Programmed death-1 (PD-1) and its ligand PD-L1 are the major members of inhibitory costimulatory molecules and express high in a variety of tumor cells and their associated cells surface,while the proportion of regulatory T cells (Tregs) are abnormally elevated in tumor infiltrating T lymphocytes cells.PD-L1 combined with PD-1 and Treg help tumors evade immune clearance,weaken immune responses and induce immune tolerance.New researches find that PD-L1 plays an important role in the development and function maintenance of inducible Treg (iTreg),and PD-L1 signal can change initial CD4 + CD25-T cells into iTreg to play a role of immunosuppression.Research on PD-1 signaling pathway can provide a new theoretical basis for the inhibition of tumor immune escape in clinical application of immunotherapy and better treatments.

Objective To evaluate the effect of lipopolysaccharide (LPS) on the viability of rat alveolar macrophages.Methods The rat alveolar macrophages were seeded in 96-well plate at a density of 4× 104/ml.After being cultured for 24 h, the cells were randomly divided into 6 groups (n =5 each) using a random number table : control group (group C), LPS 0.1 μg/ml group (group LPS0.1), LPS 1.0 μg/ml group (group LPS1.0), LPS 10.0 μg/ml group (group LPS10), LPS 5.0 μg/ml group (group LPS50), and LPS 100.0 μg/ml group (group LPS100).Phosphate buffer solution was added to the culture medium in group C, and LPS with the final concentrations of 0.1, 1.0, 10, 50.0 and 100.0 μg/ml were added to the culture medium in LPS0.1, LPS1.0, LPS10, LPS50, and LPS100 groups, respectively.At 6, 12, 24 and 48 h after addition of PBS or LPS, the cell viability was measured by methyl thiazolyl tetrazolium assay.Results Compared with group C, the viability of alveolar macrophages was significantly increased at 6 and 12 h after addition of LPS in the other five groups , and was decreased at 24 and 48 h after addition of LPS in groups LPS50and LPS100 (P＜0.05), and no significant change was found in LPS0.1, LPSL0 and LPS10 groups (P＞0.05).Conclusion Incubation with LPS 0.1-100.0 μg/ml for less than 12 h can enhance the viability of rat alveolar macrophages;incubation with LPS with the concentration ≥ 50.0 μg/ml for more than 24 h can decrease the cell viability.

Objective To evaluate the effect of electroacupuncture on endoplasmic reticulum stress in lung tissues of rats with acute lung injury (ALI) induced by endotoxin.Methods Forty healthy pathogen-free male Sprague-Dawley rats,aged 8 weeks,weighing 180-210 g,were divided into 4 groups (n=10 each) using a random number table:control group (group C),ALI group,electroacupuncture group (group E),and electroacupuncture at non-acupoint group (group NE).Lipopolysaccharide 5 mg/kg (in 0.5 ml normal saline) was injected intravenously to establish the model of endotoxin-induced ALI.Bilateral 30 min electroacupuncture stimulation of Zusanli and Neiguan acupoints was performed with the dispersedense wave (frequency 2/15 Hz,wave length 0.2-0.6 ms,intensity 1-2 mA) once a day (time for stimulation 9:30-10:30) for 4 consecutive days before and during establishment of the model in group E.Electroacupuncture was performed with the same parameters at the points 0.5 cm lateral to the acupoints of Zusanli and Neiguan in group NE.At 6 h after lipopolysaccharide injection,the rats were sacrificed,and lungs were removed for microscopic examination and for determination of wet to dry weight ratio (W/D ratio),apoptosis in alveolar epithelial cells and expression of glucose-regulated protein 78 (GRP78) and CCAAT/enhancer-binding protein homologous protein (CHOP) and caspase-12 in lung tissues (by Western blot).The pathological changes of lungs were scored.Apoptosis index (AI) was calculated.Results Compared with group C,lung injury scores,W/D ratio and AI were significantly increased,and the expression of GRP78,CHOP and caspase-12 in lung tissues was up-regulated in the other three groups (P＜0.05).Compared with group ALI,lung injury scores,W/D ratio and AI were significantly decreased,and the expression of GRP78,CHOP and caspase-12 in lung tissues was down-regulated in group E (P＜0.05),and no significant change was found in the paramneters mentioned above in group NE (P＞0.05).Conclusion The mechanism by which electroacupuncture attenuates endotoxin-induced ALI is related to inhibition of endoplasmic reticulum stress and reduction of apoptosis in alveolar epithelial cells in rats.

