The Rhei Radix et Rhizoma was one of the most widely used traditional Chinese medicine for its special biological activities. The content of rhein, one of its major compounds, was an important standard for the quantity control of Rhei Radix et Rhizoma. The major method used for the detection of rhein was instrumental analysis like HPLC, but it was complex, time-consuming and cannot detect large samples at the same time. The enzyme-linked imunmosorbent assay (ELISA) was accurate, reliable, simple, low costs, and of a high-throughout. Recently, it was widely used for the determination of those small molecule compounds in some traditional Chinese medicinal plants. In this study, an artificial antigen were synthesized by the carbodiimide (CDI) method. Rhein-bovine (rhein-BSA) conju gate and rhein-ovalbumin (rhein-OVA) conjugate, were produced as the immunogen and coating antigen, respectively. The conjugate and the hapten number in the conjugate were determined by UV-Vis spectrophotometry (UV). The conjugation ratio of Rhein and BSA was about 4.0:1, rhein acid and OVA was 2.6 : 1, respectively. Rhein-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The antiserum titer of the Rhein were higher than 8000 detected by ELISA. The successfully synthesized conjugate antigen rhein-BSA implies its feasibility in the establishment of fast immunoassay for the rhein content determination.
Animals ; Anthraquinones ; analysis ; immunology ; Antibodies ; analysis ; immunology ; Antigens, Plant ; analysis ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Mice ; Mice, Inbred BALB C ; Rheum ; chemistry ; immunology ; Rhizome ; chemistry ; immunology

Objective To investigate the effect of apolipoprotein E gene knock-out(ApoE KO) and high-fat diet on morphology and the expression of mortalin in hippocampal CA_3 neurons of mice, and to explore the impact of these factors on memory and Alzheimer's disease.Methods Ten wild-type and 10 ApoE KO mice were fed with common chow as the control group and the KO group respectively while 10 ApoE KO mice were fed with high fat diet.Twelve weeks later, the weight and the lid of these mice were measured.The brain tissues were observed using HE staining, nissl staining, protargol staining,immunohistochemistry staining and image analysis by computer.Results In the ApoE KO group, weight,total cholesterol, triglyceride, low-density lipoprotein cholesterol were higher than those in the control group,and these changes were more significant in ApoE KO high-fat diet group.The nissl was higher in the ApoE KO group (0.301±0.031) and in ApoE KO high-fat diet group (0.261±0.020) than those in the control group (0.341±0.035, F=18.068, P<0.05).The mortalin in the ApoE KO group (0.322±0.060) and in ApoE KO high-fat diet group (0.391±0.041) were higher than the control group (0.256±0.061, F=15.230, P < 0.05).Conclusions ApoE KO and high-fat diet can reduce nissl, and improve the expression of mortalin.This protein may be involved in the pathogenesis of Alzheimer's disease.

Objective To evaluate the blood supply features and effectiveness of arterial chemoembolization for pedunculated hepatocellular carcinoma.Methods Angiography and chemoembolization via supplying blood arteries of tumor were performed in five patients with pedunculated hepatocellular carcinoma.Interventional procedure was carried out with tumor vascular infusion of 350 mg hot elemene emulsion and tumor embolization by cisplantin-lipidol emulsion(cisplantin 60-80 mg+lipidol 8-15 ml)and glutin.Results Ten interventional procedures(TACE)were undertaken in 5 patients.Angiography showed that tumor blood supply mainly coming from collateral circulation adjacent to the tumors,but partially from hepatic artery.Tumor sizes decreased from 30% to 50% in 5 cases,and AFP declined in 4 cases after the treatment. Conclusion Pedunculated hepatocellular carcinoma possessing different blood supply features from intrahepatocellular carcinomas.But transarterial ehemoembolization is still an effective method of choice for this treatment.

Whether or not an abormal expression of IL-6 mRNA in PBMNCs from IDDM patientswas examined using a hihgly sensitive,specific and semiquantitative protocal,i.e.reverse tran-scription and polymerase chain reaction (RT-PCR).The relative levels of IL-6mRNA in PBM-NCs from 12 early IDDMpatients (8.20?3.85yr),29 newly diagnosed NIDDM patients(54.85?9.12yr)23 normal childrens (8.20?3.26yr) and 12 normal adults (31.92?11.22yr)weredetermined.Significantly high expresion levels of IL-6 mRNA were found in PBMNCs from pa-tients with IDDM (P

