Objective To explore the effect of chitosan on vascular smooth muscle cells inhibited proliferation from rabbit arteriovenous fistula and its mechanisms.Methods Established rabbit fistula model on carotid arteryinternal jugular vein.After 1 month cultured VSMCs with primary culture by tissue-pieces inoculation.Cultured VSMCs were divided into three groups:①normal control group.②FBS-treated group:cell were treated with 5％,10％,20％ for 48 h,respectively; established the model of rabbit VSMCs proliferation.③chitosan-treated group:VSMCs cultured with 20％ FBS were exposed to different doses of chitosan(10,100,500,1000,2000μg/ml) for 48 h.And VSMCs were treated for different time (0,12,24,48 h) with Chitosan 1000 μg/ml.Expression levels of PCNA and TLR4/ NF-κB were detected by Western blotting.RT-PCR were applied to measure the mRNA expression of PCNA and TLR4.The protein levels of TLR4 and NF-κB were detected by immunofluorescence.Results Compared with low concentration serum group,FBS-treated VSMCs exhibited a increase in mRNA and protein expression of PCNA and TLR4.FBS-induced protein expression of PCNA and TLR4/NF-κB were reduced by chitosan.Also mRNA expression of PCNA and TLR4 were reduced.They were dependent on concentration and time.In rabbit VSMCs TLR4 was mainly expressed in the cytoplasm and NF-κB expressed mainly in the nucleus.Compared with normal control group,TLR4 and NF-κB protein expression were significantly decreased by chitosan.Conclusion High concentration serum induced VSMCs proliferation.Chitosan can inhibit the proliferation of rabbit VSMCs.It is speculated that the mechanism may be related to the expression of TLR4 receptor activation,reducing expression of downstream factor MyD88 and NF-κB.It is suggest that chitosan can become potential new drugs of arteriovenous fistula prevention of intimal hyperplasia.

Objective To investigate the relationship between KRAS,NRAS and BRAF gene mutations and clinicopathological parameters in patients with colorectal carcinoma (CRC).Methods By using TagMan real-time PCR method KRAS/NRAS/BRAF hotspot mutations were detected in 260 cases of CRC.The associations between KRAS/NRAS/BRAF mutation status and clinical pathological characteristics were analysed in different groups divided by gender,age,tumor size,tumor differentiation.Results (1) The KRAS hotspot mutations were G12D,G12A,G12R,G12C,G12V,G12S in codon 12 and G13 C,G13D in codon 13.They were identified in 43.1％ CRC.KRAS mutation rate was higher in females than in males (P =0.05) and the mutation rate in patients ≥ 60 years was significantly higher than that in patients ＜ 60 years(P =0.008).The incidence of metastasis and mortality were higher in KRAS mutant than in KRAS wild type (P =0.004,P =0.037).(2)The NRAS hotspot mutations were in codon1 2,13 and 61.They were identified in 4.6％ CRC.NRAS mutation rate was significantly higher in patients ≥ 60 years and well-differentiated tumors (P =0.032,P =0.042).(3) The mutation rate of BRAF V600E in CRC patients was 4.6％.BRAF V600E mutation rate was significantly higher in patients ≥60 years,with distant metastases and tumors ＞ 5 cm (P =0.032,P =0.026,P =0.038).The incidence of metastasis and rucurrence and mortality were higher in BRAF mutant (P =0.030,P =0.002,P =0.007).Conclusions In CRC patients,KRAS mutations correlate with demographic factors,metastasis and mortality,NRAS mutations correlate with age and tumor differentiation,while BRAF mutation correlate with age,tumor size,metastasis,recurrence and mortality.

OBJECTIVETo construct and characterize the TGF-β1, induced epithelial-mesenchymal transition (EMT) model of keloid epithelial cells in vitro, and to investigate the expression of epithelial stem cells related surface markers in keloid epithelial cells during EMT induction.

METHODSThe epithelial cells from 3 keloid samples of ears were cultured in vitro and induced by transforming growth factor betal (TGF-β1, 1 ng/ml) for 5 days, which was the experimental group, the same cells untreated were considered as the negative control group. The expressions of EMT-associated markers and regulative genes were detected using immunofluorescence staining, real-time PCR and western blot analysis. Then the surface markers of epithelial stem cells were detected using real-time PCR. Statistical significance was determined using Independent-Samples t Test, a p value less than 0. 05 was considered statistically significant.

RESULTSThe mRNA expression of transcription factor snail2 and mesenchymal-specific marker vimentin increased significantly in TGF-β1, induced keloid epithelial cells (P < 0. 05), in which snail2 increasing from 0. 91 ± 0. 23 to 1. 69 ± 0. 10, and vimentin from 5. 86 ± 2. 07 to 24. 29 ± 5. 39. Whereas the mRNA expression of epithelial-specific marker E-cadherin decreased from 1. 06 ± 0. 19 to 0. 65 ± 0. 09. The mRNA expression of CD29 and Lgr6, two surface markers of epithelial stem cells, significantly increased after induction of the TGF-β1, (P < 0. 05), from 0. 55 ± 0. 14 and 1. 61 ± 0. 31 to 1. 19 ± 0. 12 and 3. 84 t 0. 62 respectively. In induced cells, the immunofluorescence results showed staining of E- cadherin became faint, but the number of positive staining cells of vimentin increased. Western blot confirmed the protein expression of E-cadherin weakened, and the vimentin and p-Smad3 enhanced (P < 0. 05).