Objective To evaluate the role of phosphatidylinositol 3-kinase/serine-threonine kinase (PI3K/Akt) signaling pathway in carbon monoxide (CO)-induced up-regulation of the mitofusin-1 (Mfn1) expression in endotoxin-challenged rat alveolar macrophages.Methods Alveolar macrophages obtained from the rats aged 12-20 weeks were subcuhured and seeded in 96 well plates at a density of 4× 104 cells/ml.After being cultured for 24 h,the cells were divided into 4 groups (n=10 each) using a random number table:control group (group C),endotoxin group (group L),lipopolysaccharide (LPS) +CO-releasing molecule-2 (CORM-2) group (group L+C) and LPS+CORM-2+PI3K inhibitor LY294002 group (group L+C+LY).Cells were cultured normally in group C.Cells were stimulated by using LPS 10 μg/ml in L,L+C and L+C+LY groups.In group L+C,CORM-2 100 μmol was given at 1 h before stimulation with LPS.In group L+C+LY,LY294002 20 μg and CORM-2 100 μ mol were given at 1.5 and 1.0 h before stimulation with LPS,respectively.The cells were continuously incubated for 24 h after the end of treatment.The concentrations of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in the supernatant were determined by enzyme-linked immunosorbent assay.The expression of PI3K,phosphorylated Akt (p-Akt) and Mfn1 in cells was measured by real-time polymerase chain reaction and Western blot.Results Compared with group C,the concentration of TNF-α was significantly increased,and the IL-10 concentration was decreased in L,L+C and L+C+LY groups (P＜0.05).Compared with group L,the concentration of IL-10 was significantly increased,the TNF-α concentration was decreased,and the expression of PI3K,p-Akt and Mfn1 was up-regulated in group L+C (P＜0.05).Compared with group L+C,the concentration of IL-10 was significantly decreased,the TNF-α concentration was increased,and the expression of PI3K,p-Akt and Mfn1 was down-regulated in group L+C+LY (P＜0.05).Conclusion PI3K/Akt signaling pathway is involved in CO-induced up-regulation of Mfn1 expression in endotoxin-challenged rat alveolar macrophages.

OBJECTIVETo study genetic difference of Cistanche tubulosa that parasites on different Tamarixs and give a reference to select host of C. tubulosa.

METHODSixteen selected primers by random amplified polymorphic DNA (RAPD) markers were used to analyze genetic distance of C. tubulosa that parasites on eight different hosts.

RESULTSixty-six point seven percent of the total bands were polymorphic, that proved the genetic diversity level in different C. tubulosa types was relatively high, especially the two that parasites on Tamarix hispida and T. chinensis. Cultural areas had more remarkable influence on genetic distance of Cistanche tubulosa than the hosts, and introduction was helpful to maintain the more genetic diversity in different C. tubulosa types. Genetic difference in different C. tubulosa types was far less than that between different species in Cistanche.

CONCLUSIONC. tubulosa types which parasite on different Tamarixs have high genetic diversity.

Cistanche ; genetics ; physiology ; DNA, Plant ; analysis ; Genetic Variation ; Host-Parasite Interactions ; genetics ; Phylogeny ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; Tamaricaceae ; classification ; genetics ; physiology

