Objective:To evaluate the effect of all-trans retinoic acid (ATRA) on the expression of epithelial-mesenchymal transition (EMT) -related molecules in human malignant melanoma A375 cells.Methods:Cultured A375 cells were divided into 4 groups: control-1 and -2 groups treated with Dulbecco′s modified Eagle medium (DMEM) for 24 and 48 hours respectively, and ATRA-1 and ATRA-2 groups treated with DMEM containing 10 μmol/L ATRA for 24 and 48 hours respectively. After the treatment, real-time quantitative PCR was performed to determine the mRNA expression of EMT-related genes E-cadherin, N-cadherin, vimentin and β-catenin in the above 4 groups, Western blot analysis to determine the relative expression of the above proteins, and direct immunofluorescence study to assess the fluorescence intensity of E-cadherin and vimentin in the ATRA-1, ATRA-2 and control-1 groups. Statistical analysis was carried out by using two-way analysis of variance, one-way analysis of variance and least significant difference- t test. Results:Real-time quantitative PCR showed that the E-cadherin mRNA expression was significantly higher in the ATRA-1 group than in the control -1 group ( F = 13.148, P < 0.05) , and higher in the ATRA-2 group than in the control-2 group ( F = 31.529, P < 0.05) ; the mRNA expression of N-cadherin, vimentin and β-catenin was significantly lower in the ATRA-1 group than in the control-1 group ( P < 0.05) , and lower in the ATRA-2 group than in the control-2 group ( P < 0.05) ; the ATRA-2 group showed significantly increased mRNA expression of E-cadherin ( F = 13.148, P < 0.05) , but significantly decreased mRNA expression of the other 3 proteins compared with the ATRA-1 group (all P < 0.05) ; there was no significant difference in the mRNA expression of the above molecules between the control-1 and -2 groups (all P > 0.05) . Western blot analysis showed that the protein expression of E-cadherin significantly increased, but the protein expression of N-cadherin, vimentin and β-catenin significantly decreased in the ATRA-1 and ATRA-2 groups compared with the control-1 group (all P < 0.05) ; compared with the ATRA-1 group, the ATRA-2 group showed significantly increased protein expression of E-cadherin ( P < 0.05) , but significantly decreased protein expression of N-cadherin, vimentin and β-catenin (all P < 0.05) . Direct immunofluorescence study showed that the fluorescence intensity of E-cadherin was significantly higher in the ATRA-1 group and ATRA-2 group (6.23 ± 0.08, 10.37 ± 0.13, respectively) than in the control-1 group (2.37 ± 0.14, both P < 0.05) , while the fluorescence intensity of vimentin was significantly lower in the ATRA-1 group and ATRA-2 group (15.17 ± 0.18, 10.29 ± 0.03, respectively) than in the control-1 group (50.16 ± 0.26, both P < 0.05) , and the cells in the ATRA-1 group and ATRA-2 group transformed from spindle- to cobble-stone-like in shape. Conclusion:ATRA can up-regulate the expression of E-cadherin, down-regulate the expression of N-cadherin, vimentin and β-catenin in A375 cells, and may inhibit the EMT of A375 cells.

Objective To discuss the role of EB virus (EBV)in the pathogenesis of systemic lupus erythematosus(SLE) in children through investigating the copies of EBV DNA and expression of EBV genes in peripheral blood mononuclear cells(PBMCs).Methods (1)PBMCs were isolated from 30 patients with SLE and 12 healthy normal controls respectively and DNA was extracted from PBMCs.(2) PBMCs were co-cultured with EBV for 12 days and RNA was extracted from PBMCs.(3)Real-time fluorescence quantitative PCR(Real-time PCR) was applied to detect the copies of EBV DNA in PBMCs.(4)Reverse transcription PCR was applied to detect expression of EBV genes.Results (1) Compared with the healthy control group [(40.1 ± 11.6) copies/μg],a significant increase of EBV DNA copies was observed in SLE group[(658.6 ± 183.6) copies/μg] (P ＜0.05).The EBV DNA copies in the active SLE group [(785.2 ± 179.2) copies/μg] were significantly higher than those in the non-active SLE group [(586.0 ± 193.1) copies/μg] (P ＜ 0.05).(2)There was no correlation between EBV DNA copies and systemic lupus erythematosus disease activity index (r =0.03,P ＞ 0.05).(3) After PBMCs got co-cultured with EBV,expression of latent EBV genes and lytic genes were both increased in the patients and healthy controls.The latent EBV genes including latent membrane protein 1 (LMP1),LMP2,EBV nuclear antigen 1 and the lytic genes including BCRF1,BLLF1 were all increased significantly in the patients compared with the healthy controls (all P ＜ 0.05).Conclusions There is a significant increase of EBV DNA copies and aberrant expression of EBV genes in SLE patients,which suggests that EBV may contribute to the pathogenesis of SLE.

