Construction and self-activation detection of yeast two-hybrid bait plasmid of human programmed cell death ligand 1 immunoglobulin variable region domain gene / 中国生物制品学杂志

@#Objective To construct a yeast two-hybrid recombinant bait plasmid of human programmed cell death ligand 1(PD-L1)immunoglobulin variable region(IgV)domain gene,detect its expression in yeast and detect the cytotoxicity and self-activation of PD-L1 IgV protein as well as the interaction between PD-L1 IgV and human thioredoxin(hTrx).Methods Human PD-L1 was analyzed by bioinformatics method,and primers were designed to amplify PD-L1 IgV domain based on the coding region of PD-L1 gene registered in NCBI GenBank database. PCR amplification was carried out with pENTERPD-L1 plasmid as template,and then cloned into yeast two-hybrid bait vector pGBKT7. The recombinant bait plasmid and pGBKT7 empty vector were transformed into Y2HGold yeast cells respectively,and the PD-L1 IgV gene and its expression were detected by PCR and Western blot;Meanwhile,the protein toxicity and self-activation of PD-L1 IgV were detected,and the interaction between PD-L1 IgV and hTrx was detected by drip plate method.Results The bioinformatics analysis results of PD-L1 were consistent with related reports. The recombinant bait plasmid pGBKT7-PD-L1 IgV was correctly constructed,and Y2HGold positive clone was obtained,in which PD-L1 IgV was stably expressed. The empty vector pGBKT7 and recombinant bait plasmid pGBKT7-PD-L1 IgV grew well on SD/-Trp and SD/-Trp/X-α-Gal plates with the same colony size and number and white colony,but they did not grow on SD/-Trp/X-α-Gal/AbA plates,which indicated that PD-L1 IgV protein had no toxicity and no self-activation effect on yeast. The results of drip plates test showed that all experimental groups grew well on SD/-Trp/-Leu plate,while only positive control group grew on SD/-Trp/-Leu/X-α-Gal/AbA plate and showed blue color,which indicated that bait protein PD-L1 IgV and hTrx did not self-activate,and there was no interaction between them.Conclusion Recombinant human PD-L1 IgV bait plasmid was successfully constructed. PD-L1 IgV protein showed no toxicity and self-activation effect on yeast cells,and there was no interaction between PD-L1 IgV and hTrx. Subsequently,hTrx can be used to construct a peptide aptamer library,from which peptide aptamers that specifically bind to PD-L1 IgV can be screened.

Comparison of arterial stiffness and ultrasound indices in patients with and without chronic obstructive pulmonary disease

SUMMARY OBJECTIVE: The purpose of this study was to compare arterial stiffness and ultrasound indices in patients with and without chronic obstructive pulmonary disease. METHODS: In our retrospective study, 83 chronic obstructive pulmonary disease patients were assigned to the chronic obstructive pulmonary disease group and 80 healthy controls were enrolled. Pearson's correlation analysis software was used to analyze the correlation between arterial stiffness (including brachial ankle pulse wave velocity and ankle-brachial blood pressure index) and ultrasound index (including resistance index, pulsatility index, and intima-media thickness) at the carotid artery in chronic obstructive pulmonary disease patients. RESULTS: The ultrasound resistance index and pulsatility index level of chronic obstructive pulmonary disease group were lower than those of control group (t=6.326, 8.321, p<0.001). Compared with the control group, the chronic obstructive pulmonary disease group had higher intima-media thickness, total plaque area, and number of plaques (t=4.574, 7.493, 5.093, p<0.001). The arterial stiffness and ankle-brachial blood pressure index level in the chronic obstructive pulmonary disease group were higher than those in the control group (t=6.392, 5.109, p<0.001). Moreover, arterial stiffness in patients with chronic obstructive pulmonary disease was negatively correlated with the ankle-brachial blood pressure index, resistance index, and pulsatility index levels (p<0.05), while it is positively correlated with intima-media thickness, total plaque area, and number of plaques (p<0.05). CONCLUSION: Our results indicated that patients with chronic obstructive pulmonary disease have stiffer arteries compared with healthy control subjects; the ultrasound index could be used as an auxiliary indicator for clinical prediction of arterial stiffness, which is helpful to improve the accuracy of prediction and thus better guide clinical interventions in high-risk groups of chronic obstructive pulmonary disease in time.

