Colorectal cancer is one of the most common malignant tumors. The majority of the patients die of metastases and recurrences after surgery. Circulating tumor cells (CTC) exist in the patients’ peripheral blood, which can be considered as the crucial step in the metastasis cascade, thus being considered as the most valuable prognostic indicator. This paper summarized the clinical research works about CTC in colorectal cancer during recent years.

Colorectal cancer is one of the most common malignant tumors. The majority of the patients die of metastases and recurrences after surgery. Circulating tumor cells (CTC) exist in the patients' peripheral blood, which can be considered as the crucial step in the metastasis cascade, thus being considered as the most valuable prognostic indicator. This paper summarized the clinical research works about CTC in eolorectal cancer during recent years.

Objective To explore the association of arachidonic acid (AA) level in gastric cancer (GC) tissue with tumor differentiation and patients' gender.Methods The contents of AA in GC tissue and adjacent matched normal mucosa were measured using gas chromatography/mass spectrometry.The relationships of AA with GC differentiation and patients' gender were analyzed.Results The level of AA significantly decreased in GC tissue (0.190％ ± 0.255 ％) compared with normal tissue (0.274％ ± 0.254％,n =30,P =0.011 ),while the level of AA had no significant difference in the tissues of matched normal mucosa and different TNM stages or among different TNM stages ( all P ＞ 0.05).The AA levels in well and moderately differentiated adenocarcinoma (0.173％ ±0.244％ ) and in poorly differentiated adenocarcinoma (0.195％ ±0.264％) were significantly decreased when compared with those in the paired normal mucosa (0.334％ ± 0.170％,P =0.018; 0.256％ ± 0.275％,P =0.043,respectively),while no significant difference was observed between the different differentiated grades (P =0.895).The level of AA significantly decreased in male patients (0.137％ ± 0.209％ ) as compared with paired normal mucosa (0.275％:± 0.238％,P =0.010),while no positive correlation was observed in female patients as compared with normal group (P=0.477) or in the comparison between male and female groups (P =0.139).Conclusions The AA level remarkably decreases in GC tissue,which may be associated with differentiated grades and patients'gender.In addition,more AA is utilized in male GC patients than female patients.

Objective To investigate the association of promoter hypermethylation of secreted frizzled-related proteins (SFRPs) in patients with colorectal cancer. Methods The promoter hypermethylation of SFRPs in 20 sporadic colorectal cancer tissues and adjacent mucosa were detected by methylation-specific PCR. The amplified DNA was subcloned into the T-A cloning vector and sequenced. Two colorectal cancer cell lines (HCT116 and SW480) were treated with 5-aza-2' deoxycytidine for demethylation. The promoter hypermethylation and protein expression of SFRPs in colorectal cancer cell lines were detected by methylation-specific PCR and Western blotting. Results It was demonstrated that the hypermethylation of SFRP 1, 2, 4 or 5 was 19/20,17/20,3/20 or 13/20in cancer tissues, respectively, whereas it was 12/20, 8/12, 1/20 or 7/20 in adjacent mucosa,respectively. SFRP 1, 2 or 5 methylation was more frequently found in cancer tissue than in adjacent mucosa (P～0.05). Methylation of SFRP 1, 2, 4 and 5 were found in HCT116 cell line, but only SFRP1 and SFRP2 were found in SW480 cell line. There was a negative correlation between protein expression and methylation of SFRPs. The Western blotting revealed that SFRP protein re-expressedafter it treated with 5-aza-2' deoxyeytidine. Conclusion Methylation of SFRP 1, 2 or 5 gene is associated with the evolution of eolorectal cancer, and is closely related to silencing expression.

Objective To determine the therapeutic effect of recombined rat insulin-like growth factory 1 gene and transforming growth factor beta-1(TGF-?_1)gene on anterior cruciate ligament transection(ACLT)- induced osteoarthritis-like changes in NZW rabbit knee joints.Methods Eighteen NZW rabbits were divided into 3 groups randomly after osteoanhritis was established by ACLT and another six rabbits were used as normal control group(group 1).Chondrocytes which had been transfected with IGF-1 gene,co-transfected with TGF-?_1 and IGF-1 gene(group 3,4)were injected into the rabbits knee joints.Experimental control group(group 2)only had ACLT bul was not transfected.After 4,8 weeks,rabbits were sacrificed and their joints were evaluated by morphological grades,histological examination,in situ hybridization examination,immunohistochemistry exami- nation,and transmission electron microscopy examination(TEM).Results The morphological grades showed that the normal control group had a very significant difference with the experimental control group(P

AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.

