Colorectal cancer (CRC) is one of the most common gastrointestinal malignancies with poor prognosis and high mortality.SEPT9 gene is a tumor suppressor gene and plays an important role in the end of cell division.Studies have shown that methylation of SEPT9 gene could be used in the early diagnosis of CRC.This article reviewed the advances in study on SEPT9 gene methylation in the screening and diagnosis of CRC.

BACKGROUND: Studies have indicated that the abnormal expression of TACC3 is closely related to the occurrence and development of many kinds of tumors, and the expression of TACC3 is up-regulated in these tumors. Therefore, in vitro specific inhibition of TACC3 expression may become an important target for the treatment or intervention of tumor growth.OBJECTIVE: To investigate the mechanism by which TACC3 gene expression regulates cell proliferation and apoptosis in oral squamous cell carcinoma.METHODS: CD133+CD44+ oral squamous cell carcinoma cells were sorted from human oral squamous cell carcinoma cell line Cal-27 by immunomagnetic beads. In experimental group, the shRNA sequence of TACC3 was designed and synthesized, which was then trasnfected into CD133+CD44+ oral cancer stem cells by LipofectamineTM 2000. Empty vector-trasnfected (negative control) and untransfected cells were used as callsed. Forty-eight hours after the transfection, effects of TACC3 gene silencing on proliferation and apoptosis in vitro in CD133+CD44+ oral squamous cell carcinoma were detected by MTT, clone formation test, and TUNEL assay. Western blot assay was used to detect the effect of TACC3 gene silencing on Ki67, Bax and Bcl-2 protein expression in CD133+CD44+ oral squamous cell carcinoma.RESULTS AND CONCLUSION: (1) Cell proliferation. The proliferation rate and expression level of Ki67 were significantly lower in the experimental group than the negative control and untransfected groups (P < 0.05). (2) Clone formation. The clone formation ability in the experimental group was significantly lower than that in the negative control and untransfected groups (P < 0.05). (3) Cell apoptosis. TACC3 gene silencing caused an obvious decrease in Bcl-2 protein expression and a significant increase in Bax protein expression. These findings further confirmed that specific interference of TACC3 gene expression could inhibit the proliferation of CD133+CD44+ cells and promote the apoptosis.

OBJECTIVEThis study aimed to explore further the mechanisms of tongue squamous cell carcinoma (TSCC) cell recurrence, metastasis, and diffusion, as well as to establish the experimental basis for the molecular biology research on TSSC. We intend to complete our objective through differential proteomics and preliminary analysis protein expression of exosomes derived from TSCC and normal mucosa cells.

METHODSWe acquired cultured supernatant fluid in vitro in the laboratory by culturing TSCC (tongue cancer Tca8113 cell line) and human normal mucosa cells (HOK cell line). The exosomes were separated and purified through differential centrifugation. Furthermore, the different protein expressions were identified through dielectrophoresis and mass spectrometry. The functions of the different protein expressions were identified through an online database search.

RESULTSTSCC and human normal mucosa cells secrete a large amount of capsule bubble structure substances in vitro, as confirmed by electron microscopy and surface markers heat shock protein-70 and major histocompatibility complex class 1. A total of 16 oral cancer cell-derived exosomes that expressed quantity more than two times, twelve that increased their expression levels, and four that cut their expressions were identified through the differential proteomics research on the two groups.

CONCLUSIONDifferential proteins that were verified through the online database serve an important function in exosome formation and in the progress of cancer.

RNF180 is a novel membrane-bound E3 ubiquitin ligase that participates in cell development,proliferation and apoptosis.It is a tumor suppressor gene that inhibits cell proliferation and induces apoptosis and may inhibit gastric cancer cell lymph node metastasis.The study found that RNF180 gene methylation and gastric cancer is closely related to the occurrence and development.Therefore,RNF180 gene methylation is expected as a tumor marker of gastric cancer for early diagnosis and prognosis of gastric cancer.In this paper,RNF180 on the diagnosis of gastric cancer research progress made a review.

