Objective To evaluate the clinical value of CTPA combined with V/Q imaging to guide the end point of anticoagulant therapy in reducing the recurrence rate of pulmonary embolism. Methods A total of 159 cases of pulmonary embolism diagnosed by CTPA were randomly divided into experimental group(n=80)and control group(n = 79). After the regular low molecular weight heparin and warfarin anticoagulation therapy ,the experimental group used the CTPA combined with V/Q imaging to evaluate the pulmonary embolism absorption ,to guide the end point of anticoagulant therapy and to evaluate the recurrence rate of pulmonary embolism at the end of 1-year treatment. But in control group ,only CTPA was used to guide the treatment and then the recurrence rate in 2 groups was compared. Results The anticoagulant treatment course of experimental group was(5.90 ± 1.80) months,which was significantly longer than that of control group(3.57 ± 1.09)months(P<0.05). The recurrence rate of experimental group was significantly lower than that of control group(7.5%vs. 22.8%)after 1-year treatment (P<0.05). However,there was no significant difference between 2 groups(8.75%vs 3.80%)in the rate of bleed-ing during anticoagulation therapy (p>0.05).Conclusions CTPA combined with V/Q imaging to guide the end point of anticoagulant therapy for pulmonary embolismhas important clinical value in reducing the recurrence rate of pulmonary embolism.

OBJECTIVE:To establish a method for the contents determination of 9 components in Guizhi decoction,and com-pare the effects of traditional decoction method and the extracting machine decoction method on these contents in Guizhi decoction. METHODS:HPLC was performed on the column of Agilent Zorbax SB-C18 with mobile phaseof acetonitrile- 0.1% phosphoric ac-id(gradient elution)at a flow rate of 1.0 ml/min,the detection wavelength was 230 nm,254 nm and 280 nm,the column tempera-ture was 25℃,and the injection volume was 10μl. RESULTS:The linear range was 0.410 2-210.0μg/ml for gallic acid(r=0.999 9), 0.994 0-254.5μg/ml for albiflorin(r=0.999 9),1.636 0-1 675.0μg/ml for paeoniflorin(r=0.999 9),0.988 3-506.0μg/ml for liquiri-tin(r=0.999 6),0.987 3-31.59 μg/ml for coumarin(r=0.999 5),0.486 8-124.6 μg/ml for cinnamic acid(r=0.999 5),2.458 0-314.6μg/ml for cinnamaldehyde(r=0.999 5),0.034 3-1.096 μg/ml for 2-methoxy cinnamaldehyde(r=0.999 8),and 1.711 0-219.0 μg/ml for glycyrrdhizic acid (r=0.999 7);RSDs of precision,stability and reproducibility tests were lower than 5%,recoveries were 93.56%-103.19%(RSD=4.00%,n=9)、101.51%-107.32%(RSD=2.21%,n=9)、95.08%-103.76%(RSD=2.87%,n=9)、100.82%-105.73%(RSD=1.85%,n=9)、85.08%-89.12%(RSD=1.40%,n=9)、92.31%-99.12%(RSD=2.71%,n=9)、99.17%-102.32%(RSD=1.24%,n=9)、100.15%-103.98%(RSD=1.18%,n=9)、99.93%-102.61%(RSD=1.03%,n=9). The content of total effective components from the extracting machine decoction method was 4 565μg/g,that from the traditional decoc-tion method was 2 742 μg/g.CONCLUSIONS:The method is simple,stable and reproducible,and can be used for the simultaneous determination of 9 componentsin Guizhi decoction. The contents of gallic acid,albiflorin and 2-methoxy cinnamaldehyde are first re-ported. The total effective components from the extracting machine decoction method are higher than that from the traditional decoc-tion method.

