Objective To explore the simplified method by examining the levels of luteinizing hormone(LH),follicle-stimulating hormone(FSH),estradid(E2) and stosterone(T) at different times in the stimutation test of luteinizing hormone-releasing hormone(LHRH).Methods Sixty patients with precocious puberty accepted LHRH stimutation test.The levels of E2,T,FSH,LH before injection and after 30,60,90 minutes were compaired.The levels of stimutation test of LHRH examined with ACS:180 chemiluminesence.Results There were 39 patients in the group of central precocious puberty(CPP).The levels of FSH and LH in CPP group significantly increased after LHRH 30 minutes injection.The ratio of LH/FSH was higher than 1.The peak level of LH was higher than 12 IU/L.There were 21 patients in group of peripheral precocious puberty(PPP).Compared with the results before injection,the levels of LH were similar to the results of 30,60 and 90 minutes after LHRH injection.Compared with the result before injection,the levels of E2 and T were similar to the result of 60 minutes after LHRH injection.The peak levels of LH and FSH in two groups all focused in 30,60 minutes.Conclusions LHRH stimutation test is mainly based on the peak level of LH and the ratio of LH/FSH,the test can be simplified to examine the levels the of LH and FSH before the test and 30,60 minutes after injection as a basis for the clinical diagnosis.

OBJECTIVETo analyze the electrical activity property changes in nucleus accumbens (NAc) of heroin-induced conditioned place preference (CPP) rats during different stages of heroin dependence and to explore NAc's roles in the formation of drug dependence.

METHODSRecording electrodes were bilaterally embedded into the NAcs of rats with the aid of stereotaxic apparatus, followed by establishment of heroin-dependent rat model. The NAc electrical activity during 3 different stages of heroin dependence, including heroin pre-exposure, immediate post-exposure and heroin withdrawal, were respectively recorded using EEG wireless telemetry techniques. The frequency distribution (ranging from 0.5 to 30 Hz) and the amplitude of NAc electrical activity were analyzed and measured.

RESULTSHeroin-dependent rat models were successfully established and their withdrawal symptoms were evident. All rats showed a conditioned place preference (CPP) for the white box after 5-10 days of heroin-exposure, and displayed a maximum withdrawal symptoms on 2d after heroin- withdrawal. During all statges of heroin-dependence, the NAc electrical activity contained the highest proportion of delta rhythm and the lowest proportion of alpha2 rhythm. The discharge frequence band was similar across different stages. There was a significantly increased ratio of low-frequency discharges (delta rhythm) and decreased ratio of high-frequency discharges (beta rhythm) in NAc of rats during the immediate post- heroin exposure stage when compared with that during pre-exposure and heroin withdrawal stages. During the withdrawal stage, the ratio of at rhythm was significantly lower than during pre- and post-heroin exposure stages (P < 0.01). Further, the mean discharge amplitude in NAcs during immediate post-exposure and withdrawal stages was significantly increased relative to pre-exposure stage. However, the mean discharge amplitude during heroin withdrawal stage was significantly lower than during immediate post-exposure stage.

CONCLUSIONThe electrical activity properties in rat NAcs showed a significant change during different stages of heroin-dependence, which suggested that neuronal activities in NAcs might contribute to the modulation of drug-dependence.

Animals ; Conditioning, Operant ; Heroin ; pharmacology ; Heroin Dependence ; physiopathology ; Male ; Nucleus Accumbens ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Telemetry

Objective To investigate the drug resistance and its distribution induced by β-lactamases and class Ⅰ integrons with the deficiency of Omp genes in Enterobacter cloacaes.Methods Totally 112 strains of Enterobacter cloacaes were isolated during January 2008 and May 2011.The identification of strains was performed by using Vitek-2 Compact automatic system; and antibiotic susceptibility was determined by K-B method.Isolates of E.cloacae were screened for carbapenemases by modified Hodge test,improved three dimensional test and EDTA-meropenem synergy test.Genes encoding AmpC β-lactamase,metallo-β-1actamases (MBLs) and OXA-like β-1actamases were screened by multiple PCR.Single PCR was used to detect ISEcpl,OmpK35/36,NDM-1 and OXA-48.The variable regions of class Ⅰ integrons were amplified and sequenced.Results Among 112 isolates,6 (5.4％) demonstrated positive in the modified Hodge test and 14 ( 12.5％ ) were positive in the improved three-dimensional test.No carbapenemases gene was found.There were 29(25.9％ ) strains positive for ESBLs genes,ISEcpl was found in the upstream of all the CTX-M-type ESBLs; OXA-1 ESBLs were detected in 2 isolates.AmpC β-lactamase genes were positive in 45 (40.2％) strains,and 82.2％ (37/45) were MIR-3 type.Twenty two isolates carried class Ⅰ integrons,and four different cassettes arrangements were identified within 16 strains:9 isolates harbored aadB-aadA2 ( 1 000 bp),5 isolates with dfrAl5 (700 bp),2 isolates with aadAl ( 1 000 bp).One isolate harbored all the above gene cassettes.The deficiency of OmpK35/36 was found in all strains.Conclusion ESBL,AmpC β-lactamase and the deficiency of OmpK35/36 are correlated with the resistance to carbapenems in Enteobacter clocace,and class Ⅰ integrons may also partly account for the multidrug-resistance.

