Objective:To identify the causative gene in patients with familial progressive hyperpigmentation (FPH) .Methods:Two families with FPH were collected in March 2005 and March 2015 respectively, and their phenotypes were observed and recorded. The causative gene was investigated by single nucleotide polymorphism (SNP) -based genome-wide linkage analysis and exome sequencing, and verified by Sanger sequencing. The candidate gene expression was determined in FPH lesions and normal skin tissues by using immunohistochemical techniques.Results:The genome-wide linkage analysis showed that the causative gene in FPH family 1 was mapped to the loci of rs1026369-rs11857925 on chromosome 15q21.1 - q22.2; a disintegrin and metalloproteinase 10 (ADAM10) gene was identified as the possible causative gene by exome sequencing; Sanger sequencing showed that a splice-site mutation c.1511+1G>A in the ADAM10 gene was co-segregated with the disease phenotype in the FPH family 1. Immunohistochemical staining demonstrated that ADAM10 was expressed in both the FPH lesions and normal skin tissues of the proband in the FPH family 1. A missense mutation c.1172C>T (p.Ser319Phe) was identified by further ADAM10 mutation analysis in another 3-generation family with FPH (family 2). Both the above mutations were not detected in 300 local healthy controls.Conclusion:ADAM10 was identified as a novel causative gene responsible for FPH.

Objective To discuss the expression and significance of neutrophil to lymphocyte ratio(NLR) and platelet to lymphocyte ratio(PLR) in peripheral blood in patients with chronic obstructive pulmonary disease(COPD).Methods One hundred and fifty cases COPD patients who were treated in Respiratory Departments of Hai'an People′s Hospital Affiliated to Nantong University from January to December 2015 were selected and divided into stable period group with 60 cases and acute exacerbation period group with 90 cases,and 50 healthy subjects were selected as control group.Clinical data were collected and white blood cell count(WBC),neutrophil count(NC),lymphocyte count(LC),platelet(PLT),NLR,PLR,forced expiratory volume in one second(FEV1%) predicted and C reactive protein(CRP) were tested.Results NLR,PLR and CRP values were significantly higher in patients in acute exacerbation period group than in stable period group and control group(NLR:(10.30±9.93) vs.(6.76±5.90) vs.(5.85±3.67);PLR:(335.88±164.94) vs.(286.80±118.98) vs.(194.11±88.14);CRP: (42.18±24.89) mg/L vs.(23.27±13.59) mg/L vs.(22.49±13.47) mg/L,F=6.637,6.808,11.007,P<0.05),but FEV1% predicted was no difference in stable period group and acute exacerbation period group((58.85±16.83)% vs.(57.47±17.76)%;t=0.228,P>0.05).There was a significant positive correlation between NLR,PLR and CRP in acute exacerbation period group(r=0.374,0.379,P<0.01),there was a positive correlation between NLR and PLR(r=0.482,P<0.01),there was a negative correlation between NLR,PLR and FEV1(r=-0.297,-0.299,P<0.05).There was a significant positive correlation between NLR,PLR and CRP in table period group(r=0.593,0.433,P<0.01),NLR and PLR were also positively correlated(r=0.618,P<0.01),NLR,PLR and FEV1 accounted for the expected value of the negative correlation(r=-0.324,-0.342,P<0.05).Conclusion Both NLR and PLR are inflammatory markers,furthermore they show a significant negative correlation to airflow limitation in patients with COPD.

