Objective To explore if B lymphocyte stimulator (BlyS) stimulates B lymphocytes from patients with chronic idiopathic urticaria (CIU) to produce anti-high affinity IgE receptor (FcεRI) or anti-IgE antibodies.Methods Totally,300 CIU patients and 300 health controls were enrolled in this study.Blood samples were obtained from these subjects.Peripheral blood B lymphocytes were isolated and cultured in vitro for 72 hours.Then,BlyS of various concentrations (2,4,8,16 ng/ml) was added to the culture medium of B lymphocytes followed by another 72-hour culture.Enzyme-linked immunosorbent assay was performed to determine the serum levels of BlyS,anti-FcεRI and anti-IgE antibodies,as well as the supernatant levels of anti-FcεRI and anti-IgE antibodies.The relationship between BlyS and anti-FcεRI and anti-IgE antibody production was assessed.SPSS software version 16.0 was used for statistical analysis.Chi-square test was performed to compare the positivity rate of antibodies,and analysis of variance and least significance difference-t test to assess numerical data.Results The CIU patients showed higher levels of serum BlyS (t =3.04,P ＜ 0.01),anti-FcεRI antibodies (t =3.51,P ＜ 0.01),and anti-IgE antibodies (t =3.29,P ＜ 0.01) compared with the health controls.The serum level of BlyS was positively correlated with that of anti-FcεRI antibodies (r =0.93,P ＜ 0.01) and anti-IgE antibodies (r =0.91,P ＜ 0.01).The levels of anti-FcεRI antibodies and anti-IgE antibodies were significantly increased in the culture supematant of patient-derived B lymphocytes treated with BlyS compared with those remaining untreated (t =3.67,3.56,respectively,both P ＜ 0.01),and the concentration of BlyS was positively correlated with the levels of both anti-FcεRI antibodies and anti-IgE antibodies (r =0.96,0.91,respectively,both P ＜ 0.01).The coincidence rate between the serum and supernatant was 94.76％ and 87.84％ in the detection of anti-FcεRI antibodies and anti-IgE antibodies respectively.Conclusions BlyS level is upregulated in the serum of patients with CIU,which may play an important role in the pathogenesis of CIU by stimulating B lymphocytes to produce anti-FcεRI antibodies or anti-IgE antibodies.

AIM: To evaluate the effects of over-expression of cellular repressor of E1A-stimulated genes(CREG) mediated by retrovirus on neointima formation in injured rat carotid.METHODS: The pluronic F127 containing pLNCX/CREG or pLNCX/GFP retroviral vectors was placed around the injured rat carotid.The neointima,media areas and the intima to media ratio were calculated.Expressions of CREG,SM ?-actin and Ki-67 were detected.RESULTS: The GFP expression was observed at day 2 in pLNCX/GFP groups.The expression of exogenous CREG was also significantly increased in arteries at day 2 after pLNCX-CREG infection.Over-expression of CREG significantly suppressed neointima formation,attenuated the expression of Ki-67 and up-regulated SM ?-actin expression.CONCLUSION: Over-expression of CREG inhibits VSMCs proliferation and promotes VSMCs differentiation after vascular injury.It suggests that modulation of CREG expression or activity may be a viable approach to treat neointimal restenosis after percutaneous coronary intervention.