Objective To investigate the underlying mechanism of bone marrow mesenchymal stem cells (BMSC) modulating the inflammatory response during the multiple organ dysfunction syndrome (MODS), especially the expression of inflammatory cytokines, which will provide new theoretical and experimental basis of MODS in clinic. Methods BMSC of Sprague-Dawley (SD) rat (female, 4 weeks) was extracted and cultivated, and the 4th passage were used in experimental study. According to the random number table, 60 female SD rats were divided into three groups (n = 20 per group): sham group, MODS group, BMSC group. MODS model in rats was induced by lipopolysaccaride (LPS, 1 mg/kg) via femoral vein injection. Sham group was injected with the sterile phosphate buffer saline (PBS) in the same volume. BMSC group, in which BMSC infusion was started at 2 hours after 0.5 mL LPS stimulation (1×106/cells) through the tail vein. The survival rate was observed after 72 hours in each group. Abdominal aortic blood was collected for routine blood and biochemical examination at 72 hours after operation. Protein microarray was used to detect the related 34 inflammatory cytokines. Signal ratio was defined as the differentially expressed factors when it was more than 2.0 or less than 0.5. And enzyme linked immunosorbent assay (ELISA) was be applied to validate the significant inflammation factor. Meanwhile, the heart, kidney, intestine tissue was harvested, then their pathological changes were observed by hematoxylin eosin (HE) staining.Results 20, 12, 16 rats lived in sham group, MODS group and BMSC group respectively at 72 hours after operation. Compared with the sham group, the indicators (routine blood, liver and kidney function, myocardial enzyme) were apparently unusual, and the heart, kidney, intestine tissue were injured obviously in the MODS group. After BMSC administration, the organ function was improved and tissue damaged was alleviated significantly. Protein microarray showed that interleukin-4 (IL-4) and receptor for advanced glycation end products (RAGE) were significantly different in 34 goal cytokines. The signal ratio change of IL-4 was 0.397, 1.124, 2.826 respectively, and the signal ratio of RAGE was 6.197, 1.552, 0.250, respectively in MODS/sham group, BMSC/sham group, BMSC/MODS group. ELISA validated the result that the expression level of IL-4 decreased significantly (ng/L:3.59±1.21 vs. 29.10±5.78) and the expression level of RAGE increased significantly (ng/L: 1.09±0.04 vs. 0.11±0.03) in MODS group as compared with sham group (bothP < 0.05). Compared with the MODS group, the level of IL-4 was obviously higher than that in BMSC group (ng/L: 9.59±2.21 vs. 3.59±1.21,P < 0.01), and RAGE decreased significantly (ng/L: 0.29±0.07 vs. 1.09±0.04,P < 0.05).Conclusions BMSC administration can regulate the expression of IL-4 and RAGE in the rats subjected to MODS. Moreover, BMSC can promote the restoration of tissue and organ function, thus improve the survival rate. BMSC may be the target in cell therapy for the inflammatory disease.

OBJECTIVETo explore the effects of stress imposed on adenoid cystic carcinoma (ACC), therefore to clarify the molecular basis and mechanism of ACC's malignant phenotype under the elevated tumor interstitial fluid pressure.

METHODSACC cells were cultured under pressure (103.74 kPa), and were divided into four groups (3 h group, 6 h group, 12 h group, 24 h group) according the pressure time. Untreated ACC2 was as negative control group, untreated ACCM was as positive control group. The level of epidermal growth factor receptor (EGFR) was detected by semiquantitative analysis of immunochemistry. Matrix metalloproteinase 9 (MMP9) and EGFR mRNA expression were assessed by reverse transcriptase polymerase chain reaction. EGFR, phosphorylation epidermal growth factor receptor (P-EGFR), MMP9, keratinocyte growth factor (KGF) and phosphorylation extracellular signal-regulated kinase (P-ERK) protein expressions were assessed by Western blot.