ObjectiveTo analyze the prognosis of 784 patients according with clinical staging of esophageal carcinoma treated with non-surgical methods,investigate the predictive value and deficiency of the clinical staging.MethodsFrom July 2003 to January 2009,784 patients with esophageal carcinoma received 3DCRT treatment.The prescribed doses ranged from 50 Gy-70 Gy with median dose of 60 Gy,1.8-2.0 Gy/fraction,1 fraction/day,5 fractions/week.65 patients received prescription dose of＜60 Gy and all the others'≥60 Gy.All the patients were divided into subgroups according to different T,N and TNM stages.Therapeutic effect was evaluated.ResultsThe follow up rate was 97.1％,503 patients were followed up for more than 3 years and 122 were followed up for more than 5 years.The 1-,3-,5-year local control rates and overall survival rates were 77.2％,54.2％,46.5％ and 69.5％,34.9％,23.9％,respectively,with median survival time of 21 months.There were significant differences of survival curves for different T stages,N stages and TNM stages.For the groups of stage Ⅰ,Ⅱ and Ⅲ,the 1-,3-,5-year survival rates were 86.4％,47.6％,45.1％ ;84.7％,46.3％,36.4％ and 64.0％,30.9％,19.1％,respectively ( x2 =29.34,P =0.000).There were 752 patients with squamous cell carcinoma ( 95.9％ )and 32 patients with non-squamous cell carcinoma (4.1％ ),the median survival time were 21 and 16 months,respectively ( x2 =4.44,P =0.035 ).There were significant difference of survival rates for the subgroups whose GTV volume ≤20 cm3,20 -40 cm3,40 -60 cm3 and ＞60 cm3 (54 months,29 months,21months and 14months,x2 =68.71,P =0.000).ConclusionsThe clinical staging of esophageal carcinoma treated with non-surgical methods could predict the prognosis accurately,for patients with different pathology and GTV volumes,there were variance in the prognosis,so we advised the complement of the two factors in the draft of clinical stages.

Objective To evaluate the efficacy of closed-loop infusion of propofol for surgery on the body Surface in preschool and school-age pediatric patients.Methods American Society of Anesthesiologists physical status Ⅰ pediatric patients of both sexes,undergoing elective surgery on the body surface,were included in the study.One hundred twenty-four preschool pediatric patients (aged 3-6 yr) were divided into 2 groups (n=62 each) using a random number table:closed-loop group (CPRE group) and openloop group (OPRE group).Thirty-eight school-age pediatric patients (aged 7-12 yr) were divided into 2 groups (n=19 each) using a random number table:closed-loop group (CSTU group) and open-loop group (OSTU group).The target bispectral index (BIS) value was set at 50 during maintenance of anesthesia.Propofol was delivered using closed-loop anesthesia delivery system,and the target plasma concentration of propofol was automatically regulated in CpRE and CSTU groups.Propofol was given using open-loop anesthesia delivery system,and the target plasma concentration of propofol was manually regulated in OpRE and OSTU groups.The adequate anesthesia time ratio (BIS40-60％) and global score (GS) of the delivery system were recorded during maintenance of anesthesia.The consumption of propofol,frequency of regulation and mean BIS value were recorded.The delivery system-related complications were recorded during the perioperative period.Results Compared with group OpRE,the consumption of propofol,GS and mean BIS value were significantly decreased,BIS40-60％ was increased (P＜0.05),and no significant change was found in the frequency of regulation in group CpRE (P＞0.05).Compared with group OSTU,GS was significantly decreased,BIS40-60％ was increased (P＜0.05),and no significant change was found in the consumption of propofol,mean BIS value or frequency of regulation in group CSTU (P＞0.05).Compared with group CpRE,the consumption of propofol and GS were significantly decreased,BIS40-60％ was increased (P＜0.05).and no significant change was found in the mean BIS value or frequency of regulation in group CSTU (P＞0.05).One pediatric patient in group CSTU developed airway spasm,and no severe complications were found in the other pediatric patients.Conclusion For surgery on the body surface in preschool and school-age pediatric patients,closed-loop infusion of propofol is safe and effective,the stability and precision of anesthesia is superior to that of open-loop anesthesia delivery system,and it provides better efficacy in school-age pediatric patients than in preschool pediatric patients.

