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 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

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.

OBJECTIVETo evaluate the effect of patients' compliance on clinical parameters in patients with chronic periodontitis during periodontal maintenance therapy period.

METHODSChronic 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.

RESULTSThe 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 [AL 4-5 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[AL 4-5 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.

CONCLUSIONSRegular periodontal maintenance enables the patients with chronic periodontitis to maintain long-term efficacy.

Chronic Periodontitis ; prevention & control ; therapy ; Dental Plaque Index ; Disease Progression ; Female ; Follow-Up Studies ; Humans ; Longitudinal Studies ; Male ; Middle Aged ; Patient Compliance ; Periodontal Attachment Loss ; prevention & control ; Periodontal Index ; Recurrence

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.

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.

Chromosomal Instability ; Colorectal Neoplasms ; genetics ; CpG Islands ; DNA Methylation ; Genome, Human ; Humans ; Microsatellite Instability ; Phenotype

OBJECTIVETo investigate the relationship between the extracellular matrix (ECM) and neoplastic progression in hamster with tongue cancer.

METHODSForty-eight specimens of hamster tongue cancer were divided into control group (n=6) and experimental group (n=42). The pathological grade of the specimens was assessed (including 3 stages, namely atypical hyperplasia, carcinoma in situ and early invasive carcinoma). The sections of the tongue were stained with Masson and aldehyde-fuchsin (AF) staining for microscopic observation of the elastic fiber and collagen fiber changes.

RESULTSWithin the connective tissue cores (CTC) of the papillae in the control group was a framework of numerous and fine Gomrori's aldehyde fuchsin-positive elastic fibers. But in the stages of dysplasia and carcinoma in situ, these elastic fibers decreased and further diminished in the CTC in early invasive carcinoma. In dysplasia and carcinoma in situ stages, most of the elastic fibers collapsed with scattered elastic fibers, and the elastic fibers decreased significantly in early invasive carcinoma. The control group showed a significantly greater number of elastic fibers in the experimental group. The collagen fiber was obviously increased and irregularly arranged in dysplasia and carcinoma in situ stage; in early invasive carcinoma, the collagen fibers became thicker with deposition in the lamina propria.

CONCLUSIONAn excessive deposition of collagen fiber and reduction of the elastic fibers is an important factor contributing to the development of tongue carcinoma in hamsters.

Animals ; Carcinoma ; pathology ; Collagen ; metabolism ; Connective Tissue ; pathology ; Cricetinae ; Elastic Tissue ; pathology ; Extracellular Matrix ; pathology ; Neoplasms, Experimental ; pathology ; Tongue Neoplasms ; pathology

OBJECTIVEIn order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes.

METHODSThe classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively.

RESULTS(1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a.

CONCLUSIONThis research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.

5' Untranslated Regions ; chemistry ; DNA Restriction Enzymes ; Genotype ; Hepacivirus ; classification ; genetics ; RNA, Viral ; analysis