To study the in vitro anti-angiogenesis effect of three curcumin pigments (curcumin, demethoxycurcumin, bisdemethoxycurcumin). In the study, the inhibitory effect of the three curcumin pigments on proliferation of HUVEC cells induced by OX-LDL and the effect on migration of HUVEC cells were detected. The effect on neovascularization was observed by chorioallantoic membrane (CAM) test. The effect on cell adhesion factors ICAM-1 and VCAM-1 of HUVECs were tested by Real-time RT-PCR. It was found that the three curcumins could inhibit the proliferation of HUVEC cells induced by OX-LDL within the dosage range 4, 8, 16 mg x L(-1), with a dose-dependence. The proliferative effect of curcumins on HUVECs was greater than the other two derivatives (P < 0.01). All of the three curcumin pigments inhibited the migration of HUVEC cells and the angiogenesis of chick chorioallantoic membrane (CAM). The migration inhibition rate of curcumins at middle and high concentrations was greater than the other two (P < 0.01). All of the three curcumin could down-regulate the expression of VEGF and ICAM-1, and curcumins showed more obvious effect in down-regulating VEGF than demethoxycurcumin and bisdemethoxycurcumin(P < 0.01); Bisdemethoxycurcumin showed the most significant effect in down-regulating ICAM-1 (P < 0.01). All of the three showed no remarkable effect on expression of VCAM-1, and only bisdemethoxycurcumin showed the down-regulating effect (P < 0.05). According to the findings, all of the three curcumin pigments could resist angiogenesis by inhibiting proliferation and migration of endothelial cells and down-regulating the expression of VEGF and adhesion molecules ICAM-1.
Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Movement ; drug effects ; Cells, Cultured ; Chorioallantoic Membrane ; drug effects ; Curcumin ; analogs & derivatives ; pharmacology ; Humans ; Intercellular Adhesion Molecule-1 ; genetics ; RNA, Messenger ; analysis ; Vascular Cell Adhesion Molecule-1 ; genetics ; Vascular Endothelial Growth Factor A ; genetics

Lonicerae Japonicae Flos was one of the most widely used traditional Chinese medicine for its special biological activities. The content of luteoloside, one of its major compounds, was an important standard for the quantity control of Lonicerae Japonicae Flos. The major method used for the detection of luteoloside was instrumental analysis. Compared with the ELISA method, instrumental analysis was time-consuming, complex pretreatment and low-throughout. Thus, it was significantly important to develop an enzyme-linked immunosorbent assay (ELISA) for luteoloside analysis. Here, the conjugates of luteoloside-bovine (LG-BSA) and luteoloside-ovalbumin (LG-OVA) were produced as the immunogen and coating antigen by the carbodiimide ( CDI) method, respectively. The conjugation ratio of carrier protein and the hapten in the conjugate were determined by UV-Vis spectrophotometry (UV). LG-BSA conjugate was used to immunize Bal b/c mice to produce antiserum. The titer and specificity of antiserum were detected by ELISA. The conjugation ratio of hapten and carries protein were 3. 7: 1 (LG-BSA) and 1. 0: 1 (LG-OVA). The antiserum titer was higher than 2 000 with the linear range of 18.4-4 852.4 μg x L(-1), R2 = 0.988 4 and IC50 = 298.7 μg x L(-1). The result showed that the conjugate antigen LG-BSA was synthesized successfully and the mice can produce specific antiserum injected with artificial antigen.
Animals ; Antibodies ; analysis ; immunology ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Immunization ; Lonicera ; chemistry ; immunology ; Mice ; Mice, Inbred BALB C

Objective: To assess the effect of balloon angiopl asty on circulating endothelin (ET) and TNF-α levels and tissue endothelin in experimental atherosclerosis in rabbits. Methods: After 20 New Z ealand rabbits had a high cholesterol diet for at least 8 weeks, successful ball oon angioplasty was performed in rihgt iliac arteries in 18 rabbits. Circulatin g levels of ET and TNF-α were measured before as well as immediately and 24 h after balloon angioplasty. Tissue endothelin immunoreactivity in atherosclerotic iliac artery wall after balloon angioplasty was assessed by immunohistochemica l technique. Results: Plasma levels of ET and TNF-α were signi ficantly increased immediately after ballon angioplasty (76.40±13.58)pg/ml vs (92.67±11.38) pg/ml and (31.35±6.23) U/ml vs (56.26±7.37) U/ml, resp ectively (P<0.05) .There was no change in plasma ET and TNF-α levels 24 h after balloon angioplasty (77.13±12.87) pg/ml vs (76.40±13.58) pg/ml and (33.41±6.79) U/ml vs (31.35±6.23) U/ml， respectively (P>0.05). T issue endothelin immunoreactiuvity was markedly increased in right iliac artery wall after balloon angioplasty than that in opposite iliac artery wall. Conclusion: The increase of plasma ET, TNF-α levels and tissue ET-IR in iliac artery wall after balloon angioplasty may be associated with the injury of l ocal vascular intima, suggesting that ET and TNF-α may take part in the corona ry constriction and the development of coronary restenosis after percutaneous tr ansluminal coronary angioplasty.

