Animals ; Antioxidants ; pharmacology ; Fatigue ; prevention & control ; Male ; Mice ; Physical Exertion ; physiology ; Random Allocation ; Recombinant Proteins ; genetics ; metabolism ; pharmacology ; Superoxide Dismutase ; genetics ; metabolism ; pharmacology ; Swimming ; physiology

Objective To investigate clinical effect of recombinant human growth hormone (rhGH)for inter-venting central precocious puberty in children,to provide reference for clinical treatment.Methods 80 patients with growth deceleration slows CCP were selected,when height growth rate below 4cm/year,40 patients on the basis of joint GnRHa used rhGH treatment were selected as the observation group,and the remaining 40 patients refused to use rhGH,continued to use of GnRHa treatment were selected as the control group.After treatment predicted height,height,bone age,growth rate and other indicators were compared.Results In observation group,the actual height Ht, predicted height PAH,the growth rate Gv,serum insulin -like growth factor -1 (IGF -1 )after treatment were (1 44.48 ±6.59)cm,(1 54.94 ±4.52)cm,(4.43 ±0.64)cm/6months,(1 32.25 ±8.84)ng/mL,compared to before treatment,the differences were statistically significant (t =6.548,P <0.01 ;t =5.734,P <0.01 ;t =28.869,P <0.01 ;t =20.65,P <0.01 ),compared with the control group,the differences were significant (t =3.943,P <0.01 ;t =4.759,P <0.01 ;t =28.247,P <0.01 ;t =20.882,P <0.01 ),there were no differences in other indices;hor-mones FSH,LH and other indicators of the two groups before and after treatment,and blood sugar,thyroid indicators showed no abnormality.Conclusion Recombinant human growth hormone in the treatment of central precocious puberty deceleration in children does not increase bone age,which will help improve the growth rate and predicted height,which worthy of clinical application.

Objective To investigate the infection rate of Legionella pneumophila(LP)in children younger than 5 years with lower respiratory infections by ELISA and PCR.Method Serum LP-IgM and IgG were measured by ELISA,and LP DNA in sputum or throat swab were detected by PCR in 300 children less than 5 years with lower respiratory infections.The data were analyzed by chi-square test and the consistency of the two methods was analyzea by NcNemar test.Results Serum antibody detected by ELISA was positive in 17 cases(5.67%),including 10 positive of IgM and 7 positiile of lgG.In these 17 eases,11 were males and 6 were females with similar positive rates(5.76%vs 5.5%).Four cases(2.53%)were positive in children aged from 27 days to 1 year,while 7(7.37%)and 6 cases(12.77%)were positive in children aged 1-3 years and 3-5 years,respectively.Seven cases(5.19%)found in the winter and spring seasons and 10 cases(6.06%)in summer and autumn seasons.Eleven children(11.83%)were from urban area and only 6(2.9%)from countryside.DNA in sputum or throat swab detected by PCR was positive in 16 cases(5.33%),included 10 males and 6 females with similar positive rates(5.24%vs 5.5%).Five cases(3.16%)were positive in children aged from 27 days to 1 year,while 6(6.32%)and 5 cases(10.64%)were positive in children aged 1-3 years and 3-5 years,respectively.Three cases(2.22%)found in the winter and spring seasons and 13 cases(7.88%)in sunmmr and alltumll seasons.Nine children(9.68%)were from urban area and only 7(3.38%)from countryside.McNemar test was used to compare the data of ELISA and PCR results with a Qm of 0.037 and a Pr of 0.8474.Conclusions LP might contribute partly to the lower respiratory infections in children less than 5 years.The infection rate of LP increases with age increasing.Urban children have a higher infection rate than those living in countryside.Both methods have a good consistency.

Objective To analyze factors influencing short-term effect of presurgical pharmacological thera-py and transsphenoidal microsurgery for somatotropinomas. Methods The clinical data of 53 patients underwent presurgical pharmacological therapy and transsphenoidal surgery for somatotropinomas were retrospectively analyzed in order to search for factors influencing effect of presurgical pharmacological therapy and transsphenoidal surgery for somatotropinomas. Results Serum GH inhibition rates decreased<50.00%from baseline in 62.26%of patients receiving presurgical pharmacological therapy. Statistical analysis concerning the influence of sex , neuropathological evaluation, tumor size and presence of invasion on presurgical pharmacological therapy effect were performed using a chi-squared test, no significant correlation was found among these factors and presurgical pharmacological therapy effect. Total remission rates were 43.40%, Statistical analysis concerning the influence of sex , neuropathological e valuation, tumor size, presence of invasion and presurgical pharmacological therapy effect on remission rate were performed using a chi-squared test, a significant correlation was found among tumor size, presence of invasion, presurgical pharmacological therapy effect and remission rate , while no significant correlation was found among the rest of the factors. Further Logistic regression analysis demonstrated a significant correlation among tumor size , presence of invasion and remission rate , while no significant correlation was found between presurgical pharmacolog-ical therapy effect and remission rate. Conclusions Presurgical pharmacological therapy effect revealed no signifi-cant correlation with sex, neuropathological evaluation, tumor size or presence of invasion. Total remission rate cor-related with tumor size and presence of invasion. A better presurgical pharmacological therapy effect may indicated a better outcome, while postoperative remission rate revealed no significant correlation with presurgical pharmacologi-cal therapy in our series.

