ObjectiveTo investigate the effects of nicotine on the expression of Th1/Th2 cytokines and transcription factor T-bet/GATA-3 in cultured CD4+ T of rat sensitized by ovalbumin.Methods Two weeks after immunization by ovalbumin,splenic CD4+ T cells of Wistar rat were purified using CD4+T cell enrichment kit.Purified CD4+ T cells of rat were cultured and divided into 4 groups:a control group,1 μg/ml nicotine stimulated group,10 μg/ml nicotine stimulated group,100 μg/ml nicotine stimulated group.These cells,in their groups,were stimulated with or without nicotine and were all challenged simultaneously with OVA.Supernatants and cell pellets were harvested after being stimulated for 24 h.The concentration of IFN-γ and IL-4 in supernatants were measured by enzyme-linked immunosorbent assay (ELISA).Real-time PCR was used to determine the mRNA expression of T-bet and GATA-3 in CD4+T cells.Results ( 1 ) IFN-γproduction was significantly decreased in all nicotine treated groups [ ( 113.78±6.06) ng/L,(70.31±7.26) ng/L,(20.00±2.14) ng/L] compared with the control group[ (142.30± 5.89) ng/L],and the level of IL-4 was significantly increased in all nicotine treated groups [ (69.49±3.91) ng/L,(93.63±4.56) ng/L,(50.97±3.07) ng/L] compared with the control group[ (36.91±3.24) ng/L].(2) Expression of T-bet mRNA in all nicotine treated groups(0.73±0.03,0.57±0.04,0.31 ±0.00) was lower than that in the control group(0.98±0.09),but expression of GATA-3 mRNA in all nicotine treated groups (4.31±0.26,5.16±0.23,1.56±0.14) was significantly higher than that in the control group(1.00±0.07).Conclusion Nicotine may play a key role in the development of Th2-type allergic inflammation in asthma by promoting over-expression of GATA-3 mRNA and downregulating the expression of T-bet mRNA.

Objective To explore the underlying mechanism of potential effect of sildenafil on porcine pulmonary artery smooth muscle cells (SMCs) proliferation induced by serotonin. Methods Porcine pulmonary artery SMCs at 3-5 passages were randomly divided into 4 groups: control group (CON), 5-HT group (HT), sildenafil group (SIL) and sildenfil-5HT combined group (S-HT). Pulmonary artery SMCs at exponential growth phase were serum starved with 0.2% FBS for 72 h, followed by sterile PBS, serotonin (10 μmol/L) and sildenafil (1 μmol/L) incubation in CON group, HT group and SIL group, respectively. In combined group (S-HT): pulmonary artery SMCs were serum starved and then exposed to sildenafil for 20 min, followed by serotonin incubation for indicated time. The phosphorylation of extracellular signal regulated kinase (ERK1/ERK2) was measured by Western blot at indicated time points. Flow active cell sorting (FACS) was used to evaluate the ratio of S phase cells in the cell cycle after 24 h of treatment. MTT color metric method was used to analyze SMCs proliferative index after 72 h of treatment. Results Compared with CON group, the phosphorylation of ERK1/ERK2, the percentage of cells in S phase, and the cell proliferation index increased remarkably after incubation with 5-HT (P<0.05). Preincubation with sildenafil followed by serotonin enhanced the phosphorylation of ERK1/ERK2 (p-ERK1/ERK2) and further elevated the percentage of cells in S phase and the cell proliferation index compared with that of HT group. While the total ERK1/ERK2 (t-ERK1/ERK2) was not statistically different among these groups. Conclusions Sildenafil potentiates the proliferative effect of serotonin on pulmonary arterial SMCs via upregulating phosphorylation of ERK1/ERK2.

