OBJECTIVE Apolyclonalantibody-basedhomogeneouschemiluminescenceimmunoas-say was developed and optimized using AlphaLISA technology for the quantitative detection of microcys-tin-LR(MC-LR)inwatersamples.METHODS Thismethodwasbasedonacompetitivemodelin which an immune complex was formed from the ingegral binding of artificial MC-LR antigen-coated lumi-nescene beads,free MC-LR standards or sa mples,antibody and biotinylated second antibody.Next sensor bead were added that approached the i mmune co mplex through biotin-streptavidin interaction. With the exciting light,the energy was passed from the sensor luminescer before a special emission light could be observed.To opti mize the reaction conditions,working dilutions of polyclonal antibody and bioti-nylated second antibody were assayed while the effect of buffer syste ms and ti me of each reaction were evaluated.RESULTS Maininfluencingfactorsoftheassaywerediscussedasworkingdilutionsofpoly-clonal antibody and biotinylated goat anti rabbit IgG,assay buffer and reacting ti me.After opti mization of reaction conditions,MC-LR AlphaLISA could be finished in 40 min,with a sensitivity of 0.006 μg·L-1 and a dynamic range of 0.006 -5 μg·L-1 .The coefficient of variation was below 10% and average recovery was 1 07.7%.Moreover,the cross reactivity rates of MC-RR and MC-RY to MC-LR were 13.2%and0.91%,respectively.CONCLUSION Thismethodishighlysensitiveandspecific,time-saving and quite suitable for high throughput determination of MC-LR water samples.

BACKGROUND:Extradural cortical stimulation combines the advantages of repetitive transcranial magnetic stimulation, transcranial direct current stimulation, subdural cortical stimulation and deep brain stimulation, which can significantly improve motor and language function after stroke. OBJECTIVE:To review the theoretical research and clinical application of extradural cortical stimulation for stroke recovery. METHODS:An online retrieval of PubMed database and CNKI database between January 1995 and April 2014 was performed for articles on theoretical research and clinical application of extradural cortical stimulation for stroke recovery, with the key words of“cortical stimulation, extradural motor cortex stimulation, extradural cortical implants, extradural cortical stimulation, stroke, rehabilitation”in English and Chinese. RESULTS AND CONCLUSION:Because of implantable cortical stimulation, the advantage of extradural cortical stimulation is its minimal invasiveness, high accuracy and transdural contact with the brain. For lack of effective treatment for the chronic phase of stroke patients with motor and language dysfunction, extradural cortical stimulation may be a new therapeutic method. Motor and language functional improvement must derive from reactivation of plasticity, local enhancement of perilesional areas, enhancement of network function and inter-hemispheric balance function, and amplification of sensory input.

Objective:To clone and sequence cDNA of MH2 domain of Smad1 gene from human dental papilla cells. Methods:Total RNA was isolated from primarily cultured human dental papilla cells and reversely transcribed into single stranded cDNA.The desired DNA product was obtained by PCR with two gene specific primers.The segment was inserted into PUC19 vector and the plasmid was transformed into E.coli JM109.The double stranded cDNA of positive clone was sequenced.Results:The sequence of MHz domain of Smad1 cloned from human dental papilla cells was consistent with that reported by Hoodless et al. Conclusion:Smad1 exists in human dental papilla cells,BMP signaling may be mediated by smad1 in human dental papilla cells.

Objective To investigate the expression of CD90,IGF1R in hepatocellular carcinoma.Methods CD90,IGF1R expression were detected by SP immunohistochemical staining in 36 cases of hepatocellular carcinoma,20 cases of normal liver tissue biopsied from patients of chronic cholecystitis undergoing cholecystectomy. Results The positive rate of CD90 (63.89％,23/36),IGF1R (52.78％,19/36) in hepatocellular carcinoma significantly increased (P ＜ 0.05 ); The positive rate of CD90 was higher in UICC Ⅲ - Ⅳ stage group (79.17％ ) than in UICC stage Ⅰ - Ⅱ group (33.33％,P ＜0.05) ;The positive rate of CD90 was higher in low differentiated group (76.92％ ) than in well-differentiated group (56.52％,P ＜0.05).The positive rate of IGF1R was higher in UICC Ⅲ - Ⅳ stage group (70.83％ ) than in UICC stage Ⅰ - Ⅱ group ( 16.67％,P ＜ 0.05 ). The positive rate of IGF1R was higher in low differentiated group (84.62％) than in well-differentiated group (37.38％,P ＜ 0.05 ).The expression of IGF1R was positively correlated with that of CD90 (P ＜ 0.05 ).The median survival of CD90+ patients (85 days) was shorter than CD90- patients ( 505 days ) ( P ＜ 0.05 ).The median survival of IGF1 R + patients (100 days) was shorter than IGF1 R- patients (408 days,P ＜ 0.05). Conclusions CD90 or IGF1R expression correlates positively with the progression of HCC.