This study was aimed to identify and distinguish Metacordyceps liangshanensis recorded by the Sichuan Province Traditional Chinese Medicine Standard from its adulterants and its relative species by combining ITS and COI barcode sequences in order to study the feasibility of this new method. After extracting DNA of 28 species of Cordyceps samples, DNA were amplified and sequenced. And then, ITS and COI sequences were received. Codon-Code Aligner V3.7.1 and Mega 5.0 were used to analyze the variable site and construct the N-J tree. The results showed that the minimum ITS inter-specific K-2P distance was relatively higher than the maximum intra-specific K-2P distance. The inter-specific sequence divergence between M. liangshanensis and its adulterants exhibited high while intra-specific sequence divergence exhibited low. And COI one was the same case. N-J tree of both ITS and COI indicated that same genus belonged together and each species belonged to relatively independent branch. It was concluded that based on the ITS and COI gene, the technology of DNA barcode can be an excellent identification of M. liangshanensis, its adulterants and its relative species. It provided technical support for the further research on species molecular identification and phylogenetics of Cordyceps .

BACKGROUND:Specific down-regulation of interleukin 1 beta (IL-1β) may al eviate the pain behaviors effectively after peripheral nervous injury. Compared with smal interference RNA (siRNA), short hairpin RNA (shRNA) could inhibit the expression of target gene more stably and efficiently. However, simple shRNA could not enter target cel s to down-regulate target gene efficiently. Adenovirus vectors have wide host range, high infection efficiency and stable expression in host cel s.
OBJECTIVE:To construct recombinant adenovirus vector expressing shRNA targeting IL-1βgene and detect its effect on the expression of target gene.
METHODS:Three siRNAs were designed on the basis of the nucleotide sequence of IL-1βobtained from NCBI and then three shRNAs (shRNA1, shRNA2 and shRNA3) were synthesized. The annealed shRNA product and adenovirus vector pHBAd/U6/GFP digested by BamH I and EcoR I were connected to construct the recombinant adenovirus vector shuttle plasmid expressing shRNA targeting IL-1β. After sequencing, HEK 293 cel s were co-transfected by the shuttle plasmid and skeleton vector, and three recombinant adenovirus vector expressing shRNA targeting IL-1β(rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3) were packaged and amplified. Rats H9C2 cel s were infected by recombinant adenovirus vector expressing shRNA targeting IL-1βand fluorescence microscope was used to observe the infection efficiency. The effect of recombinant adenovirus vector expressing shRNA targeting IL-1βon the expression of target gene was detected by western blot assay.
RESULTS AND CONCLUSION:The sequencing results showed that the sequences of three shRNAs adenovirus vector shuttle plasmid were consistent with the sequences of three designed shRNAs. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 were constructed successful y. rAd/shRNA1, rAd/shRNA2 and rAd/shRNA3 could down-regulate the expression of IL-1βin rat H9C2 cel s and the down-regulation effect of rAd/shRNA2 was the most significant.

To establish a stable mouse model of systemic Candida infection and to set up related standard operation procedure .Methods ICR mice were infected with C.albicans or C.parapsilosis by tail vein injection after immunosuppres-sion by cyclophosphamide .The quality control key points included immunosuppression , strain preparation, inoculation doses and the route of inoculation .Survival analysis , bacterial loads and pathological examination were performed to evaluate the prepared model .Results The developed model showed fugal-specific lesions in multiple organs , especially in the kidneys revealed by histopathological examination .Conclusions A stable mouse model of systemic Candida albicans infection can be successfully established by following standardized operation procedure .This mouse model may provide a useful tool for studies on pathogenesis and immune defense of fungal infection and new anti-fungal drug development and so on .