Objective To evaluate the effect of patients' compliance on clinical parameters in patients with chronic periodontitis during periodontal maintenance therapy period.Methods Chronic periodontitis patients who had completed non-surgical periodontal basic treatment were incorporated into the periodontal maintenance therapy(PMT).Clinical examination was performed in the baseline and each quarterly recall,over 3-year period.Clinical parameters including number of teeth,probing depth (PD),attachment loss(AL) level and bleeding on probing(BOP),were recorded.According to the number of PMT visit,the patients were classified into three groups:regular complier (RC) ; erratic complier (EC) ;non-complier(NC).The final parameters (three years later) were obtained by outpatient follow-up and telephone interviews.The data were calculated for the pecentage of sites with PD 4-5 mm,PD ≥ 6 mm,AL 4-5 mm,AL≥ 6 mm,BOP,and the number of tooth loss per patient and the rates of progression of periodontitis.Statistical analyses including ANOVA test and Chi-square test were performed by SPSS 16.0 software package.Results The percentage of clinical parameters in RC [AL 4-5 mm:(14.8 ± 5.0) ％,AL≥6 mm:(9.3 ± 3.1)％,BOP:(22.8 ± 4.2)％] were significantly decreased compared with that at baseline[AL4-5 mm:(19.0 ±6.0)％,AL≥6 mm:(10.6±3.1)％,BOP:(30.3 ±5.6)％] (P＜ 0.01).There was significant difference between RC and NC [AL 4-5mm:(43.3 ± 1.3) ％,AL ≥ 6 mm:(31.3±1.7)％,BOP(91.5±5.4)％](P＜0.01),and between RC and EC[AL4-5 mm:(18.9± 6.7)％,AL≥6 mm:(12.6±5.4)％,BOP:(38.4± 5.2)％] (P＜0.05).The progression rate of periodontitis [19.1％ (4/21) at subject level,0.7％ (434/61 362) at site level] and tooth loss(1.0) was significantly lower in RC compared with EC and NC patients.Conclusions Regular periodontal maintenance enables the patients with chronic periodontitis to maintain long-term efficacy.

OBJECTIVETo explore the relationship between CpG island methylator phenotype(CIMP) and genetic instability in sporadic colorectal cancer(SCRC).

METHODSSeventy-one SCRC patients were enrolled in this study. Promotor methylation status of five genes including P14(ARF ), hMLH1, P16(INK4a), MGMT and MINT1 was detected with methylation specific PCR to confirm CIMP. Microsatellite instability (MSI) status was evaluated with two microsatellite loci of BAT25 and BAT26, and the ploidy was detected with flow cytometry. The association between CIMP and MSI as well as chromosomal instability(CIN) was examined.

RESULTSThe positive rates of CIMP, MSI and aneuploidy were 21.1% (15/71), 9.9% (7/71) and 73.5% (50/68) respectively. The positive rate of MSI in positive CIMP patients was higher than that in negative CIMP ones, but the difference was not significant (20.0% vs 7.1%,P=0.158). The positive rate of MSI was 57.1% in patients with hMLH1 gene promotor hypermethylation, which was significantly higher than that (4.7%) in patients without hMLH1 gene promotor hypermethylation (P=0.001). SCRCs with positive CIMP displayed significant inclination of diploidy (P=0.003). The positive rate of diploidy among SCRCs with CIMP was 61.5% while only 18.2% of cases without CIMP demonstrated diploid.

CONCLUSIONSSCRCs with positive CIMP are significantly more likely to be diploid. Simultaneous multiple genes hypermethylation represented by CIMP may be an epigenetic mechanism competing with the genetic mechanism of CIN.

BACKGROUND: At present, finite element analysis can be used to judge intertrochanteric fractures, but mostly limited in the distribution of stress. Finite element model of various intertrochanteric fractures has not been reported in detail.OBJECTIVE: To build various types of intertrochanteric fracture models with Hypermesh 14.0 and LS-DYNA software to simulate the falling-induced external force on proximal femur, and to evaluate the effect of models, and to analyze the biomechanical mechanism of intertrochanteric fractures. METHODS: Normal side CT image data of one case of elderly intertrochanteric fracture were collected and imported into Mimics software to establish the proximal femur geometric models, were then analyzed and operated by LZ-DYNA solver after imported into Geomagic studio 2013 and Hypermesh 14.0 for smoothing and meshing. Before analysis, the material parameters were set, the boundary conditions were confirmed, and given the loading parameters. The operating results were checked in Hyper View. RESULTS AND CONCLUSION: (1) The distribution of stress of proximal femur exactly matched to the previous study. EvansⅠtype intertrochanteric fracture model was obtained under continuous shear stresses, and six types of fractures were obtained by adjusting the load. (2) These results manifest that based on the Hypermesh 14.0 and LS-DYNA software, the finite element can well simulate the intertrochanteric fractures, and shear stress plays an important role in intertrochanteric fractures, which can provide experimental basis for the prevention and treatment of intertrochanteric fractures.

This study examined the effect of Notch-1 signaling on malignant behaviors of breast cancer cells by regulating breast cancer stem cells (BCSCs). BCSCs were enriched by using serum-free medium and knocked out of Notch-1 by using a lentiviral vector. Real-time polymerase chain reaction (RT-PCR) and Western blotting were used to detect the Notch-1 expression levels in breast cancer cell lines and BCSCs, and flow cytometry to detect the proportion of BCSCs in BCSC spheres. The BCSC self-renewal, migration, invasion, and tumorigenicity were examined by the tumor microsphere-forming assay and transwell assay and after xenotransplantation. The results showed that the Notch-1 silencing reduced the number of BCSC spheres, the proportion of BCSCs, and the number of cells penetrating through the transwell membrane. It also decreased the size of tumors that were implanted in the nude mice. These results suggest that Notch-1 signaling is intimately linked to the behaviors of BCSCs. Blocking Notch-1 signaling can inhibit the malignant behaviors of BCSCs, which may provide a promising therapeutical approach for breast cancer.