Structural equation model analysis of substance addiction, T lymphocyte subsets and depression in methamphetamine withdrawal patients / 中华行为医学与脑科学杂志

Objective:To investigate the relationship between substance addiction, T lymphocytes and depression in methamphetamine (MA) withdrawal patients.Methods:A total of 105 men who met the inclusion criteria were selected from compulsory isolation drug rehabilitation center of Baini lake, Hunan Province.All participants were suveyed by desires for drug questionnaire(DDQ) and self-rating depression scale (SDS), and the function of T lymphocyte subsets were detected by flow cytometry.All data were managed and analyzed using the SPSS 21.0 and AMOS 21.0 statistical software packages.Results:(1) The respondents were mainly junior high school or below (81.9%), unmarried and divorced (67.6%), unemployed (55.2%), hot suction (47.6%), and careless friends (32.4%), and 67.7% of MA abstinence had depressive symptoms. (2) SDS was positively correlated with desire and intention(DI), negative reinforcement (NR) and CD8+ %( r=0.408-0.897, all P<0.01), while negatively correlated with medication control, CD3+ %, CD4+ %, and CD4+ /CD8+ ( r=-0.792--0.263, all P<0.01). DI had significant positive correlation with CD8 + %( r=0.216, P<0.05), and negative correlation with CD4+ /CD8+ ( r=-0.217, P<0.05). NR had significant positive correlation with CD8 + %( r=0.259, P<0.05), and had significant negative correlations with CD3+ %, CD4+ %, and CD4+ /CD8+ ( r=-0.275-0.200, all P<0.05). Medication cotrol had a significant negative correlation with CD8+ %( r=-0.363, P<0.05), and significant positive correlations with CD4+ % and CD4+ /CD8+ ( r=0.288, 0.261, both P<0.05). (3)The model fitting index showed that DI, NR and medication control had significant direct effects on T lymphocyte subsets (all P<0.05). DI, medication control and T lymphocyte subsets had significant direct effects on self-evaluation of depressive symptoms (all P<0.05). NR, DI and medication control indirectly affected depressive symptoms through T lymphocyte subsets (all P<0.05). There were three path relationships: ① DI indirectly affected SDS via T lymphocyte subsets; ② Medication control indirectly affected SDS via T lymphocyte subsets; ③ NR indirectly affected SDS via T lymphocyte subsets. Conclusion:The structural equation model suggests that the degree of addiction in MA abstinence affecting depressive mood may be related to hypofunctional T lymphocyte subsets and provide methods for the prevention and treatment of drug addiction.

Treatment efficacy of MUC1-DC vaccine in breast cancer MCF-7 cell xenografts in nude mice evaluated by monitoring tumor growth with optical molecular imaging / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：评价黏蛋白1（mucin 1，MUC1）基因转染的DC疫苗治疗乳腺癌MCF-7细胞裸鼠移植瘤的效果及其可能的机制。方法：采用GFP慢病毒转染MCF-7细胞获得GFP-MCF-7细胞，皮下种植于BALB/c裸鼠，成瘤后随机分为3组。各组裸鼠首先尾静脉注射体外活化的CIK细胞（1×108个/只），治疗组于皮下注射MUC1基因转染的MUC1-DC（MUC1-DC组）或DC（DC组）（0.2 ml，1×107个/只），对照组（Control组）注射等体积生理盐水，每天治疗1次，连续5 d；采用小动物活体光学成像系统在开始治疗前及治疗后第35天观察移植瘤荧光成像，分析荧光强度和荧光面积；并采用免疫组化法检测瘤组织中Caspase 3的表达、TUNEL法检测细胞凋亡率。结果：GFP-MCF-7接种后7 d，成瘤率100%；光学分子成像法监测结果显示，治疗前MUC1-DC组、DC组和Control组之间体内移植瘤荧光信号强度无明显差异（P>0.05）；第35天，MUC1-DC组的荧光信号强度明显低于Control组（P<0.05）；DC与Control、MUC1-DC与DC组间均无显著性差异（均为P>0.05），但MUC1-DC比DC组荧光信号更低；治疗前MUC1-DC组、DC组和Control组之间体内移植瘤荧光信号分布面积无显著差异（P>0.05）；第35天，Control组荧光信号呈多处散在分布，MUC1-DC组和DC组的荧光信号面积均明显低于Control组（均为P<0.01），MUC1-DC组的荧光信号面积低于DC组，但无显著差异（P>0.05）；Control组Caspase 3表达最少，DC组次之，MUC1-DC组呈高表达Caspase 3，3组间差异均有统计学意义（均为P<0.05）；3组细胞凋亡率分别为：Control组（4.11±2.61）%、DC组（9.63±2.27）%、MUC1-DC组（25.30±8.24）%，3组间差异均有统计学意义（均P<0.05）。结论： MUC1-DC疫苗比单纯DC免疫治疗人乳腺癌荷瘤小鼠能够更有效地抑制肿瘤的生长和扩散，发挥了更好的促进肿瘤细胞凋亡的作用。