Objective To investigate the effect of leptin on the secretion of chemokine in THP1 cells and explore its related mechanism, and to provide basis for research on the role of leptin in immune response.Methods The expressions of Ob-Rb and Ob-Rt in THP1 cells were detected by RT-PCR and flow cytometry (FCM).The THP1 cells at logarithm growth phase were selected and randomly divided into blank control group and different concentrations(10,50,100,200μg· L-1 )of leptin groups.Transwell chamber assay was performed to detect the number of invated THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group.Western blotting method was carried out to detect the expressions of p-AKT,p-ERK 1/2,and p-STAT3 in THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group,100μg·L-1 leptin+ DMSO group,100μg·L-1 leptin+50μmol·L-1 AG490 group,100μg·L-1 leptin+10μmol·L-1 LY294002 group and 100μg·L-1 leptin+ 10 mol·L-1 PD980590 group.RT-PCR and Western blotting methods were performed to detect the expression of IL-8.Results Ob-Rb and Ob-Rt were highly expressed in THP1 cells. Compared with blank control group,the number of invated THP1 cells was significantly increased in 50,100,and 200μg·L-1 leptin groups (P<0.05).Compared with blank control group,the expressions of p-STAT3,p-ERK 1/2 and p-AKT in THP1 cells were up-regulated in 100 ug·L-1 leptin group(P<0.05).Compared with blank control group,the mRNA and protein expressions of IL8 in THP1 cells in 50,100,and 200μg·L-1 leptin groups were remarkably increased(P<0.05);compared with 100μg·L-1 leptin group,the expressions of IL-8 in THP1 cells in 100μg·L-1 leptin+10 mol·L-1 PD980590 group and 100μg·L-1 leptin+10μmol·L-1 LY2 94002 group were decreased(P<0.05),while the expression of IL-8 in 100μg·L-1 leptin+ 50μmol·L-1 AG490 group had no change(P>0.05).Conclusion leptin can up-regulate the expression of chemokine in THP1 cells,which might be associated with PI3K-AKT and MAPK/ERK 1/2 signaling pathways.

AIM: To investigate the effects of auricularia auricular polysaccharide (AAP) on chronic cerebral ischemia injury in rats. METHODS: The chronic cerebral ischemia mode1 was made by permanent middle cerebral artery occlusion (MCAO) on the right side. AAP at different doses (50 mg/kg and 100 mg/kg) was intragastrically administered at the onset of ischemia and in the following days after operation, once a day for 4 weeks. After 4 weeks of MCAO, Morris water maze test was introduced to examine the learning and memory functions. Nissl staining was performed to detect the survival neurons in hippocampal slices. Level of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in brain tissue were measured. RESULTS: Rats treated with AAP showed a shorter escaping latency in spacial navigation test because the AAP treated rats spent less time to find the platform in spatial probe test. More survival neurons in hippocampal slices were observed from AAP treated rats. Also, the MDA level in brain tissue was reduced and SOD activity in brain tissue was increased in the AAP treated rats with MCAO. CONCLUSION: AAP protects rats from chronic brain ischemic injury, in which its anti-oxidative effect might be involved.

AIM: To investigate the protective effect of auricularia auricular polysaccharide (AAP) on the myocardial injury induced by ischemia and its underlying mechanism. METHODS:AAP was orally administrated to rats at the does of 50, 100 or 200 mg·kg~(-1)·d~(-1) for 20 d. Myocardial injury was induced in anesthetized Sprague - Dawley rats by left anterior descending coronary artery ligation. Myocardial infarct size, the level of lactate dehydrogenase (LDH), the activity of superoxide dismutase (SOD) , the production of lipid peroxidation malondialdehyde (MDA) and protein level of myocardial collagen of the heart were measured. RESULTS: The average myocardial infarct size in AAP groups was significantly smaller than that in ischemia group. The level of serum LDH induced by regional myocardial ischemia was significantly decreased in AAP group compared to ischemia group ( P < 0.01). AAP inhibited the production of MDA and increased the activity of SOD. Furthermore, AAP reduced the protein level of myocardial collagen after ischemia (P < 0.01).CONCLUSION: AAP prevents myocardium from ischemia injury as an antioxidant.

Objective To establish a continuous monitoring assay of serum argininosuccinate lyase (ASL) activity with automatic biochemistry analyzer, and perform methodology validation and preliminary clinical application.Methods According to the chemical reaction catalyzed by ASL and the working characteristics of automatic biochemistry analyzer, an enzyme coupled reaction system with high specificity was set up, and the methodology validation was performed.Three hundred and nine patients with various liver diseases, 269 non-liver disease patients and 40 healthy controls were enrolled in this study.Serum ASL, ALT, and AST level were determined in all subjects.Results A new kinetics assay of ASL activity was set up with automatic biochemistry analyzer.The methodological validation demonstratod that inter-assay and intra-assay coefficient of variation were 4.0% and 5.9% respectively and the mean recovery was 100.5%.The linear range was 0-167.7 U/L.The lowest detection limit was approximately 0 U/L.The interference test showed that there is no significant interferences while the concentration of bilirubin is less than 342 μmoL/L or commonly used anticoagulants is employed at their routine concentrations.However,interference was significant when Hb level is more than 0.06 g/L.Preliminary study of clinical application showed that there was no significant difference of serum ASL level between non-liver disease group and healthy group ( q = 0.027, P = 0.979 ), but there was significant differences for both serum ALT and AST levels (ALT:q =6.461,P =0.000;AST:q =6.481,P =0.000).Conclusions A continuous monitoring assay for the determination of serum ASL activity is successfully established. Serum ASL may be a good biomarker for liver injury.

