Objective:This study aimed to clarify the correlation of SPRY4-IT1 expression with the clinicopathological character-istics and prognosis of patients with esophageal squamous cell carcinoma (ESCC), as well as the role of SPRY4-IT1 in promoting ES-CC cell growth. Methods:Quantitative real-time polymerase chain reaction for SPRY4-IT1 expression was performed on 50 paired can-cerous and adjacent non-cancerous esophageal specimens. Small interfering RNA was used to suppress SPRY4-IT1 expression to fur-ther explore its role in tumor progression. Cell viability was tested in vitro by MTT assay (OD=490 nm), and cell apoptosis and cell cy-cle were investigated by flow cytometry. Results:We found markedly elevated SPRY4-IT1 expression in cancerous tissues compared with adjacent non-cancerous tissues (90%, P<0.01). Relative SPRY4-IT1 expression levels were correlated with some clinicopathologi-cal characteristics, such as tumor size (χ2=5.333, P=0.021), elevated TNM (2009) stage classi fi cation (χ2=5.556, P=0.018), and de-creased overall survival rates (χ2=5.296, P=0.021). SPRY4-IT1 expression level was not correlated with patient age, gender, smoking status, or alcohol consumption (all P>0.05). Further experiments showed that SPRY4-IT1 expression levels were significantly higher in three ESCC cell lines than in the normal human esophageal epithelial cell line Het-1A. In vitro assays of the ESCC cell line KYSE30 demonstrated that knockdown of SPRY4-IT1 expression by small interfering RNA reduced cell growth, mediated cell cycle arrest at the G0-G1 phase, and promoted cell apoptosis (all P<0.01). Conclusion:SPRY4-IT1 was overexpressed in ESCC tissues and ESCC cell lines and promoted the growth of ESCC cells. The dysregulated expression of long non-coding RNA SPRY4-IT1 may play an important role in the process of ESCC development and may be developed as a useful biomarker for the diagnosis and prognosis of ESCC.

BACKGROUND: It has been demonstrated that pedicle screw internal fixation influencing factors involve screw structural morphology, thread parameter, bone density, moment size for tightening screw during operation, and depth of screw placement. There is little known about the correlation of pullout strength of spinal pedicle screw with device for transverse traction to extraversion angle. OBJECTIVE: To assess effect of the spinal pedicle screw with device for transverse traction on pullout strength at different extraversion angles. DESIGN: Repeated measurement. SETTING: Center for Bone Joint, the First Affiliated Hospital of Nanjing Medical University. PARTICIPANTS: This study was performed at Laboratory for Material Mechanics, Hehai University between June and November 2003. A total of 18 adult dried lumbar vertebrae (L1-5) were provided by Department of Anatomy, Nanjing Medical University, and recruited for this study. The protocol was approved by the hospital's Ethics Committee. The pedicle screw was made of stainless steel. Each pedicle screw had a diameter of 5.5 nun, total length of 150 nun (thread part 50 into included), and the same thread parameter. Electrical universal material machine (EW type) was provided by Laboratory for Material Mechanics of Hehai University. METHODS: Bone density was measured with a single photon bone density determinator. According to the bone density, the lumbar vertebrae were numbered and randomly divided into 3 groups with 6 lumbar vertebrae in each: extraversion angle 5 ° group, extraversion angle 15° group, and extraversion angle 30° group. ① Installation of pedicle screw and clamping apparatus: According to Wein-Stein method, one entry-point was selected at each side of lumbar vertebra, and at the sametime, extraversion angle 5°, 15° ,and 30° were respectively defined for extraversion angle 5° , 15° ,and 30° groups. A 50 mm-depth pinhole was drilled with a drill bit with a diameter of 3.0 nun. Pedicle screw was screwed into 50 nun, and its end part was connected to the device for transverse traction. Spinal vertebrae and the device for transverse traction were fixed with a specially made clamping apparatus. ② Determination of pullout strength and observation of pedicle and vertebral injury: Spinal vertebrae, on which pedicle screw and device for transverse traction were installed, was placed on a EW electrical universal material machine together with clamping apparatus for determining the pullout strength of pedicle screw. Sensor was connected to a computer to draw strength-displacement curve. The wave crest of the curve was considered the maximum pullout strength. At the same time, injuries to pedicle and vertebra caused by pullout of pedicle screw were observed. MAIN OUTCOME MEASURES: Pullout strength and injuries to pedicle and vertebra. RESULTS:①The mean maximum pullout strength of pedicle screw was respectively 0.878 167, 1.420 333, and 2.154 167 KN for extraversion angle 5° , 15 ° , and 30° groups. There was significant difference among the 3 groups (F = 12.554 22, P < 0.01). ② In the extraversion angle 5° group, 4 patients presented with cortical bone fracture which occurred at the entrance for pedicle screw, and 2 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge; In the extraversion angle 15° group, 1 patient presented with cortical bone fracture which occurred at the entrance for pedicle screw, 4 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge, and 1 patient presented with vertebral posterior coronal fragmentation; In the extraversion angle 30° group, 1 patient presented with cortical bone fracture which occurred at the entrance for pedicle screw, 2 patients presented with fragmentation of junctional zone between pedicle and vertebral posterior edge, and 3 patients presented with vertebral posterior coronal fragmentation. There was statistical significance in the intergroup rank-sum test (P < 0.01).CONCLUSION: During application of pedicle screw with device for transverse traction, proper accrescence of extraversion angle can increase pullout strength of the screw and enhance fixative strength, and excessive extraversion angle easily injures vertebra.