RESULTSAs the extension of pressure time, the expression of EGFR, P-EGFR, MMP9, KGF, P-ERK in ACC2 gradually increased, which were positive correlation with pressure time, and were higher than that of negative control group.

CONCLUSIONUnder the stimulation of pressure, the mRNA and protein levels of adhesion molecules and metastatic relative molecules in ACC2 were sharply elevated.

Objective To report the prenatal molecular diagnosis for two gravida in a family with spondylepiphyseal dysplasia congenita(SEDC)caused by G504S mutation of COL2A1 gene.Methods DNA of the two fetuses was extracted from amniotic fluid at the 19+3 and 18+6 weeks of gestation respectively.Direct sequencing of two samples were performed after amplifying exon 23 of COL2A1 containing the potential mutation.The femur length and biparietal diameter of the first fetus were measured by sonographic scans every two weeks from 17+3 weeks to 27+3 weeks of gestation,and for the second fetus these parameters were measured from 16+1 to 19+1 weeks of gestation.Results Sequncing analysis revealed the first fetus and his mother presented the same mutation which is specifically associated with SEDC,but the second fetus did not show the mutation of COL2A1 gene.Biparietal diameters of the both fetuses were appropriate for gestational age.Femur length of the second fetus was normal for gestational age but that of the first fetus was shortened evidently after the 23 week of gestation.The parents of the first fetus determined to terminate the pregnancy.A medical termination was carried out at 27+5 weeks of gestation and a male fetus with a relatively large head and short limbs was delivered.The radiological findings of the fetus were consistent with SEDC including generalized platy spondesand shortened long bones.Conclusions Prenatal molecular diagnosis is important for the fetus with risk of SEDC and useful for genetic counseling.Genotype of fetus with risk of SEDC can be identified before sonographic scan.Molecular genetic analysis in conjunction with sonographic monitoring was helpful in prenatal diagnosis of SEDC.

Objective To report a familial dentinogenesis imperfecta type Ⅱ (DGI type Ⅱ) with a novel splicing mutation in DSPP (dentin sialophosphoprotein) gene.Methods Based on the result of linkage analysis performed previously to map the candidate gene DSPP in the family, the promoter,the first four exons and exon-intron boundaries of DSPP were directly sequenced for the members of the DGI type Ⅱ family. Denaturing high performance liquid chromatography (DHPLC) analysis was performed to confirm the results of sequencing.Results A novel splicing mutation of 23 bp deletion in intron 2 of DSPP gene was identified by DNA sequence analysis. The mutation changed acceptor site sequence from CAG to AAG, and might result in functional abolition of possible branch point site in intron 2. DHPLC result was consistent with that of sequencing. The mutation may be identified in all affected individuals, but not found in normal members of the family and 50 controls.Conclusion These results suggest the deleted mutation of DSPP gene causes DGI type Ⅱ in the family. The mutation has not been reported before.

