Objective To explore the application of high frequency ultrasonography in the rapid diagnosis of irreducible radial head subluxation in children.Methods Twenty-three patients aged 1 to 5 with unilateral radial head subluxation in children were observed by high-frequency ultrasonography and radiography,and then compared the symptomatic to the contralateral side.Results No case of radial head subluxation was found by radiography.23 cases of radial head subluxation were found by high frequency ultrasonography,10 cases with articular cavity effusion.On the diagnosis of radial head subluxation in children,the sensitivity of high frequency ultrasonography was obviously higher than that of radiography.Conclusions The sensitivity of high frequency ultrasonography in the rapid diagnosis of the radial head subluxation in children is obviously higher than that of X ray film.High frequency ultrasonography through continuous scan can determine the continuity of anatomical structure and the relationship between some views to show the location of the specific structure relations.Specifically,it can be quickly and clearly diagnosed on anterior long-axis view and the lateral coronal view of radiohumeral joint.

Objective To conduct comparative study on the analgesic and anti-inflammatory effects as well as the acute toxicity of yunaconitine and 8-deacetyl-yunaconitine isolated from the processed products of Aconiti Knsnezoffii Radix.Methods The methods of hot plate test and writhing test were used to evaluate the analgesic effect. Anti-inflammation action was observed by the models of auricle swelling caused by dimethylbenzene. LD50 was determined by the method of Bliss.Results Yunaconitine and 8-deacetyl-yunaconitine have analgesia effect on the pain caused by hot-plate, but there were no statistically significant difference. The pain caused by acetic acid had obvious analgesic action. High and low dose of yunaconitine could significantly reduce the number of mice body torsion and extend the incubation period of pain in mice. The effect of 8-deacetyl-yunaconitine was remarkable only in the high dose. Compared with solvent group, there were little differences in inhibiting effect of auricle swelling caused by dimethylbenzene, and anti-inflammatory action was not exact. The poisonousness of yunaconitine was nearly 20 times of 8-deacetyl-yunaconitine.Conclusion Yunaconitine and 8-deacetyl-yunaconitine may be the analgesic medicine for peripheral analgesic effect. The poisonousness of 8-deacetyl-yunaconitine is less than yunaconitine, the effect is remarkable to the pain caused by acetic acid, and the security is high.

Objective To establish a method for simultaneous determination of arsenic and selenium in hair by hydride generation atomic fluorescence spectrometry.Methods The A and B of the hair samples were digested via the wet method,and the arsenic and selenium contents were determined simultaneously with a dual channel atomic fluorescence spectrophotometer.Standard blank solution was continuously determined for 11 times,the detection limit was calculated according to the blank solution three times the standard deviation.Hair samples A and B were continuously determined for 11 times,and the precision was determined according to the coefficient of variation (CV).The hair samples C and D were did 11 times of standard addition experiment,the standard added was close to the sample low value.The accuracy was determined by the standard recovery rate.Results The lowest detection limits of arsenic and selenium were 0.081,0.036 μg/L,the precisions of arsenic and selenium in sample A were (0.694 9 ± 0.024 2) and (1.930 2 ± 0.098 5) μg/g,the CV were 3.54％ and 5.09％.The precisions of arsenic and selenium in sample B were (6.212 2 ± 0.137 3) and (8.282 2 ± 0.266 8) μg/g,the CV were 2.21％ and 3.22％;the recovery rates of sample C of arsenic and selenium were 96.30％ ～ 110.00％ and 94.80％ ～ 110.00％;the recovery rates of sample D of arsenic and selenium were 93.63％ ～ 108.20％ and 97.03％ ～ 108.34％.Conclusions The method using hydride atomic fluorescence for determination of arsenic and selenium in the hair has been successfully established.This method is simple and easy to operate,with high sensitivity and good accuracy and precision,high recovery,less matrix interference,and less reagent consumed etc.It is worth promoting in prevention and control of endemic diseases.