Aim To study the interference role of Bcl-2 siRNA on HL-60 cells. Methods Bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit. The synthesized siRNA was transfected into HL-60 cells with lipid siPORT transfection. Forty-eight hours after the transfection, we used MTT and immunofluorescence to detect cell proliferation and apoptosis,and used RT-PCR and immunofluorescence to detect the level of Bcl-2 mRNA and Bcl-2 protein expression. Result Bcl-2 siRNA reduced the level of Bcl-2 mRNA and Bcl-2 protein expression in HL-60 cells and induced cell apoptosis. There was no difference on the effect of other groups compared with the control. Effective Bcl-2 siRNA specifically degraded Bcl-2 expression in the levels of mRNA and protein and induced HL-60 cells apoptosis.Conclusion These results indicate that siRNA is a highly specific tool for targeted gene knockdown. siRNA-mediated gene silencing is a reliable approach for large-scale screening of gene function and drug target validation.

Objective To investigate the influence of transcriptional factor Sp1 on expression of bile acids transporters MRP3 and MRP4 so as to perfect the regulatory mechanism of MRP3 and MRP4 expression.Methods Transformed Sp1-overexpression and Sp1 siRNA plas-mids to HepG2 cell and obtained the stably cells line.Then the expression levels of bile acids transporters MRP3 and MRP4 were measured by RT-qPCR,and the change of protein levels were detected by Western blot.Results The stably cells line Sp1-OE-HepG2 and Sp1siRNA-HepG2 were successfully transformed.The mRNA expression and protein levels of MRP3 and MRP4 were significantly increased in Sp1-OE-HepG2 cells,among which the mRNA expression of MRP3 mRNA increased 2.8 times,the protein levels of MRP3 increased 3.0 times,and the mRNA expression and protein levels of MRP4 increased 3.2 times and 2.5 times respectively.Conversely,the mRNA expression and protein levels of MRP3 and MRP4 were decreased in Sp1 siRNA-HepG2 cells,among which the mRNA expression of MRP3 mRNA de-creased 52%,the mRNA expression of MRP4 mRNA decreased 58%,the protein levels of MRP3 decreased 57%,and the protein levels of MRP4 decreased 60%.Conclusion Transcriptional factor Sp1 could regulate the expression of bile acids transporters MRP3 and MRP4 in HepG2 cells.

Cinnamaldehyde was shown to have significant anti-inflammatory and anti-pyretic actions in studies from both others' and our lab. Prostaglandin E2 (PGE2) plays a key role in generation of these pathological states, while PGE, synthase-1 (mPGES-1) is one of crucial biological elements in the process of PGE2 production. And as a downstream inducible terminal prostaglandin synthase of COX-2, mPGES-1 is now regarded as a more promising novel drug target than COX-2 and is attracting more and more attention from both academia and pharmaceutical industry. The purpose of present study was to further investigate the anti-inflammatory and antipyretic molecular mechanisms of cinnamaldehyde based on the mouse macrophage cell line RAW264. 7 in vitro. The PGE2 was identified by using the method of enzyme-linked immunosorbent assay (ELISA) and the expression of COX-2 and mPGES-1 at mRNA and protein levels was detected by the Real-time PCR and Western blotting methods respectively. The experimental results suggested that cinnamaldehyde could evidently reverse the increased production of PGE2induced by IL-1beta. Moreover, the up-regulated expression levels of mPGES-1 and COX-2 were significatly decreased. Together, these results provide compelling evidence that the down-regulated actions to both the production of PGE2 as well as the expression of mPGES-I might account for, at least in part, the anti-inflammatory and anti-pyretic effects of cinnamaldehyde.
Acrolein ; analogs & derivatives ; pharmacology ; Animals ; Blotting, Western ; Cell Line ; Dinoprostone ; metabolism ; Interleukin-1beta ; pharmacology ; Intramolecular Oxidoreductases ; metabolism ; Macrophages ; drug effects ; metabolism ; Mice ; Prostaglandin-E Synthases ; Real-Time Polymerase Chain Reaction

