SUMO (small ubiquitin-related modifier) is involved in the post-translational modifications of proteins,and this process is referred to as SUMOylation.SUMOylation plays an important role in the regulation of cellular activities such as strengthening the stability of the protein,nucleocytoplasmic transport,DNA repair,DNA replication,mitotic and meiotic chromosome behavior,et al.SUMOylation is a dynamic process and can be reversed by SUMO-specific proteases (SENP).Once the balance between SUMOylation and de-SUMOylation is broken,there will be an aberrant expression of SUMO or SENPs in cells to happen,which may lead to the tumor occturtrence.

Objective To compare the difference between bone marrow stomal cell (BMSC) and adipose-derived stem cell (ADSC) of liver fibrosis in rats.Methods BMSC and ADSC of Sprague-Dawley (SD) rats were isolated and purified.The stem cell markers were detected with flow cytometry.The coculture system was set up with 0.4 μm Transwell insert semipermeable membrane.ADSC or BMSC were co-cultured with hepatic stellate cells (HSC).Normal hepatocyte cell line of rat (BRL) was co-cultured with HSC as negative control group and HSC cultured alone was blank control group.After cultured for 72 hours,the proliferation of HSC was determined by cell counting kit-8 (CCK-8) method.The expression of α-smooth muscle actin (α-SMA) of HSC was detected by Western blotting.The apoptosis of HSC was examined by flow cytometry.After BMSC,ADSC and BRL cultured alone for 72 hours,expression level vascular endothelial growth factor (VEGF),interleukin-10 (IL-10),nerve growth factor (NGF) and transforming growth factor-β1 (TGF-β1) in the culture medium were detected by enzymelinked immunosorbent assay (ELISA) method.The rats model of liver fibrosis were established.The rats were divided into BMSC treatment group,ADSC treatment group,BRL group and culture medium group,six rats in each group,which were injected with 1.5 mL BMSC,ADSC and BRL cells suspension (5 × 106) through portal vein,respectively,and same volume of culture medium was injected to the rate of culture medium group,once every two weeks for four weeks.The pathological changes of liver tissue sections were observed and liver fibrosis markers were tested.T test was performed for comparison between two samples and analysis of variance was used for comparison among multiple groups.Results BMSC and ADSC were successfully isolated and cultured.The phenotype of BMSC and ADSC was similar.Compared with blank control group and negative control group,both ADSC and BMSC could inhibit the proliferation of HSC and promote apoptosis (proliferation,2.43±0.27,2.39±0.33,1.92±0.38 and 2.18±0.31,FBMSC =25.61,FADSC =38.63,both P＜0.05 ;apoptosis rate,(5.59 ± 0.40)％,(6.82±0.57)％,(8.31± 1.03) ％ and (9.36 ± 0.54) ％,FBMSC =73.69,FADSC =97.41,both P＜ 0.05).The effects of ADSC were more significant than those of BMSC (t=5.76 and 5.18,both P＜0.05).There was difference in the cytokine levels secreted by ADSC and BMSC (NGF,(7.46 ± 0.54) pg/mL vs (3.95 ± 0.71) pg/mL,t =10.92,P＜0.05; TGF-β1,(8.79 ±0.93) pg/mL vs (6.36±0.85) pg/mL,t=7.58,P＜0.05).The cell transplantation experiment indicated that both BMSC and ADSC had significant inhibitory effect on liver fibrosis.The activity index of inflammation and degree of fibrosis in BMSC treatment group and ADSCs treatment group were 9.87±2.07,4.17 ± 0.94 and 10.13 ± 1.81,3.98 ± 0.82,which were significantly lower than those in blank control group (13.78±2.53 and 5.09±1.15)and negative control group (13.34± 1.89 and 4.95± 1.22,FBMSC=51.26 and 32.29,P＜0.05; FADSC =46.73 and 40.94,P＜0.05).The level of hyaluronic acid ((191.5±33.2) μg/L and (178.8±28.2) μg/L),type Ⅲ collagen ((19.9±5.1) μg/L and (21.7± 3.3) μg/L) and hydroxyproline ((312.6±38.8) μg/g and (325.8±28.2) μg/g) of BMSC treatment group and ADSC treatment group were significantly lower than those of negative control group and blank control group (hyaluronic acid,(282.3 ± 18.7) μg/L and (287.5 ± 26.7) μg/L),F =73.51 ; type Ⅲ collagen,(35.3± 3.3) μg/L and (32.5±4.3) μg/L,F=76.19; hydroxyproline,(458.4 ± 38.1) μg/g and (473.9 ± 63.7) μg/g,F=60.37,all P＜0.05).However,there was no difference between BMSC treatment group and ADSC treatment (all P＜0.05).Conclusions ADSC and BMSC had similar stem cell characteristics.There was difference in inhibiting the activation of HSC between ADSC and BMSC.But there was no significant difference in inhibiting liver fibrosis of rats in vivo.

