Copy number variation (CNV) is one kind of structural variations,which exists extensively in human genome.In addition,it is the crucial source of inter-individual gene differences.Recently,with the successful development of gene array and gene sequencing,molecular methods for CNV testing are more and more standardizing and automating.Therefore,ongoing attentions have been attached to the effects and mechanisms of CNV in complicated diseases like autoimmune diseases.Molecular methods for CNV testing can be divided into two categories:target screening and the whole genome scanning platforms.Currently,the studies about CNV and autoimmune diseases mainly focused on systemic lupus erythematosus and rheumatoid arthritis.Considering the limitations of measurement methods and testing costs,it is still in the infant.

A rapid resolution liquid chromatography coupled with quadrupole-time-of-flight mass spectrometric (RRLC-Q-TOF-MS) method was established and optimized for the analysis of pharmacokinetic behavior of ginsenoside Rb2 in rats by intravenous injection administration.The metabolism of ginsenosides Rb2 in vivo rat was also explored.In the experiment,Agilent SB C18 column was selected for the sample separation with 0.1％ aqueous formic acid solution as mobile phase (A) and acetonitrile as mobile phase (B) at a flow rate of 0.2 mL/min,and the injection volume was set to 5 μL.Q-TOF-MS was carried out in electron pray ionization (ESI) negative ion mode.The limit of quantification (LOQ,S/N =10) and limit of detection (LOD,S/N=3) were 0.10 and 0.08 μg/mL,respectively,and the linear range was 0.1-1.26 μg/mL.The experiment results showed that the concentration-time profile of ginsenoside Rb2 conformed to a two-compartment pharmacokinetic model after intravenous administration for rats.The mean plasma elimination half-lives were (23.58±1.10) min (t1/2α),(1306.55±147.23) min (t1/2β) for Rb2.By analyzing the urine of rats after intravenous administration and the fecal samples after oral administration of ginsenoside Rb2,it was found that the metabolites were M6,M2 (CY),F2,and C-K.

BACKGROUND: There are different methods to isolate and culture human nucleus pulposus cells, and the differences in digestive enzymes components and digestion time quite are significant. So how to rapidly and efficiently harvest human nucleus pulposus cells has become a research hotspot. OBJECTIVE: To optimize the digestive enzymes components and digestion methods for the preparation of human nucleus pulposus cells. METHODS: Nucleus pulposus tissue specimens were selected from three adult discs in the Department of Orthopedics, China-Japan Union Hospital of Jilin University. The acute traumatic disc tissues that outstanding to the spinal canal were taken under aseptic conditions, and then the peripheral white annulus and jel y-like nucleus pulposus in the center could be seen. According to different mixed enzyme concentration ratio, the samples were divided into two groups. The enzyme Ⅰ group was treated with 0.2% Ⅱ col agenase; and the mixed enzymeⅡ group was digested with 0.25% trypsin for 30 minutes, and then treated with 0.2% Ⅱ col agenase. According to digestion time, each group was divided into three subgroups: 2 hours group, 4 hours group, and overnight group. Final y, suspended cel volume was decided as 2 mL to count cells. Dulbecco’s modified Eagle’s medium containing fetal bovine serum was used for cel culture in vitro. Trypan blue staining was performed to count total cel number and ratio of living cells. Methylthiazolyldiphenyl-tetrazolium bromide assay was used to detect the growth curve of nucleus pulposus cells. RESULTS AND CONCLUSION: Based on the two digestion enzyme concentration, the number of digested cells in the enzyme Ⅰ group was larger than that in the enzyme Ⅱ group after digested for 2 and 4 hours, but the difference was not significant (P > 0.05). Overnight, cellsurvival rate was decreased in the enzyme Ⅰ group after digested for 2 and 4 hours when compared with the enzyme Ⅱ group, and the difference was significant (P < 0.05). After digested for 4 hours, tissue blocks disappeared, and the number of cells reached maximum. The results indicate that enzyme Ⅰgroup composite with Ⅱ col agenase is benefit for the separation of nucleus pulposus cells, and the digestion time is appropriate to 4 hours. This condition has the advantages of simple operation, high efficiency and low cost, and it considered that digestion of nucleus pulposus tissues with 0.2% Ⅱ col agenase for 4 hours is the best condition to obtain nucleus pulposus cells.