Objective To evaluate the application of microvascular coupler device (MVCD) in the free-tissue flap transfer for head-and-neck defects reconstruction.Methods Its a retrospective study of the clinical data of 743 patients who received 763 free-tissue flap transfer for head and neck defects reconstruction in Hunan Cancer Hospital from January,2014 to January,2016.For microvascular anastomoses,413 were done manually (the manual group) and 350 done with MVCD (the MVCD group),of which the latter included end-to-end venous anastomosis in 159,endto-side venous anastomosis in 190 and end-to-end arterial anastomosis in 1.The time for anastomosis,rate of blood leakage from the anastomosis and venous thrombosis,and the survival rate of flaps were compared between the manual group and the MVCD group.The impacts of types of anastomosis of end-to-end vs end-to-side (both with MVCD),on the formation of venous thrombosis and survival of flaps were also recorded.The analysis was performed under t-test and chi-square test using SPSS software 19.0 with P ＜ 0.05 for differences with a statistical significance.Results Time for anastomosis was significantly shorter in the MVCD group (4.43±0.51min) than in the manual group (14.75± 2.43min,P＜0.05).The rates of anastomosis leakage were 0 in the MVCD group and 9.7％ in the manual group(P＜0.01),flap necrosis were 0.57％ and 1.69％ (P=0.15),the rate of venous thrombosis were 1.15％ and 3.63％(P=0.02),respectively.There was no significant difference in the time for anastomosis,the rate of venous thrombosis and the survival rate of flaps with end-to-end anastomosis and end-to-side anastomosis with MVCD.Conclusion The application of microvascular coupler device is valuable in the reconstruction of head-and-neck defects with free-tissue flaps because it can significantly shorten the time for anastomosis,decrease the ocurrence of anastomosis leakage and venous thrombosis,reduce the risk of flap failure.

Objective To evaluate the role of interleukin?1β( IL?1β) in activation of N?methyl?D?aspartate ( NMDA) receptors in spinal dorsal horns of rats with neuropathic pain. Methods One hundred twenty?eight pathogen?free healthy adult male Sprague?Dawley rats, weighing 200-250 g, were divided into 4 groups (n=32 each) using a random number table: control group (group C), recombinant adenovirus vector group ( group rAd) , recombinant adenovirus vector shRNA group ( group rAd∕shRNA) , and recom?binant adenovirus vector shRNA plus recombinant adenovirus vector IL?1β group ( group rAd∕shRNA+rAd∕IL?1β) , receiving intrathecally injected equal volume of 0.9% sodium chloride solution, recombinant ade?novirus vector, recombinant adenovirus vector shRNA, and recombinant adenovirus vector shRNA plus re?combinant adenovirus vector IL?1β5 μl, respectively. Chronic compression of dorsal root ganglia was per?formed on 8th day after intrathecal injection. The thermal paw withdrawal latency ( TWL) was measured at 1 day before intrathecal injection ( T0 ) and at 1, 2, 3 and 5 weeks after intrathecal injection ( T1-4 ) . After measurement of the pain threshold at each time point, 8 rats selected randomly were anesthetized and sacri?ficed, and the lumbar enlargement segments of the spinal cord were harvested for determination of the ex?pression of IL?1β and phosphorylated NMDA receptor NR1 subunits at serine 896 ( pNR1S896) in spinal dorsal horns on the injured side by immunohistochemistry and Western blot. Results Compared with group C, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA (P<0.05), and no significant change was found in the parameters mentioned above at each time point in group rAd and group rAd∕shRNA+rAd∕IL?1β( P>0.05) . Compared with group rAd, the TWL was significantly prolonged, and the expression of IL?1βand pNR1S896 in spinal dorsal horns was significantly down?regulated at T2-4 in group rAd∕shRNA ( P<0.05) , and no significant change was found in the parameters mentioned above at each time point in group rAd∕shRNA+rAd∕IL?1β (P>0.05). Compared with group rAd∕shRNA, the TWL was significantly short?ened, and the expression of IL?1β and pNR1S896 in spinal dorsal horns was significantly up?regulated at T2-4 in group rAd∕shRNA+rAd∕IL?1β ( P<0.05) . Conclusion IL?1β is involved in the activation of NM?DA receptors in spinal dorsal horns of rats with reuropathic pain.

Objective To investigate the safty and feasibility of simultaneous minimally invasive technique for congenital heart disease (CHD) and pectus excavatum (PE).Methods From July 2006 to June 2011,6 children with PE associated with CHD were treated by simultaneous minimally invasive technique.They were 4 boys and 2 girls,aged from 4 years to 6 years 5 months ( average 5 years and 4 month),.The body weight were from 16 kg to 20 kg [ average ( 18.00 ± 1.79kg ].CT Hailer index were 3.9 - 5.0 ( average 4.35 ± 0.43 ).4 patients with ventricular septal defect (VSD) were treated by minimally invasive closure device,including 3 membranous VSD and 1 subaortic VSD.The defect diameter was 4 - 5 mm.2 cases with atrial septal defect (ASD) were treated using the same technique.the diameter of ASD was 12 - 16 mm.After minimally invasive heart surgery,Nuss operation was carried out in all 6 patients for their PE.Results All operations were successful and patients were extubated 5 - 11 h after operation [ mean ( 8.17 ± 2.04) h ].The pericardial mediastinal drainage was removed at 48 h postoperatively.There were no operative mortality and severe postoperative complications.1 patient had delayed wound healing and was successfully discharged.The plate was removed in 3 cases,and they all had satisfactory outcomes.Conclusion Simultaneous minimally invasive technique is a safe and effeictive mothod for treating pectus excavatum and congenital heart disease.