Objective To determine the association of subclinical hypothyroidism (SCH) with body composition (measured by body impedance analysis) and insulin resistance (IR) in overweight/obese patients with polycystic ovary syndrome (PCOS).Methods A retrospective analysis was conducted of the clinical records of 109 overweight or obese [body mass index (BMI) ≥ 24 kg/m2] PCOS patients who visited the Department of Endocrinology and Fertility Center of Nanjing Drum Tower Hospital between April 2013 and April 2016 for menstrual disorder or infertility and were referred to receive weight management.24 of the patients had SCH,and 85 had euthyroid (EU).We determined the body composition of the patients with Biospace Inbody 720 body composition analyzer,measured the patients' serum lipid profiles,conducted in each patient the 75 g oral glucose tolerance test and the insulin release test,and calculated the homeostasis model assessment of insulin resistance (HO-MA-IR) indices.Results Patients with SCH and autoimmune thyroiditis (AIT) accounted for 22.02％ and 24.36％ of the total.The obesity level [(145.19±13.75)％ vs.(153.31±18.15)％,t=-2.032,P=0.045],VFA [(132.48±20.85) cm2vs.(147.35±24.26) cm2,t=-2.730,P=0.007],body fat (BF) [(31.91±5.88) kgvs.(35.43±6.89) kg,t=-2.274,P=0.025],body fat percentage (BFP) [(40.92±3.701)％ vs.(43.07±4.26)％,t=-2.241,P=0.027],body mass index (BMI) [(30.49±2.88) kg/m2 vs.(32.19±3.81) kg/m2,t=-2.026,P=0.045] and waist circumference (WC) [(98.34±7.13) cm vs.(102.86±8.74) cm,t=-2.324,P=0.022] of SCH group were significantly lower than those of euthyroid,with significant statistical difference.The levels of serum thyroid hormone (TSH) in patients with EU were positively correlated with the degree of obesity,the BF,the BFP,the BMI and the hip circumference (P=0.019,0.042,0.005,0.019,0.039),but not with the VFA (t=1.797,P=0.076).There were no statistically significant differences (P＞ 0.05) between patients with SCH and those with EU in their HOMA-IR indices,insulin levels,blood glucose,blood lipid,and ratio of IR (defined as HOMA-IR ≥2.69).Conclusions Obesity related body composition parameters were lower in PCOS patients with SCH than in those with EU.However,there was no significant difference between the two groups in blood lipid level and the ratio of IR.TSH levels in the EU group were correlated with the BMI and the BFP,but not with the VFA.A larger sample is needed to identify how and why thyroid hormones may affect the body composition and glycolipid metabolism of females with PCOS.

Objective To evaluate the relationship between p38 mitogen-activated protein kinase (p38MAPK) signaling pathway and lipopolysaccharide (LPS)-induced mitochondrial fission in alveolar epithelial cells using an in vitro experiment.Methods The cultured alveolar epithelial cells were subcultured and seeded in 96-well plates at the density of 2 × 105 cells/ml (200 μl/well).The cells were divided into 4 groups (n=10 each) when cell confluence reached 80％ using a random number table method:control group (group C),LPS group,LPS+SB203580 group (group LPS+SB) and LPS+dimethyl sulfoxide (DMSO) group.Cells were incubated with LPS 10 μg/ml for 24 h in group LPS.Cells were incubated with p38MAPK inhibitor SB203580 10 μmol (dissolved in DMSO) for 1 h and then with LPS 10 μg/ml for 24 h in group LPS+SB.Cells were incubated with the equal volume of DMSO for 1 h and then with LPS 10 μg/ml for 24 h in group LPS+DMSO.Malonaldehyde (MDA) content and superoxide dismutase (SOD) activity were measured.The expression of phosphorylated p38MAPK (p-p38MAPK),heme oxygenase-1 (HO-1),dynamin-related protein 1 (DRP1) and fission protein 1 (FIS1) was determined by Western blot.Results Compared with group C,the MDA content was significantly increased,the SOD activity was decreased,and the expression of p-p38MAPK,HO-1,FIS1 and DRP1 was up-regulated in LPS,LPS+SB and LPS+ DMSO groups (P＜0.05).Compared with group LPS,the MDA content was significantly increased,the SOD activity was decreased,the expression of p-p38MAPK and HO-1 was down-regulated,the expression of FIS1 and DRP1 was up-regulated in group LPS+SB (P＜0.05),and no significant change was found in the parameters mentioned above in group LPS+DMSO (P＞0.05).Conclusion The p38MAPK signaling pathway activation can up-regulate the expression of HO-1,thus reducing LPS-induced mitochondrial fission in alveolar epithelial cells.