BACKGROUND:Advancement in bioengineering based upon tissue engineering techniques may offer the possibility of repairing degenerative intervertebral disc.
OBJECTIVE:To summarize the research progress in the scaffolds of tissue engineered intervertebral disc.
METHODS:A computer-based retrieval was performed to search manuscripts describing tissue engineered intervertebral disc scaffolds published between January 1st, 1900 and December 31st, 2012 in PubMed database with the key words of“tissue engineering, intervertebral disc, scaffold”in English.
RESULTS AND CONCLUSION:Scaffold is an important part of tissue-engineered research. There are three kinds of materials for intervertebral disc scaffolds:natural biomaterials, synthetic materials, and composite materials. A variety of scaffold materials have their own advantages and disadvantages. Up to now, none of these scaffold materials is accepted as the most suitable one. The selection of scaffold materials is stil to be further studied. The study and development of nanoscale biomaterials is an inevitable trend. Otherwise, with the help of bionics, improving scaffolds is also an inexorable trend in progress of simulating human intervertebral disc. Furthermore, injectable scaffold is also an research hot spot, and the selection range of injectable scaffold materials mainly focuses on chitosan, typeⅡcolagen,hyaluronic acid,fibrin,elastin,and alginate.C urrently, studies on chitosan as a scaffold material are relatively more.

ObjectiveInvestigate the value and feasibility of image registration with reverse rotation in lateral cerebral DSA.Methods ( 1 ) Experimental study:the target images were subtracted directly,and subtracted again after reverse rotation.Software of registration and subtraction with reverse rotation edited by the author utilizing Visual Basic.The function of the automatic angle detection by the software were evaluated to see whether it detected the angle of line.The subtraction function of DSA by the software was evaluated.(2) Clinical retrospective study:the untreated mask and target inages of 15 patients with motion along vertical axis during lateral cerebral DSA were uploaded to the software.The target images were processed with and without the software to get two sets of images.( 3 ) Evaluation:four experienced radiologists read and compared the two sets of the images,and graded their findings.Results ( 1 ) The automatic detection by the software suggested that the target images should be rotated counterclockwise 1.3°. The subtraction result of the software was satisfactory.(2)In the 15 sets of images,there were only three sets of images deemed optimal after traditional subtraction.After reverse rotation,artifacts were significantly reduced and the image sharper. There were ten cases with significant artifacts after traditional subtraction,and those images were sharper and showed more peripheral vessels after reverse rotation. The traditional subtraction images of two sets could not be interpreted,the reverse rotation registration images reached the diagnostic quality.(3)Subjective evaluation: there were more information and less noise and distortion in the registration images with reverse rotation than in the traditional subtraction. But the image resolution decreased slightly after reverse rotation registration.ConclusionThe registration of digital angiography with reverse rotation can improve the image quality in lateral cerebral DSA.

Objective In order to evaluate that whether Pluronic P85 coated poly(butylcyanoacrylate) nanoparti-cles was able to deliver antiepileptic drug phenytoin into the brain va bypassing mesial temporal lobe epilepsy (MTLE)-induced Pgp in a rat model of MTLE. Methods The rat model of MTLE, induced by li-pilocarpine, was divided in-to two groups (6 for nanoparticle drug group and 7 for PHT drug group). Immunohistochemistry assay was performed to detect Pgp expression at the hippocampus. Nanoparticles were prepared by interfacial polymerization method. Dialysate samples of brain were collected at 30, 60, 120, 180, 240 and 300 min after drug administration by microdialysis tech-nology. Samples were analyzed by high performance liquid chromatography (HPLC). Results The area under the curve (AUC) ratio of brain/plasma in Nanoparticle drug group was 0.370.10 which was significantly higher compared with 0.190.06 in conventional PHT drug group (P<0.05). The Pgp immunopositive area, as assessed by analysis of labeled surface area, was higher in the DG, CA3 and CA1 sector in the hippocampus of MTLE rats when compared to the normal rats. Conclusions Pluronic P85 coated PBCA nanoparticles can significantly deliver PHT into brain via bypassing MTLE-induced Pgp in a rat model of MTLE.

