Interleukin-17 (IL-17) is a newly found cytokine secreted by activated T lymphocytes. From preliminary study, IL-17 was found to play a role in the relationship between T lymphocytes and hematopoiesis, but many of its characteristics remain unknown up to now. To investigate its biological functions and related mechanisms, we cloned the complete coding region of human IL-17(hIL-17) from activated human peripheral blood mononuclear cells (PBMC) by RT-PCR and constructed an integrative vector pLCM182-hIL-17 for cloning, expressing and sequencing the hIL-17 gene. By DNA sequencing, the cloned hIL-17 is identical to the reptorted, and with temperature inducing, the hEL-17 was highly expressed in E. coli up to 30% of bacterial protein. The expressed hIL-17 protein could stimulate primary human fibrob-lasts to secrete IL-6 and GM-CSF after initial purification. This results indicate that the expressed hIL-17 has biological activity.

Objective: To investigate the effects of DC derived from IL-12-induced erythroleukemia cells on the induction of CTL and protective antituinor immunity. Methods: The cytotoxicity of CTL was assessed by 4h 51 Cr-release assay. HE staining and eleclromicroscopy were used to observe the histological changes after immunized with erythroleukemia-derived DCs. Results: After incubation with naive T cells, the IL-12-induced FBL-3 cells could apparently induce the generation of CTL which exihibit the specific cytotoxicity against wild-type FBL-3 cells in vitro. We further assayed the cytotoxicity of CTL in vivo after immunized with erythroleukemia-derived DCs. CTL cytotoxicity of mice immunized with IL-12-in-duced FBL-3 cells was higher than that of mice immunized with IL-I2-uninduced FBL-3 cells. C57BL/6 mice immunized with IL-12-induced FBL-3 cells could resistant the subsequent rechallenge with the wild-type FBL-3 cells. The tumor growth was efficiently inhibited. Histological observation showed that more inflammatory cells infiltrated into the tumor tissue and necrosis of leukemia cells existed. By transmission electron microscopy, apoptositic phenomena were observed in the tumor of group immunized with IL-12-induced FBL-3 cells. However, these changes were not observed in the group immunized with IL-12-uninduced FBL-3 cells. Conclusion: These data indicate that erythroleukemia cells-derived DCs could induce specific CTL efficiently in vitro and in vivo, and may be used as new vaccine to activate antituinor immune responses.

Objective To study risk factors of cardiovascular disease (CVD) and status of bone mineral density (BMD) in women with hypoestrogenism.Methods From Jul 2011 to April 2013,a total of 256 women with hypoestrogenism in the First Affiliated Hospital of Shanxi Medical University were enrolled in this retrospective study,which were divided into four groups:133 women in ppausal group,25 women in premature ovarian failure (POF) group,67 women in menopausal transition group and 31 women in premature ovarian failure transition group.General statue,CVD risk factors and BMD were compared among four groups.General statue include menopausal period,menopausal symptoms (Kupperman Index),CVD risk factors include body mass index,blood pressure,waist circumference,waist-hip ratio,blood lipids and glucose,BMD include left hip,lumbar spine bone mineral density and T or Z value.Results (1) The median menopausal period were 3.4 years in postmenopausal group and 3.6 years in premature ovarian failure group,which did not show no statistical difference (P ＞ 0.05).Kupperman Index in four groups were 12 in postmenopausal group,9 in POF group,9 in menopausal transition group and 8 in premature ovarian failure transition group,which reached statistical difference (P ＜ 0.05).(2) The difference of body mass index (BMI),waist circumference,waist-hip ratio,diastolic blood pressure were no statistically significant among four groups(P ＞ 0.05) ; the systolic blood pressure in four groups were 120,110,110,110 mm Hg (1 mm Hg =0.133 kPa),their differences were statistically significance (P ＜ 0.05); the high-density lipoprotein (HDL-C) was 1.6 mmol/L in postmenopausal group,and 1.3 mmol/L in premature ovarian failure transition group,their differences were all statistically significance (P ＜ 0.05) ; the difference of the fasting plasma glucose (FPG) was not statistically different in 4 groups (P ＞0.05).(3) The abnormal rate of lower bone mass in lumbar spine were 57％ (46/81) postmenopausal group,8/15 in POF group,32％ (9/28) in menopausal transition group,12/19 in premature ovarian failure transition group,and osteoporosis was 9％ (7/81),3/15,1％ (3/28)and 0 respectively,their differences were statistically different (P ＜ 0.05) ; the abnormal rate of BMD of left hip and lumbar spine of 11/15 and 12/16 in POF group was higher than 65％ (53/81) in postmenopausal group.In the mean time,the abnormal rate of BMD of left hip and lumbar spine were,12/19 and 10/20 in premature ovarian failure transition group,which were significantly higher than 43％ (12/28) and 39％ (12/31) in the menopausal transition group.Conclusions The menopausal symptoms resulting from hypoestrogenism in natural postmenopausal women are mostly remarkable.The decrease of BMD in lumbar spine is more significant than that of left hip among postmenopausal women.Women with earlier menopause was prone to cause the changes of blood fat and abnormal of BMD,especially HDL-C decreased significantly compared with those natural postmenopause,it is more likely to cause CVD and osteoporosis.

