Objective To construct a lentivirus-based expression plasmid and gene knockout plasmid of human interleukin(IL)-26 so as to lay a foundation of studying the function of IL-26 gene in cell signaling pathway and autophagy.Methods IL-26 gene sequence was amplified from human peripheral blood mononuclear cells by RT-PCR and cloned into pCDH-CMVMCS-EF1-copGFP eukaryotic expression vector to construct overexpression plasmid;Four knockout targets,Exon1sgRNA1,Exon1sgRNA2,Exon3sgRNA1 and Exon3sgRNA2,were designed based on the exon sequence of IL-26,and constructed into lentiCRISPRv2 vector by CRISPR/Cas9 technology to construct gene knockout plasmid. The overexpression plasmid and gene knockout plasmid were transiently transfected into HEK293T cells respectively,and the expression of IL-26 was verified by RT-qPCR and Western blot. In addition,amino acid sequence analysis,structure prediction and subcellular localization observation of IL-26 were performed.Results The results of restriction digestion,sequencing and bioinformatics analysis showed that IL-26 was 516 bp in length,encoding 171 amino acids. The IL-26 mRNA level and protein level of HEK293T cells transfected with IL-26 overexpression plasmid increased by 656. 789 times and 1. 978 times respectively with significant differences as compared with the normal control group(t = 17. 976 and 7. 859,P < 0. 000 1 and < 0. 001,respectively). With the transfection of 4 knockout targets Exon1sgRNA1,Exon1sgRNA2,Exon3sgRNA1 and Exon3sg-RNA2 into HEK293T cells,the expression of IL-26 decreased by 0. 930,0. 980,0. 523 3 and 0. 316 9 times,respectively,among which Exon3sgRNA2 significantly down-regulated the expression of IL-26(t = 7. 440,P < 0. 001). IL-26protein showed signal peptide structure and certain transmembrane function in the first 22 amino acids,which existed in cytoplasm.Conclusion IL-26overexpression and gene knockout plasmids were successfully constructed,which laid a foundation of the follow-up study of the function of IL-26.

LCH-2011 regimen)was referenced,and the targeted drug Dabrafenib was added to treat those with the BRAF-V600E mutation.The clinical characteristics,BRAF-V600E mutation status,and treatment response between the single-system LCH(SS-LCH)group and the multisystem LCH(MS-LCH)group were summarized.Survival curves were plotted using the Kaplan-Meier method,and the Log-Rank test was used to compare the survival rates between the two groups.Results This study included a total of 31 cases,with 18 males and 13 females.The median age of onset was 10 months(ranging from 1 to 84 months).9 cases were SS-LCH,and 22 cases were MS-LCH,with 5 cases experiencing pituitary involvement/diabetes insipidus.Among the 27 cases that underwent BRAF-V600E mutation testing,20 were positive(3 cases in the SS-LCH group,with a positivity rate of 37.5%;17 cases in the MS-LCH group,with a positivity rate of 89.5%).The difference in the BRAF-V600E mutation positivity rate between the two groups was statistically significant(P = 0.011).The median follow-up time was 24 months(ranging from 3 to 62 months).The effective rate after 6 weeks of induction chemotherapy was 88.9%in the SS-LCH group(8/9)and 81.8%in the MS-LCH group(18/22).The observed progression-free rate at the end of the observation period reached 29.0%(9/31).All three deaths occurred in the MS-LCH group with involvement of high-risk organs.There was no statistically significant difference in the overall survival rate between the SS-LCH and MS-LCH groups,as well as between the BRAF-V600E mutation positive and negative groups(χ2 = 1.156,0.437;P = 0.282,0.508).Conclusion LCH in children is more common in infants and young children,with a high incidence of BRAF-V600E gene mutation in affected children,and is often seen in MS-LCH.Dabrafenib may help improve the prognosis of children with BRAF-V600E mutation.