Objective:To lay background for studying rejection mechanisms in xenotransplantation, class II DQA genes of swine leukocyte antigens (SLA) of three Chinese pig strains GXP, GZP and YNP were cloned and sequenced.Methods:RT-PCR was performed to proliferate SLA-DQA genes (GXPDQA, GZPDQA and YNPDQA),which were cloned into pGEM-T Easy vector for sequencing and analysis.Results:All three allelic sequences examined, are not identical to those reported, which allows the sequences receiving their accession numbers from GenBank as AY102473, AY102474 and AY102475, which encompass an open reading frame of 765, 768 and 768 nucleotides respectively.Conclusion:Three new alleles of SLA-DQA genes were obtained. Analysis of the amino acid sequences of swine, human and mouse DQA (or equivalent) also indicated that GXPDQA, GZPDQA and YNPDQA were more similar to human DQA than mouse H-2-A?, which might provide an understanding of how the immune system evolved.

Objective To investigate the role of PI_3 K/Akt signal pathway in Ephrin-Al gene mediated invasion,metastasis of Huh-7 cells.Methods Western blot was used to test the protein expression of phosphatidylinositol 3-kinase(PI_3 K)and mitogen-activated protein kinase(MAPK)after Huh-7 cells were treated with Ephrin-A1/Fc fusion protein.According to the protein expression,LY294002 was used to block PI_3 K/Akt pathway specifically,then p-Akt protein expression,mobility and invasive ability of Huh-7 cells were examined.Results In Huh-7 cells actived by Ephrin-Al/Fc fusion protein,p-Akt expression was higher than that in control group(t=4.564,P＜0.05),but there was no difference of p-p38MAPK expression between Ephrin-Al/Fc fusion protein group and IgG/Fc fusion protein group(P＞0.05).PI_3 K/Akt pathway was specifically blocked by LY294002,the p-Akt protein expression decreased in Huh-7 cells,and the mobility and invasive ability mediated by Ephrin-Al in Huh-7 cells decreased(P＜0.05).Conclusions PI_3 K/Akt pathway effects an important role in mobility and invasive ability of Huh-7 cells mediated by Ephrin-A1.

0.05),however,it can shorten curative time obviously on Ⅱ type dental ulcer.Simultaneously,the general reaction and focal mucosa of borneolum and borax pellicle group had no obvious variation before and after the test.Conclusions Chitosan has no effect on min-pigs' nerve,cardiovascular and respiratory system;and it is relatively safe given by mouth or peritoneal injection.The borneolum and borax pellicle can shorten curative time obviously on Ⅱtype dental ulcer.