Current status of surgical treatment for traumatic arthritis / 解放军医学杂志

Traumatic arthritis (TA) is one of the common diseases of bone and joint caused by trauma. The main pathological changes are degeneration of articular cartilage and secondary hyperplasia and ossification of cartilage. The main clinical manifestations are joint pain and dysfunction of movement. Its pathogenesis is still unclear. At present, the treatment of TA is based mainly on symptoms rather than etiology, including physical therapy, drug treatment, surgical treatment, etc. Conservative treatment (physical therapy, drug treatment) can only alleviate short-term pain, and the long-term effect is not satisfactory. Thus, patients with middle and late TA tend to choose surgical treatment. At present, the surgical treatment of TA includes arthroscopic debridement, arthrodesis, cartilage repair, osteotomy, artificial joint replacement, 3D printing technology, etc. There are differences in the postoperative efficacy. This article reviews the current situation of surgical treatment for TA.

Correlation between the ability of pull-up and the composition of upper limbs among male college students in Guangxi / 中国学校卫生

Objective@#To explore the correlation between pull-up ability and upper body composition of male college students in a college in Guangxi, and to provide scientific guidance for college students’ exercise.@*Methods@#A total of 685 male college students were randomly selected from a college in Guangxi.Pull up tests were implemented according to the national physical health test standards. Measurements of muscle mass, fat mass, fat percentage, etc. of the upper limbs were conducted by using the Ogilvy Body Composition Meter (TANITA MC-180). Data entry and analysis were performed by using SPSS 23.0.@*Results@#The pass rate of male college students in the region was 21.7%; There was a statistically significant difference in the fat mass and percentage of body fat between males with different pull-up ability (F=11.30，14.18，12.91，15.22，P<0.01).After controlling age, height, weight and BMI, partial correlation analysis showed that there was a negative correlation between the pull-up ability of male college students and the fat mass and limb fat rate of both upper limbs(r=-0.22, -0.33, -0.31, -0.38, P<0.01).@*Conclusion@#The ability of male students in Guangxi to pull up is needed to be improved. Pull-up exercises can reduce fat mass and fat percentage in the upper limbs and improve body composition.

Analysis of mental state of allergic rhinitis patients in Chengdu city by symptom check list 90 (SCL-90) scale / 中华耳鼻咽喉头颈外科杂志