Objective:To investigate relationships between serum chemerin and bone mineral density (BMD) in patients with newly diagnosed Graves disease (GD).Methods:A total of 120 newly diagnosed GD patients with a course more than 3 months were enrolled from the Department of Endocrinology between June 2013 and June 2015.Sixty age-and sex-matched healthy people served as a normal control.Serum levels of chemerin,β-crosslaps (β-CTX),and N-MID-osteocalcin (N-MID-OT) were measured by ELISA.Fat mass and BMD were evaluated by dual energy X-ray absorptiometry (DEXA).Results:Compared with the normal control,the fat mass,lean weight,fat mass index (FMI) and body mass index (BMI) in the GD group were decreased,and BMD in all skeletal sites was decreased.There was a positive correlation between them (all P<0.05).Serum level of chemerin was increased and it was positively correlated with β-CTX or N-MID-OT level and negatively correlated with fat mass,FMI or BMI in the GD group.There was a negative correlation between chemerin level and BMD in femoral neck,total hip,lumbar or right forearm distal 1/3 (rs=-0.352,-0.279,-0.379,-0.289,-0.394;P<0.05).After adjusting for age,fat mass or BMI,the correlation of chemerin with total hip or bone mineral density remained significant (rs=-0.273,-0.378;P<0.05).Multiple linear regression analysis revealed that chemerin or BMI was correlated with BMD (P<0.05).Conclusion:The decrease of bone mineral density in patients with GD is not only related to the direct or indirect effect of excessive thyroid hormones on systemic and osteoblastic cells,but it is also related to the negative regulation of bone metabolism due to the elevated chemerin level.

The patient is an 11-year old boy, who was born with universe alopecia as well as dry and coarse skin. When he was 3 months old, photophobia was noticed, and since then, upper respiratory tract infection had occurred twice a month complicated by frequent diarrhea. He had short stature with slight conjunc- rival congestion, corneal vascularization, opacity, coarseness and poor vision. No abnormality was found in the teeth, sweating ability, or hearing. He had universal alopecia; his skin was dry and rough with generalized rhombus- or polygon-shaped scaly patches. Particularly thick brown scales were observed on the upper limbs. Moreover, there were spiny follicular papules on the abdomen and axillae, hyperkeratosis of palm and sole, and dystrophic nails. Hyperextensibility of proximal interphalangeal joints of the third, fourth and fifth fingers was noticed. He also suffered from mental retardation, the verbal intelligence quotient being 52, performance intelligence quotient lower than 40, full intelligence quotient lower than 40, but no abnormality was found in the heart, lung, liver or spleen. Histopathology of skin on the abdomen suggested a change characteristic of ichthyosis. Chromosome analysis revealed a karyotype of 46, XY. This is the first diagnosed case of ichthyosis follicularis with atrichia and photophobia syndrome in China.

Objective To investigate the eitect oi Yiqi Fuzhi Granule in improving learning and memory of rats with mul- ti-infarct dementia (MID). Methods MID rat models were established by embolus injection through the common carotid artery. Effects of Yiqi Fuzhi Granule(24. 84 g/kg, 12.42 g/kg, 6.21 g/kg, ig)on rat learning and memory and cerebral structure were observed. Results Yiqi Fuzhi Granule could obviously increase the scores of maze test and protect cerebral tissue from damage of multi-infarction rats. Conclusion Yiqi Fuzhi Granule has good effect in improving intellect and treating dementia.

Objective To investigate the influences of grape seed proanthocyanidin extract (GSPE) on cardiovascular system, nervous system and respiratory system of experimental animals, and provide general pharmacological data for further research and application. Methods The influences of GSPE on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs after duodenal administration were observed, the impacts on spontaneous activity, coordinated motion, and the sleep situation with threshold dose and subthreshold dose of pentobarbital sodium in mice after intragastric administration were observed. Results GSPE showed no side effects on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs at the dosage of 857.00, 214.29, 42.86 mg/kg (P>0.05). At the dosage of 428.57, 214.29, 42.86 mg/kg, GSPE had no obvious influence on spontaneous activities and coordinated movements in mice (P>0.05). GSPE did not evidently change the number of sleeping animals, the sleep latency and the sleeping duration with subthreshold dose and threshold dose of pentobarbital sodium (P>0.05). Conclusion GSPE has no evident adverse effects on central nervous system, cardiovascular system and respiratory system in animals.

