Objective:To study the changes in long non-coding RNA C2dat1 expression in kidney tissues of rats at different stages of diabetic kidney disease (DKD) and its relationship with renal interstitial fibrosis.Methods:Forty-eight male SD rats were randomly divided into two groups with 24 rats in each group: control group and DKD group. The rats in the control group were fed with ordinary diet, while those in the DKD group were fed with high-fat diet and drank water freely. After eight weeks of feeding, the rats were fasted for 12 h with free access to water. Then, the DKD group was given a one-time intrabitoneal injection of streptozotocin and the control group was given an equal dose of sodium citrate buffer. After 72 h, the random peripheral blood glucose concentration (≥ 16.7 mmol/L for three consecutive days) and urine sugar (positive) were tested to assess the establishment of the diabetes model. Urine, blood and kidney samples were collected at 3, 6, 9 and 12 weeks. The urinary protein excretion rate within 24 h, urinary creatinine and serum total cholesterol were measured by automatic biochemical apparatus. Pathological changes in kidney tissues were observed by HE staining. The expression of calcium/calmodulin-dependent protein kinase Ⅱ delta (CaMK2D), p65, p50, α-SMA and E-cardherin was detected by immunohistochemistry. Quantitative real-time PCR (qPCR) was used to detect the expression of lncRNA C2dat1 and CaMK2D. The relationship of lncRNA C2dat1 with α-SMA, E-cardherin and CaMK2D was analyzed by correlation analysis. In in vitro experiment, renal tubular epithelial cells HK-2 were induced by high glucose. The expression of lncRNA C2dat1 and CaMK2D in HK-2 cells was detected by qPCR after 24, 48 and 72 h of intervention. Results:The rats in the DKD group showed typical symptoms such as polydipsia, polyphagia, significant weight loss and increased blood glucose as compared with the rats in the control group. Results of the biochemical tests revealed that compared with the control group, the DKD group had increased 24 h excretion rate of urinary protein, decreased urinary creatinine and up-regulated total cholesterol. HE staining showed that the rats in the control group had intact glomeruli, normal basement membrane and no mesangial hyperplasia or inflammatory cell infiltration. However, enlarged glomeruli and evenly thickened basement membrane were observed in the DKD group. Immunohistochemistry indicated that the expression of CaMK2D, p50 and α-SMA was higher in the DKD group than in the control group, while the expression of E-cardherin was lower in the DKD group. qPCR results showed that the expression of lncRNA C2dat1 and CaMK2D at mRNA level was higher in the DKD group than in the control group. In in vitro experiment, the expression of lncRNA C2dat1 and CaMK2D at mRNA level was also higher in HK-2 cells induced by high glucose than in the control group. Correlation analysis indicated that lncRNA C2dat1 was positively correlated with α-SMA and CaMK2D, but negatively correlated with E-cardherin. Conclusions:During the progression of DKD, the high expression of lncRNA C2dat1 might promote diabetic renal interstitial fibrosis by regulating the expression of CaMK2D to activate the NF-κB signaling pathway.

ObjectiveTo screen out the mRNAs involved in the resistance of hepatoma cells to anlotinib using ceRNA microarray. MethodsHigh-dose shock combined with low-dose induction was used to culture hepatoma cells resistant to anlotinib, and CCK8 assay was used to verify the difference in the proliferation of drug-resistant hepatoma cells treated by anlotinib. The ceRNA microarray was used to screen out the differentially expressed genes between drug-resistant hepatoma cells and normal hepatoma cells, and real-time PCR was used to verify the differentially expressed genes detected by some microarrays. the independent samples t-test was used for comparison of continuous data between two groups, and the Kaplan-Meier method was used to analyze the overall survival of hepatoma cells samples, and the log-rank test was used to compare survival rates. Fisher’s exact test was used for chip screening. ResultsThere was a significant difference in gene expression between drug-resistant hepatoma cells and normal hepatoma cells, and 10 genes with the greatest difference were screened out for analysis by reducing the range. There were 4 genes associated with drug resistance and tumor growth, i.e., BIRC2, BIRC7, ABCC2, and MAPK8. There were significant reductions in the expression levels of BIRC2, ABCC2, and MAPK8 (P=0001 4, 0001 2, and 0.011 8), and there was a significant increase in the expression of BIRC7 (P＜0.001). The results of real-time PCR were consistent with those of microarray (t=10.74,32.65,18.34, and 2.80; P=0.000 4, 0.000 1, 0.000 1, and 0.044 8). The high expression of BIRC7 and the low expression of MAPK8 were associated with the significant reduction in survival time (P=0.022 0 and 0.005 6). ConclusionBIRC2, BIRC7, ABCC2, and MAPK8 are differentially expressed between anlotinib-resistant hepatoma cells and normal hepatoma cells and may be involved in the resistance of hepatoma cells to anlotinib.