Objective To investigate the rationality and efficacy of enhanced recovery after surgery (ERAS) in liver transplant recipients. Methods Clinical data of 465 liver transplant recipients were retrospectively analyzed. All recipients were divided into the ERAS group (n=163) and control group (n=302) according to whether they received ERAS. The severity of disease in the ERAS group was worse than that in the control group. Operation situations including the operation time, anhepatic phase and intraoperative blood transfusion volume of the liver transplant recipients were observed and recorded. Postoperative recovery conditions including the length of intensive care unit (ICU) stay, total length of hospital stay, total ventilator removal time at postoperative 28 d and postoperative re-intubation rate were recorded. The survival rates at 90 d, 180 d and 1 year after liver transplantation were calculated. The influencing factors of survival rate of liver transplant recipients were analyzed. Results The anhepatic phase in the ERAS group was 45 (39, 53) min, significantly longer than 40 (32, 48) min in the control group (P < 0.05). The volume of erythrocyte infusion in the ERAS group was 10 (7, 13) U, significantly less than 18 (10, 28) U in the control group (P < 0.05). The length of postoperative ICU stay and total length of hospital stay in the ERAS group were 135 (84, 212) h and 24 (18, 33) d, significantly shorter than 154 (103, 253) h and 34 (20, 50) d in the control group (both P < 0.05). Total ventilator removal time at postoperative 28 d was 26 (25, 27) d, significantly longer than 26 (23, 27) d in the control group (P < 0.05). The postoperative re-intubation rate in the ERAS group was 11.0%, significantly lower than 20.8% in the control group (P < 0.05). The 90 d, 180 d and 1-year survival rates in the ERAS group were 92.8%, which were significantly higher than 81.1%, 78.1% and 75.7% in the control group (all P < 0.05). ERAS and operation time were the independent influencing factors of survival rate of liver transplant recipients (both P < 0.05). Conclusions ERAS after liver transplantation can improve the survival rate of recipient, shorten the length of hospital stay, reduce the re-intubation rate and accelerate the rehabilitation after liver transplantation.

Objective To investigate the effect of human umbilical cord mesenchymal stem cells (HUC-MSCs) on CD4+T cells in liver after hepatic ischemia-reperfusion injury (HIRI) in mice. Methods Two hundred and twenty-five mice were randomly divided into sham group, control group and MSC group, with 75 mice in each group. HIRI model mice were used in MSC group and control group. HUC-MSCs were injected in MSC group through inferior vena cava. Normal saline was injected in control group through inferior vena cava. Only laparotomy and abdominal closure were performed in sham group without blood vessel clipping. At 6, 12 and 24 h after operation, 15 mice of each group were randomly selected to sample eyeball blood and liver tissues, and the 30 mice left in each group were used to extract intrahepatic mononuclear cells. The number of intrahepatic mononuclear cells, percentage, number and positive rate of CD4+T cells in the mice of various groups at different time points were compared. The content of interleukin (IL)-17 in serum and liver tissue as well as expression levels of costimulatory molecules B7-1 and B7-2 messenger RNA (mRNA) in liver tissues of the mice at different time points were compared. Results At 12 and 24 h after operation, the number of intrahepatic mononuclear cells of control group was significantly higher than that of sham group, while the number of intrahepatic mononuclear cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the percentage, number and positive rate of CD4+T cells of control group were significantly higher than those of sham group (all P<0.01), while the percentage of CD4+T cells of MSC group was significantly lower than that of control group (P<0.01-0.05). At 12 and 24 h after operation, the number and positive rate of CD4+T cells of MSC group were significantly lower than those of control group (P<0.01-0.05). At 6, 12 and 24 h after operation, the IL-17 contents in serum and liver tissues of control group were higher than those of sham group (all P<0.01), while the IL-17 contents in serum and liver tissues of MSC group were lower than those of control group (all P<0.01). At 6 h after operation, the mRNA expression level of B7-2 of control group was higher than that of sham group (P<0.05). At 12 and 24 h after operation, the mRNA expression levels of B7-1 and B7-2 of control group were higher than those of sham group (all P<0.01), while the mRNA expression levels of B7-1 and B7-2 of MSC group were lower than those of control group (all P<0.01). Conclusions HUC-MSCs inhibits the number of CD4+T cells and the secretion of IL-17 in liver after HIRI, as well as decreases the number of intrahepatic mononuclear cells and the mRNA expression of B7-1 and B7-2, thereby alleviating HIRI.