CONCLUSIONSTGF-β1, initiated EMT in keloid epithelial cells by inducing the up-regulation of snail2, and TGF-β1,/Smad3 signaling pathway was involved in EMT. EMT could change the phenotype of epithelial stem cells in keloid.

Biomarkers ; metabolism ; Cadherins ; genetics ; metabolism ; Epithelial Cells ; drug effects ; physiology ; Epithelial-Mesenchymal Transition ; drug effects ; physiology ; Humans ; In Vitro Techniques ; Keloid ; pathology ; RNA, Messenger ; metabolism ; Signal Transduction ; Smad3 Protein ; genetics ; metabolism ; Snail Family Transcription Factors ; Transcription Factors ; genetics ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; pharmacology ; Up-Regulation ; Vimentin ; genetics ; metabolism

Objective To observe the outcome of free TAAP flap in the reconstruction of defect after oral tumor radical resection.Methods From June,2010 to April,2015,12 patients with oral tumor underwent radical resection,including 4 cases of gingival carcinoma,3 cases of tongue carcinoma and 5 case of buccal cancer.The cause ranged from 2 to 12 months.The radical resection left defects with size ranged from 4.5 cm×3.5 cm to 6.0 cm× 4.5 cm which were reconstructed by free TAAP flaps with size ranged from 7.0 cm×4.0 cm to 12.0 cm×8.0 cm.The length of TAAP flap was (6.5±0.6) cm.The width of flap was (4.3±0.5) cm.And the thickness of flap was (1.1±0.3)cm.The length of pedicle was(8.4±0.2) cm.All the arteries of TAAP were anastomosed with superior thyroid arteries,while the venae comitans were anastomosed with superior thyroid venae or internal jugular venae.Results The perforators existed consistently.All 12 flaps survived uneventfully.The donor sites were closed directly in all cases.All patients were followed up for 14-38 months with satisfied esthetic and functional results in reconstructed tongue.No local recurrence happened.The shoulder function was not affected.Conclusion The TAAP flap has consistent blood supply,good color match and texture,while leaving minimal morbidity at donor site,is an ideal choice for buccal tumor reconstruction.

Objective: To investigate the application of free medial thigh chimeric myocutaneous perforator flap in the reconstruction of tongue defect after radical resection of tongue carcinoma. Methods: From June 2013 to November 2015, 17 cases with tongue carcinoma underwent radical resection and tongue and mouth floor defects after surgery were reconstructed with medial thigh chimeric myocutaneous perforator flaps at the same stage. These 17 cases included tongue carcinoma on lingual margin (n=9), ventral tongue (n=7) and mouth floor (n=1). Appearance and functional results were evaluated with follow-up. Results: Medial thigh chimeric myocutaneous perforator flap was (9.0±0.3) cm in length, (4.5±0.3) cm in width , and (1.8±0.4) cm in thickness. The length of pedicle was (7.4±0.6) cm. All of the 17 flaps survived uneventfully, and the donor sites were closed directly in all cases. All patients were followed up for 10-28 months with satisfied esthetic and functional results in reconstructed tongue. Aside from linear scar in the donor sites, the functions of thighs were not affected. No local recurrence occurred. Conclusion The free medial thigh chimeric myocutaneous perforator flap is an ideal choice to reconstruct the tongue and mouth floor defect after radical resection of tongue carcinoma.

Objective To design an air-cushioned belt for skin traction of lower extremities to reduce the incidence of deep vein thrombosis and pressure ulcers in patients with lower limb fractures. Methods The belt was composed of upper and lower leggings.The legging had an air cushion at its interior surface,and the air cushion consisted of several chambers.There was a connecting tube between every two chambers, and each tube was equipped with a pressure non-return valve. The chamber on the top of the cushion had a charging mechanism for inflation.Totally 100 patients from June to December 2016 were selected and divided equally into an experiment group and a control group.The experiment group used the developed air-cushioned belt,and the control group applied the traditional one.The incidence rates of deep venous thrombosis of lower limbs were compared in the two groups,and χ2test was carried out on the results.Results There were no cases of pressure ulcers and deep venous thromboses of lower limbs occurred in the experiment group,while 5 cases of deep venous thromboses of lower limbs and 4 cases of pressure ulcer happened in the control group,and there were significant differences between the incidence rates in the groups(P<0.05).Conclusion The air-cushioned belt has easy operation,relieves the patients'pains and the nurses'workload when used to prevent deep venous thrombosis of lower limbs and pressure ulcer,and thus is worthy promoting clinically.