Objective:To evaluate the mid- and long-term outcomes of Dynesys hybrid surgery (in some segments act as a non-fusion device, in other segments act as an alternative of rigid fixation in combination with interbody fusion) in the treatment of multi-segmental lumbar degenerative disease (LDD).Methods:The data of 27 patients who received Dynesys hybrid surgery (hybrid group) for the treatment of LDD from May 2011 to September 2016 and completed the follow-up were retrospectively analyzed. Among them, there were 8 males and 19 females; their average age was 59.1±11.9 years (23-78 years). Main diagnosis: 13 cases of lumbar spinal stenosis, 14 cases of lumbar disc herniation; 4 cases of combined lumbar dynamic position instability, 7 cases of combined lumbar spondylolisthesis. There were 15 cases of two-segment disease, 11 cases of three-segment disease, and 1 case of four-segment disease. Segments distribution: 9 cases of L 3-L 5, 6 cases of L 4-S 1, 7 cases of L 3-S 1, 4 cases of L 2-L 5, and 1 case of L 2-S 1. Midline incision was used to exposure, followed by bilateral pedicle screws implantation, and interbody fusion cage with bone grafting were performed at the fusion level. Twenty-seven patients who underwent TLIF+rigid internal fixation during the same period were included as the control group. Clinical outcomes were measured by visual analog scale (VAS) for low back pain and leg pain, and Oswestry disability index (ODI). Radiological outcomes included fusion rate, intervertebral disc height (DH) of surgical segments and the proximal adjacent segment, range of motion (ROM) of non-fusion segments and the proximal adjacent segment. At the same time, the occurrence of complications was observed. Results:Patients of Hybrid group and control group were followed up for an average of 83.8±20.9 months (48-112 months) and 87.3±16.2 months (53-114 months), respectively. Baseline data of the two groups (average follow-up time, age, gender, surgical level, diagnosis) showed no significant difference. The operation time (183.0±27.8 min) and intraoperative blood loss (301.9±178.9 ml) in the hybrid group were significantly lower than those in the control group (operation time t=2.337, P=0.023; blood loss t=2.706, P=0.01). At the final follow-up, the VAS scores of low back pain and leg pain (low back pain t=12.164, P<0.001; leg pain t=20.603, P<0.001), as well as ODI were significantly improved ( t=22.827, P<0.001). A total of 32 segments received TLIF+Dynesys stabilization and 35 segments received Dynesys non-fusion stabilization in the hybrid group, with 28 segments (87.5%) achieved solid fusion at 1-year follow-up. There were 67 fusion segments in the control group, and the fusion rate at 1-year follow-up was 85.1%. DH of non-fusion segments were lower than that before surgery with statistical significance at final follow-up ( t=2.647, P=0.012), while DH of the fusion segments in the hybrid group and the surgical segments in the control group increased compared with that before surgery at the final follow-up. A certain degree of ROM (2.4°±1.5°) was retained of the non-fusion segments at the final follow-up; the ROM of proximal adjacent segments of non-fused segments was significantly smaller than that of proximal adjacent segments of fused segments ( t=2.126, P=0.044). In the hybrid group, screw loosening occurred in 4 patients (8 screws) and adjacent segment degeneration (ASD) occurred in 5 patients. In the control group, screw loosening occurred in 3 patients (6 screws), while ASD occurred in 8 patients. No screw fracture was observed during the follow-up period and no patients received reoperation. Conclusion:Hybrid surgery of Dynesys stabilization combined with interbody fusion is a safe and effective method for the treatment of multi-segmental LDD. Compared with multi-segmental fusion, this lumbar hybrid surgery has the advantages of less trauma and retaining partial segmental ROM.