A 53-year-old male patient presented with hypopsia of his right eye for 2 months and lower extremities weakness for 8 days. Thoracic MRI demonstrated a lesion at T3 level appearing as hyperintense on T2-weighted images with non-enhancement by contrast medium and demyelinating lesion was considered. Aquaporin-4-Ab was positive and the antibody titer was 1:320 in serum. The diagnosis of neuromyelitis optica spectrum disorders was made. In addition, systemic lupus erythematosus and thymoma coexisted in this patient. After methylprednisolone impact treatment, plasma exchange and immunosuppressive therapy, the right vision and lower extremities weakness of the patient were improved.
Anti-Inflammatory Agents ; therapeutic use ; Antibodies ; blood ; Aquaporin 4 ; immunology ; Humans ; Lupus Erythematosus, Systemic ; complications ; Male ; Methylprednisolone ; therapeutic use ; Middle Aged ; Neuromyelitis Optica ; complications ; diagnostic imaging ; drug therapy ; Thymoma ; complications ; Treatment Outcome

[Objective] To investigate the mesenchymal stem cells (MSC) in treating acute lung injury (ALI) via ALI mouse model.[Methods] By monoclonal antibody Anti-GD2 of specific antigen ganglioside (GD2) only expressed on MSC as a carrier,new fluorescent molecule probe were synthesized through covalently coupling Anti-GD2 and fluorescent group CyDye mono-reactive NHS Esters (Cy7).Synthetic Anti-GD2-Cy7 and MSC were labeled by the specific binding of antigen and antibody in vitro.Total 84 balb/c male mice were selected and randomly selected 48 mice were divided into three groups:sham group (n =16),MSC+ ALI group (n =16),NS + ALI group (n =16).The lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature were examined at 24 h,48 h after ALI mouse model.Other 36 mice were randomly divided into three groups:normal group (n =12),sham group(n =12),MSC +ALI group(n =12).Labeled MSC-GD2-Cy7 were transplanted into MSC+ALI group and sham group mice via tail vein injection.At 30 min,1 d,3 d,and 7 d post-MSC-GD2-Cy7 injection,the mice were sacrificed after anesthesia and then the lung was removed.Excised lung was detected on small animal fluorescent imager.[Results] Contrast to NS+ ALI group,the lung histopathology and scores,lung W/D ratio and permeability of lung microvasculature of MSC +ALI group were more greatly improved at both 24 h and 48 h.Fluorescent results showed that the signal intensity in thc lung of MSC +ALI group was significantly higher than that of sham group at each time point [(3.37 ± 0.02)× 10-4 vs (2.05 ± 0.04) × 10-4 scaled counts/s;(35.54 ± 0.47)× 10-4 vs (25.29 ± 1.48) × 10-4 scaled counts/s;(11.17 ± 0.75)×10-4 vs (6.09 ± 0.62)× 10-4 scaled counts/s;(3.10 ± 0.14) vs (0.00 ± 0.00)× 10-4 scaled counts/s;all P ＜ 0.05].[Conclusion] Our study showed that a proportion of cells migrated into normal and injured lungs 30 min after cell transplantation,and the cells started to recruit and largely gather in injured lungs at day 1 and persisted to day 7,these results suggest that MSC possess the ability to home into injured tissues.

