Objective To discuss the management and selection of donor and recipient veins in the transfer of vascularied autogeneous submandibular gland (SMG).Methods The SMGs of 48 patients with severe keratoconjunctivitis were transfered to the temporal region by microsurgery from June,2002 to June,2013.The secreted saliva was used as the substitute of tear.Donor and accepting-site vessels,vessels crisis and managements,survival of grafts were retrospectively analysed.Results Transplantation succeeded in 45 patients and failed in 3.For donor veins,39 were facial veins,12 were venae comitantes of facial artery,1 was vein near the duct.For revipient veins,41 were superficial temporary veins,6 were deep temporary veins and 5 were veins in the upper neck.For revipient artery,except superficial temporary artery,deep temporary artery was also a good selection.After surgery,2/5 venous crisis cases were rescued by reanastomosising veins.TC99m examination suggested that the 49 TSMGs were survived,and the ducts were unobstructed.Follow up lasted for 6 months to 10 years,the symptoms of photophobia and anemophobia were alleviated,the symptoms of corneal xerosis disappeared.Good clinical efficacy was obtained after transplantion.Conclusion During SMGs transplantion,facial veins,venae comitantes of facial artery or vein near the duct can be used for donor vein.For recipient veins,except the superficial temporary veins as major,deep temporary veins or veins in the upper neck is also a secection.Correct selection and microsurgical management of donor and revipient veins are keys to successful SMGs transplantion.

Objective To investigate whether 17β-estradiol (E2) can stimulate the proliferation,migration,and secretion of trefoil factor family 1 (TFF1) in papillary thyroid cancer K-1 cells and explore the molecular mechanisms.Methods ELISA was used to detect the content of TFF1 in the supernatant of K-1 cells after the treatment of E2,propylpyrazoletriol (PPT,ERα agonist) or diarylpropionitrile (DPN,ERβ agonist).The expression of ERα and ERβ in the untreated cells was measured by Western blotting.ERα siRNA and ERβ siRNA by RNA interference were designed and synthesized,and the change of TFF1 was measured by ELISA again after the transfection.The interaction between TFF1 promoter and ER was evaluated by chromatin immunoprecipitation analysis (CHiP).The proliferation and migration were detected in the K-1 cells after E2 treatment by MTT assay and Transwell chamber test respectively.Festults After E2 treatment,the TFF1 content in the supernatant of K-1 cells was increased gradually,reached peak at 24 h,and then declined slowly.PPT treatment enhanced the secretion of TFF1 but DPN decreased it in the K-1 cells.Transfection of ERα siRNA obliterated the inductive effect of E2 on the secretion of TFF1,but that of ERβ siRNA increased the inductive effect in the K-1 cells.Western blotting showed that the expression level of ERα was higher than that of ERβ in the K-1 cells.ChIP results confirmed that ERα protein was bound to the promoter of TFF1 gene in K-1 cells.E2 treatment promoted cell proliferation and improved cell migration in the K-1 cells.Conclusion E2 induces the expression and secretion of TFF1 in K-1 cells through ERα-dependent manner,and thus promotes the proliferation and migration of the cells.

AIM:To observe the characterization in neural cells derived from the hippocampus of embryonic rats and to examine the effect of myelin-associated glycoprotein(MAG) on the proliferation,differentiation and neurite growth of neural stem cells(NSCs).METHODS:The hippocampus cells of embryonic rats were isolated and cultured in vitro.The expressions of nestin and doublecortin,the marks of NSCs,were observed by immunocytochemical method.The rate of proliferating cells was examined by BrdU immunocytochemistry.The average neuronal neurite length and the percentage of differentiated neurons were detected by immunocytochemistry staining.RESULTS:The hippocampus cells of 16 days old embryonic rats had the characteristics of NSCs.The percentage of differentiated neurons(?-tubulin Ⅲ-positive cells) was 18.17%?2.79% and the average neuronal neurite length was(136.27?33.66)?m,seven days after the differentiation initiated in vitro in control group.After NSCs were treated with MAG-Fc(200 ?g/L),the percentage of differentiated neurons and the average neurite length were decreased,respectively,to 10.05%?3.42%(P0.05).CONCLUSION:MAG-Fc inhibits the differentiation and neurite growth of the NSCs,but has no effect on the proliferation.