The present study examined the role of Wnt/β-catenin signaling pathway in the degeneration of nucleus pulposus cells and the protective effect of DKK1 on nucleus pulposus cells. The model of nucleus pulposus cell degeneration was induced by intra-disc injection of TNF-α, and the expression of β-catenin protein was detected by Western blotting. The cultured rabbit nucleus pulposus cells were divided into 4 groups. In group A, the cells were cultured with normal medium and served as control group. In group B, the cells were cultured with TNF-α and acted as degeneration group. In group C, the cells were cultured with TNF-α and transfected with Adv-eGFP and was used as fluorescence control group. In group D, the cells were cultured with TNF-α and transfected with Adv-hDKK1-eGFP, serving as intervention group. The expression of type II collagen, proteoglycan, β-catenin, and MMP-13 in each group was detected by immunocytochemistry and RT-PCR. The result showed that TNF-α increased the expression of β-catenin and MMP-13, and significantly inhibited the synthesis of type II collagen and proteoglycan, which resulted in the degeneration of nucleus pulposus cells. This effect could be obviously reversed by DKK1. We are led to concluded that TNF-α could activate the Wnt/β-catenin signaling pathway, and increase the expression of MMP-13, thereby resulting in disc degeneration. Specifically blocking Wnt/β-catenin signaling pathway by DKK-1 could protect the normal metabolism of intervertebral disc tissue. The Wnt pathway plays an important role in the progression of the intervertebral disc degeneration.

Objective To investigate the prevalence and clinical significance of different subtypes (IgG,IgM and IgA) of anticardiolipin antibodies (aCL) and anti-β2-glycoprotein Ⅰ antibodies (aβ 2GP1),as well as lupus anticoagulant (LA) in systemic lupus erythematosus (SLE).Methods IgG/IgM/IgA,IgG,IgM,IgA aCL and anti-β2GP1 were tested by enzyme-linked immunosorbent assay (ELISA) in 100 patients with SLE (42 patients were diagnosed as secondary antiphospholipid syndrome),44 healthy controls and 32 patients with other connective tissue diseases excluding SLE and antiphospholipid syndrome (APS).Meanwhile,LA was tested by modified Dilute Russell's viper venom time (dRVVT).The correlation between antiphospholipid antibodies and clinical manifestation was analyzed by Spearman correlation analysis.The postiverate of antiphospholipid antibodies in SLE patients,health controls and patients with other connective tissue diseases were compared by chi square test.The concentrations of antiphospholipid antibodies in different groups were compared using independent sample Kruskal Wallis test.The diagnostic efficacy of antiphospholipid antibodies in SLE patients was analyzed by crosstable using clinical diagnosis of APS as gold standard.P ＜ 0.05 was considered statistically significant.Results The prevalence of IgG aCL (x2 =15.031,P ＜ 0.001),IgA/G/M (x2 =11.678,P =0.003) and IgA (x2 =6.17,P =0.036) antiβ2GP1 were significantly higher in patients with SLE than in the other two groups.IgA/G/M (r =0.207,P=0.039),IgG (r=0.230,P=0.021) and IgA (r=0.217,P=0.030) aCL,IgA/G/M (r=0.218,P=0.029) and IgA (r =0.255,P =0.01) anti-β2GP1,as well as LA (r =0.233,P =0.02) were associated with thrombotic events.IgA/G/M anti-β2GP1 (r =0.22,P =0.029) and LA (r =0.254,P =0.011) were associated with pathological pregnancy.23.1％ (6/26) aCL positive SLE patients were IgM and/or IgA aCL positive.53.6％ (15/28) anti-β2GP1 positive SLE patients were IgM and/or IgA antiβ2GP1 positive.In SLE patients,the specificity and sensitivity of IgA/G/M aCL for APS were 98.3％ and 26.2％,respectively.The specificity and sensitivity of IgA/G/M anti-β2GP1 were 84.5％ and 40.5％,respectively.The specificity of at least two isotypes positive for APS (both aCL and anti-β2GP1 were 98.3％),was higher than IgG aCL (94.8％) or anti-β2GP1 (93.1％).The sensitivity of at least one isotype of aCL (47.6％) or anti-β2GP1 (42.9％) positive for APS were higher than IgG aCL (40.5％)and anti-β2GP1 (21.4％).Conclusions IgG and IgM aCL together would be better than IgA/G/M aCL for APS screening.IgA/G/M anti-β2GP1 would be better for APS screen due to higher sensitivity and strong association with thromboembolic events and pathologic pregnance.IgA aCL or anti-β2GP1 was associated with thromboembolic events.IgA aCL or anti-β2GP1 would be useful for APS diagnosis in IgG and IgM aCL or anti-β2GPl negative patients.

