Objective To investigate the impact of stable expression of perlecan shRNA lentiviral particles on proliferation of NIH3T3 cells. Methods Mouse fibroblasts were cultured.Lentiviral particles-green fluorescent protein (LV-GFP) was used to transfect the cultured NIH3T3 cells with multiplicity of infection (MOI) of 10,30 and 50.The GFP expression was observed with fluorescence microscopy after transfection for one week to estimate the proper MOI and the time of GFP expression needed.The transfection efficiency of LV-GFP with the proper MOI by fluorescence-activated cell sorting was detected.The stably transfected cell lines were developed by puromycin screening for more than 2 weeks.The third generation HFF in good condition was randomly divided into 3 groups:GFP group,shRNA group and control group.RT-PCR,Western blot and MTT assays were used to detect the expressions of perlecan mRNA and protein and cell proliferation in the 3 groups. Results Perlecan mRNA and protein showed high expressions in the control and GFP groups but low expressions in the shRNA group,with significant differences respectively between the shRNA group and the other 2 groups ( P ＜ 0.05).There was no significant difference between the 3 groups in the optical density at the first 2 days ( P ＞ 0.05).On 3 to 6 days the cells in the control and GFP groups grew normally while the cells in the shRNA group proliferated in a weak manner.the transfected cells in the shRNA group showed a significantly reduced proliferation rate compared with the other 2 groups ( P ＜ 0.05 ). Conclusion The growth of NIH3T3 cells can be inhibited significantly by transfection with perlecan shRNA lentiviral particles.

Objective To investigate the effects of Huangqi Sijun Decoction on the levels of thyroxin and cAMP and cGMP in model rats with spleen deficiency syndrome and to explore its therapeutic mechanism.Methods Male SD rats aged three months were randomly divided into 3 groups:normal control group,spleen deficiency model group and Huangqi Sijun Decoction group.The rat models of spleen deficiency syndrome were established by gavage with the decoction of Rhubarb.The serum levels of thyroxin(T4),3,5,3'-triiodothyronine(T3),thyrotropin(TSH)and plasma levels of cAMP,cGMP were determined.Results T3 and T4 were significantly lower but the levels of cAMP and cAMP/cGMP were significantly higher in the spleen deficiency models than those in the normal control group(P

Objective To explore the associations between vitamin D receptor ( VDR) Fok- Ⅰ and Apa- Ⅰ polymorphisms and myasthenia gravis (MG) in Chinese Han population.Methods Polymorphisms of VDR Fok- Ⅰ and Apa-Ⅰ were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.The frequencies of genotypes and hyplotypes were compared among 286 normal controls and 267 MG patients in different subgroups classified by gender,age of onset,presence of thymoma,and Osserman classification sat maximal severity in the follow-up.The association between the genotypes and maximal severity of MG and short-term glucocorticoid treatment were also investigated.Results There were no significant differences in frequencies of genotypes and hyplotypes of both Fok-Ⅰ and Apa-Ⅰ between MG group and control group,and among subgroups of MG.The Fok- Ⅰ showed no statistical difference between the patients with better and less improvement after short-term treatment of glucocorticoid.The frequency of Apa-Ⅰ alleles in the patients with better improvement (55/186,29.6% ) significantly differed from the less improved group ( 7/48,14.6%,OR = 2.46,95% CI 1.04-10.43,x2 = 4.400,P = 0.036).The patients with the genotype A A/Aa were more likely to improve better after the treatment(48/93,51.6%) than in the worse group(7/24,29.2%,OR =2.59,95% CI 0.98-14.60,x2 =3.858,P= 0.049).Conclusions Alleles and genotypes of VDR-Fok- Ⅰ and Apa-Ⅰ were not found to be related with MG onset and severity.MG patients with VDR-Apa-A allele may have better improvement short-term treatment of glucocorticoid.

Objective To develop new scales on the severity of myasthenia gravis ( MG) which are suitable for clinical practice in China .Methods A final version of new comprehensive scale was developed by item optimization from a preliminary scale established by combination of Delphi method and previous evaluation of items from source scales .With the original quantitative data from 60 MG patients, items were selected on the basis of interobserver and test-retest reliability of items , the contributions of items on internal consistency and construct validity of the preliminary scale .Evaluation of this final scale includes internal consistency, interobserver and test-retest reliability, construct validity and correlation with currently used scales.A self-rating version was simplified from the final scale by including items that can be measured by patients, which was then evaluated in 57 generalized MG patients.Results One item ( palpebral fissure ) was deleted from the preliminary scale due to poor interobserver and test-retest reliability.The remaining nine items constituted the final comprehensive scale which covered six commonly involved muscle groups in MG.All items in the comprehensive scale had fair to excellent interobserver reliability ( K=0.43-0.80, P<0.01) and fair to excellent test-retest reliability ( K =0.41 -1.00, P<0.01) .The comprehensive scale had acceptable internal consistency (Cronbach ɑ=0.63), excellent interobserver reliability (intraclass correlation coefficient (ICC) =0.88, P<0.01) and test-retest reliability (ICC=0.86, P<0.01).The comprehensive scale had four common factors and good construct validity ( cumulative dedication rate 68.46％).There was high correlation between the comprehensive scale and Myasthenia Gravis Composite ( MGC ) , Absolute and Relative Score of MG ( ARS-MG ) and Quantitative Myasthenia Gravis score (QMGs) ( r=0.87 -0.90, P<0.01), and moderate correlation between the comprehensive scale and Myasthenia Gravis Activities of Daily Living (MG-ADL) (r=0.68, P<0.01).A self-rating scale was developed by deleting ocular and facial items and keeping the remaining six items , which covers the bulbar , limb, neck and respiratory muscles.Internal consistency (Cronbach ɑ=0.69), interobserver reliability and test-retest reliability (ICC=0.90 for both, P<0.01) of the self-rating scale were better than those of the comprehensive scale.The self-rating scale had two common factors and good construct validity ( cumulative dedication rate 64.24％) .There was high correlation between the self-rating scale and the comprehensive scale, MGC and ARS-MG (r=0.74-0.86, P<0.01), and moderate correlation between the self-rating scale and QMGs and MG-ADL ( r=0.55 -0.68, P<0.01 ) .Conclusions Item optimization was performed from a preliminary scale established by Delphi method .A new comprehensive scale and a self-rating scale were developed .The comprehensive scale and self-rating scale had good reliability and validity , which can be used to assess the severity of MG reliably and effectively .

ViewRay magnetic resonance (MR) guided radiotherapy system not only solves the problem of imaging dose,but also can set up accurately,online adaptive radiotherapy and gated irradiation according to magnetic resonance imaging (MRI).The development of this system provides a new technical means of accurate radiotherapy.This review describes the main structure of the ViewRay system,and summarizes quality assurance (QA),dosimetric comparison,respiratory motion management,online adaptive radiotherapy,and preliminary treatment effect.