0 05).Conclusion C kit level may be more closely related to the clinical parameters in SLE patients,which may reflect the clinical status of SLE patients.

Background Dimethyl sulfoxide(DMSO) is a commonly used adjuvant to promote testing drug solubility to prepare multi-levels testing drug concentrations.DMSO is cell type-dependent cytotoxic and its toxicity can interfere the testing drug evaluation.Determining its safe concentration on commonly used cell types is important for ocular drug development.Objective This study was to determine the minimal toxic concentration of DMSO for in vitro ocular cell lines in a simulated drug screening setting.Methods Retinal pigment epithelial (RPE) cells were isolated from one pigmented rabbit and primarily cultured.Human RPE cell strain (ARPE19),scleral fibroblasts line (S75-Fron),human Müller cell line (MIO-M1),human lens epithelial cell line (HLEC),human choroidal melanoma cell line (OCM-1),human umbilical endothelial cell (HUVEC) and human HeLa cell line (HELA) were cultured.Different concentrations of DMSO (1.6％,1.0％,0.8％,0.4％,0.2％ and 0.1％) were prepared with 160 μl DMSO solution and 9.84 ml RPMI1640 (or DMEM/F12 or DMEM) containing 2％ fetal bovine serum.Different concentrations of DMSO were added in medium for 96 hours,and the and viability (absorbance) of the cells was detected using MTS to evaluate the cytotoxicity of DMSO.Results Rabbit primary RPE cells showed the yellow-green fluorescence for cytokeratin(CK) and HMB45 red fluorescence for S100.The viability of the cells was gradually declined as the increase of DMSO dose,showing significant differences in ARPE19,S75-Fron,HLEC,OCM-1,HUVEC and primary RPE cells (all at P＜0.05),and when DMSO concentrations were ≥ 0.8％,the cell viabilities were significantly lower.But no significant difference was found in MIO-M1 cells among different doses of DMSO (F=0.830,P=0.547).The minimal toxic concentration of DMSO for ARPE19,HUVEC,HELA,HLEC,MIO-M1,OCM-1,primary RPE cells and S75-Fron was 0.8％,0.1％,0.8％,＞1.6％,＞1.6％,0.2％,0.2％,0.2％,respectively,and HUVEC was more sensitive to the cytotoxicity of DMSO (P=0.02),and MIO-M1 was the least sensitive to DMSO (P =0.39).The viability of HUVEC and primary RPE cells went down with the increase of DMSO dose,and S75-Fron viability started to decline in 0.1％ DMSO and then stabilize with the higher concentrations until 1.6％ DMSO at which the viability showed further decline.Conclusions The tolerability of ocular cells in vitro to DMSO varies depending on the cell types.The minimal toxic concentration ranged from 0.1％ to 1.6％.The result suggests that a concurrent DMSO control should be set up along with the testing compound.

Objective:To investigate the effects of 17β-Estradiol (E2) on mesenchymal stem cells (MSC) and to evaluate the effects of MSC treated with E2 on the maturation and function of dendritic cells (DC).Methods: We first isolated and cultured MSC from the human fetal lung.The MSC were treated with E2 for 24 hours at various concentrations ( 10~(-9),10~(-8) and 10~(-7) mol/L).After cell counting,proliferation,adherent ability and immunophenotypes of MSC were detected by flowcytometry.The gene expressions of cytokine (IL-6,TGF-βand VEGF) were measured by RT-PCR.The effects of MSC treated with E2 on the maturation and function of DC were determined.Results:After treated with E2,the proliferation and adherent ability of MSC were increased,while the immunophenotypes of MSC were not affected.When MSCs co-cultured with DC,MSC could inhibit the immuophenotypes and function of DC.However,when DC co-cultured with E2-pretreated MSC,the immunophenotypes (MHC Ⅱ,CD80 and CD86) of DC had been reconstructed.After treated with the high concentration of E2 for 24 hours,MSC secreted lower level of TGF-β than that in the control group,while IL-6 and VEGF expressions were increased compared with those in the control group.Conclusion: Estrogen may alter the immuno-suppressive effects of MSC on DC via modulating the cytokine secretion of MSC.