Objective:To explore clinical significance of sinus heart rate turbulence (HRT) phenomenon in aged pa‐tients with stable angina pectoris (SAP) .Methods :A total of 120 aged SAP patients ,who received 24h DCG in our hospital from Jan 2013 to Oct 2015 ,were selected as SAP group .Meanwhile ,another 144 aged patients ,who re‐ceived 24h DCG examination simultaneously and coronary angiography results were normal ,were regarded as nor‐mal control group .According to coronary lesion severity ,SAP group was further divided into single vessel coronary disease group (single vessel group ,n=35) ,double‐vessel coronary disease group (double‐vessel group ,n=48) and multi‐vessel coronary disease group (multi‐vessel group ,n=37) .The 24h DCG ,HRT indexes ,including turbulence onset (TO) and turbulence slope (TS) ,were measured and compared among all groups .Results:Compared with normal control group ,there was significant rise in TO [(0.77 ± 0.37)% vs .(1.26 ± 0.92)% ] and significant reduc‐tion in TS [(5.45 ± 4.02) ms/RR interval vs .(1.53 ± 0.70) ms/RR interval] ,P<0.01 both ;significant rise in ab‐normal rates of TO (19.44% vs .42.50% ) ,TS (15.97% vs .31.67% ) and TO + TS (11.11% vs .30.83% ) in SAP group ,P<0.01 all .Compared with single vessel group ,there was significant rise in TO [(0.66 ± 0.22)% vs .(1.28 ± 1.11)% vs .(1.46 ± 1.20)% ] and significant reduction in TS [ (2.04 ± 0.82) ms/RR interval vs .(1.66 ± 0.38) ms/RR interval vs .(1.29 ± 0.58) ms/RR interval] in double‐vessel group and multi‐vessel group ,and TO of multi‐vessel group was significantly higher than that of double‐vessel group ,TS of multi‐vessel group was significantly low‐er than that of double‐vessel group , P<0.01 all .Conclusion:Sinus heart rate turbulence can be used as risk predic‐tor for aged patients with stable angina pectoris ,which can provide basis for clinical effective treatment and progno‐sis assessment .

Objective To explore the change of glyoxalase I in type 2 diabetic ocular muscles palsy (DOMP) and its associations with advanced oxidation protein products (AOPP) and oxidative stress. Methods 58 DOMP patients, 50 T2DM and 30 normal controls were enrolled in this study. Levels of blood lipids, fasting blood glucose, hemoglobin A1c, insulin, serum glyoxalase I, AOPP, malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) were measured. Homeostasis model assessment was performed to evaluate the status of insulin resistance (IR). Results Levels of high-density lipoprotein cholesterol, SOD and T-AOC were positively correlated with glyoxalase I and inversely associated to AOPP. Levels of triglycerides , low-density lipoprotein cholesterol , fasting blood glucose , hemoglobin A1c , IR and MDA were negatively correlated with glyoxalase I and positively related to AOPP. AOPP had an inverse association with glyoxalase I (r = -0.823, P < 0.001). Multivariate regression analysis showed that serum levels of glyoxalase I (Sβ = 0.554) and AOPP (Sβ= -0.469) were influencing factors of groups. Conclusion Serum glyoxalase I levels were significantly decreased in DOMP and correlated with AOPP and levels of oxidative stress , which suggest that glyoxalase I could play crucial roles on the development of DOMP.

Objective To investigate the effects of advanced oxidation protein products (AOPP) in type 2 diabetic ocular muscles palsy (DOMP). Methods 58 DOMP patients and 50 type 2 diabetes patients were included in the research. Hemoglobin A1c (HbA1c), blood glucose (FPG), fasting insulin (FINS) and triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL) were measured and recorded. Homeostasis model assessment was performed to evaluate the status of insulin resistance (HOMA-IR), basal insulin secretion (HOMA-β) and the insulin sensitivity index (ISI). Serum AOPP was measured by enzyme linked immunosorbent assay. Unconditional logistic regression model was used to evaluate the influencing factors of DOMP. Results The DOMP group showed higher levels of plasma AOPP, TG, LDL, FPG, FINS, HbA1c and HOMA-IR, but lower levels of HDL, HOMA-β and ISI than those of the T2DM group. Unconditional logistic regression analysis revealed that AOPP was an independent risk factors for DOMP (OR =3.01, P = 0.002). Conclusion AOPP may be involved in the pathogenesis of DOMP. AOPP could be a useful indicator for monitoring the development of DOMP and for evaluating its severity.

Objective To explore the effect of transitional care model (TCM) on the prevention of complications in patients with severe cerebral infarction. Methods Sixty patients with severe cerebral infarction were randomly divided into the experiment group and the control group, 30 cases in each group. The control group was given conventional nursing and the observation group received TCM. The two groups were compared in terms of the incidence of complications and re-admission rate. Result The incidence of complications and the rate of re-admissions in the experiment group were significantly lower than those of the control group (P<0.05). Conclusion TCM can reduce the incidence of complications in patients with severe cerebral infarction, lower the incidence of hospital admission, relieve the pain of patients and improve the quality of life of patients.