Animals ; Extinction, Psychological ; physiology ; Fear ; physiology ; Male ; Mental Recall ; physiology ; Rats ; Rats, Sprague-Dawley ; Sleep Deprivation ; complications ; physiopathology ; Sleep, REM ; physiology

The aim of this study is to compare the contents of five types of anthraquinones which mainly includes chrysophanol, physcion, emodin, rhein and physcion and phenolic acids in ten different processed products from Rheum officinale, and to investigate the effect of different initial processing method on the contents of anthraquinones and phenolic acids. Principal component analysis (PCA) was carried out by SPSS software to evaluate the quality of different processed products from Rh. officinale. In conclusion, the contents of anthraquinones in different processed products from Rh. officinale assume the certain regularity. Whether fresh-cut Rheum officinale Bail and how to dry it are derectly effect the contents of anthraquinones and phenolic compounds. The content of anthraquinones in rheum officinale of drying is obviously higher than smudging, and was more abundant in branch root than tap roots. Rh. officinale of drying which growed in Fengjie gained the highest score in PCA. Meanwhile, the procedure of wetting also help to increase the content of anthraquinones and decrease the content of phenolic acids.
Anthraquinones ; chemistry ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; Hydroxybenzoates ; chemistry ; Rheum ; chemistry

Objective To explore the association of COX-2 Gly587Arg polymorphism with the risk of primary liver cancer.Methods Two hundred and seventy patients with primary liver cancer and 540 health people were selected as our subjects.DNA were extracted from peripheral blood lymphocytes,and genotypes were measured by polymerase chain reaction-restriction fragment length polymorphism method.Odds ratios(OR) and 95％ confidence intervals(CI) were estimated by logistic regression.Results Two kinds of genotype (587Gly/ Gly and Gly/Arg) were found in all participants.No one carried 587Arg/Arg genotype.Among primary liver cancer patients,91.5％ (247/270,) 8.5％ (23/270) of individuals carried 587Gly/Arg and Gly/Arg genotype,which was significantly higher than that of controls (96.5％ (521/540,) 3.5％ (19/540)).Multivariate Logistic regression analysis showed that individual carried 587Gly/Arg genotype had an increased risk of developing primary liver cancer (OR =2.56,95％ CI =1.37-4.79,P =0.003) compared with 587Gly/Gly carriers.Conclusion COX-2 Gly587Arg polymorphism is a risk factor for primary liver cancer in Han.

Objective:To investigate the effect of carvedilol on expression of cardiac matrix metalloproteinases(MMPs)and tissue inhibitors of metalloproteinases(TIMPs)after myocardial infarction in rats.Methods:An animal model of acute myocar- dial infarction(AMI)was established by descending left coronary artery ligation in 24 rats and they were divided into carvedilol (10 mg?kg~(-1)?d~(-1))group(n=12)and normal saline group(n=12).Sham operated group(n=9)received the same proce- dure but with no ligation.All animals were treated for 6 weeks via a gastric lavage.Heart function and hemodynamic parame- ters were determined after 6 weeks.The protein expression of cardiac MMP-2,MMP-9 and TIMP-2 was detected by immuno- histoehemical analysis in AMI groups,and the MMPs activities were assessed by zymography.Gene expression of myocardial MMPs/TIMPs(MMP-2,9 and TIMP-1,2)and cytokines(TNF-?,IL-1?)were measured by real-time quantitative PCR.Re- suits:Compared with Sham-operated group,earvedilol group had significantly higher left ventricular end-diastolic pressure(LV- EDP)and lower LV upstroke velocity(+dp/dt_(max))and LV descent velocity(-dp/dt_(max))(P

OBJECTIVETo identify the biomechanical feasibility of the thoracic extrapedicular approach to the placement of screws.

METHODSFive fresh adult cadaveric thoracic spine from T1 to T8 were harvested. The screw was inserted either by pedicular approach or extrapedicular approach. The result was observed and the pullout strength by pedicular screw approach and extrapedicular screw approach via sagittal axis of the vertebrale was measured and compared statistically.

RESULTSIn thoracic pedicular approach, the pullout strength of pedicle screw was 1001.23 N+/-220 N (288.2-1561.7 N)ls and that of thoracic extrapedicular screw approach was 827.01 N+/-260 N when screw was inserted into the vertebrae through transverse process, and 954.25 N+/-254 N when screw was inserted into the vertebrae through the lateral cortex of the pedicle. Compared with pedicular group, the pullout strength in extrapedicular group was decreased by 4.7% inserted through transverse process (P larger than 0.05) and by 17.3% inserted through the lateral cortex (P less than 0.05). The mean pullout strength by extrapedicular approach was decreased by 11.04% as compared with pedicular approach (P less than 0.05).

CONCLUSIONSIt is feasible biomechanically to use extrapedicular screw technique to insert pedicular screws in the thoracic spine when it is hard to insert by pedicular approach.