BACKGROUND: Recently, it is investigated that shell of plantain seed is a soluble dietary fiber which can be added into foods to regulate content of cholesterol.OBJECTIVE: To investigate the interventional effect of plantain seed on lipid and its lipid peroxidation in rats with hyperlipidemia.DESIGN: Completely randomized grouping design and controlled animal study.SETTING: Laboratory of Pharmacology and Toxicology for New Drug in Hebei Province.MATERIALS: ① A total of 24 healthy SD rats, of grade I, aged 60-70 days, weighting (210±22) g, of either gender, were selected in this study. ② Basic feed was provided by Experimental Animal Center in Hebei Province,and the fractional mass of each component was mentioned as following:flour 0.25, bran 0.1, corn dust 0.22, bean cake 0.22, fish dust 0.02, bone dust 0.02, grass dust 0.05, salt 0.01, yeast dust 0.02, and sunflower seed 0.03. High fat feed was provided by Experimental Animal Center in Hebei Province, and the fractional mass of each component was mentioned as following: basic 0.9, cholesterol 0.015, lard 0.08, and hyocholic salt 0.003.③ Lipid kit was provided by Baoding Changcheng Clinical Reagent Company, and kits of superoxide dismutase (SOD), catalase (CAT),glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) were provided by Nanjing Jiancheng Bioengineering Institute.METHODS: The experiment was completed at the Laboratory of Pharmacology and Toxicology for New Drug in Hebei Province from June to December 2004. ① All 24 rats were randomly divided into 3 groups:normal control group, model group and plantain seed group with 8 in each group. Rats in the normal control group were fed with basic feed. Rats in plantain seed group were fed with high fat feed + 15 g/kg plantain seed and drank routinely. Experimental rats were fed in cages, respectively.Each one was fed with 25 g/d food and drunk freely. The experimental cycle was 12 weeks. ② At the end of experiment, rats were anesthetized to assayed levels of serum triacylglycerol (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), serum SOD and MDA, activities of CAT and SOD in myocardial tissue, content of MDA, and activities of CAT and GSH-Px in hepatic tissue with related kits. ③ Measurement data were compared between each two group with t test.MAIN OUTCOME MEASURES:Comparison of serum lipid level and anti-oxidation among groups at 12 weeks after modeling.RESULTS: All 24 rats were involved in the final analysis. ① At 12 weeks after modeling, activities of SOD in serum and myocardial tissue were lower in model group than those in normal control group and plantain seed group (P ＜ 0.05), but levels of MDA in serum and myocardial tissue were higher in model group than those in normal control group and plantain seed group (P ＜ 0.05). ② At 12weeks after modeling, activities of CAT and GSH-Px in serum and myocardial tissue were lower in model group than those in normal control group and plantain seed group (P ＜ 0.05). ③ At 12 weeks after modeling, levels of TC and TG in serum were higher in model group than those in normal control group and plantain seed group (P ＜ 0.05), but level of HDL-C and ratio between HDL-C and TC in serum were lower in model group than those in normal control group and plantain seed group (P ＜ 0.05).CONCLUSION: Plantain seed at dosage of 15 g/kg can decrease content of lipid and strengthen anti-oxidation of economy in rats with hyperlipidemia.

