Objective To investigate the protective effect of fructose 1,6-diphosphate (FDP) on the brain against ischemia-reperfusion (I/R) injury and the possible mechanism. Methods One hundred and eighty SD rats weighing 275-325 g were randomly divided into 3 groups (n = 60 each): Ⅰ gham operation group; Ⅱ I/R group and Ⅲ FDP group. Global cerebral I/R was produced by 4-vessel technique. Bilateral vertebral arteries were coagulated and bilateral common carotid arteries were occluded for 5 min and then released for reperfusion. In sham operation the four vessels were exposed but not occluded. In FDP group FDP 1.5 mg?kg-1 was given Ⅳ when reperfusion was started, while in sham-operation group and I/R group normal saline (NS) 1.5 ml?kg-1 was given Ⅳ instead of FDP. The animals were killed at 2, 6, 12, 24, 48 and 72 h of reperfusion ( n = 5 each) for determination of cerebral SOD activity and MDA contents, the number of apoptotic neurons (TUNEL) and expression of P38 and Ref-1 in the brain (immuno-histochemical method) .Results The MDA content was significantly higher whereas the SOD activity and P38 and Ref-1 expression were significantly lower at all time points in I/R group than in sham operation group ( P

Objective To investigate the therapeutic mechanism of umbilical cord mesenchymal stem cell(UC-MSCs)trans-plantation for the graves disease(GD)mice .Methods Thirty two mice were divided into 3 groups as following :normal control group (G0) ,GD control group (G1) ,UC-MSCs group(G2) .Enzyme linked immunosorbent assay(ELISA)was used to measure the level of TSAb in blood serum and the expression of FT4 was measured by chemiluminescence .Thyroid sections were stained with hema-toxylin and eosin(HE)for histological examination .Splenocytes were stained with multicolor immunofluorescence and detected by flow cytometry to analyze the percentages of CD1dhiCD5+CD19+ regulatory B cells(Bregs) .Expressions of IL-10 and TGF-βmR-NA in spleen organization were measured by Real-time PCR .Results At 26 weeks ,the level of TSAb in blood serum in G2 was more significantly decreased than in G1(P<0 .05) ,and the level of CD19+ B in spleen in G2 was also more significantly decreased than in G1(P<0 .05) ,however ,the percentage of CD1dhiCD5+CD19+ Bregs splenocytes and the levels of IL-10 and TGF-βmRNA in spleen organization were more significantly increased than in G1(P<0 .05) .The concentration differences of TSAb in serum was negatively correlated with the percentage differences of CD1dhi CD5+ CD19+ Bregs ,however ,positively correlated with the expres-sion differences of IL-10 and TGF-βmRNA in spleen before and after transplantation .Conclusion Activation of Bregs may be one of the mechanisms of UC-MSCs therapeutic effect on GD mice .

Objective To explore the role of OAZ gene in the pathogenesis of systemic lupus erythe-matosus (SLE) by using RNA interfering technique. Methods Peripheral blood mononuclear cells (PBMC) from SLE patients were collected. Each sample was equally divided into four groups for cell culture in 96 well plates. Specific siRNA for OAZ and GAPDH were concordantly added to the experimental group and the positive control group, while nonspecific siRNA was added to the negative control group and only culture medium was added to the Mock control group. Cells and supernatants were harvested after culturing for 72 hours, then RNA was extracted and reverse transcripted to cDNA. OAZ, Id1, Id2, Id3, Id4 mRNA expression levels were analyzed by using real-time PCR. Levels of IFN-γ, IL-4, IL-10, IL-12, IL-21, CCL2, ANA in the supernatant were tested by ELISA. Relationships between the expression levels of OAZ mRNA with levels of cytokines and ANA were analyzed. Results OAZ, Id1, Id2, Id3 gene mRNA expression levels (△Ct: 12.5±1.4, 8.9±1.5, 4.3±0.8, 8.04±1.1) in the experimental group were significantly decreased comparing to those in the negative control group (△Ct: 10.2±1.1, 6.5±1.2, 2.4±1.3, 6.2±1.2 respectively, P<0.05). Levels of IFN-γ, IL-10, IL-12, IL-21 and ANA in the experimental group were significantly lower than those in the negative control group (P< 0.05), but level of CCL2 was higher than the negative control group (P<0.05). Difference of OAZ mRNA expression levels (△△Ct) between the experimental group and the negative control group was negatively correlated with changes of ANA, IL-21 levels, but positively correlated with changes of Th1/Th2, CCL2. Conclusion OAZ siRNA can effectively reduce the expression of genes involved in the OAZ signaling pathway in SLE. OAZ may lead to abnormal production of ANA via regulating Id genes and cytokines.