[This corrects the article DOI: 10.1016/j.apsb.2019.03.006.].

Nanomedicine is charactered with a high specific surface area, diversified structure and function, and charged surface. It can realize the targeted therap by functional modification of surface or introducing the stimuli-responsive unit. Therefore, nanomedicine is increasingly being concerned. Because nanomedicine can accumulate efficiently in the lungs, drug delivery systems based on nanotechnology have broad prospects in the field of the diagnosis, prevention, and treatment in pediatric lung diseases. Herein, we reviewed the research progress of nanomedicine in pediatric lung diseases, especially in respiratory syncytial virus infection and cystic fibrosis.

Recently, considerable attention in the field of cancer therapy has been focused on the mammalian rapamycin target (mTOR), inhibition of which could result in autophagic cell death (ACD). Though novel combination chemotherapy of autophagy inducers with chemotherapeutic agents is extensively investigated, nanomedicine-based combination therapy for ACD remains in infancy. In attempt to actively trigger ACD for synergistic chemotherapy, here we incorporated autophagy inducer rapamycin (RAP) into 7pep-modified PEG-DSPE polymer micelles (7pep-M-RAP) to specifically target and efficiently priming ACD of MCF-7 human breast cancer cells with high expression of transferrin receptor (TfR). Cytotoxic paclitaxel (PTX)-loaded micelle (7pep-M-PTX) was regarded as chemotherapeutic drug model. We discovered that with superior intracellular uptake and more tumor accumulation of micelles , 7pep-M-RAP exhibited excellent autophagy induction and synergistic antitumor efficacy with 7pep-M-PTX. Mechanism study further revealed that 7pep-M-RAP and 7pep-M-PTX used in combination provided enhanced efficacy through induction of both apoptosis- and mitochondria-associated autophagic cell death. Together, our findings suggested that the targeted excess autophagy may provide a rational strategy to improve therapeutic outcome of breast cancer, and simultaneous induction of ACD and apoptosis may be a promising anticancer modality.

Objective To investigate the effects of tumor necrosis factor-related ligand-1A(TL1A)on activation of T helper 9(Th9)cells of colonic tissues in chronic experimental colitis mice.Methods The chronic experimental colitis mice model was established with drinking dextran sulfate sodium salt(DSS).A total of 32 lymphocytes TL1A highly expressed mice and wild type(WT)mice were divided into WT control group, transgene control group,WT modeling group and transgene modeling group.The mice of control groups were administrated with distilled water. The mice of modeling groups received 3% DSS in drinking water discontinuously.The mice were sacrificed on 29 days after modeling.Body mass was measured,length of colon was recorded,scores of gross colon and the disease activity index(DAI)were calculated.The colonic morphological changes were observed by hematoxylin-eosin(H-E)staining.The lamina propria mononuclear cells(LPMC)were isolated and the number of Th9 cells was tested by flow cytometry.The levels of interleukin-9(IL-9)in serum and LPMC were detected by enzyme-linked immunosorbent assay(ELISA).The expressions of IL-9 protein and mRNA of the colonic tissues were measured by Western blotting and real-time polymerase chain reaction(PCR),respectively.T test and single factor analysis of variance were performed for statistical analysis.Results The percentage of body mass loss of WT modeling group was lower than that of transgene modeling group(16.2% ± 1.0% vs 18.9% ± 1.2%),and the difference was statistically significant(t=4.90, P<0.05).The scores of gross colon,DAI and pathology of transgene modeling group were all higher than those of WT modeling group(2.80 ± 0.64 vs 1.60 ± 0.31,2.55 ± 0.20 vs 1.58 ± 0.17,and 11.85 ± 0.86 vs 9.50 ± 0.79),and the differences were statistically significant(t=4.77,10.45 and 5.69,all P<0.05).The number of LPMC in transgene modeling group was higher than that of WT modeling group(3.70×106± 0.28×106vs 2.65×106± 0.32 × 106)and the difference was statistically significant(t= 6.98,P< 0.05).The percentage of Th9 in total CD4+T cells of LPMC in colonic tissues of transgene modeling group was higher than that of WT modeling group(0.54% ± 0.04% vs 0.23% ± 0.03%),and the difference was statistically significant(t= 17.54,P< 0.05).The serum IL-9 level of transgene modeling group was higher than that of WT modeling group((170.23 ± 5.69)pg/mL vs(150.62 ± 6.45)pg/mL),and the difference was statistically significant(t= 6.50,P< 0.05).The level of IL-9 secreted by LMPC of transgene modeling group was higher than that of WT modeling group((265.21 ± 8.76)pg/mL vs (237.58 ± 10.24)pg/mL),and the difference was statistically significant(t= 5.80,P< 0.05).The expressions of IL-9 protein and mRNA of transgene modeling group were higher than those of WT modeling group(1.31 ± 0.09 vs 1.18 ± 0.03,and 8.26 ± 1.13 vs 2.25 ± 0.29,respectively),and the differences were statistically significant(t=3.88 and 14.57,both P< 0.05).Conclusion TL1A high expression in lymphocytes can promote Th9 cells differentiation and IL-9 secretion which involved in the genesis of chronic experimental colitis.