Background To suppress vascular endothelial growth factor (VEGF) is a researching hot topic for the treatment and prevention of retinal neovascularization.Some detectable efficacy of VEGF small interference RNA (VEGF siRNA) in anti-tumor neovascularization has been well-known.But relevant study on VEGF siRNA on retinal neovascularization is seldom.Objective Present study was to investigate the inhibiting effect of VEGF siRNA on retinal neovascularization.Methods The 48 clean C57BL/6J mice aged 7-day-old were randomly divided into normoxia group,hypoxia control group,vector group and VEGF siRNA group and 12 mice for each.Hypoxia models were established by raising the pups with mother mice in the airtight oxygen-cabin for 5 days.The lipofectamineTM 2000 (LF2000)-mediated vector plasmids or VEGF siRNA recombinant plasmids were then injected intravitreally in 12 12-day-old pup mice respectively.The animals were sacrificed in 1 week after intravitreal injection,and the numbers of vascular endothelial cell nuclei extending beyond the internal limiting membrane (ILM) were counted by hematoxylin-eosin stain.The expressions of VEGF protein and mRNA in retina were assayed by immunoinfluorescence technique and RT-PCR.Results The numbers of vascular endothelial cell nuclei extending beyond the ILM were 0.19±0.09,24.89±2.03,23.65±2.15 and 8.83±1.12 in normoxia group,model control group,vector group and VEGF siRNA group separately,showing significant decrease in VEGF siRNA group compared with model control group or vector group (q=5.67,q=4.97,P＜0.01).RT-PCR revealed that VEGF mRNA was faintly expressed in mouse retina in normoxia group.However,in model control group and vector group,the level of VEGF mRNA was 52.3 times and 36.7 times more than that of normoxia group respectively and only 3.5 times in VEGF siRNA group,presenting a inhibitory rate of 43.39% of VEGF siRNA on VEGF.Immunofluorescence showed that the expression of VEGF was weaker in normoxia group and strong positive response in model control group and vector group,but the expression intensity of VEGF protein was significantly weaker in VEGF siRNA group.Conclusion VEGF siRNA recombinant plasmids can efficiently inhibit retinal neovascularization in oxygen-induced retinopathy mouse model through intravitreal injection.

BackgroundFlap creation is one of the most important steps during laser in situ keratomileusis (LASIK). As the microkeratome blade technology is developing, the accuracy, uniformity and reproducibility of corneal flaps created by the microkeratome blade are of high clinical concern. ObjectiveThe aim of this trial was to compare the features of corneal flaps created using the Moria M2 microkeratome 110 μm-knife with regular blade versus the Med-Logics O blade. MethodsA pilot and prospective study was designed. Two hundred and four eyes of one hundred and two patients were enrolled in this clinical trial. The patients were divided into the Moria M2 microkeratome 110 μm-knife with Med-Logics 0 blade group ( 110-0 group) ( 102 eyes) and Moria M2 microkeratome with 110 μm-knife with regular blade group (110 group) (102 eyes),with the matched demography. Fourier-domain optical coherence tomography ( RTVue OCT) was used to measure flap thickness using 28 settings on the 204 corneas at one week postoperatively. The features of the LASIK flaps were analyzed on the basis of the outcomes. Written informed consent was obtained from each patient prior to LASIK. Results There was no statistically significant difference in uncorrected visual acuity and the mean spherical equivalent between the 110-0 group and 110 group ( Z =-0. 375,P =0. 708 ; u =0. 056, P =0. 956 ) one week after LASIK. The mean flap thickness of the 110-0 group was considerably thinner than that of the 110 group ( 133.28+15.41μ m versus 142.81 ±10. 07μm) ( u =-5. 227,P＜0. 001 ). The corneal flaps in both the 110-0 group and in 110 group showed a meniscus shape. The nasal flap thickness of the right eyes was not evidently different from that of temporal ( P＞0. 05 ) , but in the left eyes, nasal flap thickness was obviously thicker than the temporal flap thickness (P＜0. 05) in both groups. The mean deviation between the achieved and attempted flap thickness ( 130 μm) were (17.46±2.28) μm in the 110-0 group and ( 16. 82±6. 12) μm in the 110 group, showing a significant difference between them ( u ==0. 517, P=0. 608 ).ConclusionsThe shape of flaps created using the Moria M2 110-0 is more uniform and closer to the expected thickness of 130 μm than the ones created using the Moria M2 110 microkeratome.