Objective To examine the effects of gross tumor volume (GTV) and radiation dose on the prognosis of hepatocellular carcinoma (HCC) patients treated with whole body gamma knife.Methods The clinical data of 69 HCC patients who underwent body gamma knife treatment from January 2012 to June 2015 in the Radiotherapy Center of the PLA General Hospital were retrospectively reviewed.Based on a 50% or 60% isodose coverage of the planning target volume (PTV), patients were treated with a radiation dose of 4-5 Gy per fraction, and a total marginal dose of 36-50 Gy (median dose 45 Gy).Short-term efficacy, overall survival (OS), and the adverse effect of the treatment were evaluated.The optimal cut-off tumor volume was identified using the receiver operating characteristic curve, and survival was determined by the Kaplan-Meier method.Univariate and multivariate analyses were performed using the log-rank test and Cox proportional hazards regression model, respectively.Results The overall short-term response rate of the 69 patients was 67%.The 1-and 2-year OS rates were 62% and 40%, respectively, with a median survival of 18.6 months.The multivariate analysis showed that gross tumor volume (GTV)<93 cm3(P=0.013) and short-term efficacy of radiotherapy (P=0.000) were significant independent prognostic factors for survival.When GTV was<93 cm3, prognosis was significantly better in patients treated with a dose of ≥45 Gy than in those with<45 Gy (P=0.019).In contrast, radiation dose had no significant effect on survival among patients with GTV>93 cm3(P=0.665).Conclusions GTV is an independent prognostic factor for overall survival of HCC patients.Although high-dose radiotherapy provides survival benefits to patients with small GTV, it is not necessarily suitable for patients with large GTV.

Objective: To analyze the ratio of pulmonary valve ring to aortic valve ring (GA ratio), and to explore the application value of GA ratio in predicting the need of transannular patch in the radical operation of children with tetralogy of Fallot (TOF). Methods: A retrospective analysis was performed in 355 children (182 males and 173 females) with TOF and underwent radical operation in the Pediatric Cardiac Center of Beijing Anzhen Hospital Affiliated to Capital Medical University from January 2016 to December 2017.They were divided into transannular patch group and non-transannular patch group.The values of pulmonary valve ring and aortic valve ring in two groups were collected, and the Z-score of pulmonary artery and GA ratio were calculated respectively for statistical analysis. Results: Among the 355 patients, 156 children (43.9%) required a transannular patch, and 199 patients (56.1%) did not receive transannular patch.The GA ratio and the Z-score of of pulmonary artery in the transannular patch group were lower than those in the non-transannular patch group [0.45±0.12 vs. 0.54±0.15, (-1.75±0.98) scores vs.(-0.86±1.39) scores], and the differences were statistically significant (t=5.29, 6.32, all P<0.01). Receiver operating characteristic curve analysis showed that in the case of the Z-score of pulmonary artery of -2 for the children received transannular patch, the area under the curve (AUC) was 0.702 (95%CI: 0.64-0.76), sensitivity was 69%, and specificity was 83%; in the case of the GA ratio of 0.58 for the children received transannular patch, the AUC was 0.712 (95%CI: 0.66-0.77), sensitivity was 70%, and specificity was 87%. Conclusions The GA ratio is a simple and effective predictor of transannular patch in the radical operation of TOF and can be applied in clinical practice.

Objective To evaluate the methods in early warning of schistosomiasis and elimination of snails in an ecological public forest in Yexie Town of Songjiang District, and provide the basis for monitoring and controlling snails in forest land in the future. Methods The monitoring data of snails, schistosomiasis and snail elimination in the public forest were collected to evaluate the effect of controlling snails. Results The area with existence of snails was found to be 10.47 hm², and it was decreased by 98.28% after three times of snail elimination.A total of 1 904 serological tests was performed (test rate of 88.76%) with the positive rate of 0.16%.Pathogenic test result was negative in 3 subjects examined.Serological samples of 11 dogs and 1 sheep were all negative for the test.None of the 1 480 live snails tested was found to be positive in infection.The awareness rate of schistosomiasis control knowledge and behavior formation in 275 students were 98.2% and 95.3%, respectively. Conclusion The snails in the public forest in Yexie Town have been effectively controlled, but there are still scattered living snails.The measures in eliminating and monitoring of snails should be continued and strengthened.

Objective To explore the role of regulatory T-lymphocytes(Treg) in the immune pathogenesis of suba-cute thyroiditis (SAT). Methods The proportion of Treg in CD4+T cells in peripheral blood of 46 SAT patients and15 controls was detected using flow cytometry. And the concentration of interleukin-10(IL-10), transforming growth factor-beta1(TGF-β1) and prostaglandin E2(PGE2) in serum of 46 SAT patients and 15 controls was measured with ELISA. In addition, the Forkhead box protein 3 (Foxp3) positive cells in thyroid tissue of 29 SAT patients and20 controls was detected by immunohistochemistry. Results The proportion of Treg in peripheral blood of SAT pa-tients was significantly lower than that of controls (P＜0.05). And the concentration of TGF-β1 in serum of SAT patients was apparently higher than that of controls(P＜0.05). Additionally, the positive rate of Foxp3 in thyroid tissue of SAT patients was markedly higher than that of controls(P＜0.05).Conclusions The decrease of Treg may play an important role in the immune pathogenesis of SAT.