Objective To investigate the effect of Zidan decoction containing serum on endometrial receptivity in uterine immunity environment,and then to discuss if it's effect on endometrial receptivity through its controlling the expression of cell factors which related to endometrial receptivity.Methods Mice's endometrium tissue samples and uterine Natural Killer (UNK) cells were obtained from pregnant mice on gd 4.5 days and gd 12.5 days.After endometrial glandular epithelial cells,endometrial stromal cells and UNK cells were separated through primary culture,epithelial cells and endometrial stromal cells at rate of 1 ∶ 1 a certain concentration UNK were mixed together and highly purified-cultured.Different concentrations of Zidan decoction was acted on the culture solution.FBS,black and aspirin control group were setup for contrast.The expression of LIF and VEGF which influenced by Zidan decoction in the immunity environment was assayed by Realtime-PCR.Results The expression of LIF and VEGF was significantly increased in the immunity environment of uterine (in 10％ Zidan decoction group,10％ aspirin group and 10％ blank control group:LIF was 2.10±0.20,1.98±0.14,0.90±0.05 separately,and VEGF was 3.17±0.31,2.52±0.09,0.92±0.06respectively,P＜0.05).Conclusion Zidan decoction could improve endometrial receptivity by increasing the expression of cell factor which related to endometrial receptivity with the help ofUNK paracrine.

Objective To determine the level of interleukin (IL)-10 in sera and culture supernatants of CD4+CD25+T cells from children with atopic dermatitis (AD),and to evaluate its relationship with clinical course and severity of AD.Methods Totally,46 children with AD and 31 healthy controls were included in the study.All the patients were divided into 3 groups,i.e.,mild (n =10),moderate (n =16) and severe (n =20) group,according to severity scoring of atopic dermatitis (SCORAD) score.Venous blood samples were obtained from the patients and healthy controls.CD4+CD25+ regulatory T cells were separated from the blood samples by magnetic cell sorting (MACS) system in two steps and cultured in vitro.Enzyme linked immunosorbent assay (ELISA) was conducted to quantify the level of IL-10 in sera and culture supernatants of CD4+CD25 + T cells from these subjects.Analysis of variance was carried out to compare the level of supematant and serum IL-10 between the patients and controls,and Pearson correlation analysis to assess the relationship between the level of IL-10 and SCORAD score.Results The patients with mild,moderate and severe AD showed a similar serum IL-10 level compared with the healthy controls ((43.10 ± 25.07) pg/ml,(68.40 ± 36.65) pg/ml and (55.55 ± 41.97) pg/ml vs.(58.27 ± 36.84) pg/ml,all P ＞ 0.05).The level of supernatant IL-10 secreted by CD4+CD25+ T cells from the controls was significantly higher than that from the patients with severe AD ((55.15 ± 11.15) pg/ml vs.(27.25 ± 7.01) pg/ml,P ＜ 0.05),but similar to that from the patients with mild and moderate AD ((52.96 ± 11.69) pg/ml and (49.86 ± 9.18) pg/ml,respectively,both P ＞ 0.05).The level of secreted IL-10 was negatively correlated with SCORAD score (r =－0.757,P ＜ 0.01),whereas the serum level of IL-10 showed no statistical correlation with SCORAD score.Conclusion CD4+CD25+ T cells and IL-10 may be implicated in the development of AD.