Objective To identify seven species of Gynostemma BI.,including G.pentaphyllum,G. pentagynum,G.cardiospermum,G.longipes,G.yixingense,G.laxiflorum,and G.guangxiense,by in- ter-simple sequence repeat(ISSR)markers.Methods General DNA was isolated from leaves of the seven species in Gynostemma B1.by CTAB,57 primers constituted by ISSR were tested for PCR and sepharose electrophoresis.Results Fourteen primers amplified polymorphic bands,the amplification patterns of primers UBC-873 and UBC-895 were higher in terms of polymorphic and amplified band ratio.They are used to distinguish all the examined seven species.Conclusion ISSR-PCR Method provides a quick,reli- able molecular marker technique for the identification of different species of Gynostemma B1.

ObjectiveTo explore the application of diffusion-weighted magnetic resonance imaging (DWMRI) in precise radiotherapy of esophageal carcinoma.MethodsThirty-seven patients with biopsy proven esophageal cancer from March 2010 to January 2011 were included.To delineate the gross tumor volume (GTV) using CT and DWMRI images,each patient was examined by DWMRI and CT scan using the same position before radiotherapy.To compare the maximum diameters and volumes of tumor between CT and DWMRI. The tumor lengths measured by esophagogram,esophagoscope,CT and DWMRI were compared.ResultsTumor lengths under esophagogram,esophagoscope,CT and DWMRI were 5.70 cm,6.06 cm,7.97 cm and 5.79 cm respectively. The lengths between CT and esophagogram,CT and esophagoscope,CT and DWMRI had statistical significance respectively (F=4.88,P=0.003).The maximum diameters of tumor shown on CT and DWMRI were 3.79 cm and 3.81 cm respectively ( t =-0.32,P=0.751 ).The GTV were 45.75 cm3 and 38.05 cm3 in CT and DWMRI respectively (t=5.30,P =0.001 ).53 lymph nodes were assessed positive on both CT and DWMRI.DWMRI excluded 25 positive lymph nodes assesed by CT; also confirmed 15 negative lymph nodes excluded by CT,6 of which were paraesophageal lymph nodes.The addition of DWMRI information altered the clinical stage in 6 patients.ConclusionsTumor lengths measured on DWMRI and esophagogram had the optimal approximation.It was easy to find paraesophageal lymph nodes via DWMRI.With the addition of DWMRI information,the target range and clinical stage were alerted in some patients.

Objective To observe the relationship between computed tomography-gross tumor volume (GTV) and non-surgical T stage in patients with esophageal squamous cell carcinoma (ESCC) and the survival rates of patients with different GTVs,and to investigate the impact of GTV on the prognosis of ESCC after three-dimensional radiotherapy.Methods A retrospective analysis was performed on 223 ESCC patients without lymph node metastasis and distant metastasis who were hospitalized from July 2003 to January 2009.The prescribed doses of three-dimensional radiotherapy ranged from 50-70 Gy.These patients were divided into 3 or 4 groups according to different percentile intervals of GTVs.The Spearman rank correlation analysis was used for investigating the relationship between non-surgical T stage and GTV.The Kaplan-Meier method was used for calculating survival rates,and the log-rank test was used for survival difference analysis.Results The follow-up rate was 98.2％.A total of 163 patients were followed up for at least 3 years.The median GTVs of patients with T1 +2 ESCC,T3 ESCC,and T4 ESCC were 19.31 cm3,33.69 cm3,and 41.25 cm3,respectively,exhibiting a positive correlation between non-surgical T stage and GTV (P =0.000).The 5-year survival rates were 59％,43％,and 24％ (P =0.000) in 3 GTV-based groups and were 55％,51％,31％,and 24％ (P =0.004) in 4 GTV-based groups.The primary cause of death for the patients with GTVs of ≤35 cm3 and ＞35 cm3 was failure of local control (57.9％ vs 52.1％) ; 21.9％ and 13.8％ of the patients with a GTV of ＞ 35 cm3 died of uncontrol and excessive bleeding,versus 9.4％ and 3.1％ of the patients with a GTV of ≤ 35 cm3 (P =0.046 ; P =0.029) ;2 of the patients with a GTV of ＞ 35 cm3 died of grade 5 radiation pneumonitis.Conclusions GTV is positively correlated with nonsurgical T stage in esophageal cancer patients who receive radiotherapy.The 3-level and 4-level grading of GTV can be used for prognostic evaluation,and the 3-level grading of GTV is more closely related to prognosis.The patients with a larger GTV have higher incidence of uncontrol and bleeding and risk of treatment-related death than those with a smaller GTV after radiotherapy.