This research is for the purpose of comparative analysis of the microbial flora structure in the chilled beef with no packing and cling film, which under the same terms of sale. It was used the V3 area fragment of 16S rDNA to carry on PCR-DGGE, Meanwhile used the 16S rDNA sequence to analysis the microbial flora structure of the two samples, according to the technology of clone .The research discovered that the flora structure displays a biggish difference; there was 6 OTU in the chilled beef with cling film, mainly was that Lactococcus(28%), Lactobacillus (26%), Carnobacterium(18%) and Brochothrix (10%); but there was 18 OTU in the chilled beef with no packing, mainly was that Lactococcus(28%), Brchothrix(18%), Acinetobacter (11%). The result indicates that cling film played a certain inhibitory action regarding the Staphylococcus as well as the cold pole bacteria and such bacterium. And it can provide a certain theory ba-sis for the meat processing in the department of microorganism’s control.

It is used the method of pure culture,Selected 32 strains,which were obvious difference in the shape,color and so on common characteristic,From the chilled beef with no packing and cling film on sale in this research;and it was included 12 strains from the chilled beef sample packed with cling film;20 strains from the chilled beef sample with no packing.Simultaneously selected 4 strains which were predominant in each bacterium from the two samples to conduct the further research,8 strains serial numbers are:S01～S08,S01～S04 from the chilled beef sample with no packing;S05～S08 from the chilled beef sample packed with cling film.Through ARDRA(Amplified ribosomal DNA restriction analysis) as well as 16S rDNA to clarify the bacterium's classified status.The physiological and chemical tests were done to determine the various bacteria respective genus.The experiment indicated:S01 is Pseudomonas putida;S02 is Shewanella cincia stain;S03 and S05 are the same Shewanella putrefaciens;S04 is Stenotrophomonas mal-tophilia;S06 is Psychrobacter;S07 is Staphylococcus sciuri;S08 is Microbacterium-laevaniformans.It was proved that two samples altogether have the same predominant bacterium.It can provide certain theory basis for the chilled meat processing craft as the preliminary investigation in the cultured microorganism situation in two samples.

ObjectiveTo study the efficacy of using multileaf collimators with different position and different degree in the treatment of nasopharyngeal carcinoma (NPC) using intensity-modulated radiotherapy techniques.Methods Ten patients withNPC were administered andanalyzed.Thepenumbra characteristics, dose of target, and radiation conformal indexes (CI) of mode T1 and mode T2 were measured and compared using dose volume histogram generated by Varian Eclipse three-dimensional planning computer system. Mode T1 :The angles of seven coplanar beams were 0°, 52°, 106°, 160°, 212°, 258°and 308°,respectively. There were no restriction on the position and degree of multileaf collimators. Parameters were set and optimized. Mode T2 :The beam angles and the parameters were as same as mode T1. According to the actual situations, the position and the degree of the multileaf collimators were changed. Then thedose optimization was performed. Results Target dose coverage in both mode T1 and T2 could be clinically accepted, and the CI were 0. 82 and 0. 83(t = -0. 25, P =0. 815). The maximum dose reductions in the lens, eyes, optic nerves and corneas were 28. 7% (t = 4. 80, P = 0. 000), 2. 7% (t = 2. 99, P = 0. 021),1.4%(t= 1.05,P=0.032), and 30.5% (t=2.99,P=0. 020), respectively. However, the mean dose and V35 of the parotid were increased by 0. 6% (t = - 2. 82, P = 0. 043) and 9.9% (t = - 2. 05, P =0. 038). ConclusionsOpimization of multileaf collimators can reduce the scattering and leaking rays. Compared with mode T1 ,controlling the position and degree of multileaf collimators could reduce the radiation dose to the eyes and optic-nerves, especially to the lens.

Objective To investigate the effects of acyl and deacyl ghrelin on the expressions of PI3Kp85α,Akt/PKB and GLUT4,the key factors in insulin receptor signaling pathway of insulin resistance in skeletal muscle cells.Methods Insulin resistance models were made by palmitic acid induced rat L6 myoblasts.Successful models were divided into acyl ghrelin group,deacyl ghrelin group,PI3K inhibitor(LY) + acyl ghrelin group,LY + deacyl ghrelin group and control group with corresponding interventions for 24h.The glucose uptake of all group was measured through laser confocal microscopy and flow cytometry.Expressions of phosphorated/total PI3Kp85α,phosphorated/total Akt and cell membrane/total GLUT4 of skeletal muscle cells were measured by Western blot,and PI3Kp85α,Akt,GLUT4 mRNA expression were detected by real-time PCR.Results Induced L6 myoblasts differentiation and insulin resistance model were successfully established.Acyl and deacyl ghrelin could increase glucose uptake for 1.25 and 1.28 folds compared to control group,and the phosphorated and total PI3Kp85α expressions were 1.78 and 1.89 folds to control group,and phosphorylated/total Akt and cell membrane/total GLUT4 were 1.84 and 1.80 folds to control group.The PI3Kp85α,Akt/PKB and GLUT4 mRNA expression were also upregulated compared to control group.The above indexes of LY + acyl or deacyl ghrelin group decreased significantly compared to acyl and deacyl ghrelin group without LY.Conclusions Acyl and deacyl ghrelin can both improve insulin resistance in skeletal muscle cells,increasing the glucose uptake under insulin-stimulation,and up-regulated the phosphorated PI3Kp85α,phosphorated Akt/PKB,and cell membrane GLUT4 relative protein expressions and mRNA expressions.PI3K inhibitor,LY294002,can inhibit the above improvement effect of acyl and decyl ghrelin.