OBJECTIVETo explore the myeloid-derived suppressor cell (MDSC) expression in the peripheral blood and lesions of 4NQO-induced tongue carcinoma in mouse.

METHODSThe established 4NQO mouse model was used to analyze the distribution of MDSC and T cell subsets in the peripheral blood by flow cytometry. The relations of MDSC with T cell subsets and CD4⁺/CD8⁺ changes were evaluated. The distribution of MDSC in the lesions of tongues was analyzed by immu- nohistochemistry, and the expression of arginase 1 (ARG-1) in tongue tissues was detected by real-time polymerase chain reaction.

RESULTSDuring tumor progression, a significant increase was observed in the frequency of MDSC in the peripheral blood of 4NQO treated mice (P < 0.01). The frequency of MDSC was positively correlated with systemic CD3⁺CD8+T cells but negatively correlated with the CD4⁺/CD8⁺ ratio. Squamous cell carcinomas were extensively infiltrated with MDSC, whereas dysplastic area and normal tongue mucosa had only sparse MDSC infiltration. The majority of MDSCs were located in the stroma, particularly along the tumor invasive front. Moreover, 4NQO-treated mice showed significantly higher ARG-1 mRNA levels in the tumor site (P<0.01).

CONCLUSIONMDSC may contribute to oral tumor progression and represents a potential target for immunotherapy of oral cancer.

4-Nitroquinoline-1-oxide ; Animals ; Arginase ; Cell Count ; Flow Cytometry ; Mice ; Models, Animal ; Myeloid-Derived Suppressor Cells ; immunology ; Real-Time Polymerase Chain Reaction ; T-Lymphocyte Subsets ; immunology ; Tongue Neoplasms ; immunology

[Objective] To investigate the inhibitory effect of CTLA-4Ig fusion protein on atherosclerosis in the mice with an apolipoprotein-E gene defect fed on cholesterol diet.[Methods] Twenty-five male 10-week-old ApoE-/- mice were selected and fed on cholesterol diet for 4 weeks,5 out of which were executed at random as control group and their pathological sections were kept to observe the early fatty streaks.The other 20,divided into CTLA-4Ig treatment group,PBS group,IgG1 group,and blank group at random,5 in each.Three groups were given intraperitoneal injection of CTLA-4Ig (10 μg per time),PBS (100 μL per time),Rat-IgG1 (10 μg per time) respectively,twice a week,for 8 weeks.The blank group has no treatment.Followed by 8-week treatment,the whole aorta from the root to crotch of iliac artery was separated after anesthesia with the intraperitoneal injection of 1 % pentobarbital.Subsequently,the area ratio of plaque and lumen,the thickness ratio of endangium and tunica media,the lipid-soaking extent intra-plaque and the content of collagen fibrils and smooth muscle cells intra-plaque were analyzed by image-processing soft.[Results] After fed on cholesterol diet for 4 weeks,there were obviously atherosclerosis in the aorta in the ApoE-/- mice.There were typical atherosclerotic plaque in ApoE-/- mice fed on cholesterol diet after another 8 weeks.The area ratios of plaque and lumen in CTLA-4Ig group,PBS group,IgG1 group,and blank group were 0.27 ± 0.08,0.40 ± 0.08,0.43 ± 0.08,and 0.46 ± 0.10,and obviously increased than those in control group (0.05 ± 0.01,P ＜ 0.05).The thickness ratios of endangium and tunica media in four groups were 2.6 ± 0.6,6.0 ± 0.9,5.7 ± 0.8,and 5.9 ± 0.6 and obviously increased than those in control group (0.5 ± 0.1,P ＜ 0.05).The lipid-soaking extent intra-plaque in experimental groups were 26.0 ± 3.0,40.8 ± 5.7,40.6 ± 3.0,and 43.2 ± 5.7,and were obviously increased than those in control group (7.2 ± 1.4,P ＜ 0.05 ).It was found that the area ratio of plaque and lumen,the thickness ratio of endangium and tunica media,and the lipid-soaking extent intra-plaque in CTLA-4Ig group were significantly lower than those in PBS group,IgG1 group,and blank group (P ＜ 0.05),but there was no significant difference in those between the PBS group,IgG1 group,and blank group (P ＞ 0.05).The content of collagen fibrils in CTLA-4Ig group were 16.0 ± 1.1 and higher than those in PBS group,IgG1 group,and blank group (8.6 ± 1.2,9.2 ± 1.5,and 9.0 ± 1.3,P ＜ 0.05).The content of smooth muscle cells in plaque in CTLA-4Ig group were 11.8 ± 1.0 and higher than those in PBS group,IgG1 group,and blank group (7.8 ± 0.8,7.5 ± 0.9,and 7.3 ± 0.7,P ＜ 0.05).There was no significant difference in content of collagen fibrils and smooth muscle cells between the PBS group,IgG1 group,and blank group (all P ＞ 0.05).[Conclusion] CTLA-4Ig fusion protein could evidently inhibit the atherosclerosis progression and enhance the stability of plaque through increasing the content of collagen fibrils produced by smooth muscle cells intra-plaque in ApoE-/- mice fed on cholesterol diet.