ObjectiveTo investigate the effect of kaempferol on the proliferation, migration, invasion, and apoptosis of human hepatoma Bel-7402 cells and related molecular mechanism. MethodsHepatoma Bel-7402 cells cultured in vitro were randomly divided into control group and low-, middle-, and high-concentration experimental groups. The experimental groups were treated with low-, middle-, and high-concentration kaempferol (25, 50, and 100 μmol/L), and the control group was treated with an equal volume of dimethyl sulfoxide. CCK-8 assay was used to observe the effect of kaempferol on the viability of Bel-7402 cells; plate colony formation assay was used to evaluate the effect of kaempferol on cell colony formation ability; wound healing assay and Transwell chamber were used to observe the effect of kaempferol on cell migration and invasion; Western blot was used to measure the expression of apoptosis- and cycle-related proteins. A one-way analysis of variance was used for comparison between multiple groups, and the least significant difference t-test was used for further comparison between two groups. ResultsAfter 24 hours of treatment, the cell viability was 100.00%±2.72% in the control group and 75.70%±2.42%, 62.79%±2.45%, and 43.41%±2.11%, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell viability (all P＜0.05). The number of cell colonies was 923.3±35.2 in the control group and 682.7±24.4, 464.0±22.0, and 327.3±14.0, respectively, in the low-, middle-, and high-concentration experimental groups, and compared with the control group, the experimental groups had a significant reduction in cell colony formation ability (all P＜0.05). After 24 hours of treatment, the relative migration rate was 100.00%±1.11% in the control group and 63.33%±1.16%, 51.72%±3.23%, and 37.18%±2.71%, respectively, in the low-, middle-, and high-concentration experimental groups, and the number of transmembrane cells was 212.0±3.0 in the control group and 134.0±2.0, 71.0±2.0, and 34.0±1.0, respectively, in the low-, middle-, and high-concentration experimental groups; compared with the control group, the experimental groups had significant reductions in relative migration rate and number of transmembrane cells (all P＜0.05). After 48 hours of treatment, compared with the control group, the low-, middle-, and high-concentration experimental groups had a significant reduction in the expression of the anti-apoptotic protein Bcl-2 (all P＜0.05), a significant increase in the expression of the pro-apoptotic protein Bax (all P＜0.05), and a significant reduction in the expression of C＜italic/＞yclinD1 (all P＜005). ConclusionKaempferol can inhibit the proliferation, migration, and invasion of human hepatoma Bel-7402 cells and promote the apoptosis of such cells, possibly by regulating the apoptosis proteins Bax and Bcl-2 and downregulating the expression of CyclinD1.

Objective To investigate the clinical value of combined detection of serum carcinoembryonic an-tigen(CEA) ,carbohydrate antigen-125 (CA125) ,cytokeratin 19 fragment antigen (CYFRA21-1) ,squamous cell carcinoma(SCC) antigen ,neuron-specific enolase(NSE) and plasma progastrin-releasing peptid(ProGRP) in the diagnosis ,pathological typing and clinical staging in lung cancer .Methods The serum CEA ,CA125 , CYFRA21-1 ,SCC ,NSE levels and plasma ProGRP level were detected by adopting the chemiluminescent mi-croparticle immunoassay method in 378 cases of lung cancer ,200 cases of benign lung diseases and 200 people undergoing healthy physical examination .Results The levels and positive rates of CEA ,CA125 ,CYFRA21-1 , SCC ,NSE and ProGRP in the patients with lung cancer were significantly higher than those in the patients with benign lung diseases and healthy control group ,the difference was statistically significant (P<0 .05);in the single index detection ,the biomarkers of highest positive rate in adenocarcinoma ,squamous cell carcinoma and small cell lung carcinoma were CEA ,CYFRA21-1 and SCC ,NSE and ProGRP respectively .The sensitivi-ty ,specificity ,accuracy ,negative predictive value and positive predictive value of combined detection of these 6 indexes were 92 .86％ ,85 .00％ ,88 .17％ ,92 .64％ and 85 .40％ respectively ,except the specificity and positive predictive value were slightly decreased ,the levels of other indexes were higher than those of single index de-tection .The aresa under the receiver operating characteristic (ROC) curves of CEA ,CA125 ,CYFRA21-1 ,SCC ,NSE and ProGRP were 0 .775 ,0 .778 ,0 .891 ,0 .585 ,0 .710 and 0 .620 respectively .The area under ROC curves of the combined detection was 0 .950 .The positive rates of CA125 ,CYFRA21-1 ,NSE and the combined detection in the patients with stage Ⅲ ,Ⅳof lung cancer were obviously higher those in the patients with stageⅠand Ⅱof lung cancer ,the difference was statistically significant (P<0 .05);the combined detection obviously improved the positive rates for the diagnosis in the patients with different stages of lung cancer .Conclusion The combined detection of CEA ,CA125 ,CYFRA21-1 ,SCC ,NSE and ProGRP is conducive to improve the di-agnosis performance and early detection rate for lung cancer ,differentially diagnosing different pathological types of lung cancer and judge the clinical stage of lung cancer .The combined detection of 6 tumor biomarkers is an ideal diagnosis index of lung cancer .