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 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.

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.

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

Objective To investigate the diversity of mosquito-borne viruses in Xinjiang , China, and to identify mosquitos-borne viruses of medical importance rapidly .Methods The virus-derived RNAs in mosquitos captured in wild were screened and confirmed by using Illumina deep sequencing approach and reverse transcription PCR , respectively .The alignment analysis was performed by using gene sequences from GenBank.Results One hundred and forty-four Culex Flavivirus ( CxFV, Flavivirus genus, Flaviviridae) specific sequences were identified .The overlapping reads were assembled into 7 uncontinuous viral genomic contigs.The gaps between the contigs were further filled by RT-PCR products, which resulted in reconstruc-tion of viral genomic 5′and 3′terminus (687 nt and 411 nt).Phylogenetic analysis showed that the newly identified CxFV belonged to America/Asian genotype , which specifically clustered into a clade with other CxFV strains from China mainland ,sharing 98.2%-99.5%homologies in nucleotide sequences and 99.5%in amino acids sequences among them .Conclusion Illumina deep sequencing approach was successfully applied to arthropod-borne virus surveillance .The recently emerged Culex Flavivirus was detected for the first time in Xinjiang, China.

ObjectiveTo investigate the application of intraoperative Computed Tomograph (CT) using in surgery for complex acetabular fractures.MethodsFrom June 2008 to December 2010,14 patients (9 males,5 females; with the mean age of 45.1 years; range,28-62 years) with complex acetabular fractures were operated using intraoperative CT.Preoperative radiotherapy and CT scan were adopted to evaluate the fractures.Three dimensional reconstruction based on CT scan was used to mimic surgery.The surgery approach and the type of internal fixators were noted.Intraoperative C-arm and CT scan were used to evaluate the fractures reduction respectively.Decision of additional reduction was made by surgeons according to above mentioned methods respectively and the results were noted.Comparing to preoperative design,the change of surgery plan were noted.Overall time,frequency and radiation dose of intraoperative CT scan were also noted.ResultsAll patients in this study received average 2.7 times of intraoperative CT scan.Mean time of CT scan was 40.4 min and the overall dose of radiation was 47.2 mGy.Decision of additional reduction was made in 3 cases according to C-arm radiography and 4 cases according to CT scan (above mentioned 3 cases were included).The change of surgery plan was made in one case.In postoperative radiography evaluation according to Matta's score system,anatomical reduction were achieved in 8 cases,imperfect reduction in 3 cases and poor reduction in 3 cases.ConclusionIntraoperative CT scan increases the radiation time and dose of patients dramatically.When used to evaluate fracture reduction intraoperatively,it can't take the advantage of traditional C-arm radiography.When delicate preoperative plan is made with radiography and three dimensional reconstruction based on CT data,the efficiency of intraoperative CT scan for complex acetabular fractures are to be discussed.

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.