Objective To explore the expression levels of interferon-inducible genes in patients with systemic lupus erythematosus ( SLE) , and to validate these gene expressions as potential biomarkers for the differentiation of disease flare and infection.Methods Peripheral blood was obtained from 48 SLE, 16 rheumatoid arthritis ( RA) patients and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expressions of MX1, OASL,OAS1, ISG15 and LY6E at transcription level.Univariate logistic regression analysis and multivariate conditional logistic regression model were applied to analyze 5 related factors for infection or activity.Results ( 1 ) The expression levels of MX1, OASL, 0AS1, ISG15, and LY6E mRNA in SLE patients were significantly increased as compared with normal controls ( P all < 0.01 ) , while the expression levels of OASL,OAS1 ,ISG15 and LY6E mRNA in SLE patients were also higher than those in RA patients (P all <0.05 ).(2)There were no significant difference between male and female patients of the 5 gene expression in SLE patients.(3) By logistic regression analysis, ISG15 and LY6E were independent risk factors for active SLE patients (P <0.01) , OASL expression was an independent risk factor for SLE patients with infection ( P = 0.003 ).Conclusion All the 5 interferon inducible genes are highly expressed in SLE patients, in which ISG15 and LY6E are independently associated with disease flare, while OASL may be helpful for the evaluation of infection in SLE patients.

Objective To study the rule of lower-cervical lymphatic metastasis in thoracic esophageal carcinoma,and make evaluation about the reasonable extent of lymphadenectomy. Methods One hundred and eight cases of thoracic esophageal carcinoma through chromatic ultrasound and CT before operation were divided into different groups selectively,while three fields lymphadenectomy (3-FL) was adopted in 31 cases,and two fields lymphadenctomy (2-FL) was adopted in 77 cases. Results The rate of lower-cervical lymphatic metastasis was 87.1%(27/31) through chromatic ultrasound and CT,and that was 25.0%(27/108) before the two up-mentioned examinations (P<0.05). In all cases, the rate of lower-cervical lymphatic metastasis in the upper pectoral esophageal carcinoma was 47.6% (10/21),that in the middle pectoral esophageal carcinoma was 21.3%(13161),and that in the middle and lower pectoral esophageal carcinoma was 19.5%(17187) ,P<0.05. Through 3-FL, the rate of lower--cervical lymphatic metastasis in the upper pectoral esophageal eareinoma was 57.1%(12/21), that in the middle pectoral esophageal carcinoma was 23.0%(14/61), and that in the middle and lower pectoral esophageal carcinoma was 21.8%(19/87), P<0.05. Conclusions The regionality metastasis is the main fashion in thoracic esophageal carcinoma with lower-cervical lymphatic metastasis. It is feasible to judge lower-cervical lymphatic metastasis through chromatic ultrasound and CT before operation.The 3-FL of the upper pectoral esophageal carcinoma is recommended. The 3-FL is selectable in the middle and lower pectoral esophageal carcinoma according to the result of chromatic ultrasound and CT.