Objective: To set a quantified diagnostic standard for large intestinal cancer of spleen qi deficiency syndrome. Methods: The spleen qi deficiency syndrome was identified by experts on the basis of clinical epidemiological investigation of 311 patients suffering from large intestinal cancer. Corresponding points were assigned to the correlative factors (traditional Chinese medicine symptoms) on the basis of symptom differences between spleen qi deficiency syndrome and non-spleen-qi-deficiency syndrome. The best threshold was determined by receiver operating characteristic curve (ROC) according to syndrome differentiation from expert team, and the quantified diagnostic standard was established. The syndrome identification from the expert team which was regarded as golden standard was tested retrospectively. Results: All the traditional Chinese medicine symptoms possibly related to spleen qi deficiency syndrome were analyzed based on the opinions of experts, and 28 symptoms were confirmed as candidate correlative factors. The occurrence of 11 symptoms between spleen qi deficiency syndrome and non-spleen-qi-deficiency syndrome showed statistical differences by means of crosstabs analysis (P<0.05). The 11 symptoms were filtered by logistic regression analysis, and tiredness, fatigue, loose stool, and poor appetite were finally determined as the symptoms relative to large intestinal cancer. These four symptoms were analyzed with conditional probability conversion and endowed with 16, 11, 4 and 8 points respectively. The diagnostic standard of spleen qi deficiency syndrome of large intestinal cancer was over 13 points. The sensitivity, specificity and accuracy of retrospective examination were all above 80%, and its positive likelihood ratio was 9.89. Conclusion: The quantified diagnostic standard for spleen qi deficiency syndrome of large intestinal cancer is in accordance with clinical characteristics of large intestine cancer and the characteristics of TCM syndrome diagnosis.

Objective To investigate the effect of expectant management on the maternal and/or infant out-comes of early onset severe preeclamp sia in different gestafional weeks. Methods A retrospective study was carried out on 158 patients with early onset severe preeclampsia. They were divided into three groups according to their onset gestation ago:group A( <28weeks,n=28) ,group B(≥28、<32weeks,n=51) and group C(≥32、< 34weeks,n =79). Results The rates of complications declined along with the postponement of the onset gestation age, but there was no statistical significant difference among these three groups. The neonatal asphyxia rate and perinatal infant mor-tality of these three groups declined along with the postponement of gestation age, and there were statistical significant differences among these three groups ( P<0.05 ). Expectant treatment time of group B was significantly longer than that of the other two groups ( P<0.05 ), and cesarean section was a main method of pregnancy termination for the groups B and C. Conclusion The smaller the gestational ages in the early onset severe preeelampsia,the higher the maternal and/ or infant complication rates, neonatal morbidity and mortality.

Objective To investigate the effects of Xueshuantong injection on levels of inflammatory factors and homocysteine (Hcy) in patients with acute cerebral infarction.Methods 128 patients of acute cerebral infarction who received therapy in the hospital from January 2014 to May 2015 were collected as research objects to retrospectively analyze their clinical materials.According to therapeutic schemes, those patients were divided into control group (n=62) and observation group (n=66).The control group were given conventional treatments, such as anti-platelet aggregation, lipid regulation, maintaining balance of water-electrolyte and acid-base, controlling blood glucose, controlling blood pressure and so on.Besides that, the observation group were given Xueshuantong injection.After two weeks treatment, the curative efficacy, levels of inflammatory factors as high-sensitivity C reactive protein (hs-CRP), tumor necrosis factor α(TNF-α), interleukin 8 (IL-8), Hcy and adverse reactions were compared.Results The total efficacy of observation group was 93.9%, which was statistically higher than that of 80.6%in control group (χ2 =5.166,P=0.023).After treatment, hs-CRP, TNF-α, IL-8 levels in observation group were significantly lower than those in control group (all P<0.05).After one and two weeks treatment, The Hcy level in observation group was statistically lower than that in control group ( all P<0.05 ).The adverse reactions in observation group was 8 cases ( 12.1%), 6 cases ( 9.7%) in control group, with no significant difference between two groups (χ2 =0.196, P =0.658 ). Conclusion Xueshuantong injection is effective in treatment of acute cerebral infarction, which could significantly increase curative efficacy, alleviate inflammatory reaction and reduce level of Hcy with less adverse reactions.

Objective To investigate the muhilineage differentiation potential of bone marrow-derived mesenchymM stem eels (MSCs) in patients with systemic lupus erythematosus (SLE).Methods Density gradient centrifugation and plastic adherence methods were used for isolation of marrow-derived MSCs.Then tIIeir differentiation potentiality to lipoblasts and osteoblasts waft tested.MSCs loading on hydroxyapatite were elnbedded in the nude mouse's subcutaneous tissues.Eight weeks later.osteogenesis was evaluated by HE staining.PPA Rγ2,LPL,Runx2/CBFA1,osteocalcin gene expression in MSCs after differentiation were examined by RT-PCR.Results The positive rates of lipoblasts stained by oil red O and optical density in SLE were decreased than in the control group[(35±7)% vs (80±5)%] (0.14±0.04 vs 0.27±0.04),and the positive rates of osteoblasts stained by Alizarin Red S in SLE were decreased than those in the control group [(35±4)% vs (45±4)%].Osteoblast differentiation in the SLE group was less than that of the contro]group.The mRNA expression of LPL (0.369±0.020 vs 0.481±0.038).Runx2/CBFA1 (0.371±0.000 vs 0.563±0.069).osteoealcin (0.819±0.023 vs 0.962±0.049) of MSCs after difierentiation in the SLE group was decreased than that of the control group.There was no significant difference in the expression of PPARγ2 mRNA between SLE and controI group (0.421±0.052 vs 0.441±0.012).Conelusion MSCs from SLE have abnormalities in osteogenie and adipogenic differentiation potential.