Objective To examine the expressions of FHIT gene and HER_2 in non-small cell lung cancer(NSCLC)and the relationship between the expressions and the clinicopathological features of NSCLC, and to investigate the relationship between FHIT protein and HER_2 expression. Methods FHIT protein and HER_2 expressions in 50 lung cancer cases and 21 adjacent non-cancer tissues were performed by immunohistochemistry. And the positive rates of FHIT and HER_2 were measured. Results The positive rates of FHIT protein were 30.00% in lung cancer tissues. The expression levels of FHIT were significantly lower in lung cancer tissues than that in non-cancer lung tissues (P

Objective To analyze the expression of T lymphocyte activation antigen in peripheral blood of aged patients with non-small cell lung cancer . Methods The lymphocytes from peripheral blood in 45 aged patients with non-small cell lung cancer were immunologically labeled in double fluorescence and CD3-FITC, CD25/HLA-DR and CD69-PE and determined by flow cytometry. Normal aged donors, young patiens with lung cancer and aged benign lesion group were used as controls. Results Peripheral blood CD3 +/CD25 +, CD3 +/HLA -DR + and CD3 +/CD69 + in T lymphocyte with 45 aged lung cancer(7.24?1.85,28.46?5.39 and 7.78?2.63, respectively) were significantly lower than those in normal aged controls(10.35?2.54,37.16?5.51,11.02?2.18, respectively)and aged benign lesion (9.53?3.02, 35.33?5.23, 10.67?2.45, respectively)( P 0.05). Significant differences were found among them in stages Ⅲ, Ⅳ(7.15?1.13, 25.32?5.23, 7.14?2.81, respectively) and stagesⅠ,Ⅱ(8.06?1.21, 30.27?6.05, 8.43?2.67, respectively)( P 0.05). Conclusions Detection of CD3 +/CD25 +, CD3 +/HLA -DR + and CD3 +/CD69 + levels by flow cytometry might be helpful for reflecting the human immune function and the prognosis evaluation in patients with aged non-small cell lung cancer.

Objective To observe the uhrasenographical characters of liver and spleen of residents and their changes in endearic areas of schistosomiasis japonica in Poyang Lake region.Jiangxi Province and to explore the value of ultrasonography for assessment of the morbidity of the disease.Methods All permanent residents aged above 3 years old were examined by ultrasonography and Kato-Katz method.Results The schistosome positive rates of fecal examinations decreased obviously from 16.29%in 1995 to 8.54%in 2007(P<0.01).However,the rates of hepatomegaly and splenomegaly between 1995 and 2007 were not significantly changed(P>0.05),with the rates of 8.82% and 20.33% in 1995 and 8.54% and 21.34% in 2007,respectively.The abnormal rate of portal vein diameter decreased significantly.from 32.47%in 1995 to 6.50% in 2007.The abnormal rate of liver parenehyma increased remarkably(P<0.01),from 34.85% to 51.83%.The changes of liver parenchyma Grade I showed a bidirectional trend,29.90% of them chased into Grade 0(normal image on ultrasonngraphy),and 34.02% changed into Grade 2 and above.The abnormality of various indices of uhrasonography examinations were related to age,occupation and schistesome infection status.Conclusions Ultrasonography can show the damages of liver and spleen of patients infected with Schistosoma japonicum directly.but it is necessary to study further on the sensitive indices that reflecting early pathological changes and the best combination of the indices for the assessment of schistosomiasis-related morbidity.

The recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene was constructed by using AdEasy-1 system and its expression was detected in human breast cancer cell line MDA-MB-468. Human PTEN cDNA was cloned into adenovirus shuttle plasmid pAdTrack-CMV to generate a recombinant plasmid pAdTrack-CMV-PTEN, then homologeous recombination was carried out in the E. coli BJ5183 by contransforming linearized shuttle vector with adenovirus backbone plasmid pAdEasy-1. The newly recombined defective adenovirus vector Ad-PTEN containing green fluorescent protein (GFP) was packaged and propagated in 293 cells. After being purified by cesium chloride gradient centrifugation, the adenovirus was transfected into human breast cancer cell line MDA-MB-468 in vitro. The expression of PTEN mRNA and protein in infected human breast cancer cell line MDA-MB-468 was detected by RT-PCR and Western blot respectively. The recombinant defective adenovirus vector carrying PTEN gene was constructed successfully. The viral titer of purified adenovirus was 2. 5 X 10(10) pfu/mL, and about 70% breast cancer cells were infected with Ad-PTEN when multiplicity of infection (MOI) reached 50. The exogenous PTEN mRNA and protein were expressed in MDA-MB-468 cells infected with Ad-PTEN by RT-PCR and Western blot. The recombinant defective adenovirus vector of PTEN gene was constructed successfully using AdEasy-1 system rapidly, which paved a sound foundation for gene study of breast cancer.