Objective To study the role of the expression levels of 5 type Ⅰ interferon (IFN)-inducible genes (LY6E, OAS1, OASL, MX1, and ISG15) in the diagnosis and disease activity evaluation of systemic lupus erythematosus (SLE). Methods Peripheral blood was obtained from 68 SLE patients, 50 patients with other connective tissue diseases and 26 normal controls, and total RNA was extracted and reverse transcribed into complementary DNA. Real-time PCR technique was used to determine gene expressions at transcription level. An IFN score for each individual was calculated according to the expression of 5 1FN genes. Comparisons of gene expression and IFN score were made among groups. The genes expression levels in patients with SLE were analyzed using receiver operative characteristic curve. The association between IFN scores and disease activity, as assessed by the SLEDAI scores and 24 h proteinuria, was analyzed using Spearman correlation analyses. Results ① The expression levels of MX1, OASL, OAS1, ISG15 and LY6E mRNA in SLE patients were significantly increased as compared with normal controls and disease controls (P all＜0.01 ).② IFN scores in SLE patients (17.9±29.1) were significantly increased as compared with normal controls (0±3.3)and disease controls (3.0±8.1) (P all＜0.01 ). ③ IFN scores area under the ROC curve (AUCROC) was 0.846. When The IFN scores reached 2.56, its sensitivity and specificity for the diagnosis of SLE were 93.1%and 78.3%, respectively. ④ Levels of IFN score was positively correlated with SLEDAI scores (r=0.256,P＜0.05) and 24 h proteinuria (r=0.337, P＜0.05). Conclusion The 5 IFN-inducible genes are highly expressed in SLE patients. IFN score level is valuable for the diagnosis of SLE and a high IFN score is usually associated with an elevated disease activity.

Objective To investigate the in vivo effects of allogeneic mesenchymal stem cells transplantation (MSCT) on OAZ signaling pathway in patients with systemic lupus erythematosus (SLE).Methods Isolated and expand human MSCs from bone marrow cells or umbilical cord of healthy donors were infused into SLE patients. Peripheral blood cells were collected from 10 pre-MSCT patients as well as 1 week and 4 week post-MSCT, and RNA was extracted and reverse transcripted to cDNA. mRNA expression levels of OAZ and Id1-3 were measured by using real-time PCR. Serum levels of IL-10, IL-12, IL-21, CCL2 and ANA were tested by ELISA. Relationships of the gene expression levels with levels of cytokines and ANA were analyzed. Results mRNA expression levels of OAZ, Id1 and Id3 gene in patients with SLE were significantly decreased at week 1(△C:12.4±1.1, 9.7±1.9, 9.7±1.9, 2.1±1.0) and at week 4 (△Ct:13.3±1.2, 10.4±1.5,10.8±1.2, 2.1±1.2) after MSCT when compared to those of the pre-MSCT (△Ct:11.0±0.9, 7.4±2.1, 7.8±2.1, 0.1±1.5 respectively, P all＜0.05). Levels of IL-10, IL-21 and ANA were significantly lower 4 week after MSCT than those before (P＜0.05); while level of CCL2 was higher than pre-MSCT (P＜0.05). Cytokines and ANA levels 1 week after MSCT were not differentially changed comparing to those of the pre-MSCT. Alteration of OAZ mRNA expression levels pre- and post-MSCT were negatively correlated with changes of ANA, IL-21levels and positively correlated with changes of IL-12/IL-10 and CCL2 levels. Conclusions The expression of genes involving in the OAZ signaling pathway is effectively reduced along with the alteration of several cytokines and ANA after allogeneic MSCT in SLE patients. OAZ signaling pathway may play an important role in MSCT treatment for SLE.

Objective To investigate the effect of tea polyphenols on global cerebral ischemia reperfusion injury in rats.Method Forty-five pathogen-free male SD rats weighing 180-220 g were randomly divided into 3 groups( n =15 each):sham operation group (group S),cerebral ischemia reperfusion group (group IR) and tea polyphenols group (group TP).Global cerebral ischemia reperfusion injury was establish by four-vessel occlusion method.At 24 h of reperfusion,five rats were chosen and Evan's blue(EB) was injected iv,and then sacrificed and brain was removed for determination of EB content; another five rats were sacrificed and brain was removed for determination of water content; five rats were chosen for Morris water maze test.Result Compared with group S,EB content and water content in brain tissue were increased in groups IR and 'rP,and escape latency was prolonged,frequency of crossing the original platform was reduced in group IR ( P ＜ 0.05 ).Compared with group IR,EB content and water content in brain tissue were decreased,and escape latency was shortened,frequency of crossing the original platform was increased in group Tp ( P ＜ 0.05).Conclusion Tea polyphenols can attenuate global cerebral ischemia reperfusion injury in rats.