This study was aimed to investigate whether the extracts of Celastrus orbiculatus enhanced the invasion function of maspin tumor inhibitor gene through the construction of maspin overexpression human gastric carcinoma MGC803 cell line. Maspin was cloned into plasmid GV208-EGFP eukaryotic expression vector. And then, the recombinant plasmid GV208-maspin-EGFP was transfected into human gastric carcinoma MGC803 cells. After the maspin overexpression MGC803 cell were treated with Celastrus orbiculatus extracts in different concentrations (10, 20, 40 μg·mL-1), the invasion effects were detected by Transwell chamber assay. The results showed that after the successful construction of maspin overexpression cell line, the number of cells invading through Matrigel was obviously decreased in the Transwell chamber assay. It also showed drug concentration dependency. It was concluded that maspin gene can inhibit invasion of gastric carcinoma MGC803 cells. Simultaneously, the extracts of Celastrus orbiculatus can enhance the function of maspin gene.

This study was aimed to investigate the effect of Nan-She-Teng (Celastrus orbiculatus) extract on epithe-lial-mesenchymal transition(EMT) in HepG2 cells. Except the control group, human hepatocellular carcinoma HepG2 cells in other groups were treated with Celastrus Orbiculatus extract in different concentrations (10, 20, 40, 80, and 160 μg·mL-1). The protein expression levels related to EMT were detected by western blotting. At 48 h after fertiliza-tion, the zebrafish embryos were randomly assigned to 7 groups as follows: untreated control group (E3 buffer), DMSO group (E3 buffer with 1% DMSO), and different dosages treatment of C.orbiculatus extract (10, 20, 40, 80, and 160μg·mL-1) for 24 h. The protein expressions of mTOR signaling pathways were detected by western blotting. The re-sults showed that compared with the control group, C.orbiculatus extract significantly increased E-cadherin protein expression. Simultaneously, C.orbiculatus extract inhibited vimentin and mTOR signaling pathways at protein levels. It was concluded that to a certain extent, C.orbiculatus extract prevented EMT in HepG2 cells by modulating the mTOR signaling pathway. Therefore, it suggested that mTOR can be chosen as a new therapeutic target for clinical treatment of hepatic carcinoma.

OBJECTIVETo study the effects of the ingredients from Chinese herbs with the nature of cold or hot on the expression of TRPV1 and TRPM8.

METHODThe effects of ingredients from herbs on primary culture DRG neurons are observed in vitro. The expression quantity of gene is detected by the method of real time PCR. the 2 (-deltadeltaCT) method is applied to analyze the data.

RESULTIngredients from herbs with the nature of cold up-regulate the expression level of TRPV1 and down-regulate that of TRPM8, especially under the temperature condition of 39 degrees C; while ingredients from herbs with the nature of hot up-regulate the expression level of TRPM8 and down-regulated that of TRPV1, which is more significant under the temperature condition of 19 degrees C.

CONCLUSIONThe regulatory changes of TRPV1 and TRPM8 mRNA expression induced by the chemical ingredients might be related to the cold and hot natures of the herbs from which the ingredients are extracted. And this could be one of the therapeutic mechanisms for the treatment of Chinese herbal medicines to cold- and heat-related diseases.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; Gene Expression ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley ; TRPM Cation Channels ; genetics ; metabolism ; TRPV Cation Channels ; genetics ; metabolism

To analyze the clinical data of 2 neonates with Langerhans cell histiocytosis (LCH). The rashes appeared in both cases shortly after birth.Case 1 had both rashes and neonatal sepsis, and no other tissues and organs were involved.After anti-infective treatment, the rashes gradually disappeared.Case 2 had secondary pneumonia, abnormal coagulation function and gastrointestinal bleeding.Both cases were positive for CD1a and S-100 by immunohistochemical staining of skin biopsy, and they were diagnosed as multi system-LCH.The early diagnosis of LCH is particularly important.The detection methods of skin or lymph node biopsy like immunohistochemistry, need to be performed as early as possible.Because the course of the disease is not clear, a close monitoring and follow-up are needed.