Objective@#To analyse the mental state of patients with allergic rhinitis (AR) in Chengdu.@*Methods@#One thousand five hundred and thirty-six AR patients from Sichuan Provincial Integrated Traditional Chinese and Western Medicine Hospital, West China Hospital of Sichuan University, Affiliated Hospital of Chengdu University of Traditional Chinese Medicine, Sichuan People′s Hospital, Sichuan Second Hospital of Traditional Chinese Medicine were selected from July 2013 to January 2018. Eight hundred and twenty-seven patients were screened into study group by inclusion and exclusion standards. The symptom check list 90 (SCL-90) was used to group and score the mental state of these patients according to nine classification criteria: gender, BMI, age, marital status, monthly salary, disease duration, living environment, education level and working environment. Then, the scores were compared within groups. Inter-group comparison was made between the study group and the Chinese norm, and the positive factors for psychological disorders were extracted. Four symptoms in the study group, i.e. nasal itching, sneezing, clear discharge and nasal congestion, were scored on the visual analogue scale (VAS). SPSS 19.0 software was used to carry out statistical analysis. Partial correlation analysis was performed between the positive factors and the symptom scores by multiple regression statistical method.@*Results@#The total score of SCL-90 in the study group was 2.64±0.25, which was accorded with mild to moderate mental health impairment. There were 124 (15.0%) without mental health damage, 176 (21.3%) with mild damage, 474 (57.3%) with mild to moderate damage, 41 (5.0%) with moderate to severe damage and 12 (1.4%) with severe damage. The in-group comparison showed that the top three categories of different items were the living environment, gender and working environment. The scores of somatization, obsessive-compulsive symptoms, interpersonal sensitivity, depression, anxiety, psychosis, other (sleep, diet) and total average score of urban residents were higher than that of country residents (3.29±0.61 vs 2.65±0.50, 2.81±0.77 vs 2.05±0.38, 3.10±0.19 vs 2.49±0.67, 3.40±0.84 vs 2.49±0.70, 3.04±0.64 vs 2.33±0.51, 3.02±0.55 vs 2.40±0.77, 3.40±0.41 vs 2.52±0.77, 2.91±0.11 vs 2.29±0.40, Z value was 4.88, 5.25, 4.57, 5.91, 5.09, 4.63, 5.55, -4.55, respectively, all P<0.05). Women scored higher than man for somatization, interpersonal sensitivity, depression and others (2.66±0.51 vs 2.00±0.45, 3.37±0.47 vs 2.63±0.51, 3.44±0.57 vs 2.85±0.52, 3.47±0.36 vs 2.76±0.45, Z value was -5.10, -5.51, -4.86, -5.28, respectively, all P<0.05). The scores of somatization, interpersonal sensitivity, psychosis and other (sleep, diet) were higher in the indoor group than those in the outdoor group (3.49±0.64 vs 2.78±0.46, 3.33±0.30 vs 2.56±0.68, 3.28±0.60 vs 2.67±0.31, 3.50±0.85 vs 2.85±0.37, Z value was 5.31, 5.79, 4.89, 5.00, respectively, all P<0.05). The outdoor group scored higher on obsessive-compulsive symptoms, anxiety and hostility (3.44±0.40 vs 2.83±0.35, 3.40±0.50 vs 2.57±0.93, 3.34±0.88 vs 2.69±0.56, Z value was 4.96, 6.22, 5.08, respectively, all P<0.05). The inter-group comparison found that depression, anxiety, psychosis and other (sleep, diet) could be partially correlated with VAS scores as 4 positive factors. The results of partial correlation analysis showed that depression was positively correlated with sneezing and nasal runny discharge, anxiety was positively correlated with nasal itching and nasal obstruction, psychosis was positively correlated with nasal itching and sneezing, and other (sleep, diet) was positively correlated with nasal runny discharge and nasal obstruction.@*Conclusion@#AR patients have mild to moderate mental health impairments, which are correlated with AR symptoms.

Pulmonary vein parameters are similar or better predictors than left atrial diameter for paroxysmal atrial fibrillation after cryoablation