Objective To quantitatively assess the virucidal activities of three commercial disin-fectants against human infected highly pathogenic avian influenza viruses subtype H5. Methods The 50%tissue culture infective dose ( TCID50 ) of avian influenza viruses was calculated. Quantitative suspension test was performed to evaluate the efficacy of three disinfectants. In that test, 105 TCID50 of avian influenza viru-ses were exposed to different disinfectants at different concentrations for different times with or without the in-terference with fetal bovine serum ( FBS) simulating the contaminated condition. The residual infectivity was determined by endpoint titration in Madin-Darby canine kidney ( MDCK) cells. The detail steps were that the mixture of viruses and disinfectants was inoculated at 37℃ with 5% CO2 for 1 hour. Then, it was re-placed by virus dilution medium and further incubated for 18 to 20 hours. ELISA was performed for the cal-culation of TCID50 . The titers of residual viruses were calculated according to Reed and Muench method. The low pathogenic avian influenza virus H9N2 was chosen as the control in this study. Results The re-mained infectivities of three viruses after 1 minute exposure to 1% Virkon solution were below the limit of de-tection (1. 0 lgTCID50/100 μl). Exposing to 0. 5% Virkon solution decreased the viral titers of H5N1 and H9N2 viruses below the detection limit and reduced the titer of H5N6 virus to 1. 75 lgTCID50/100 μl. The virucidal efficacy of 0. 25% Virkon solution against some of the detected viruses was achieved by increasing the exposure time to 5 minutes. The 84 Disinfectant solutions at concentrations of 10%, 5% and 2. 5% low-ered the viral titers of three viruses below the detection limit of 1. 0 lgTCID50/100 μl, but the 1. 25% 84 Disinfectant solution only lowered the viral titers to 1. 25-2. 5 lgTCID50/100 μl. The similar results were ob-served in groups treated with SOLARSEPT solutions. 1% 84 Disinfectant solution didn′t show any virucidal activity against the three viruses after 1 minute of exposure even when the exposure time was extended to 5 minutes. Under the contaminated condition, 1% Virkon solution, 10% and 5% 84 Disinfectant solutions as well as 100% and 50% SOLARSEPT solutions lowered the viral titers below 1. 0 lgTCID50/100μl. Conclu-sion The three commercial disinfectants (1% Virkon solution, 10% 84 Disinfectant solution and SOLAR-SEPT solution) were efficient virucides for highly pathogenic avian influenza viruses subtype H5 even under the contaminated condition. Increasing the exposure time had no significant effects on the efficacy of three disinfectants after the virucidal activities were neutralized by enough viruses. No significant differences in vi-rucidal activities of three disinfectants against HPAI H5 viruses and LPAI H9 virus were observed.

