?Corneal collagen cross-linking ( CXL ) could increase the mechanical strength, biological stability and halt ectasia progression due to covalent bond formed by photochemical reaction between ultraviolet - A and emulsion of riboflavin between collagen fibers in corneal stroma. Corneal melting is an autoimmune related noninfectious corneal ulcer. The mechanism of corneal melting, major treatment, the basic fundamental of ultraviolet- A riboflavin induced CXL and the clinical researches status and experiment in CXL were summarized in the study.

Objective To study the effect of hypoxia on the mRNA expression of osteoprotegerin(OPG) and receptor activa‐tor for nuclear factor‐κB ligand(RANKL) in human periodontal ligament cells(hPDLCs) and to explore the role of hypoxia in the orthodontic bone resorption in pressure side .Methods The primary hPDLCs were cultivated with enzyme digestion assay and tis‐sue cultivation .The 3-5 generations of hPDLCs were respectively cultured 3 ,6 ,12 and 24 h in normoxia condition (20% O2 ) or in hypoxia condition (2% O2 ) .The mRNA expression of OPG and RANKL were detected with RT‐PCR .The experimental data was analyzed by one way ANOVA using SPSS15 .0 .Results After cultivated in hypoxia condition for 3 h or 6 h ,the mRNA expression of OPG and RANKL in hPDLCs didn′t change significant(P> 0 .05) .After cultivated in hypoxia condition for 12 h or 24 h ,the mRNA expression of OPG in hPDLCs decreased while the RANKL increased .Thus the ratio of RANKL/OPG increased and the difference was significant(P<0 .05) .Conclusion Hypoxia can regulate the mRNA expression of OPG and RANKL in hPDLCs and will promote the orthodontic bone resorption in pressure side .

Objective Benzoquinone ansamycin antibiotic, geldanamycin (GA), is a new anticancer agent that could inhibit Hsp90 by occupying its NH2-terminal ATP-binding site. This study was to investigate the antitumor efficacy of GA on Her2/neu tyrosine kinase overexpressing human breast cancer cell line SKBr3. Methods The degradation of Her2/neu tyrosine kinase was analyzed by Western blotting, the proliferation index was determined by MTT assay,cell cycle distribution was detected by flow cytometry, Cyclin D1 mRNA transcription was measured by RT-PCR and real-time PCR, and cell motility was evaluated by the cell culture insert model. Results GA induced a dose- and a time-dependent degradation of the Her2/neu tyrosine kinase protein and concurrently, the inhibition of cancer cell proliferation. The antitumor effects mediated by GA included: GA treatment decreased the survival rates of cancer cells,and led to a dase-dependent G1 arrest. Furthermore, this antitumor effect was proved to be related to declined transcription of Cyclin D1. Concurrently, the motility of cancer cells was reduced by GA. Conclusion GA treatment could induce the degradation of Her2/neu tyrnsine kinase efficiently, inhibit cancer cell proliferation and reduce motility in Her2/nen tyrosine kinase overexpressed human breast cancer cell line SKBr3.

Objective: To establish analytical method for the determination of vanadium ( V ), chromium ( Cr ), nickel ( Ni ), selenium ( Se ) and arsenic ( As ) in calcium based toothpastes by microwave digestion-inductively coupled plasma tandem mass spectrometry ( ICP-MS/MS ). Methods: The 21 calcium based toothpaste samples from supermarkets and shops in the urban areas of Hangzhou were digested by 6 mL HNO3 and 1 mL H2O2 in microwave digestion system. Then He-SQ mode and O2-MS/MS mode of ICP-MS/MS were respectively used for the determination of Ni and V, Cr, Se, As. Indium ( In ) was used as internal standard for calibration. Results: Good linear relationships were obtained for these five elements from 1.0 to 32.0 μg/L, with the correlation coefficients ranged from 0.999 3 to 1.000 0. The detection limits of the method ranged from 0.000 25 to 0.006 08 mg/kg. The recovery rates of standard spiking were 80.7%-105.7% when set at 0.4, 0.8 and 1.6 mg/kg, the recovery of standard reference material was 102.2%, and the relative standard deviations were 2.6%-4.8%. The concentrations of V, Cr, Ni, Se and As in 21 calcium based toothpaste samples were 0.024-1.935 mg/kg, 0.085-5.759 mg/kg, 0.090-3.673 mg/kg, <0.002 72-0.016 mg/kg and <0.006 08-0.321 mg/kg. Conclusion Microwave digestion-ICP-MS/MS can effectively reduce the interferences of polyatomic ions and doubly charged ions from the matrix, which is suitable for the determination of V, Cr, Ni, Se and As in calcium based toothpaste.