Adult ; Biomechanical Phenomena ; Bone Screws ; Female ; Humans ; Male ; Middle Aged ; Thoracic Vertebrae ; surgery

Antigens, CD ; metabolism ; Antigens, CD34 ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Arteriovenous Malformations ; complications ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; Magnetic Resonance Imaging ; Middle Aged ; Multiple Sclerosis ; Spinal Cord Diseases ; complications ; metabolism ; pathology

AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

Objective To discuss how to protect the intracranial vertebral artery and posterior inferior cerebellar artery by observing and measuring the intracranial vertebral artery in the surgery adopt far lateral approach. Methods Mimicking far lateral approach, 20 adult cadaveric heads connected to neck fixed with 10% formalin were dissected. Intracranial segment of the vertebral arteries and their main branches were exposed and measured under operating microscope. Results The intracranial vertebral artery joined with the contralateral one into the basilar artery after traveling through the atlanto-occipital sulcus. The relationship between the vertebral artery and the hypoglossal nerve is close. Thirty sides (75%) of the vertebral arteries traveled to pons medulla sulcus in front of the hypoglossal nerve roots and 2 sides (5%) behind the hypoglossal nerve roots, while 8 sides (20%) traveled among the hypoglossal nerve roots; 70% of the vertebral arteries were contacted to the hypoglossal nerve roots, 30% of which compressed the hypoglossal nerve. The main branches of intracranial segment of the vertebral arteries were the posterior inferior cerebellar arteries, the anterior spinal arteries, the posterior meningeal arteries,and some perforating arteries. Posterior inferior cerebellar arteries all originated from the intracranial vertebral artery were the largest vertebral artery's branches; their trip was mostly loop-shaped and they had close relationship with Ⅸ, Ⅹ, Ⅺ cranial nerves. The starting points of the posterior inferior cerebellar arteries were different, even in the same specimen, but most of them originated from the upper 1/3intracranial vertebral artery. No anterior inferior cerebellar artery was noted originated from the vertebral artery in our specimen. Anterior spinal arteries originated from the vertebral arteries joined with the branches of the bilateral vertebral arteries and traveled down through the tortuous anterior median fissure to supply the spinal cord. Conclusion Being familiar with the characteristics and anatomic vertebral arteries variations of the intracranial vertebral artery and its branches can contribute to identify and protect the intracranial segment of the vertebral artery and its main branches in the surgery adopt far-lateral approach.

Serum used widely in mammalian cell culture is also a potential source of bacterial, mycoplasmal and viral contaminations. In addition, the complex biological components in serum make harder the subsequent product recovery process. High cost, high batch variation and potential source limitation are among the other shortcomings. So serum-free or even protein-free medium are preferable for recombinant protein production. However, without serum to provide essential components such as hormones, growth factors and binding proteins, cells are easy to die. In this study, CHO-dhfr- cells were genetically engineered to make them adapted to IMEM, a protein-free medium, and resistant to apoptosis. The genes in choice are insulin-like factor (Igf-1), Bcl-2 and cyclin E. Bcl-2 is a mitochondrial membrane-integrated protein. It can block the release of cytochrome c by maintaining the integrity of mitochondrial membrane, and thus inhibit apoptosis. Igf-1 is similar both in structure and function to insulin, a growth factor added to serum-free medium to promote cell growth and is the only protein component in many currently used serum-free media. cyclin E is a cell cycle protein expressed continuously in G1 phase. When cyclin E accumulates to certain amount, cell cycle was driven to S phase. So cyclin E is a proliferation-promoting protein. By co-express Igf-1/Bcl-2 or Bcl-2/ cyclin E in CHO-dhfr- cells with a dicistronic expression vector, we constructed two cell lines: CHO-IB and CHO-BC. The high expression of each protein was confirmed by Western blot and flow cytometry. Apoptosis was analyzed by flow cytometry and DNA ladder detection, and the two cell lines were both found much more resistant to apoptosis induced by withdrawal of serum or addition of actinomycin D than the CHO-dhfr- parent cell. Cell proliferation assay by MTT method showed that the two cell lines proliferated much faster than CHO-dhfr- in IMDM medium without serum. Continuously culture assay proved that the two cell lines grow very well in IMEM protein-free medium supplemented with fibronectin and vitronectin to ease adherence. When compared to CHO-dhfr-, the two cell lines exhibited much more viable cell numbers and faster growth rate.
Animals ; Apoptosis ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Culture Media ; Cyclin E ; genetics ; Genes, bcl-2 ; Insulin-Like Growth Factor I ; genetics

OBJECTIVETo label the primary articular chondrocytes overexpressing human insulin-like growth factor (hIGF 1) with green fluorescent protein (GFP) for repair of articular cartilage defects in rabbits.

METHODSGFP cDNA was inserted into pcDNA3.1 hIGF 1 to label the expression vector. The recombinant vector, pcGI, a mammalian expression vector with multiple cloning sites under two respective cytomegalovirus promoters/enhancers, was transfected into the primary articular chondrocytes with the help of lipofectamine. After the positive cell clones were selected by G418, G418-resistant chondrocytes were cultured in medium for 4 weeks. The stable expression of hIGF 1 in the articular chondrocytes was determined by in situ hybridization and immunocytochemical analysis and the GFP was confirmed under a fluorescence microscope. Methyl thiazolyl tetrazolium (MTT) and flow cytometer methods were employed to determine the effect of transfection on proliferation of chondrocytes. Gray value was used to analyze quantitatively the expression of type II collagen.

RESULTSThe expression of hIGF 1 and GFP was confirmed in transfected chondrocytes by in situ hybridization, immunocytochemical analysis and fluorescence microscope observation. Green articular chondrocytes overexpressing hIGF 1 could expand and maintain their chondrogenic phenotypes for more than 4 weeks. After the transfection of IGF 1, the proliferation of chondrocytes was enhanced and the chondrocytes could effectively maintain the expression of type II collagen.

CONCLUSIONSThe hIGF 1 eukaryotic expression vector containing GFP marker gene has been successfully constructed. GFP, which can be visualized in real time and in situ, is stably expressed in articular chondrocytes overexpressing hIGF 1. The labeled articular chondrocytes overexpressing hIGF 1 can be applied in cell-mediated gene therapy as well as for other biomedical purposes of transgenic chondrocytes.

Animals ; Cartilage, Articular ; metabolism ; Cells, Cultured ; Chondrocytes ; metabolism ; Flow Cytometry ; Genetic Markers ; Genetic Therapy ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Luminescent Agents ; RNA, Messenger ; analysis ; Rabbits