OBJECTIVETo investigate the dynamic changes of the epididymal size 1 year after vasectomy.

METHODSFifty male volunteers received vasoligation. Before and at 1, 2, 3, 6, and 12 months after operation, we measured the size and detected the internal echoes of the epididymis using color Doppler ultrasonography.

RESULTSThe bilateral epididymides were both thickened post-operatively in all the 50 cases, with statistically significant differences between the baseline and the 1st month, the 1st and the 2nd month, the 2nd and the 3rd month, or the 3rd and the 6th month after surgery (all P < 0.01), but not between the 6th and the 12th month (P > 0.05).

CONCLUSIONWithin 6 months after vasectomy, the bilateral epididymides manifested a progressive thickening, but basically restored their balance of secretion-absorption after 6 months.

Epididymis ; diagnostic imaging ; pathology ; physiology ; Humans ; Male ; Organ Size ; Postoperative Period ; Time Factors ; Ultrasonography, Doppler, Color ; Vasectomy

Omics docking study for polygenic inheritance tumors has become an important strategy in oncology research. This review focuses on the conceptions and technologies of omics, and puts forward the central contents and omics docking for polygenic inheritance tumor to reveal the role of molecular changes at different stages of polygenic inheritance tumor at multidisciplinary and multilayer level. It is a new strategy to explore the mechanism of tumor carcinogenesis, and to regulate the network, key molecules, and drug target by combined biology effects.
Carrier Proteins ; biosynthesis ; genetics ; Genomics ; methods ; Glycoproteins ; biosynthesis ; genetics ; Humans ; Membrane Proteins ; biosynthesis ; genetics ; Multifactorial Inheritance ; genetics ; Neoplasms ; genetics ; metabolism ; Phosphoproteins ; biosynthesis ; genetics ; Proteomics ; methods ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVEStudying the gene expression profiling of different stage hairy root of Salvia miltiorrhiza, in order to find functional genes.

METHODThe contents of second metabolites were determined by HPLC and gene expression profiling was detected by cDNA microarray. cDNA labeled with a fluorescent dye (Cy5 and Cy3-dCTP) was produced by Eberwine's linear RNA amplification method and subsequent enzymatic reaction. The microarrays were scanned with a ScanArray Express scanner using ScanArray 2.0 software and quantified by signal intensities of individual spots from the 16-bit TIFF images using GenePix Pro 4.0. The linear normalization method was used for data analyze. Northern blot was used to test the gene expression results obtained by microarray. Different expressed genes were sequenced and analyzed by gap4 software, and then they were analyzed with BLASTX, BLASTN, GO and KEGG.

RESULTGrowth rate and second metabolites analysis indicated that the stage from 30 d to 45 d was the growth stage, while the stage from 45 d to 60 d was the second metabolites accumulation stage. Accordingly 30 d hairy root was chosen as a reference, which was hybridized with 45 d and 60 d hairy root separately. Total 203 different expressed genes were obtained. Northern blot showed that the result was identical with the microarray result. After sequenced, there were 172 genes clustered into 114 clusters (Unigenes). Among them, 62 unigenes had known functions, 34 unigenes were hypothetical protein, 9 unigenes were homologues with no similarity and 9 unigenes were unidentified protein with low similarity. Total 67 genes were classified into cellular component ontology, molecular function ontology and biological process ontology based on GO analysis. Total 26 genes, which represented 29 metabolic-related enzymes, were located in metabolic maps based on KEGG pathway classification.

CONCLUSIONSeveral important functional genes related to second metabolite synthesis were cloned such as P450 and copalyl diphosphate synthase genes. cDNA microarray was a useful tool for functional genomics of traditional Chinese medicine.

Alkyl and Aryl Transferases ; genetics ; metabolism ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation, Developmental ; Gene Expression Regulation, Plant ; Genomics ; methods ; Oligonucleotide Array Sequence Analysis ; Plant Proteins ; genetics ; metabolism ; Plant Roots ; genetics ; growth & development ; metabolism ; Plants, Medicinal ; genetics ; growth & development ; metabolism ; Salvia miltiorrhiza ; genetics ; growth & development ; metabolism