OBJECTIVETo verify the existence and significance of calcium/calmodulin dependent serine protein kinase/inhibitors of differentiation 1 (CASK/Id1) pathway in fibroblasts of human keloid.

METHODSImmunofluorescence laser was used to confirm CASK and Id1 protein expression and localization in fibroblasts of the keloid and normal skin. RT-PCR and Western-blot were adopted to analysis the CASK and Id1 expression and differences between keloid and normal skin fibroblasts. The natural combination of CASK and Id1 protein of keloid fibroblasts was tested by immunoprecipitation.

RESULTSCASK and Id1 protein expression were both found in fibroblast cells of keloid and normal skin under normal circumstances. Most of CASK and Id1 were distributed in the cytoplasm and nucleus of fibroblasts. The results of RT-PCR showed that the expression of CASK mRNA in the keloid group was 0.658 +/- 0.024, which was lower than that in the normal control group (1.076 +/- 0.008, t = 11.159, P < 0.05). The expression of Id1 mRNA was 0.497 +/- 0.014, which was higher than that in the normal control group (0.307 +/- 0.017, t = 15.148, P < 0.05). The results of Western-blot showed that the expression level for CASK protein in the keloid group was 0.057 +/- 0.006, which was lower than that in the normal control group (0.168 +/- 0.012, t = 13.524, P < 0.05); the expression of Id1 protein was 0.812 +/- 0.035, which was higher than that in the normal control group (0.368 +/- 0.031, t = 16.356, P < 0.05). The results of immunoprecipitation showed that Id1 could be detected in the CASK precipitate, while CASK also could be detected in the Id1 precipitate. There was a natural binding of CASK and Id1 in keloid fibroblasts.

CONCLUSIONCASK/Id1 signal pathway may be existed and involved in the proliferation of keloid fibroblasts, which is related with the occurrence of keloid.

Cell Proliferation ; genetics ; Cyclin-Dependent Kinase Inhibitor Proteins ; genetics ; metabolism ; Fibroblasts ; metabolism ; Humans ; Inhibitor of Differentiation Protein 1 ; genetics ; metabolism ; Keloid ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction

Site-directed mutagenesis method was used to introduce two desired mutations, which were confirmed by DNA sequencing, into mouse complement receptor Type II gene(MCR2). Then the constructed eukaryotic expression vectors containing wild type mouse CR2/1(wtMCR2/1), mutant type mouse CR2/1 (mtMCR2/1) and human CR2 (hCR2) cDNA were transferred into mouse SP2/0 cells by electroporation. After two-week screening by G418, the stably transfected clones were obtained. Several ways including PCR, RT-PCR, and immunohistochemistry were utilized to screen those clones with interesting genes integrated and expressed. Then Epstein-Barr virus(EBV) was used to infect these transfected cells and EBER-1 (EBV encoded RNAs) hybridization results showed that only hCR2 and mtMCR2 transfected SP2/0 cells could be infected by EBV, but positive rate of the former was much higher than the latter. This study sets groundwork for elucidating the mechanism by which EBV enters the cells and for establishing the animal model of EBV-related nasopharyngeal carcinoma (NPC).

Objective To explore the risk factors for clinical anastomotic leakage after resection of rectal cancer in China. Methods By meta-analysis we made a comprehensive analysis of the risk factors for clinical anastomotic leakage after resection of rectal cancer based on 19 articles published in China between January 1999 and January 2009. Results The anastomotic leakage rate was higher in the patients aged 60 years old and above than in those younger, with the combined odds ratio (OR) value being 0.50 (95% CI: 0.33-0.76) (P<0.01). The incidence rate was higher in the male patients than in the female ones, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate in the patients with the distance of tumor from the lower margin to anal verge being 7cm and shorter was higher than that with longer distance, with the combined OR value being 1.79 (95% CI: 1.37-2.35) (P<0.01). The incidence rate in the patients who had received radiotherapy preoperatively was higher than that in those who had not, with the combined OR value of 3.66 (95% CI: 2.19-6.09) (P<0.01). The incidence rate in the patients who had received stapler anastomosis was higher than that in the patients who had received manual anastomosis, with the combined OR value being 0.70 (95% CI: 0.47-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients with diabetes mellitus than in the healthy ones, with the combined OR value being 3.16 (95% CI: 2.27-4.39) (P<0.01). The incidence rate was lower in the patients with Dukes A and B stages than in those with Dukes C and D stages, with the combined OR value being 0.61 (95% CI: 0.45-0.83) (P<0.01). The incidence rate in the patients with high malignance degree in clinicopathological types was higher than that with low malignance degree, with the combined OR value being 2.17 (95% CI: 1.38-3.42) (P<0.01). The incidence rate was lower in the patients who had received preventive colostomy than in those who had not, with the combined OR value being 0.39 (95% CI: 0.14-1.05), but there was no significant difference between them (P>0.05). The incidence rate was higher in the patients who had got selective operation than in those who had got emergency operation, with the combined OR value being 0.27 (95% CI: 0.13-0.56). Conclusion The risk factors of anastomotic leakage after resection of rectal cancer are as follows: 60 years old and above, male patients, diabetes mellitus, preoperative neo-adjuvant radiotherapy, the distance of tumor from the lower margin to the anal verge being shorter than 7cm, Dukes C and D stages, high malignance degree in clinicopathological types, and emergency operation.