Objective To evaluate the correlation between the types of vertebral artery occlusion and their compensatory hemodynamic changes and posterior circulation ischemia using color Doppler flow imaging combined with transcranial color-coded sonography.Methods From June 2015 to June 2016,A total of 108 patients with vertebral artery occlusion confirmed by vascular sonography,digital subtraction angiography (DSA) or CT angiography (CTA) were enrolled retrospectively.According to the magnetic resonance imaging (MRI)-diffusion weighted imaging (DWI) findings,they were divided into posterior circulation infarction (n=78 in infarction group) and non-posterior circulation infarction (n=30 in TIA group).Color Doppler flow imaging and transcranial color Doppler ultrasonography were used to examine the contralateral vertebral artery extracranial diameter, peak systolic velocity(PSV) and end diastolic velocity(EDV) of bilateral extracranial and intracranial vertebral arteries.The differences of the vertebral artery occlusion types,establishment of collateral circulation and hemodynamic changes of the contralateral vertebral artery were compared between the two groups.Results The patients with single vertebral artery occlusion in the infarction group and TIA group were 69 (88.5%) and 26 (86.7%) respectively;those with bilateral vertebral artery occlusion were 9 (11.5%) and 4 (13.3%) respectively.There was no significant difference in the number of vertebral artery occlusion between the two groups (χ2=0.000,P=1.000).The proportion of patients with vertebral artery occlusion in intracranial segment in the infarction group was higher than that in the TIA group (70.5% [55/78] vs.36.7% (11/30);χ2=10.444,P=0.001).The proportion of patients with the establishment of collateral circulation in the infarction group was lower than that in the TIA group (14.1% [11/78] vs.43.3% (13/30);χ2=10.711,P=0.001).The peak systolic velocity (PSV) and the end diastolic velocity (EDV) of contralateral extracranial vertebral artery in patients with single vertebral artery occlusion in the TIA group were higher than those in the infarction group (65±21 cm/s vs.57±15 cm/s,25±8 cm/s vs.20±7 cm/s,t=2.043 and 2.606 respectively,all P<0.05).Conclusion The establishment of collateral circulation and hemodynamic compensation of the contralateral vertebral artery after vertebral artery occlusion were closely associated with the occurrence of posterior circulation ischemia.

AIM:ToinvestigatetheexpressionofFoxp3+regulatoryTcells(Foxp3+Tregs)andprogrammed death receptor 1 (PD1) in gastric cancer tissues and their association with clinicopathological factors and prognosis of the patients.The correlation between the 2 molecules was also analyzed at the same time.METHODS: The tumor sections from 111 gastric cancer patients were stained for Foxp3 and PD1 by the method of immunohistochemistry.The associations of the expression levels of these 2 molecules with clinicopathological factors involved in the disease progression and progno-sis were statistically analyzed .The relationship of their expression was detected.RESULTS:Foxp3 +Tregs and PD1 were expressed in the gastric cancer tissues, and PD1 was expressed in the tumor infiltrating lymphocytes ( TILs) .The expres-sion of Foxp3 and PD1 was correlated with lymph node metastasis, clinicopathological stage and prognosis of gastric cancer patients.The expression of these 2 determinants in the patients with lymph node metastasis and an advanced clinicopatho-logical stage was distinctly higher ( P<0.05 ) .The patients with positive expression of the 2 indexes presented a lower overall survival rate and worse prognosis (P<0.05).A significantly positive correlation between the infiltration of Foxp3 +Tregs and the expression of PD1+TILs was also observed (P<0.01).CONCLUSION:Foxp3 +Tregs and PD1 +TILs co-infiltrate in the gastric cancer tissues, which can be used as biological markers to predict the disease progression and prog-nosis.

Laryngopharyngeal reflux is an independent disease that affects the quality of life in children,but its pathogenesis is not clear and its diagnostic criteria is not unified.With the reports of laryngopharyngeal reflux continuing to emerge in recent years,this study aims to review the progress in pathogenesis,diagnosis and treatment of laryngopharyngeal reflux in children.