Objective To investigate the expression and significance of Epstein-Barr virus (EBV) genes in children with systemic lupus erythematosus (SLE).Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 20 children with SLE and 12 healthy human controls.Enzyme-linked immunosorbent assay (ELISA) was conducted to detect anti-EBV viral capsid antigen (VCA) IgG/IgM antibodies.The culture supernatants of cells from patients with anti-EBV VCA IgG/IgM antibodies were collected,and PBMCs from the patients and controls were co-cultured with the supernatants respectively for 12 days.RNA was extracted from PBMCs before and after the coculture,and reverse transcription-PCR was performed to detect the expression of EBV genes,including LMP1,LMP2,EBNA1,BCRF1,BLLF1 and BILF1 genes.Results LMP1 gene was detected in fresh PBMCs from 10 out of 20 patients and 1 out of 12 controls (P ＜ 0.05).No significant differences were observed between the patients and controls in the detection rate of LMP2 gene (4/20 vs.1/12),EBNA1 gene (13/20 vs.3/12),BCRF1 gene (3/20 vs.1/12) or BLLF1 gene (5/20 vs.2/12) in fresh PBMCs.After co-culture with the supernatants of cells from patients with anti-EBV VCA IgG/IgM antibodies,the expressions of EBV genes in these PBMCs were increased to different degrees,and there was a significant difference in the expressions of EBV latent genes LMP1,LMP2 and EBNA-1 as well as EBV replicative genes BCRF1 and BLLF1 between the patient-derived and control-derived PBMCs (all P ＜ 0.05).Conclusions There is an aberrant expression of EBV genes in children with SLE,and EBV genes may contribute to the development of SLE.

Objective To investigate a new method of improving design of the skin flap pedicled with descending hranch of lateral femoral circumflex artery,in order to increase the accuracy of preoperative Doppler location.Methods Six fresh cadavers underwent a whole body,intra-arterial injection of a lead ox- ide and gelatine preparation.Observe the perforators of anterolateral thigh by dissection,measured their diam- eter;course,branches and location for there were showed up by angiography and photography.A specific pro- gram,3D-doctor,was used for the detection of the regions of interest in angisome and its 3D-reconstructive.In addition to the average area supplied by a single perforator within that territory was calculated from digital an- giograms using Scion Image.Results There were 16 perforators that the diameter≥0.5 ram,20%were septocutaneous,80%were musculocutaneous.Their average external diameter was 0.8 ram.The average ped- icle length from the deep fascia was(3.15?1.43)ram;the perforators from descending branch of lateral cir- cumflex femoral artery walk length average about 2.63 cm in superficial fascia.The area of each perforator supplied blood was average 45.61 cm~2.Conclusion Modified lead oxid-gelatine technique provides high quality angiograms for the study of cutaneous artery and perforator falp.Based on our data,the maximum di- mension of the anterolateral thigh perforator flap can be 30 cm?20 cm in with only a single dominant perfora- tor.The perforator flaps deviced by perforators from the anterolateral thigh are transplanted to the lower limbs and the other part of the body.

Objective:To conduct a systematic review and Meta-analysis of the efficacy and safety of Apatinib and tegafur in colonic cancer.Methods:With "Apatinib" "Tegafur" "Colonic cancer" as keywords, PubMed, Embase, Web of Science, Cochrane Library, WanFang Data, VIP, CNKI and CBM were searched from inception to December 2020 to collect randomized controlled trail about treatment for colonic cancer with Apatinib and Tegafur. Evaluated the portion remission and stable duration and progression-free survival. Meta-analysis was performed by using RevMan 5.3 software.Results:Meta-analysis showed that in colonic cancer patients, the portion remission and stable duration, tumor progression of Apatinib were not inferior to those of Tegafur ( RR=1.10, 95% CI: 0.71-1.71, P=0.640; RR=0.51, 95% CI: 0.28-1.32, P=0.205). But for progression-free survival, Apatinib was superior to Tegafur in overall patients( SMD=0.90, 95% CI: 0.42-1.37, P<0.000 1). Conclusion:In the treatment of colon cancer, compared with Tegafur, Apatinib can effectively improve the progression-free survival and has better overall survival.