Objective To evaluate the value of high-frequency echocardiography in assessing Bax gene transfer influence on survival of heterotopic cardiac allograft in rats.Methods Thirty rat models of heterotopic heart transplantation were established.Group A received heart transplantation only; Group B received cyclosporin (CsA) after operatiom Group C received ultrasound targeted microbubble destruction (UTMD) combined with Bax-shRNA.All rats were tested by high-frequency echocardiography at day 1,3,6 after transplantation.The ultrasound parameters included left ventricular internal dimension diastole (LVIDd),left ventricular internal dimension systole(LVIDs),left ventricular ejection fraction(LVEF),left ventricular thickness (LVT),left ventricular thickening (LVTR) and so on.The pathologic examinations were carried out on five rats at day 6 after echocardiography exam.The other rats in each group were observed for the survival time of cardiac allograft.Results ①The left ventricular long axis view and the apical four chamber view could be displayed clearly by high-frequency echocardiography.②The survival time of allografts in group C was (16.21 ± 5.01)d,which was longer than that in group B [(11.14 ± 1.72)d,P ＜ 0.05] and group A [(7.26 ± 1.50)d,P ＜0.01].③LVT index got higher after transfection.At day 6,LVEF of group A was lower than group B and C markedly(P ＜0.05) while no significant difference was found between group B and C(P ＞0.05).At day 3 and 6,LVT of group A and B were higher than group C (P ＜0.05) while LVTR was lower(P ＜0.05).Conclusions The Bax gene transfer influence on survival of heterotopic cardiac allograft in rats could be evaluated accurately by high-frequency echocardiography which could assess cardiac structure and function.LVT and LVTR can be considered as early evaluation indexes for its high sensitivity over LVEF.

Objective To investigate the changes of cholesterol efflux,the scavenger receptor class B type Ⅰ(SR-BI) protein expression in THP-1 maerophage derived foam cells treated with Lippolysaecharide (LPS), and to discover the role of NF-κB pathway in this process.Methods The foam cells were treated with LPS along or treated with N-p-Tosyl-L-phenylalanine chloromethyl ketone(TPCK) for 24 h.The protein levels of SR-BI and intranuclear NF-κB p65 were measured by Western blotting.Cellular lipid accumulation was determined by high performance liquid ehromatograpby analysis.Cholesterol efflux was determined by FJ-2107P type liquid scintillator.Results The expression of SR-BI was decreased after treated with LPS,while the intranuclear NF-κB p65 protein level was increased by LPS.The results also showed that cellular lipid accumulation was increased ,while the cellular cholesterol efflux was decreased in THP-1 maerophage derived foam cells after exposed to LPS for 24 h and these changes can be reversed partly by pretreatment with TPCK.Conclusion LPS could down-regulate the expression of SR-BI, promote the accumulation of lipid and decrease cellular cholesterol efflux in THP-1 maerophage derived foam cells ,which should be related to the TLR4/NF-κB dependent pathway.