This study was aimed to compare the antioxidant activity of puerarin and 3 other flavonoid compounds, and to investigate their structure-activity relationship. The intragastric administration of 4 kinds of typical flavonoids compounds (soybean element, puerarin, quercetin and rutin) were given to mice, respectively. The model mice of acute hepatic injury were established with intraperitoneal injection of 0.1% carbon tetrachloride (CCl4) after 7 days. After 18 h fasting, liver tissues were removed. The histomorphology was observed after paraffin sectioning and hematoxylin-eosin staining. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transpeptidase (GGT) and alkaline phosphatase (ALP) in serum were detected with automatic biochemical analyzer. The content of malondialdehyde (MDA), the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in liver tissues were detected with homogenization. The pathological results of liver tissues showed that hepatic damages were decreased in all 4 medicine treatment groups compared with the model group, but there were no significant differences among these treatment groups. The results of blood serum bio-chemical analysis showed that compared with the model group, puerarin and quercetin could decrease the activities of ALT, AST and GGT in serum significantly (P < 0.05, orP < 0.01). There were no content changes of ALP. In the soybean element group, only the activities of ALT and AST decreased obviously (P < 0.05, orP < 0.01). There was no obvious change in the serum of mice in the rutin treatment group. The homogenate detection results of liver tissues showed that compared with the model group, quercetin and rutin significantly lowered MDA (P < 0.05), increased SOD and GSH-Px (P < 0.05, orP < 0.01); while soybean element and puerarin only improved GSH-Px levels (P < 0.05, orP < 0.01). It was concluded that the antioxidant capacity of quercetin was better than that of soybean element, puerarin and rutin, which may be related to its structure. Compared with 3 other chemical compounds, quercetin had more polyhydroxies and its polyhydroxies were not glycosylated, which suggested that the structure of quercetin may be closely related to its antioxidation activity.

ObjectiveTo investigate the targeting infection of single chain antibody againstAFP (scFv anti-AFP) directed lentivirus and the inhibitory effects of a dual-growth inhibition systemon hepatocarcinoma cells.MethodsPlasmids WtP53-pPRIME-miR30-shRNA-IGF1R,pMD2G-Anti-AFP,and psPAX2 have previously been constructed to cotransfect to the packaging cell line 293Tusing Lipofectamine2000.The infection results were observed through fluorescence microscopy.PCRand Western blotting were used to demonstrate the successful transduction and transcription of theWtP53-pPRIME-miR30-shRNA-IGF1R gene.The effects of reconstructed lentivirus infected liver cellgrowth were assessed by the cell growth curve of CCK8 cells. Apoptosis was evaluated by theTUNEL assay.ResultsRecombined lentivirus was successfully constructed with the functional PFUtiters of recombined lentivirus at 4.58× 109PFU/ml.This positive result was confirmed by PCR andWestern blotting.ConclusionsThe targeted therapy mediated by anti-AFP scFv could significantlyinhibit the proliferation of HEP3B cells and promote the apoptosis.

10.3969/j.issn.1000-8179.2013.12.002

Exploring the influence of extract of Ginkgo biloba (EGB) on the proliferation, apoptosis of ACC-2 cell in lacrimal adenoid cystic carcinoma and analyzing the influence of EGB on the gene expression of Survivin and TIP30 based on the levels of the gene and protein. ACC-2 cell in human with ACC of lacrimal gland disposed by EGB of different concentration was in vitro cultured. MTT method was used for cell proliferation detection. Annexin V/PI double-staining flow cytometer was used to detect cell apoptosis and cell cycle. Survivin and TIP30 gene expression together with protein expression were analyzed by RT-PCR and Western blotting. And it is indicated that EGB has inhibitory effect on the proliferation of ACC-2 cell in vitro. Furthermore, the dose-effect relationship was significant. Compared with the control group, it had statistical difference (P <0.01). The inhibitory concentration 50% (ICso) is 88 mg . L-1. By flow cytometer examination, it was indicated that EGB can gradually increase ACC-2 cell in G0-G1 stage and decrease it in G2-M and S stage. With the increase of dose, the apoptosis rate of ACC-2 cell obviously increased (P <0.05 or P <0.01). Both of the expression results of RT-PCR and Western hybrid proteins have showed that the concentration of EGB increased, it could be seen a significant decrease in Survivin gene expression (P <0.01). Meanwhile, the TIP30 gene expression got a significant increase. Therefore, EGB can effectively inhibit ACC-2 cell Survivin gene expression in human with adenoid cysistic carcinoma of larcrimal gland as well as promoting TIP30 gene expression, inducing the ACC-2 cell apoptosis and inhibiting tumor cell proliferation, which provided a certain theoretical and experimental basis for the application of Chinese herbal medicinal ingredient in the treatment of tumors.
Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Carcinoma, Adenoid Cystic ; drug therapy ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; Gene Expression ; Ginkgo biloba ; chemistry ; Humans ; Lacrimal Apparatus ; drug effects ; Plant Extracts ; chemistry ; isolation & purification ; pharmacology

Acupuncture Therapy ; methods ; Adult ; Aged ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Tennis Elbow ; therapy

With references of historical materials and modern literature regarding acupoint characteristic, a secondary analysis on the concept, origin, related factors and research methods in present research of acupoint characteristic is performed. The acupoint characteristic should be considered as an acupoint inherent attribute that could explain physiological and pathological manifestations at the same time, including location attribute and function attribute, which is related with time and treatment method. Some re-thinking on acupoint characteristic is proposed as well as advice on further research method and direction, hoping to promote the research development of acupoint characteristic.
Acupuncture Points ; Acupuncture Therapy ; history ; China ; History, Ancient ; Humans ; Medicine in Literature ; Meridians