Objective To investigate the changes of thyroid hormone in peritoneal dialysis patients and analyse its impact factors, as well as the therapeutic effects of small dose of thyroxine.Methods 150 uremic patients in Hunan Provincial People’s Hospital from December 2013 to December 2014 were selected, 70 cases of uremia non-dialysis patients were divided into group A, while 80 uremia peritoneal dialysis for more than half a year were divided into group B.70 cases healthy examinees during the same period in our hospital were selected as control group ( group C ) . The total triiodothyronine (T3), total thyroxine (T4), free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH), hemoglobin (Hb), serum albumin (ALB), total cholesterol (TC), triglyceride (TG), serum creatinine (SCr), C reactive protein (CRP) and left ventricular ejection fraction ( LVEF) , subjective global assessment of nutritional act ( SGA) and other indicators were detected in three groups.Patients in group B were divided into two sub-groups according to thyroid hormone levels: B1 group had normal thyroid level while B2 abnormal.And the administration of small dose of thyroid hormone was given to patients in group B2, and the effect of the administration was evaluated by the above indexes.Results The FT3 in group A and B were significantly lower than that in group C (P<0.01).There were significant differences of levels of ALB, CRP, SGA between group B1 and group B2, and the FT3 level in group B was significant correlated with SGA, ALB, LVEF(r=0.815,P<0.001;r=0.780,P<0.001;r=0.953,P<0.001).After treated with small dose of thyroid hormone, FT3 and LVEF were improved while FT4, TSH, ALB, SGA, CRP were not improved in group B2.Conclusion The thyroid hormone level in patients with continuous ambulatory peritoneal dialysis decreases which is dominated with FT3.The decreased thyroid level is significantly correlated with nutrition ( ALB, SGA) and left ventricular function.The administration of small dose of thyroid hormone can improve the left ventricular systolic function.

OBJECTIVETo observe the effects of rukangyin (RKY) on the lymphangiogenesis and lymph node metastasis of breast cancer transplantation tumor mice, thus exploring its anti-tumor metastasis mechanisms.

METHODSHuman breast cancer cell line MDA-MB-435S were in situ implanted into the mammary fat pad of 30 female nude mice to establish breast cancer transplantation tumor spontaneous metastasis model. They were randomly divided into six groups, i.e., the model control group, the 5-FU control group, the small, medium, large dose RKY groups, and the medium dose RKY +5-FU group, 5 in each. Normal saline was given to mice in the model control group at the daily dose of 0.4 mL/kg by gastrogavage. 5-FU was given to mice in the 5-FU control group at the daily dose of 30 mg/kg by peritoneal injection. RKY was given to mice in the small, medium, large dose RKY groups at the daily dose of 18, 45, and 90 g/kg by gastrogavage. 5-FU 30 mg/kg (by peritoneal injection) + RKY 45 g/( kg x d) (by gastrogavage) was given to mice in the medium dose RKY +5-FU group. All medication was carried out once daily for 6 successive weeks. The tumor volume, the tumor inhibition ratio, and the inhibition ratio of axillary lymph node metastasis were detected after medication. The lymphatic microvessel density (LMVD) and expressions of vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor receptor-3 (VEGFR-3) of the breast cancer tissues were detected using immunohistochemical assay.

RESULTSCompared with the model control group, the tumor volume was markedly reduced in the small, medium, large dose RKY groups, and the medium dose RKY +5-FU group, the expressions of VEGF-C and VEGFR-3 were significantly down-regulated and LMVD were obviously lowered, showing statistical difference (P < 0.05, P < 0.01). The inhibition rates of tumor and axillary lymph node metastasis were highest and the LMVD was the lowest in the medium dose RKY +5-FU group, showing statistical difference when compared with other medication groups (P < 0.05, P < 0.01).

CONCLUSIONRKY might inhibit the lymph node metastasis of breast cancer possibly through intervening the expressions of VEGF-C and VEGFR-3, and suppressing lymphangiogenesis.