BACKGROUND:Embryonic stem cells (ESCs) serve as a major cell source for smooth muscle cells,but the heterogeneity of cells derived from ESCs result in difficulty to obtain high purity smooth muscle cells.OBJECTIVE:To construct a double expression vector of puromycin resistance (pac) gene and enhanced green fluorescence protein (EGFP) gene driven by smooth muscle specific SM22α promoter (pSM22α-PAC-IRES2-EGFP),in addition,to detect its availability and specificity in ESCs.DESIGN,TIME AND SETTING:The observational experiment of gene level was performed at the Cardiovascular Institute,General Hospital of Shenyang Military Region from April 2007 to September 2008.MATERIALS:ESCs line R1 with number SCRC-1011TM was purchased from American ATCC Company.The pSM22α-EGFP vector was constructed by our laboratory.And the pIRES2-EGFP,pSM2C and pSuper.basic vectors were purchased from Invitrogen Company.METHODS:SM22α promoter was cloned from pSM22α-EGFP by polymerase chain reaction.CMV promoter of pIRES2-EGFP vector was replaced by SM22 promoter to establish pSM22α-IRES2-EGFP.Pac gene,excised from pSM2C by HindⅢ/Clal digestion,was sub-cloned into pSuper.basic to establish pSuper-PAC.After BgⅢ/Accl enzyme digestion of pSuper-PAC,pac gene fragment was obtained,which was further sub-cloned into pSM22α-IRES2-EGFP to produce pSM22α-PAC-IRES2-EGFP.ESCs were transfected with pSM22α-PAC-IRES2-EGFP using lipofectamine.Positive clones were selected by G418 and induced to differentiate and further identified by amplification of pac gene by RT-PCR.Differentiated cells were immunostained by SM α-actin,and expression of SM α-actin and EGFP was observed simultaneously under fluorescence microscope.MAIN OUTCOME MEASURES:Sequencing result of pSM22α-PAC-IRES2-EGFP;Amplification of pac gene;EGFP expression;as well as SM α-actin immunostaining.RESULTS:Three segments of 261 bp,664 bp,and 5000 bp were obtained by HindⅢ/Clal digestion,which was coincident with expectation,and the sequencing results showed that pSM22α-PAC-IRES2-EGFP vector was successfully constructed.Amplification of pac gene identified 4 ESCs clones successfully transfected.After induction of differentiation,partial portion of differentiated cells expressed EGFP,accompanied by positively stained by SM α-actin antibody.CONCLUSION:pSM22α-PAC-IRES2-EGFP vector was successfully constructed.ESCs clones transfected with this vector expressed pac gene and EGFP gene,and the expression of EGFP is smooth muscle specific.

Objective To study the dosimetry for copper compensator in IMRT.Methods 10 patients (3 nasopharyngeal carcinoma,4 esophageal cancer,3 colorectal cancer) from IMRT plans made in Oncentra Planning System enrolled in this study.The fluence matrices of each plan were converted into compensator thickness matrices based on attenuation coefficient of copper under 6 MV radiation ray which was measured with ion chamber.While irradiating the homogenous phantom with the intensity modulated plans performed by copper compensator,film was used to measure the planar dose.In the end,γanalysis was carried out between the measured and calculated planar doses with the criteria of 3％ DD/3 mm DTA.Paired t test to MU differences of MLC plans and copper compensator plans.Results The γanalysis results showed the planar dose pass rates of 10 plans lie between 90.2％ and 98.2％,which means the IMRf technique based on copper compensator can meet the clinical needs.The MU for copper compensator IMRT plans is less than the MLC IMRT plans (873.9 vs.975.1,P =0.005).Conclusions Compared to conventional IMRT based on MLC,copper compensator has the advantage of lower requirement of hardware and lower scattering dose in patient.Copper compensator can meet the needs of clinical practice.

Objective: To clone cDNA encoding Ⅴ-Ⅶ extra-cellular domains of VEGF receptor KDR and investigate biological activity of the recombinant protein. Methods: Total RNA from the umbelical venous endothelial cells was obteined. From it, KDR cDNA was cloned into the BamH1 and Kpn 1 sites of pQE31 and made expressed with induction of IPTG. The recombinant protein was purified, refolded and determined for its biological activities. Results: The expression system could express the human recombinant extra-cellular protein of KDR with expression level accounting for 5% of the total bacterial protein. After renaturation, the recombinant protein could bind to 125 I-VEGF and suppress endothelial cell proliferation and angiogenesis of chick chorioallantoic membrance induced by VEGF. Conclusion: Human KDR extra-cellular domain can be expressed in E coli and after being refolded, has an action of binding to VEGF.