Aim To explore the best way of calculating antihypertensive effect of nifedipine GITS on trough to peak ratio (T/PR), and smoothness index (SI) of the drug from ambulatory blood pressure monitoring (ABPM). Methods 32 cases of mild to moderate essential hypertension patients were enrolled and each was given 30 mg of nifedipine GITS once daily. ABPM was repeated for four weeks. ABPM data were analyzed statistically and T/PR calculated by both individual and whole group way. Results The casual blood pressure(CBP) and ABP were lowered by (24?12)/(12?8) mmHg and ( 14.5 ? 3.9 )/( 11.2 ? 3.0) mmHg .The T/PR by individual way was 0.65 ? 0.23 for SBP and 0.66 ? 0.25 for DBP, while by whole group way 0.62 for SBP and 0.68 for DBP. The smoothness index (SI), a new method for assessing the homogeneity of 24 hour blood pressure reduction by antihypertensive therapy, was 3.74 for SBP and 3.77 for DBP after treatment. Conclusion Nifedipine GITS lowers blood pressure effectively and smoothly for 24 hours long. Antihypertensive effects can be reflected by T/PR and SI.

Objective To investigate the variation of biomarker of coagulation, anti-coagulation, fibrinolysis in elderly critical patients and find out whether they are related to the disease severity. Methods Sixty-seven patients were no less than 60 years old. Eligible criteria: coincidence with the diagnostic criteria of systemic inflammatory response syndrome (SIRS) and APACHE Ⅱ score was no less than 10 scores. Blood sample was drawn from the venous for the test of biomarker (APTT, PT, TT, D-D, Fib, AT-Ⅲ , PC, PAI-1). According to the existent status,all the patients were divided into two groups:survival group (43 cases) and death group(24 cases) ,meanwhile,according to the diagnostic criteria of MODSE,all the patients were divided into MODSE group (30 cases) and non-MODSE group (37 cases). Results There were significant differences in APACHE Ⅱ score between MODSE group and non-MODSE group, survival group and death group [(25.83 ± 1.19) scores vs(18.1±20.73) scores and(18.81±0.72) scores vs(26.50 ± 1.42) scores](P <0.01). The PT and D-D in MODSE group anti death group were higher than those in non-MODSE group and survival group, the differences were significant (P <0.05),while the activity of AT-Ⅲand PC in MODSE group and death group were lower than those in non-MODSE group and survival group, the differences were significant (P <0.05). The PT,D-D and PAI-1 were positively correlated to APACHE Ⅱ score (related coefficients were 0.328, 0.308, 0.335,P <0.05). The AT-Ⅲ and PC were negatively correlated to APACHE Ⅱ score (related coefficients were -0.469, -0.559,P <0.01). Conclusions The abnormality of eoagnlation-fibfinolysis system exists in elderly critical patients. The extended PT, elevated D-D and PAI-1 ,descended PC and AT-Ⅲ are the hints of disease severity and poor prognosis.