Objective To investigate the effectiveness and safety of intranasal different dose of dexmedetomidine for pediatric echocardiography sedation and to discuss the factors concerning recovery.Methods In a single-blinded randomized clinical trial,183 children were studied with a range of 2months and 33 months of age,and American Society of Anesthesiologists(ASA) physical status Ⅰ to Ⅱ.Those children were divided randomly into one of three groups.Groups D1,D2,and D3,which were received intranasal dexmedetomidine 1.0,1.5,and 2.0 μg/kg,respectively.The induction time,recovery time,examination time,and total sedation time were compared.The success rate of sedation and the occurrence of any side-effects with the drug were compared.Sex,age,weight,dose,induction time,and examination time were used as independent valuations,the recovery time was used as dependent valuation,and then the multiple linear regression analysis was performed to filtrate and formulate the valuable factors influencing recovery time.Results The induction time had no significantly difference among groups (P ＞ 0.05).The recovery time of group D3 was longer than group D1 and group D2 (P ＜ 0.05).The total sedation time of group D3 was longer than group D1 (P ＜ 0.05).The success rate of sedation and the incidence of sideeffects had no significantly difference among groups (P ＞0.05).Children's weight and medicine dose were found to affect recovery time.Conclusions Intranasal dexmedetomidine 1 ～ 2 μg/kg could be used effectively and safely in children undergoing echocardiography examination.Weight and dose were considered as key indexes to predict recovery time.

Objective To investigate the diagnostic value of ADC in the evaluation of small round cell malignant tumors(SRCMT) of nasal and paranasal sinus.Methods This study included 143 patients with surgically confirmed SRCMT and Non-SRCMT of nasal and paranasal sinus between 2008 and 2015, all patients underwent diffusion weighted MRI at 3.0 T with a b factor of 0 and 1 000 s/mm2.Quantitative analysis of ADC values was performed.Difference in ADC values between SRCMT and Non-SRCMT was evaluated using the independent samples t test.One-way analysis of variance(ANOVA) test was performed to compare the ADC values of SRCMT.Receiver operating curves (ROC) were developed to determine the cutoff points to differentiate SRCMT from Non-SRCMT.Results There were 98 SRCMT, of which 20 lesions were rhabdomyosarcoma(RMS), 19 lesions were non-Hodgkin's lymphoma(NHL), 4 lesions were malignant melanoma(MM), 14 lesions were neuroendocrine carcinoma(NEC), 12 lesions were Ewing sarcoma or primitive neuroectodermal tumor(EWS or PNET), 11 lesions were extramedullary plasmacytoma(EMP), and 8 lesions were olfactory neuroblastoma(ON).There were 45 Non-SRCMT, of which 28 lesions were squamous cell carcinoma(SCC) and 17 lesions were adenoid cystic carcinoma(ACC).The mean ADC value of SRCMT[(0.66 ± 0.12) × 10-3mm2/s] was significantly different (t=14.97, P＜0.01) from Non-SRCMT [(1.02± 0.16)× 10-3mm2/s].All of 7 kinds of SRCMT were divided into 3 groups according to ADC values: NHL,MM, NEC,EMP;RMS,EWS,PNET;ON.There was statistically significant difference among all 3 groups(F=39.743, P＜0.01), and the differences between any 2 groups were still statistically significant.The area under the ROC of ADC values diagnosing SRCMT was 0.975.Compared with pathological results, an ADC value of 0.82 × 10-3mm2/s was used as the threshold for diagnosing SRCMT with a sensitivity of 97.8％ (44/45),specificity of 89.8％(88/98), and accuracy of 92.3％ (132/143).ADC value had high correlations compared with pathological results (Kappa value was 0.831).Conclusion The ADC value is a non-invasive imaging parameter that can be used to effectively assess SRCMT of nasal and paranasal sinus.