Objective The objective of this study was to provide a way to assess the nutritional status of patients and to afford targeted nutritional supports during the radiotherapy on the basis of the laboratory parameters related to nutrition and chest muscle size in lung cancer patients at the different time.Methods The laboratory parameters were obtained in a cohort of 160 lung cancer patients who received thoracic radiotherapy in our department from March 2012 to November 2015.Fourteen patients who had complete chest CT scan images during radiotherapy were selected to evaluate chest muscles volume.The Chest muscles and its volume were delineated and calculated by CT scan images.Results The levels of(Hemoglobin)HGB,lymphocyte,total protein and albumin were decreased in different degrees during and after radiotherapy,which had the positive correlation with the number and doses of radiotherapy(P<0.05).The changes of hemoglobin and lymphocyte were more obviously among them;these indicators began to recover after 5 weeks of radiotherapy.Chest muscle volume was also the same as the trend of hematological indicators(P>0.05).Conclusion Cancer patients were prone to suffer from malnutrition during radiotherapy.The intake of energy and protein was less than the requirements.We should always take the nutritional status of patients into account and provide targeted nutritional support to improve treatment tolerance and quality of life of patients during radiotherapy.

Objective To investigate the reference values and Z scores regression equations of newborn undergoing echocardiography. Methods Two hundred and eighty-eight newborns (aged 0-28 days) of Shenzhen Children′s Hospital underwent echocardiography examination, including M-mode, two-dimensional (2D) and real-time three-dimensional (3D) echocardiography, color Doppler lfow imaging (CDFI) and tissue Doppler imaging. The correlation between echocardiography results and weight were analyzed and Z scores were calculated. Results The normal values of right ventricular diameter (RV) and left ventricular end-diastolic diameter (LVEDD) measured by M-mode, the mitral annulus diameter in four chamber view (MV-D1), mitral annulus diameter in two chamber view (MV-D2), mitral annulus diameter in longitudinal view (MV-D3), aortic ring diameter (ARD), aortic sinus diameter (ASD), ascending aorta diameter (AAO), transverse aorta diameter (TA), aortic isthmus diameter (AI), aorta diaphragm diameter (AO-Dia), tricuspid annulus diameter in four chamber view (TV-D1), tricuspid annulus diameter in right ventricular inlfow tract view (TV-D2), right ventricular outlfow tract diameter (RVOT), pulmonary valve diameter (PVD) and main pulmonary artery diameter (PA) measured by 2D echocardiography and the normal values of mitral valve inflow Doppler component during early diastole (MV-E), mitral valve inlfow Doppler component during atrial contraction (MV-A), tricuspid valve inlfow Doppler component during early diastole (TV-E), tricuspid valve inflow Doppler component during atrial contraction (TV-A), aortic valve peak velocity (AV-max), aortic valve velocity-time integral (AV-VTI), pulmonary valve peak velocity (PV-max), pulmonary valve velocity-time integral (PV-VTI) measured by pulse Doppler, the mitral annular tissue Doppler component during systole (MV-s′), mitral annular tissue Doppler component during early diastole (MV-e′), mitral annular tissue Doppler component during atrial contraction (MV-a′), tricuspid annular tissue Doppler component during systole (TV-s′), tricuspid annular tissue Doppler component during early diastole (TV-e′), tricuspid annular tissue Doppler component during atrial contraction (TV-a′), interventricular septum annular tissue Doppler component during systole (IVS-s′), interventricular septum annular tissue Doppler component during early diastole (IVS-e′), interventricular septum annular tissue Doppler component during atrial contraction (IVS-a′) measured by tissue Doppler, the normal values of left atrial volume (LAV), left ventricular end-systolic volume (LVEDV), stroke volume (SV) and cardiac output (CO) measured by bi-plane method and the normal values of LVEDV, SV and CO measured by real-time tri-plane method, together with the normal values of left ventricular (LV) mass, left ventricular mass index [LV mass/BSA, LV mass/H2.7, body surface area (BSA) and height (H)], all showed nonlinear positive correlations with body weight (all P<0.01). The values of MV-E/A, PV-E/A, MV-e′/a′, TV-e′/a′, IVS-e′/a′, MV-E/IVS-e′, LV mass/LVEDV and left ventricular ejection fraction (LVEF) showed no correlations with body weight (all P>0.05). Except for RV, MV-D1, MV-D2, MV-D3, TV-D1, TV-E, MV-s′, IVS-a′, TV-s′and TV-e′, all R2 obtained by nonlinear regression method (lnY=a+bX+cX2+dX3) were larger than those obtained by linear regression method (Y=a+bX). The Z score showed a normal distribution and no correlation with body weight. Conclusions The normal reference values of newborn undergoing echocardiography reflect the variation in weight. The Z scores can be obtained by the predicted nonlinear regression equations and show standard normal distribution. The echocardiography normal reference values have important significance for the diagnosis and treatment of neonatal heart disease.