Animals ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Lymphangiogenesis ; drug effects ; Lymphatic Metastasis ; Lymphatic Vessels ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor Receptor-3 ; metabolism

OBJECTIVETo investigate the prognostic factors of lymph node-negative advanced gastric cancer patients in order to guide adjunctive therapy and surveillance tragedy.

METHODSA total of 236 advanced gastric cancer patients with no less than 12 retrieved lymph nodes and without lymph node metastasis from Fujian Provincial Hospital between 1998 and 2008 were collected retrospectively. Univariate and multivariate prognostic analysis were performed.

RESULTSTwo hundred and twenty-four patients(94.9%) were followed up and 5-year overall and disease-free survival rates were 75.2% and 66.4% respectively. Univariate prognostic analysis showed that depth of infiltration, Lauren histotype and retrieved lymph nodes were associated with 5-year overall survival(all P<0.05). Multivariate prognostic analysis testified that depth of infiltration was independent prognostic predictor(P<0.05). Recurrent rates of T2 and T3 patients were 5.8%(8/138) and 14.0%(12/86),5-year overall survival rates were 82.5% and 59.0%, 5-year disease-free survival rates were 70.4% and 52.2% respectively. These differences were all statistically significant (all P<0.05).

CONCLUSIONST2N0 gastric cancer patients have a better prognosis than T3N0 patients. Depth of infiltration should be considered to stratify lymph node-negative gastric cancer patients for an adjunctive treatment and follow-up scheduling.

Female ; Follow-Up Studies ; Gastrectomy ; Humans ; Lymph Node Excision ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery

Objective To predict the incidence of birth defects in Xi'an using the auto-regressive integrated moving average product seasonal model.Methods In Xi'an,the trend of the incidence of birth defects was analyzed and tested from October 2009 to August 2015.Using the data from September to December 2015,the actual birth defects were compared with the model fitting data to evaluate the predictive performance of the model.Multiple seasonal ARIMA model was then fitted under time series to predict the incidence of birth defects in 2016.Results Seasonal effect was seen in the incidence of birth defects in Xi'an.A multiple seasonal ARIMA(0,0,1) (0,1,1)12 was established.The mean of absolute error and the relative error were 9.5 and 0.084,respectively,when compared to the simulated number of patients from September to December in 2015,suggesting that ARIMA (0,0,1) (0,1,1)12 has a better predictive ability.Results under the prediction of multiple seasonal ARIMA model showed that the number of patients in 2016 was similar to that of 2015 in Xi'an,with a slight increase and a decrease in the peak value.Conclusion Multiple seasonal ARIMA(0,0,1)(0,1,1)12 model could be used to successfully predict the incidence of birth defects in Xi'an.

Objective To investigate the dynamic epidemiological characteristics of perinatal birth defects in Xi'an so as to provide scientific evidence for the priority of future birth defect prevention.Methods We made a statistical description of the data of perinatal birth defects from 2010 to 2015 reported by all the maternity hospitals in Xi'an.Results The incidence of birth defects in Xi'an from 2010 to 2015 was 89.62/104 (87.60/104,91.74/ 104) and had an annual ascending tendency (x2-157.13,P＜0.01).The top five main birth defects in a descending order were congenital heart disease (26.53/104),polydactyly (13.10/104),total cleft lip (11.42/104),neural tube defects (6.06 /104),and congenital hydrocephalus (5.08/104),accounting for 69.39％ of the total number of birth defects.And the incidence of congenital heart diseases showed a year-by-year ascending trend (x2=837.65,P＜ 0.01).The incidence of birth defects was higher in urban areas than in rural areas (OR 1.108,95％ CI:1.056-1.162).The birth defects were more frequently seen in males than in females (OB=1.151,95％ CI:1.097-1.208).Mother's age ＜20 (OR=1.764,95％ CI..1.532-2.032) and mother'sage≥35 (OR=1.283,95％ CI:1.179-1.395) also were risk factors for birth defects.Up to 72.54％ of the defects were live births.There were 52.83％ cases diagnosed by ultrasound and 46.24％ ones by clinical examination.The prenatal diagnosis rate was 33.20％.Conclusion The incidence of perinatal birth defects in Xi'an from 2010 to 2015 showed an annual ascending trend.We should improve the prenatal diagnosis level of all monitored hospitals in order to reduce incidence of birth defects and increase the quality of population.