We developed a scalable AAV5/5 vector packaging system by using replication competent recombinant herpes simplex type 1 virus as helper virus. The fragment containing rep and cap genes of AAV5 was inserted into the non-necessary gene (UL2) of HSV1 genome, resulting in the helper virus rHSV1-rep5cap5. An AAV5/5 vector pAAV5neo carrying two AAV5 ITRs was constructed by inserting a neo gene expression cassette into the plasmid backbone of pAV5CMV-GFP. pAAV5neo-EGFP was constructed by inserting EGFP gene into pAAV5neo. BHK21 cell was transfected with pAAV5neo-EGFP and cultured in the presence of G418. EGFP expression positive monoclonal cells were picked up, and one that produced rAAV5/5-EGFP with the highest efficiency under the help of rHSV1-rep5cap5 was chosen as the production cell line named as C020. rAAV5/5-EGFP was produced by infecting C020 cells with rHSV1-rep5cap5, and crudely purified by our previous method of 'chloroform treatment-PEG8000/NaCl precipitation- chloroform extract'. rAAV5/5-EGFP preparation with high purity was obtained by ultrafiltration with molecular weight cut-off value of 100 kDa. SDS-PAGE stained with Coomassie brilliant blue R250 showed clearly specific pattern of three bands of AAV capsid proteins. rAAV5/5-EGFP was also assayed using negative stain transmission electron microscopy and the majority of the virus particles were found solid. About 30% green fluorescent cells could be seen after infecting HEK293 cells with rAAV5/5-EGFP 24 h at 1 x 10(5) vg/cell. In conclusion, we have established an efficient AAV5/5 vector production system and could produce recombinant AAV5/5 virus in large amounts for gene therapy research.
Dependovirus ; genetics ; physiology ; Genetic Therapy ; Genetic Vectors ; HEK293 Cells ; Herpesvirus 1, Human ; genetics ; physiology ; Humans ; Reassortant Viruses ; genetics ; Recombination, Genetic ; Viral Proteins ; genetics ; Virus Assembly

We chose Gaussia luciferase (Gluc), a secreted luciferase gene as reporter to real-time detect and dynamically monitor hydrodynamic injection gene expression. First, we constructed an expression vector pAAV2neo-Gluc. Then Huh7 and HepG2 cells were transfected with pAAV2neo-Gluc and the activity of Gluc in the supernatant and cell lysates were assayed. Results showed that the Gluc activity in the supernatant was about 100 higher than that in cell lysates, indicating the expressed Gluc existing mainly as a secreted form as reported. Live bioluminescence imaging of mice hydrodynamic injected pAAV2neo-Gluc showed whole body distribution, while the pAAV2neo-Fluc primarily located in the liver. Then we injected different doses of pAAV2neo-Gluc into mice by tail-vein hydrodynamic injection, took minor amount of blood from mice tails at different time points and measured the luciferase activity to investigate dynamic changes of Gluc expression and secretion in vivo. The results suggested that the time courses of Gluc expression were highly consistent among each dose groups. The luciferase activity in blood could be detected as early as 2 h after injection, reached the peak at about 10 h and gradually decreased from then on. The expression level of Glue was positively correlated with the dose of injected plasmid DNA. To further detect the assay sensitivity of the ex vivo Gluc measurement method, we investigated three additional groups of mice injected with lower doses of 0.001 microg, 0.01 microg and 0.1 microg pAAV2neo-Gluc respectively. Results revealed that activity of Gluc in blood could be detected even at dose as low as 0.001 microg DNA, suggesting the assay sensitivity was extremely high. In conclusion, a real-time ex vivo detection method of dynamically monitoring of gene expression in vivo by hydrodynamic injection can be a valuable means for the study of gene expression regulation in vivo.
Animals ; Gene Expression Regulation ; Genes, Reporter ; genetics ; Genetic Therapy ; Hep G2 Cells ; Humans ; Hydrodynamics ; Injections, Intravenous ; Liver ; metabolism ; Luciferases ; administration & dosage ; biosynthesis ; genetics ; pharmacokinetics ; Male ; Mice ; Mice, Inbred BALB C ; Transfection

OBJECTIVEThis study aims to establish a stable animal model with altered anterior guidance and investigate histological changes on temporomandibular joint (TMJ) synovium in rats.

METHODSThirty-two rats were divided into four groups, namely, control group, anterior-guidance-deficient group (T1), and anterior guidance inclined up to 15 degree (T2) and 30 degree (T3) groups. Metal crowns that add 15 and 30 degrees to the palatal side were cemented to the incisors of maxilla to increase the inclination of anterior guidance. The low-speed diamond wheel shorts for both upper and lower incisors of rats were applied to avoid contact between anterior teeth in all direction, thus establishing an anterior guidance deletion model. TMJ joints on one side of two animals in each group were harvested after 3, 7, 14, and 28 days. Pathological changes were investigated in the TMJ synovium using hematoxylin-eosin (HE) staining.

RESULTSThe weight of T1 decreased briefly at first, and then increased slightly (P<0.05). The weight of T3 obviously reduced and then slowly increased, but remained below the initial weight (P<0.05). T3 manifested obvious synovial lining proliferation and vascular dilation (P<0.05).

CONCLUSIONSThe rat model of anterior guidance inclined up to 30 degree could imitate the pathological features of TMJ synovium with excessive increase in anterior guidance. A higher inclination of anterior guidance might cause more damage of TMJ synovium. The effect of anterior guidance deletion on TMJ synovium is yet to be determined.
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Animals ; Dental Cementum ; Disease Models, Animal ; Incisor ; Maxilla ; Rats ; Synovial Membrane ; Temporomandibular Joint ; Temporomandibular Joint Disorders