Objective To investigate the clinical value of coronary artery Z-scores on echocardiography in diagnosing coronary artery abnormalities. Methods The echocardiography results of 612 patients with Kawasaki disease (KD) at the acute and recovery phase were retrospectively studied. Coronary artery luminal diameters were converted to body-surface-area-adjusted Z-scores. According to coronary Z-scores classiifcation, all the subjects were divided to four groups:415 cases with no dilation (ND), 133 cases with small coronary artery abnormalities (SCAAs), 47 cases with large coronary artery abnormalities (LCAAs), and 17 cases with giant coronary artery abnormalities (GCAAs). Clinical features (gender, age, typical clinical manifestations, fever duration) and laboratory results (CRP, ESR, WBC, PLT) were compared among all the four groups. Coronary artery diameters and the Z-scores were compared between acute and convalescence phase. Results Along with the increase of coronary Z-score, fever duration was prolonged [ND group:(7.75±3.12) d, SCAAs group (8.50±4.12) d, LCAAs group: (8.57±3.58) d, GCAAs group: (11.88±4.33) d, F=22.375, P<0.05]. With coronary Z-score increasing, PLT also increased (F=22.029, P=0.000), and the highest PLT was observed in GCAAs group. There were no significant differences in the CRP, ESR and WBC among all the four groups (F=0.236, 1.116, 0.121, all P>0.05). No significant different coronary diameters were found in ND cases between recovery and acute phase [(2.24±0.34) mm vs (2.33±0.36) mm, t=1.926, P > 0.05]. But there were significant difference in the coronary Z-scores of ND patients between recovery and acute phase (0.41±0.82 vs 1.17±0.75, t=8.332, P < 0.05). The coronary Z-scores in SCAAs group (1.32±0.89 vs 3.40±0.62, t=11.073, P < 0.05), LCAAs group (3.12±2.27 vs 6.20±1.28, t=4.579, P<0.05) and GCAAs group (11.88±6.77 vs 20.4±9.70, t=3.480, P<0.05) at recovery phase were smaller than values at acute phase. Conclusions The KD coronary Z-scores are the body-surface-area-adjusted standard value, and not subject to the influence of children growth and development. Therefore, it may accurately evaluate the severity of coronary artery abnormalities and its recovery process. Accurate quantitative of the coronary artery luminal dimensions is important in KD clinical management and prognosis prediction.

Objective To investigate the expressions of Wnt3a and Wnt5a in papillary thyroid carcinoma (PTC) and their clinical significances. Methods Immunohistochemical SABC method was used to detect the expressions of Wnt3a and Wnt5a proteins in PTC tissues and their paracancerous tissues collected from 79 patients in Dandong First Hospital from January 2014 to June 2018, and the relationships between the expressions of Wnt3a and Wnt5a proteins and clinicopathological features of PTC patients were analyzed. The expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues and paracancerous tissues were detected by Western blot. Results The results of immunohistochemistry showed that the positive expression rates of Wnt3a and Wnt5a proteins in PTC tissues were significantly higher than those in paracancerous tissues [69.6％ (55/79) vs. 25.3％ (20/79), 60.8％ (48/79) vs. 20.2％ (16/79)], and the differences were statistically significant (χ 2 values were 31.092 and 26.894, both P < 0.01). The results of Western blot showed that the expressions of Wnt3a and Wnt5a proteins in 10 pairs of fresh PTC tissues was significantly higher than those in paracancerous tissues(1.61±0.40 vs. 0.43±0.14, 1.38±0.291 vs. 0.36±0.13), and the differences were statistically significant (t values were 16.234 and 13.493, both P < 0.01). The expressions of Wnt3a and Wnt5a in PTC tissues were correlated with TNM stage, differentiation, extramembranous invasion and lymph node metastasis (Wnt3a: χ2 values were 6.645, 15.945, 8.783 and 11.220; Wnt5a: χ2 values were 21.525, 7.611, 17.880 and 12.581, all P < 0.05), but not with patients'age, sex and tumor diameter (all P > 0.05). There was a positive correlation between Wnt3a and Wnt5a proteins expressions in PTC (r = 0.597, P < 0.01).Conclusion The abnormal expressions of Wnt3a and Wnt5a proteins in PTC may be related to the development of PTC.