Objective Recently, The incidence of fatty liver is increasing , with the improvement of people′s living standard. We here established an available model with non-alcoholic fatty liver disease in rabbits and then studied its sonographic findings . Methods Forty male Japanese rabbits were randomly divided into 2 groups:control group and model group.The rabbits in model group were fed with high-fat diet for the establishment of the model of non-alcoholic fatty liver disease.The rabbits in control group were fed with standard diet. At the baseline, 12th, 16thand 20th week , all the livers in 2 groups were scanned by ultrasonography, and at the end of 20 th week, all the rabbits in 2 groups were killed for pathological analysis. Results Both the ultrasonography and in pathology demonstrated the successful establishment of non-alcoholic fatty liver models.The result of study demonstrated significant differences (P<0.05).At the 12th week, all of the 19 livers in model group showed fatty livers in sonography:8 low-, 9 middle-and 2 high-grade.The degree of steatosis aggravated pro-gressively with modeling time.Most of livers showed middle-grade fatty at the 16th week, and at the 20th week, they all demonstrated middle-or high-grade fatty liver:10 middle-and 9 high-grade, furthermore, ascites occurred in 3 cases.The pathological results were consistent with the findings of sonography, and fibrosis were observed in pathology. Conclusion Animal model with non-alcoholic fatty liver disease in rabbits can be established by high-fat diet.Besides, ultrasonography is a good method to monitor the establishment of the model .

Objective To investigate the in vivo effects of artesunate (ART) treatment on OAZ gene expressions in MRL/lpr lupus mice.Methods Twenty-four 12-week-old female MRL/lpr lupus mice were randomly divided into two groups by the random number table,i.e.,the lupus control group and the ART treatment group,12 in each group.Besides,another 8 BABL/C mice were recruited as the normal control group.Peripheral blood mononuclear cells (PBMC) were collected from lupus mice before treatment as well as 4 week and 8 week after treatment,and RNA was extracted and reverse transcripted to cDNA.mRNA expression levels of OAZ and Id1-3 were measured by using real-time polymerase chain reaction (PCR),and the data between the two groups were analyzed by t test,a plurality of samples were compared with the single factor analysis of variance.Serum levels of IFN-γ,IL-4,BAFF and ANA were tested by enzyme-linked immunosorbent assay (ELISA).Relationships of the gene expression levels with levels of cytokines and ANA were analyzed.Statistical analysis were uising t test,ANOVA and LSD-t test.Results mRNA expression levels of OAZ,Id1 and Id3 gene (△Ct:12.9±0.8,12.0±0.4,10.2±0.8) in lupus mice were significantly different from 8 weeks after the treatment comparing to those in the lupus control group (△Ct:9.8±1.0,9.3± 1.1,8.1±0.8) and the normal control group (△Ct:13.9±1.2,11.4±0.7,4.7±0.8,10.3±1.0)(F=7.46,P=0.008; F=6.37,P=0.032; F=5.63,P=0.042),and alteration of OAZ mRNA expression levels before and after the treatment were positively correlated with changes of Id1,Id3 levels (r=0.867,0.947; P＜0.05),and levels of IL-4 and ANA were significantly lower 8 weeks after the treatment than those in the lupus control group (P＜0.05); while level of IFN-γwas higher than those in the lupus control group (P＜0.05).Alteration of OAZ mRNA expression levels before and after the treatment were negatively correlated with changes of ANA,BAFF levels (r=-0.955,r=-0.937; P＜0.05) and positively correlated with changes of Th1/Th2 levels (r=0.976,P＜0.01).Conclusion The expression of genes involving in the OAZ and downstream gene are effectively reduced along with the alteration of several cytokines and ANA after ART treatment.OAZ signaling pathway can play an important role in ART treatment for SLE.