Objective To stuay the serum levels of sCD40L,hsCRP,ICAM-1 and VCAM-1,and the expression of CD40L of the CD4+T cells in patients withacute coronary syndrome(ACS),and to explore the relationship between CD40L and inflammatory factors and the effects of CD40/CD40L on ACS.Method Thirty-two coronary heart disease patients without history of other discernible systemic disease and medicine of steroids or immunosuppressants taken were divided into acute myocardial infarction group(AMI,n=11),unstable angina pectoris group(USP,n=14)and stable angina pectoris group(SAP,n=7).The control group was composed of eight healthy volunteers(CON group).Theexpression of CD40L Was determined by flow cytometry(FCM).Serum sCD40L.ICAM-1 and VCAM-1 were determined by using ELISA.The serum hsCRP was assayed by using immunoturbidimetry.Data were analyzed with SPSS 11.0 software for windows.Results The expression percentage(%)of CD40L of the CD4+T cells,and the serum levels of sCD40L,hsCRP,ICAM-1 and VCAM-1were sifnificantly higher in patients of AMI group than those in patients of other groups(P＜0.05 or P＜0.01).Similarly,those biomarkers in patients of UAP group were usually higher than those in patients of CON or SAP groups(P＜0.05).There Was a positive correlation between the expression of CD40L and the serum level of VCAM-1 in paients of AMI group(P＜0.05),and likewise,a positive correlation also existed between the serum level of sCD40L and other factors,hsCRP,ICAM-1 as well as VCAM-1,in patients of AMI group(P＜0.05).Conclusions The enhanced expression of CD40L of the CD4+T cells and high serum level of sCD40L are present in patients with acute coronary syndrome.The hsCRP,ICAM-1 and VCAM-1 play roles in the pathogenesis of ACS,and they have correlation with enhanced expression of CD40L and high serum level of sCD40L.Therefore,CD40L and sCD40L may be used as indicators of risk in coronary heart disease.

Aim To explore new ways for developing anticancer drugs by the separation of pigment from Fu-sarium species JN158 ( Fusarium sp JN158 ) , the iden-tification of its structure, the screening of anticancer components and the study of its partial mechanism. Methods Pigment separation was done by HPLC, structural analysis by UV, IR, NMR, the screening of anticancer activity by MTT. Western blot was used to analyze the protein expression of CyclinD1, NF-κB, VEGF in tumor cells. Results The results showed that the pigment from Fusarium produced a total of six different peaks, of which peak Ⅵ was the anthocya-nins. Its molecular weight is about 382, molecular for-mula is C17 H18 O10 . According to investigation, this pig-ment was probably a new compound, which could in-hibit the proliferation of MCF-7 cells markedly ( IC50:0.011mmol·L-1 ,P<0.05;the control medicine ube-nimex IC50:10 mmol · L-1 ) in a concentration-de-pendent manner, and had no effect on human umbilical cord intravenous endotheliocyte ( HUVEC ) . The influ-ence on the gene expression of CyclinD1, NF-κB, VEGF in MCF-7 cells varied with the concentration of this compound. The Western blot results showed that VI pigment compound inhibited CyclinD1, NF-κB, VEGF gene expression (P<0.05 or 0. 01),compared with the control group. Conclusion The Ⅵ pigment compound from Fusarium sp JN158 could inhibit MCF-7 proliferation by inhibiting CyclinD1, NF-κB, VEGF gene expression. The compound may be a promising compound against breast cancer.

Objective To determine the levels of soluble thrombomodulin(sTM) in patients with acute pulmonary embolism (PE), and evaluate sTM clinical significance.Methods The sTM levels were determined with enzyme linked immunosorbent assay in PE patients, and compared with healthy control group. Eighteen PE patients were divided into massive PE group and non - massive PE group, non - respiratory failure group and respiratory failure group, and compared the sTM level in the groups. Results Level of sTM in PE patients was higher than that of control group (P