Objective To explore whether receptor interacting protein (RIP)1/RIP3 pathways participate in glutamate induced cell death in HT-22 neuronal cells and investigate the potential neuroprotection ofnecrostatin-1 in glutamate induced cell death in HT-22.Methods (1) In vitro cultured mouse hippocampal neuronal HT-22 cells were divided into control group,zVAD group,necrostatin-1 (Nec-1) group,glutamate group,glutamate+zVAD group,glutamate+zVAD+Nec-1 group and glutamate+Nec-1 group;they were treated with zVAD,Nec-1 and glutamate at the final concentrations of 20 μmol/L,30 μmol/L and 3 mmol/L for 24 h.Cell viability was detected using a luminescence-based commercial kit Cell Titer-Glo (CTG).Necrotic cell death was measured by propidium iodide (PI) and HE stainings.(2) HT-22 cells were divided into control group Ⅰ,glutamate group Ⅰ and glutamate+Nec-1 group Ⅰ;MitoSox Red was used to detect mitochondrial reactive oxygen species (ROS) level.(3) HT-22 cells were divided into control group Ⅱ,glutamate group Ⅱ and glutamate+tertiary butyl-hydroxyanisole (BHA) group;the final concentration of BHA was 100 μmol/L;necrotic cell death was measured by PI and HE stainings after 24 h of treatment.(4) HT-22 cells were divided into RIP3 siRNA and control group Ⅲ,and then,they were transfected with RIP3 siRNA or negative siRNA,respectively;the RIP3 protein expression was determined by Westem blotting after 72 h of treatment.(5) HT-22 cells were divided into negative siRNA+Control,RIP3 siRNA,negative siRNA+glutamate and RIP3 siRNA+glutamate groups;the cells were transfected with RIP3 siRNA or Negative siNRA,respectively;48 h later,the glutamate groups were treated with 3 mmol/L glutamate;PI positive cells and cell viability were measured by PI and HE stainings and CTG at 24 h after glutamate treatment.Results (1) As compared with control group,percentage of PI positive cells was greatly increased and cell viability was decreased in glutamate group and glutamate+zVAD group,with statistically significant differences (P＜0.05);as compared with those in the glutamate group,percentage of percentage of PI positive cells was was significantly decreased and cell viability was statistically increased in glutamate+Nec-1 group (P＜0.05).(2) ROS level in HT-22 cells of the glutamate group was significantly increased than that in the control group Ⅰ (P＜0.05);however,ROS level in HT-22 cells of glutamate+Nec-1 group Ⅰ was significantly decreased than that in glutamate group Ⅰ (P＜0.05).(3)Percentage of PI positive cells in the glutamate group Ⅱ was significantly higher than that in the control group Ⅱ (P＜0.05),and that in the glutamate+BHA group was statistically lower than that in the glutamate group Ⅱ (P＜0.05).(4) The RIP3 protein expression in the RIP3 siRNA group was obviously down-regulated as compared with that in the control group Ⅲ.(5) As compared with those in the negative siRNA group,percentage of PI positive cells was statistically increased and cell viabilities were statistically decreased in glutamate group (P＜0.05);however,percentage of PI positive cells was significantly decreased and cell viability was significantly increased in RIP3 siNRA+glutamate group as compared with those in the glutamate group (P＜0.05).Conclusion RIP1/RIP3 pathway and ROS might mediate glutamate induced cell death in HT-22 cells.