Objective To investigate ectopic expression of key elements of Wnt/?-catenin signaling pathway,including E-cadherin,?-catenin and Cyclin D1,as well as the relationship between ectopic expression of ?-catenin and overexpression of Her2 in human breast carcinoma tissues.Methods Immunohistochemical technique(Elivision) was used to detect the expression of Her2 and three elements of Wnt/?-catenin signaling pathway(?-catenin,E-cadherin,Cyclin D1) in 90 samples of human breast carcinoma.Results In E-cadherin normally-expressed group,the ectopic expression rate of ?-catenin(27.3%) was significantly lower than that(75.0%) in E-cadherin negative breast carcinoma;in ?-catenin ectopic expression group,overexpression rate of Cyclin D1(83.3%) was significantly higher than that(40.7%) in the other group.In Her2 overexpressed-breast carcinoma tissue,the ectopic expression rate of ?-catenin(73.5%) was significantly higher than that(19.6%) in Her2 negative breast carcinoma.Her2 overexpression could coexist with ectopic expression of Wnt/?-catenin in promoting axillary lymph node metastasis in human breast carcinoma.Conclusion Ectopic expression of Wnt/?-catenin signaling pathway does exist in some human breast carcinoma tissues,and the abnormal expression correlates with overexpression of Her2.Furthermore,the instantaneous activation of them could present synergism in promoting metastasis of axillary lymph nodes.

AIM: To establish nasopharyngeal carcinoma(NPC) cell lines stable expressing NPC-derived latent membrane protein 1(LMP1) gene. METHODS: General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed by using recombinant techniques, then transfected these vectors into a poor differentiated NPC cell line named CNE-2 ,integration and expression of N-LMP1 in CNE-2 cells were detected by PCR,RT-PCR and Western blot. RESULTS: (1) General expression vector and epithelium-specific expression vector of NPC-derived LMP1 gene were constructed successfully.(2) It showed that N-LMP1 gene expressed in CNE-2 cells correctly. CONCLUSION: The first NPC cell lines which stable express NPC-LMP1 were established. The cell lines obtained will provide important basis for exploring the role of NPC-LMP1 in nasopharynx carcinogenesis.

OBJECTIVETo explore the value of multidetector CT on aiding intraoperative judgement of serosal invasion of gastric cancer.

METHODSClinical data of 206 cases of gastric cancer undergoing radical surgery in the First Hosptial of China Medical University from August 2009 to June 2011 were analyzed retrospectively. Preoperative CT findings and intraoperative judgement of serosal invasion in gastric cancer were compared with pathological results in order to investigate their values.

RESULTSThe sensitivity and specificity of preoperative CT findings and intraoperative judgement of serosal invasion were 88.5%, 81.5% and 98.9%, 61.3%, respectively. The accuracy of preoperative CT in diagnosing serosal invasion was higher than that of intraoperative judgement, while the difference was not statistically significant (84.5% vs. 77.2%, P=0.060). The rates of serosal invasion of normal type, reactive type, nodular type, tendonoid type and color-diffused type were 0 (0/29), 2.5% (1/40), 40.5% (15/37), 59.2% (29/49), and 82.4% (42/51) respectively. The accuracy of preoperative CT in diagnosing serosal invasion of gastric cancer with tendonoid type was higher than that of intraoperative judgement, and the difference was statistically significant (61.2% vs. 87.8%, P=0.002).

CONCLUSIONSMultidetector CT plays an important role in aiding intraoperation judgement of serosal invasion of gastric cancer. For gastric cancer with tendonoid and color-diffused macroscopic serosal appearance, surgeons should pay attention to the value of preoperative CT findings.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Intraoperative Care ; Male ; Middle Aged ; Neoplasm Invasiveness ; diagnostic imaging ; Retrospective Studies ; Sensitivity and Specificity ; Serous Membrane ; diagnostic imaging ; pathology ; Stomach Neoplasms ; diagnostic imaging ; pathology ; Tomography, Spiral Computed ; methods