Left atrial diameter (LAD) has been considered an independent risk factor for atrial fibrillation (AF) relapse after pulmonary vein isolation (PVI). However, whether LAD or other factors are more predictive of late recurrence in patients with paroxysmal AF remains unclear. We aimed to evaluate the value of pulmonary vein (PV) parameters for predicting AF relapse 1 year after patients underwent cryoablation for paroxysmal AF. Ninety-seven patients with paroxysmal AF who underwent PVI successfully were included. PV parameters were measured through computed tomography scans prior to PVI. A total of 28 patients had recurrence of AF at one-year follow-up. The impact of several variables on recurrence was evaluated in multivariate analyses. LAD and the time from first diagnosis of AF to ablation maintained its significance in predicting the relapse of AF after relevant adjustments in multivariate analysis. When major diameter of right inferior pulmonary vein (RIPV) (net reclassification improvement (NRI) 0.179, CI=0.031-0.326, P<0.05) and cross-sectional area (CSA) of RIPV (NRI: 0.122, CI=0.004-0.240, P<0.05) entered the AF risk model separately, the added predictive capacity was large. The accuracy of the two parameters in predicting recurrence of AF were not inferior (AUC: 0.665 and 0.659, respectively) to echocardiographic LAD (AUC: 0.663). The inclusion of either RIPV major diameter or CSA of RIPV in the model increased the C-index (0.766 and 0.758, respectively). We concluded that major diameter of RIPV had predictive capacity similar to or even better than that of LAD for predicting AF relapse after cryoablation PVI.

Ginsenoside Rb1 attenuates diabetic retinopathy in streptozotocin-induced diabetic rats

Abstract Purpose: To investigated the effects of ginsenoside Rb1 on diabetic retinopathy in streptozotocin-induced diabetic rats. Methods: Diabetes was induced by a single intraperitoneal injection of streptozotocin (80 mg/kg) in male Wistar rats. Ginsenoside Rb1 (20, 40 mg/kg) was injected (i.p.) once a day for 4 weeks. Then, using fundus photography, the diameter and vascular permeability of retinal vessels were investigated. Retinal histopathology was undertaken. Contents of malondialdehyde (MDA) and glutathione (GSH) in retinas were assayed. Levels of nuclear factor erythroid 2-related factor 2 (Nrf2), glutathione cysteine ligase catalytic subunit (GCLC), and glutathione cysteine ligase modulatory subunit (GCLM) were measured. Results: Treatment with ginsenoside Rb1 attenuated the diabetes-induced increase in the diameter of retinal blood vessels. Ginsenoside Rb1 reduced extravasation of Evans Blue dye from retinal blood vessels. Ginsenoside Rb1 partially inhibited the increase in MDA content and decrease in GSH level in rat retinas. Nrf2 levels in the nuclei of retinal cells and expression of GCLC and GCLM were increased significantly in rats treated with ginsenoside Rb1. Conclusion: These findings suggest that ginsenoside Rb1 can attenuate diabetic retinopathy by regulating the antioxidative function in rat retinas.

Immunosuppressive mechanism of Hypoderma lineatum secreted serine esterase, a potential modulatory method used to inhibit transplant rejection

Background: Although immunosuppressive therapies have made organ transplantation a common medical procedure worldwide, chronic toxicity has a major issue for long-term treatment. One method to improve therapies and methods is the application of immunomodulatory agents from parasites such as Hypoderma lineatum. Hypodermin A (HA) is a serine esterase secreted by the larvae of Hypoderma lineatum, several studies demonstrated its immunosuppressive mechanism in vitro, and recently we discovered that HA inhibits the expression of interferon (IFN)-γ and interleukin (IL)-2 and activates IL-10 expression. Therefore, we hypothesized that it might be a potential agent used to block allograft rejections. However, most studies of the immunosuppressive mechanisms associated with HA were undertaken at the cellular level. In order to augment these studies, we evaluated the immunosuppressive effects of HA in vivo using an HA transgenic mouse model. Result: Our results revealed similar findings to those reported by in vitro studies, specifically that HA induced prostaglandin E2 expression, downregulated IFN-γ and IL-2 expression, and promoted IL-10 secretion via E-type prostanoid receptor 4. Additionally, we observed that HA overexpression inhibited lipopolysaccharide-induced TLR4 activation. These findings provide insight into a new potential agent capable of blocking graft rejection. Conclusion: Our founding suggested that HA-related treatment could be a promising option to improve the viability of grafts in human.

extract of Perilla stem inhibits Src homology phosphatase-1 [SHP]-1 and influences insulin signaling