Objective To investigate the inhibitory effects of periplocin from cortex periplocae (CPP) on human lung cancer cell line QG56 and to discuss its mechanism. Methods QG56 cells were cultured in vitro. The final concentrations of CPP in control group were 1.25, 2.50, 5.00, 10.00 and 20.00μg/L. QG56 cells were treated with ascending concentration of CPP for 24 h, 48 h and 72 h. The cell proliferation was measured using MTT method. The morphological changes of QG56 cells were observed under inverted microscope. Flow cytometry (FCM) was used to detect the effects of CPP on cell cycle and cell apoptosis. The expression of apoptosis associated gene bax mRNA in QG56 cells was detected by RT-PCR. The expres-sion of bax protein before and after treatment of CPP was examined by SP immunocytochemistry. Results The inhibitory ef-fect of CPP on the proliferation of QG56 cells was increased with the increasing concentrations of CPP and the prolonged du-ration of treatment. The morphological changes were displayed in QG56 exposed to CPP. The results of FCM showed that CPP caused cell cycle arrest at G0/G1 phase. The apoptotic rate of QG56 cells was significantly increased after CPP treatment for 48 h (P<0.05). The expression of bax mRNA was increased in QG56 exposed to CPP. The result of immunocytochemis-try indicated that CPP up-regulated the expression of bax protein. Conclusion CPP showed significant inhibitory effect on human lung cancer cell lines QG56 through inducing cell cycle arrest and apoptosis.

The study is aimed to establish a method to determine the content of Vinorelbine in liposomes by HPLC.The experiment was carried out on Kromasil C18(4.6 mm×250 mm,5 μm) colum with the mobile phase consistedof acetonitrile-0.06 mol·L-1 KDP buffer(pH 3.0,35:65) at a flow rate of 1.0 mL·min-1.The detection wavelength was at 215 nm and the column temperature was at 30℃.The result showed that the linear range of Vinorelbine was from 2.08 μg·mL-1 to 20.8 μg·mL-1(r=0.9999).The average recovery rate was 98.64％(RSD =1.09％).It is con cluded that the method used in this analysis is simple,precise and replicable with high recovery and accuracy.It can be used to determine the content of total Vinorelbine in liposomes.

Objective To evaluate the effect and complication of inductio n chemot herapy combined with three-dimensional conformal radiation therapy (3DCRT) for l ocally advanced non small cell lung cancer (NSCLC). Methods Ninety-two such pa t ients were randomized into radiation therapy alone group(RT-, 50 patients) and i nduction chemotherapy combined radiotherapy group (CMT-, 42 patients). The indu c tion chemotherapy consisted of 2-4 cycles of platinum-based regimen. Results Th e overall median survival time was 15 months with 12 months in the RT group and 18 months in the CMT group(P=0.014)respectively. The 1-year o verall survival rates were 48.6% and 71.2% in RT and CMT group,respectively (P=0.004). The 2-year survival rates w ere 20.8% and 37.6% in RT and CMT group, respectively (P=0.0 41). Treatment was w ell tolerated and the toxicities were similar in either group. C onclusion The ad dition of induction chemotherapy to 3DCRT takes a survival advantage over 3DCRT alone for Stage Ⅲ NSCLC without increasing toxicities.

OBJECTIVETo investigate the differences in the condylar position of subjects with skeletal class I and skeletal class II. To provide a basis of diagnosis and treatment.

METHODSGroup A was composed of 50 subjects with skeletal class I (27 males and 26 females; age range = 18 years to 30 years; mean age=26 years). Group B comprised 50 subjects with skeletal class II (24 males and 26 females; age range = 18 years to 28 years; mean age=25 years). The condylar position and the shapes of the condyle and the glenoid fossa were linearly measured on the sagittal and coronal sections by cone-beam computed tomography (CBCT). Data were analyzed by SPSS 19.0.

RESULTSNo statistically significant differences were found in the measurements of the condylar position between the sides of each group on the sagittal plane and the coronal plane (P > 0.05). There were significant differences on the anterior space and the posterior space between group A and B (P < 0.05). The A/P joint space ratio of group A was larger than that of group B (P < 0.05).

CONCLUSIONThe subjects of skeletal class I show an anterior condyle position. The subjects of skeletal class II show a posterior condyle position.