Objective To understand the antigenic and genetic characteristics of highly pathogenic avian influenza H5N1 viruses isolated from poultry related environments of surrounding areas of Qinghai Lake.Methods Poultry and wild birds related environments from surrounding areas of Qinghai lake were collected and handled.Viruses were isolated with embryonated chicken eggs.The virus subtyping was performed by real-time RT-PCR.The highly pathogenic avian influenza H5N1 viruses were chosen to conduct next generation sequencing and hemagglutination inhibition ( HI) assay.Results In total of 19 H5N1 virus strains were isolated, they were all from poultry related environments and collected in January-March of year 2013.Seven representative highly pathogenic avian influenza H5N1 viruses were sequenced and analyzed for antigenicity.According to phylogenetic analysis of HA gene, 6 viruses were belong to clade 2.3.2.1c except A/Environment/Qinghai/XN02032/2013 which belongs to clade 7.2.Antigenic analysis showed that 6 viruses were antigenically similar to A/Barn swallow/Hong Kong/D10-1161/2010, while A/Environment/Qinghai/XN02032/2013 presented low reaction with all reference antisera.Conclusions The existence of highly pathogenic avian influenza H5N1viruses in poultry related environments of surrounding areas of Qinghai Lake was confirmed.The status of highly pathogenic avian influenza H5N1 viruses with evolution diversity emphasized the necessary of strengthening the avian influenza surveillance in these areas.

Objective To investigate the effects of total nutrient admixture (TNA) on the recovery of patients with gastric cancer after radical gastrectomy.Methods The clinical data of 50 patients with gastric cancer who were admitted to the Affiliated Hospital of Luzhou Medical College between March 2013 and March 2014 were retrospectively analyzed.Among 50 patients receiving radical gastrectomy,26 patients receiving TNA were allocated to the experimental group and 24 patients receiving conventional fluid infusion were allocated to the control group.Patients in the experimental group received the nutritional support therapy using TNA at preoperative day 5 and at postoperative days 1-5,and patients in the control group received the postoperative intravenous rehydration including water,glucose,electrolyte,vitamins and micro elements.The nutritional indexes [albumin (Alb),prealbumin,transferrin and hemoglobin (Hb)],time to anal exsufflation,incidence of complications (wound infection,anastomotic leakage,blooding and intestinal obstruction) and duration of hospital stay were observed before nutritional support therapy and at postoperative day 8.The count data were analyzed using the chi-square test.The chi-square value of correction for continuity was used when 1 ≤ minimum theoretical frequency ≤ 5.The measurement data with normal distribution were presented as (x) ±s and analyzed using the t test or repeated measures ANOVA.The ordinal data were analyzed by the analysis of variance.Results The Alb,prealbumin,transferrin and Hb in the experimental group were (38.6 ± 2.0) g/L,(281 ± 33) mg/L,(2.5 ± 0.9) g/L and (111 ± 20) g/L before nutritional support therapy and (38.2 ± 1.9) g/L,(277 ± 16) mg/L,(2.3 ± 1.1) g/L and (112 ± 37) g/L at postoperative day 8,respectivley.The Alb,prealbumin,transferrin and Hb in the control group were (38.3 ±2.4) g/L,(287 ± 34) mg/L,(2.4 ± 1.1) g/L and (107 ± 21) g/L before nutritional support therapy and (30.3 ±2.3) g/L,(190 ± 41) mg/L,(1.6 ± 0.3) g/L and (93 ± 22) g/L at postoperative day 8,respectivley.There were significant differences in the nutritional indexes at postoperative day 8 between the 2 groups (F =174.042,95.637,9.529,4.919,P ＜ 0.05).The time to anal exsufflation in the experimental group were (52 ± 11) hours,which was significantly different from (70 ± 12) hours in the control group (t =-5.176,P ＜ 0.05).The incidence of complications was 15.4％ (4/26) in the experimental group,which was significantly different from 58.3％ (14/24) in the control group (x2=6.460,P ＜0.05).Patients with complications in the 2 groups were cured by anti-infective or symptomatic treatment.The duration of hospital stay was (9 ± 3) days in the experimental group and (12 ± 4) days in the control group,with a significant difference between the 2 groups (t =-2.912,P ＜ 0.05).Conclusion TNA can improve the nutritional status of patients after radical gastrectomy in a short time.It could help patients to get through the perioperative period smoothly,and enhance the postoperative recovery.