Protein tyrosine phosphatases [PTPs] are enzymes that catalyze protein tyrosine dephosphorylation of which Src homology phosphatase-1 [SHP-1] is one of the best-validated, a widely distributed intracellular tyrosine phosphatase that contains two SH2 domains. Down regulation of SHP-1 tyrosine phosphatases was significantly increased sensitivity to insulin in insulin signaling pathway. Through in vitro enzymatic reaction kinetics experiment, we found that the extract of Perilla stem was a potential inhibitor to [delta]SHP-1, the catalytic domain of SHP-1 protein tyrosine phosphatase, and its IC[50] was 4ug/ml, and was more sensitive towards SHP-1than other PTPs, which indicated that SHP-1 might be a target of the extract of Perilla stem. It can strengthened the level of tyrosine phosphorylation of insulin receptor [IR] and extracellular signal-regulated protein kinase [ERK] in HepG2 cells, and then activated the insulin signaling pathway through inhibiting the protein phosphorylation of SHP-1. These results demonstrated that the extract of Perilla stem could play an important role for diabetes treatment through inhibiting the level of SHP-1 in insulin signaling pathway

Effect of ω-3 Polyunsaturated Fatty Acid on Toll-like Receptors in Patients with Severe Multiple Trauma / 华中科技大学学报（医学）（英德文版）

This study examined the effects of ω-3 polyunsaturated fatty acid (ω-3PUFA) on the expression of toll-like receptor 2 (TLR2),toll-like receptor 4 (TLR4) and some related inflammatory factors in peripheral blood mononuclear cells (PBMCs) of patients with early-stage severe multiple trauma.Thirty-two patients who were admitted to the Department of Traumatic Surgery,Tongji Hospital (Wuhan,China) between May 2010 and November 2010,and diagnosed as having severe multiple trauma with a injury severity score (ISS) no less than 16,were enrolled in the study and divided into two groups at random (n=16 in each):ω-3PUFA group and control group in which routine parenteral nutrition supplemented with ω-3PUFA or not was administered to the patients in two groups for consecutive 7 days.Peripheral blood from these patients was collected within 2 h of admission (day 0),and 1,3,5 and 7days after the nutritional support.PBMCs were isolated and used for detection of the mRNA and protein expression of TLR2 and TLR4 by using real-time PCR and flow cytometry respectively,the levels of NF-κB by quantum dots-based immunofluorescence assay,the levels of TNF-α,IL-2,IL-6 and COX-2 by ELISA,respectively.The results showed that the mRNA and protein expression of TLR2 and TLR4 in PBMCs was significantly lower in ω-3PUFA group than in control group 5 and 7 days after nutrition support (both P＜0.05).The levels of TNF-α,IL-2,IL-6 and COX-2 were found to be substantially decreased in PBMCs in ω-3PUFA group as compared with control group at 5th and 7th day (P＜0.05 for all).It was concluded that ω-3PUFA can remarkably decrease the expression of TLR2,TLR4 and some related inflammatory factors in NF-κB signaling pathway in PBMCs of patients with severe multiple trauma,which suggests that ω-3PUFA may suppress the excessive inflammatory response meditated by the TLRs/NF-κB signaling pathway.

Gene transfection efficiency and destination of recombinant adenovirus gene transfer through internal jugular vein injection in rats / 中南大学学报(医学版)

OBJECTIVE@#To investigate the adenovirus-mediated LacZ gene expression and the destination in different organs of SD rats after the intravenous injection in rats.@*METHODS@#Recombinant adenovirus vector containing LacZ was transferred to SD rats by injecting into the internal jugular vein. To identify the sites and periods of LacZ gene expression, X-gal staining was used to detect beta-gal level and period of LacZ gene expression of different organs in the transfected and non-transfected rats at different time intervals.@*RESULTS@#On the 1st day after the injection, the lung, liver, kidney, and spleen expressed some beta-gal; on the 3rd day after the injection, the lung, liver, kidney, and spleen expressed beta-gal obviously; their peak levels were on the 7th day; the beta-gal level decreased on the 14th day; beta-gal expression disappeared in the most organs except the lungs on the 28th day. In all animals, the brain did not express any beta-gal.@*CONCLUSION@#The adenovirus-mediated exogenous gene transfer in the internal jugular vein may be an effective approach of gene therapy in some diseases in the lung, liver, and kidney.