Objective To evaluate the anatomic changes and dosimetric variations of patients with nasopharyngeal carcinoma (NPC) during the course of simultaneous integrated boost intensity-modulated radiotherapy (SIB-IMRT) by comparison of the dosimetric differences with or without replanning.Methods Twelve cases with NPC treated with SIB-IMRT underwent repeated CT scans after 20- 25 fractions of the initiation of therapy.The original treatment plan ( Plan1 ) based on the first CT scan ( CT1 ) and the second IMRT plan (Plan 2) based on the second CT scan (CT2) were calculated with an inverse planning system (Pinnacle3,Philips Medical System).In addition,the hybrid IMRT plan,Planl (CT2),was generated by deformable registration with MIMVISTA software,and the doses in Plan 1 ( CT1 ) and Plan 2 ( CT2 ) were accumulated based on CT2.The dosimetric differences were compared among the Plan 1 ( CT1 ),Plan 1 (CT2) and Plan 1 + 2(CT2).Results Compared with CT1,the mean volumes of the right and left parotid glands in the CT2 were significantly smaller by ( 24.6 ± 11.9 ) ％ and ( 35.1 ± 20.1 ) ％,respectively.Compared with Plan 1 ( CT1 ),the dose received by 95％ of the target ( D9s ) to PGTV,PTV1 and PTV2,and mean dose (D ) to PGTV,and PTV2 were all significantly lower in the Plan 1 (CT2),indicating that the doses to targets decreased without replanning.With repeated CT and replanning after 25 fractions as shown in Plan 1 + 2 (CT2),the doses to targets would be improved.The doses to normal tissue were increased without replanning,although no statistical significance was observed.In 5 of 12 cases,the doses to the spinal cord and brainstem exceeded the constraint without replanning,while the corresponding values decreased with replanning.Conclusions During the course of IMRT for cases with NPC,the volumes of the targets and parotid glands decrease significantly.Mid-treatment CT scanning and replanning should be recommended to ensure adequate doses to the targets and safe doses to the normal tissues.

Objective To evaluate the dose distribution of target volume and normal tissues in forward intensity modulated radiotherapy (fIMRT) and inverse intensity modulated radiotherapy (iIMRT) modes for breast cancer after radical mastectomy.Methods Both fIMRT and iIMRT plans were developed for 10 patients with breast cancer after radical mastectomy.On each patient's CT images the supraclavicular area, chest wall, and internal mammary area were delineated.The prescription dose was 50 Gyin 25fractions.In the fIMRT plan X-ray irradiation at the dose of 6 MV was adopted for the supraclavicular and the chest wall areas and electron irradiation at the dose of 9 - 12 MeV was adopted for the internal mammary area, and the doses of cold and hot spots were adjusted according to the fitting doses of these 3 regions.In the iIMRT plan the supraclavicular area, chest wall, and internal mammary area were taken asa whole target, 6 MV X-rays was used, and inverse optimal design was performed.The dose distribution oftarget volume and normal tissues, conformal index (CI) , and heterogeneous index (HI) , and acceleratormonitor unit (MU) were analyzed using dose-volume histogram (DVH)for the two intensity modulated modes.Results The maximum dose of PTV of the iIMRT plan was significantly lower than that of the fIMRT plan(t = -3.23,P ＜0.05), the minimum dose and V95% of PTV of the iIMRT were significantly higher than those of the fIMRT plan(t = 4.08, -2.69, both P ＜0.05).The CI level of the iIMRT plan was significantly higher than that of the fIMRT plan and the HI level of the iIMRT plan was significantly lower than that of the fIMRT plan (t = -3.13, 2.74, both P ＜0.05).There were not significant differences in V10, V20, V25, V30, and Dmean of the ipsilateral lung between these 2 groups.However, the V15 of ipsilateral lung of the iIMRT group was significantly lower by 4.2% than that of the fIMRT group (t= 3.2, P ＜ 0.05).There were not significant differences in the mean dose (Dmean) and V30 of heart, and Dmean of contralateral lung and contralateral breast between these 2 groups.Conclusions Compared with fIMRT, the iIMRT plan results in more PTV coverage, higher conformity index, and more homogeneous dose distribution, with lower dose upon the lung at the affected side, and better protection of the contralateral lung, heart, and breast.

The L-ATC hydrolase and L-SCC amidohydrolase which convert L-ATC to L-cysteine in Pseudomonas sp.TS-1138 are purified about 83.9 and 90.3 fold by salting-out method, Sephadex G-75 gel chromatography, DEAE-cellulose 52 ion exchange and Sephadex G-100 gel chromatography, etc. The purified enzyemes are both demonstrated by SDS-PAGE to be a homogeneous protein. Their molecular weight are about 37.5kD and42.8kDa respectively. The optimum reaction temperature are both 35℃, and the optimum pH are 7.0 and 8.0 respectively. The Km of the two enzymes are 0.67 mmol/L and 0.15 mmol/L, and the Vmax are 0.39?10 -3mmol/L?min and 0.42?10 -3mmol/L?min respectively.

The L-cysteine desulfhydrase gene (cd) of Pseudomonas sp.TS1138 was amplified by PCR,and the amplified gene was recombined in the cloning vector pBluescript SKII.The 1.2kb DNA fragment containing cd was sequenced,and its homology with other desulfhydrases was blast; then the cd was cloned into the expression vector pET-21a(+), and afterward expressed by IPTG inducement.The expression protein was purified by Ni-NTA His-Bind Resin.Then the expression protein was identified by the method of activity staining of desulfhydrase, and the characterization of L-cysteine desulfhydrase and the critical role it played in the L-cysteine biosynthetic pathway were discussed.