Adolescent ; Adult ; Cone-Beam Computed Tomography ; Face ; Female ; Humans ; Male ; Mandibular Condyle ; Temporomandibular Joint ; Young Adult

Objective To explore the regulating effects of Egr-1 promoter activated by ionizing radiation (IR) and doxorubicin (ADM) on the expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) genes. Methods The human GM-CSF cDNA and enhanced green fluorescent protein (EGFP) cDNA were linked together with IRES(internal ribosome entry site) and then inserted into the expression vector pCIneo under control of the Egr-1 promoter(Egr-EG). The vector was transferred into human bone marrow stromal cell line HFCL by liposome transfection. And the cells were exposure to ADM and IR. The activity of EGFP in HFCL/EG cells were detected by FACS. The effect of N-acetylcysteine on the expression of EGFP following exposure to ADM and IR was examined. The amounts of GM-CSF in HFCL/EG after chemotherapy or radiation were measured with ELISA. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM derived from cord blood were also studied. RT-PCR analysis for the expression of GM-CSF mRNA in HFCL/EG after exposure to ADM or IR. Results The percentage of EGFP+ HFCL/EG cells exposed to ADM and IR was increased compared with non-treatment group (1.2 % and 15.2 % vs 18.2 %, t = 5.11, P < 0.01). The levels of secreted GM-CSF in HFCL/EG cells exposed to ADM and IR was increased (P < 0.01), but no difference between ADM group and IR group (P 0.05). The expression of EGFP by HFCL/EG treated with ADM and IR was significantly decreased by N-acetylcysteine. The effects of GM-CSF in HFCL/EG cultural supernatants on expansion of CFU-GM in ADM group and IR group were significantly higher than that in HFCL group and non-treatment group. However, The CFU-GM count of IR group was higher than that of ADM group. The expression of GM-CSF mRNA in HFCL/EG cells exposed to ADM and IR was significantly increased(t = 4.37, P < 0.01). Conclusions GM-CSF gene expression regulated by Egr-1 promoter induced by ADM and IR could help the recovery from hematopoietic injury.

Objective To investigate the clinical features of internal carotid steal syndrome. Methods The clinical manifestations, CT or MRI, digital subtraction angiography, and blood flow compensation in 6 patients with internal carotid steal syndrome were analyzed. Results Of the 6 patients, 2 had unilateral internal carotid artery stenosis, 4 had severe stenosis (in which 2 were on the left side, 1 was on the right side, and 1 was on both sides). The clinical manifestations of the patients with internal carotid steal syndrome were watershed infarction and transient ischemic attack. Four patients had posterior circulation ischemia and 2 had anterior circulation ischemia. Digital subtraction angiography demonstrated that collateral circulation was established in all the 6 patients. The anterior communicating artery, posterior communicating artery, and pial artery were the common compensatory vessels. Conclusions Internal carotid artery steal syndrome can be presented as anterior or posterior circulation ischemia, and the collateral circulation plays an important role in the compensation.

Objective To investigate the potential of adult mesenchymal stem cells (MSCs) derived from human bone marrow to undergo cardiomyogenic differentiation after exposure to 5-azacytidine (5-aza) in vitro. Methods A small bone marrow aspirate was taken from the iliac crest of human volunteers, and hMSCs were isolated by 1.073g/mL Percoll and propagated in the right cell culturing medium as previously described. The phenotypes of hMSCs were characterized with the use of flow cytometry. The hMSCs were cultured in cell culture medium (as control) and medium mixed with 5-aza for cellular differentiation. We examined by immunohistochemistry at 21 days the inducement of desmin, cardiac-specific cardiac troponin I (cTnI), GATA 4 and connexin-43 respectively. Results The hMSCs are fibroblast-like morphology and express CD44+ CD29+ CD90+ / CD34- CD45- CD31- CD11a. After 5-aza treatment, 20-30% hMSCs connected with adjoining cells and coalesced into myotube structures after 14days. Twenty-one days after 5-aza treatment, immunofluorescence showed that some cells expressed desmin,GATA4, cTnI and connexin-43 in 5,10 μmol/L 5-aza groups, but no cardiac specific protein was found in neither 3μmol/L 5-aza group nor in the control group. The ratio of cTnI positively stained cells in 10 μmol/L group was higher than that in 5 μmol/L group (65.3 ± 4.7% vs 48.2 ± 5.4%, P ＜ 0.05). Electron microscopy revealed that myofilaments were formed. The induced cells expressed cardiac-myosin heavy chain (MyHC) gene by reverse transcription-polymerase chain reaction (RT-PCR). Conclusions Theses findings suggest that hMSCs from adult bone marrow can be differentiated into cardiac-like muscle cells with 5-aza inducement in vitro and the differentiation is in line with the 5-aza concentration. (J Geriatr Cardiol 2004;1(2) :101-107. )