and to determine the key genes closely related to the sensitivity of rectal cancer to radiotherapy and chemotherapy.Results:Network modules including black, blue, green, yellow and purple were obtained by WGCNA, and five key genes including SLC22A14, SIDT2, CABP4, EPHB6 and RAB11B were screened out.Conclusions:Five gene co-expression network modules and five key genes related to chemoradiotherapy resistance of rectal cancer were screened by weighted gene co-expression network analysis, which provided clues for finding molecular markers and potential therapeutic targets for neoadjuvant chemoradiotherapy resistance evaluation.

Objective TGF-β1 can promote EMT,then strengthen the invasion and metastasis ability of cancer ceils.However,the mechanism for TGF-β1 in gastric cancer still keeps unclear.Aim of this study was to investigate the expression of epithelial to mesenchymal transition (EMT)marker,LMO1 and metastasis related genes on the human gastric cancer cell cell line MKN28 treated with TGF-β1,and test whether down-regulate LMO1 expression can affect the pro-EMT and pro-metastatic roles of TGF-β1 in MKN28 cells.Methods Primary human gastric cancer cell line MKN28 was cultured in vitro.Cells were treated with TGF-β1 to induce cells to undergone EMT.Cells were divided into four groups:control group (5 ％ BSA),TGF-β1 induced group (10 μg/L),negative transfect group (TGF-β1 +negative transfect siRNA),and LMO1-siRNA transfect group (TGF-β1+ LMO1-siRNA).Real time-PCR and Western blot was used to examine the difference of EMT marker (E-cadherin and N-cadherin),LMO1 and metastasis related genes (MMP-9 and VEGF)expression.Transwell assays were performed to identify the differences and changes of invasive and metastatic ability in gastric cancer cell line MKN28.Western blot was used to examine the expression levels of MMP-9 and VEGF.Results TGF-β1 stimulation induced classical EMT morphological change,as was confirmed by E-cadherin decrease and N-cadherin,LMO1,MMP-9,VEGF increase (P＜0.01).Accompanied with the EMT,cell invasion and migration ability was markedly increased (P＜0.01).However,Down-regulation of LMO1 expression reversed the pro-migratory effect of TGF-β1 to a great degree (P＜0.01).Conclusion LMO1 played a central role in coordinating TGF-β1 induced EMT and pro-migratory effects in gastric cancer MKN28 cells.Using siRNA to downregulate the expression of LMO1 can inhibit the invasion and metastasis ability of gastric cancer MKN28 cells.