OBJECTIVE: To establish a supplementary diagnostic indicator (infrared radiant intensity) in tongue diagnosis of traditional Chinese medicine (TCM) in patients with hyperplasia of mammary glands through correlation analysis of infrared radiant intensity between hyperplastic breast tissue and tongue surface. METHODS: Infrared radiant intensity of the hyperplastic breast tissue and different points on tongue surface in 20 cases of hyperplasia of mammary glands with liver-energy stagnation and phlegm retention syndrome and 16 cases of hyperplasia of mammary glands with irregular thoroughfare and conception vessels syndrome were measured with external infrared spectrometer PHE-201 made by Shanghai Institute of Technical Physics. Correlation of infrared radiant intensity between the hyperplastic breast tissue and the different points on tongue surface was assessed by using bivariate correlation analysis. RESULTS: The results showed that the numbers of positive correlated wave bands of infrared radiant intensity between the hyperplastic breast tissue and different detected points on tongue surface in the patients with liver-energy stagnation and phlegm retention syndrome and irregular thoroughfare and conception vessels syndrome were 127 (83.55%) and 71 (46.71%), respectively. Infrared radiant intensity between the hyperplastic breast tissue and the tongue surface had a positive correlation. CONCLUSION: Infrared radiant intensity can be used as one of supplementary diagnostic indicators in TCM tongue diagnosis of hyperplasia of mammary glands.

Objective To evaluate the contraception efficacy, mode of bleeding, side effects and other positive effects of drospirenone-ethinylestradiol (Yasmin) in healthy Chinese women. Methods This was a multicenter, randomized, control study of 768 healthy Chinese women who consulted about contraception. The subjects were randomized into Yasmin group (30 μg ethinylestradiol plus 3 mg drospirenone, 573 cases) or desogestrel group (30 μg ethinylestradiol plus 150 μg desogestrel, 195 cases) with the ratio of 3: 1. Each individual was treated for 13 cycles. Further visits were required at cycle 4, cycle 7, cycle 10 and cycle 13 of treatment. Weight, height, body mass index were evaluated at each visit. The menstrual distress questionnaire (MDQ) was given to the women at baseline, visit 3 (cycle 7) and visit 5 (after cycle 13). Results The values of basal features were similar between two groups (P> 0.05). The Pearl index (method failure) of Yasmin was 0. 208/hundred women year which was lower than that of desogestrel (0. 601/hundred women year). The mode of bleeding was similar between two groups after trial without showing any significant difference. According to MDQ subscale, the improvement of water retention and increasing appetite during inter-menstrual period and water retention and general well-being during menstrual period in the Yasmin group ( -0. 297, -0. 057, 0. 033, 0. 150 respectively) was more obvious than that in the desogestrel group ( - 0. 108, 0. 023, 0. 231, - 0. 023 respectively) with a significant difference (P < 0. 05 ). Some other values which improved in beth two groups, especially the improvement of breast tenderness and pain and skin abnormality in Yasmin group (18.0%, 89/494; 12. 6%, 62/494) was more distinct than that in desogestrel group (11.3%, 19/168; 5.4%, 9/168). The mean weight increased in desogestrel group (0. 57 kg) while it decreased in Yasmin group ( -0. 28 kg) with a significant difference (P < 0. 01 ). Conclusions Both Yasmin and desogestrel have good efficacy on contraception and similar modes of menstrual bleeding. Yasmin is better than desogestrel in terms of weight control and premenstrual syndrome of oral contraceptive.