Objective To compare the clinical efficacy between continuous and intermittent androgen deprivation in high risk prostate cancer.Methods Sixty-four patients with high risk prostate cancer were treated from January 2008 to April 2009,36 cases who accepted goserelin and bicalutamide were taken as intermittent hormonal therapy (intermittent treatment group),while 28 cases who accepted bilateral orchiectomy in addition to flutamide were regarded as continuous hormonal therapy (continuous treatment group).The comparison of tumor specific mortality,time of prostate specific antigen (PSA) to nadir,tine to PSA recurrence,serum testerone and quality of life score were assessed between the two groups.Results In continuous treatment group and intermittent treatment group,follow-up period was (26.4 ± 10.3) and (28.1 ± 8.7) months,the time of PSA to nadir was (3.8 ± 2.1 ) and (4.0 ± 3.6) months,the time to PSA recurrence was (20.1 ± 12.3) and (24.5 ± 14.6) months,respectively.There was no significant difference between the two groups.At the time of 18,24 and 30 months after therapy,serum testerone was 0.85,0.88,0.89 μg/L in continuous treatment group,while 1.21,1.36,1.48 μg/L in intermittent treatment group,respectively (P ＜ 0.05 ).Similarly,quality of life score was 38.7,40.5,39.8 scores in continuous treatment group,while 49.2,51.4,52.3 scores in intermittent treatment group at the time of 12,18 and 30 months after therapy,respectively (P ＜ 0.05 ).Conclusions Clinical efficacy could not been found between continuous and intermittent endocrinic therapy of prostate cancer.During intermittent,quality of life seems to be better and increases in accordance with serum testerone recurrence at given time.

OBJECTIVETo evaluate clinical results of early total care (ETC) treatment for elderly patients with intertrochanteric femur fractures.

METHODSClinical data of 106 elderly patients with intertrochanteric fracture treated from January 2012 and February 2015 were retrospectively studied. According to whether receiving the early total care mode, the patients were divided into 2 groups, 34 cases were diagnosed and treated with early total care pattern (ETC group), including 14 males and 20 females with an average age of (74.88 ± 4.38) years old ranging from 70 to 86. According to Evans types, 4 cases were type I, 5 cases were type II, 13 cases were type III, 11 cases were type IV, 1 case was type V. Seventy-two patients were treated with conventional trauma method (conventional group), including 35 males and 37 females with an average age of (74.46 ± 3.63) years old ranging from 70 to 85. According to Evans type, 8 cases were type I ,13 cases were type II, 25 cases were type III, 25 cases were type IV, and 1 case was type V. All fractures were treated with proximal femoral nails anti-rotation (PFNA). Operative time, hospital stays, leaving bed time, complications, cases of death at 1 year after operation, postoperative Harris score at 12 months were observed and compared.

RESULTSAll patients were followed up, the time of ETC group ranged from 9 to 18 months with an average of 13.29 ± 1.51, and the time in conventional group ranged from 12 to 16 months with an average 12.93 ± 1.15, while there was no significant difference between two groups in time of following-up (t = 1.368, P = 0.174). There was no significant meaning in cases of death between ETC group (2 cases) and conventional group (8 cases). Three cases occurred complications in ETC group, and 20 cases in conventional group,there was obvious meaning between two groups (χ² = 0.739, P = 0.318). Operative time,hospital stays,leaving bed time in ETC group respectively was (2.03 ± 0.67) d, (15.41 ± 2.87) d and (3.62 ± 0.74) d; while in conventional group respectively was (4.17 ± 1.59) d, (20.11 ± 4.24) d and (5.35 ± 1.22) d; there were significant differences between two groups in operative time, hospital stays, leaving bed time. Postoperative Harris scores at 12 months in ETC group was (82.32 ± 4.56), and (79.24 ± 5.52) in conventional group, there was obvious meaning between two groups (t = 2.833, P = 0.006).