Objective To develop a flow cytometrie assay to for diagnosis of X-Linked Hyper.1gM Syndrome(XHIM).Methods Heparinized peripheral blood obtained from patient,mother and a healthy control was diluted with RPMI- 1640(unstimulated control)or with RPMI-1640 containing 15μl PMA(1 rig/μl)and 6 trl ionomycin(50 ng/μl)(stimulated cell).Using directly labeled antibodies,we have examined CD40 ligand levels on CD3+ CD8- lymphocyte surface,and CD69 levels on CD;lymphocyte Surface to determine whether the cells were activated.Results CD69 levels on CD3+ lymphocyte surface from stimulated group and from unstimulated group were above 96% and below 3%,respectively.CD40L levels Oil CD3 CDs-T lymphocyte surface from stimulated group were 0.8% (patient),60.04%(mother) and 62.87%(healthy contr01).CD40L levels on CD3+ CD8-T lymphocyte surface from unstimulated group were 0.88% (patient),4.15%(mother)and 5.51%(healthy contr01).Conclusion This flow cytometric assayis accurate and convenient,which Can be used in neonatal screening.

Aged ; Axilla ; Axillary Vein ; surgery ; Blood Vessel Prosthesis ; Breast Neoplasms ; complications ; pathology ; surgery ; Female ; Histiocytoma, Malignant Fibrous ; complications ; pathology ; surgery ; Humans ; Jugular Veins ; surgery ; Male ; Neoplasm Recurrence, Local ; Upper Extremity Deep Vein Thrombosis ; etiology ; surgery ; Vascular Grafting ; methods

Objective To evaluate the effect of hemorrhagic shock factor on the pharmacokinetics of rocuronium in pigs.Methods Sixteen pathogen-free Bama miniature pigs of both sexes,aged 3-5 months,weighing 22-25 kg,were divided into 2 groups (n=8 each) using a random number table:control group (group C) and hemorrhagic shock group (group HS).In group C,rocuronium 3.78 mg/kg was injected via the auricular vein.In group HS,the animals were subjected to volume-controlled hemorrhage,about 40％ of blood volume was withdrawn from the left femoral artery over 15 min (30 ml/kg),and rocuronium 3.78 mg/kg was injected via the auricular vein after the model was successfully established.At 0,2,4,7,10,15,20,30,60,120,180,240,300,360 and 420 min after rocuronium injection,blood samples were collected from the internal jugular vein for determination of the plasma concentration of rocuronium by high-performance liquid chromatography-tandem mass spectrometry.The pharmacokinetic parameters of rocuronium were calculated.Results Compared with group C,the plasma concentration of rocuronium was significantly increased at 20 and 60-420 min after rocuronium injection,the elimination half-life and mean residence time were prolonged,and the plasma effect-site equilibration rate constant was decreased in group HS (P＜0.05).There was no significant difference in the maximal concentration and area under the concentration-time curve between the two groups (P＞ 0.05).Conclusion The elimination of rocuronium is slower in a pig model of hemorrhagic shock.

To observe different expression patterns of β-catenin and its clinical significance in colorectal cancer (CRC).Methods A total of 181 cases of CRC tissues and 30 cases of normal colorectal tissue were investigated by immunohistochemistry for the expression of β-catenin.Results The expression rate of β-catenin was 56.9％ (103/181) in CRC,and higher than that in normal colorectal tissue (P ＜ 0.05).The overexpression of nuclear β-catenin was significantly correlated with histological differentiation,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05),and no relationship with other pathological parameters,such as age,gender and the depth of infiltration.The incomplete membranous expression of β-catenin was significantly correlated with histological differentiation,the depth of infiltration,lymph node metastasis and Dukes' stage in CRC (P ＜ 0.05).The high expression of nuclear β-catenin related to histological differentiation and Dukes' stage in CRC (P ＜ 0.05).In the follow-up data of 82 cases of CRC,the expression of nuclear β-catenin was associated with poor prognosis,and the 5-year survival rate was significantly lower than that of self-control groups (P ＜ 0.05).Conclusion β-catenin plays important roles in colorectal carcinogenesis.Abnormal expression of β-catenin was related to the aggressive progression of CRC and may be helpful for evaluating the prognosis of patients with CRC.β-catenin is expected to become a new target for diagnosis and treatment of CRC in future.