BACKGROUND: Great concerns haven been given increasingly on inhibition of nutrient with antioxidation efficacy on lipid peroxidation and its effect on prevention of cardiac vascular disease.OBJECTIVE: To probe into the effects of plantain seed that acts on eliminating oxygenic free radical and antioxidation on lipid metabolism and antioxidation in rats.DESIGN: Randomized control experiment was designed.SETTING: Experimental Room of Pharmacology and Toxicology of New Drug in Hebei Province.PARTICIPANTS: The experiment was performed in Experimental Room of Pharmacology and Toxicology of New Drug in Hebe Province from January to December 2004, in which, 40 SD rats were employed, provided from Hebei Experimental Animal Center, of healthy grade I, mass weighted (210±22) g and of either sex. They were randomized into 5groups, named, blank control, positive control, low dosage experiment group, moderate dosage experiment group and high dosage experiment group, 8 rats in each one.METHODS: In blank control, the rats were bred everyday with basic forage that was tallied with AoAc animal nutrient criteria and they were free of drinking. In positive control, the rats were bred with high-lipid forage and free of drinking. In the groups of low, moderate and high dosages of plantain seed, the rats were bred with 2.5 g/kg, 5 g/kg and 15 g/kg plantain seed successively besides high-lipid forage and they were free of drinking. The weight was measured and the food intake was recorded every week. Fasting blood was collected to check total cholesterol in serum once every two weeks. The experiment was end in 12 weeks. Under anesthesia,the blood was collected from hypogastric aorta to check the level of serum blood lipid and the activities of superoxide dismutase (SOD) and catalase.After blood collection, the heart and liver were extracted immediately for management to measure SOD activity and content of lipid peroxide (LPO)in myocardial tissue and the activities of catalase and glutathione peroxidase in liver tissue.MAIN OUTCOME MEASURES: Level of blood lipid and activities of SOD and catalase in rats.RESULTS: Forty rats were employed and all entered result analysis. [1]Serum total cholesterol: It was lower significantly in high dosage group compared with positive control [(1.40±0.13, 1.83±0.13) mmol/L, P ＜ 0.05].[2] Serum SOD activity: It was lower remarkably in positive control compared with blank control [(174.29±10.33, 193.19±.7813) NU/mg, P ＜ 0.05].[3] LPO content in serum: It was higher significantly in positive control compared with blank control [(3.64±0.26, 2.91±0.50) mmol/mg, P ＜ 0.05]and it was lower significantly in moderate dosage group compared with positive control (3.13±0.26, 3.64±0.26, P ＜ 0.05). [4] Activity of catalase in liver tissue: It was lower remarkably in positive control compared with normal control (34.64±3.26, 44.72±2.67, P ＜ 0.05) and it was higher remarkably in moderate dosage group compared with positive control (44.84±3.79,34.96±3.64, P ＜ 0.05).CONCLUSION: Plantain seed reduces the levels of total cholesterol,triglycerin (TG) and LPO in serum and increases SOD activity. At the concentration of 15 g/kg, plantain seed acts most remarkably on eliminating oxygenic free radical and antioxidation and alleviates lipid metabolic disturbance.

Objective To investigate the effects of total saponins of Panax ginseng(TSPG) combined with erythropoietin(EPO) or granulocyte macrophage colony stimulating factor(GM CSF) on the differentiation of K562 cells. Methods Cells were cultured in vitro . Morphologic changes of cells were observed with light microscope and electron microscope. The levels of hemoglobin, peroxidase, non specific lipase ? NAE in cells were detected respectively by benzidine staining, peroxidase(POX) staining and ? NAE cytochemical staining. Levels of cell surface differentiation antigens CD15, HIR2, EPO R and GM CSF R were determined by immunocytochemistry. Results TSPG combined with EPO or GM CSF resulted in more mature morphological features of K562, increased expressions of hemoglobin and myeolperoxidase in K562 cells, and enhanced expressions of cell surface differentiation antigens CD15, HIR2, EPO R and GM CSF R. Conclusion TSPG combined with cytokine EPO or GM CSF can induce more mature differentiation of K562 cells.

Object To clarify the mechanism for total saponins of Panax ginseng C.A.Mey. (TSPG) inducing K562 cells to apoptosis, and to provide the theoretical basis and the experimental evidence of TSPG's clinical application. Methods By using in vitro cell culture, morphometry, flow cytometry, morphological investigation and immunocytochemistry, the effects of TSPG on apoptosis in K562 cells were studied. Results The results indicated that TSPG could inhibit the proliferation of K562 cells significantly, and induce K562 cells to apoptosis. It was also showed in the experiments that after induced by TSPG, the ratio of positive K562 cells expressing C-MYC and BCL-2 is decreased. Conclusion The mechanism for TSPG to induce K562 cells to apoptosis may be related to the expression of oncogene in K562 cells.