CONCLUSIONETC pattern is a novel method for diagnosis and treatment of intertrochanteric femur fractures in elderly, it could shorten operative time, hospital stays, leaving bed time, decrease complications and promote recovery of function.

Aged ; Aged, 80 and over ; Female ; Femur ; injuries ; surgery ; Follow-Up Studies ; Fracture Fixation, Intramedullary ; Hip Fractures ; surgery ; Hip Joint ; surgery ; Hospitalization ; Humans ; Male ; Operative Time ; Postoperative Care ; Retrospective Studies

Critical Care ; Critical Illness ; Emergencies ; Female ; Humans ; Infant ; Infant, Newborn ; Intensive Care Units, Pediatric ; Male ; Retrospective Studies ; Transportation of Patients

Fragmentation behavior of diterpenoids was investigated by ESI/MSn and the qualitative analysis of diterpenoids in the bark of Pseudolarix kaempferi was performed using high-performance liquid chromatography/ multi-stage mass spectrometry (HPLC-ESI/MSn). The characteristic fragmentation behaviors of the diterpenoids are the cleavages of the lactone ring and C4-O bond. Furthermore, the eliminations of substituent groups at C-18, C-7 and C-8 can also be observed in the MS" (n = 3-4) spectra. For C-4 acetoxy subsititued diterpenoids, [M+Na-60]+ and [M-H-104] are the base peaks of MS2 spectra in the positive and negative ionization modes, respectively. For C-4 hydroxyl subsititued diterpenoids, [M+Na-44]+ and [M-H-62] are the base peaks of MS2 in the positive and negative ionization modes, respectively. For C-18 glucosylated or esterized diterpenoids, [M+Na-44]+ is the base peak of MS2 spectra in positive ionization mode. These fragmentation rules were successfully exploited in the identification of diterpenoids in methanol/water (6:4) extract of P. kaempferi by LC-MS in positive ionization mode. A total of 9 diterpenoids were identified or tentatively characterized, and one of them is reported here for the first time. The described method could be utilized for the sensitive and rapid qualitative analysis of P. kaempferi.

Objective To evaluate the clinical significance of CT angiography(CTA)in the diagnosis of arterioportal shunts(APS)associated with hepatocellular carcinoma(HCC).Methods One hundred and twenty-seven consecutive HCC patients accepted both dynamic enhancement CT and DSA examinations.The interval between CT and DSA exam was from 3 to 15 days.Based on transverse CT images in hepatic artery phase,CTA was performed for all the patients.By contrast with DSA results,the capabilities of transverse CT and transverse images combined with CTA in APS diagnosis were analyzed. Results In all 127 HCC cases,52 cases with APS were confirmed by DSA(40.94%),33 with central type of APS and 19 with peripheral type.Diagnostic sensitivity of APS based on transverse CT and combined CTA with transverse CT images were both 94.23%(49/52).However,specificity was 84.00%(63/75) and 97.33%(73/75),respectively,accuracy was 88.19%(112/127)and 96.06%(122/127),the predictive value of positive cases was 80.33%(49/61)and 96.08%(49/51),and the predictive value of negative cases was 95.45%(63/66)and 96.05%(73/76).Combined with CTA,false positive cases of 4 central type of APS and 6 peripheral type of APS were excluded which were demonstrated by transverse CT images.By contrast with DSA,the coincidence rate of the type of APS diagnosed by transverse images combined with CTA was 88.46%(46/52),including 90.91%(30/33)of central type of APS and 84.21%(16/19)of peripheral type.The supplying arteries of central type of APS were intuitively displayed by CTA in 23 cases,19 from proper hepatic artery and 4 from gastro-duodenal artery.Conclusion CTA techniques based on the dynamic enhancement CT exams could effectively promote the specificity and the accuracy of APS diagnosis.