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1.
China Journal of Chinese Materia Medica ; (24): 3722-3729, 2023.
Article in Chinese | WPRIM | ID: wpr-981504

ABSTRACT

This paper aims to compare the difference of growth and quality between wild and cultivated Artemisia stolonifera, thereby providing references for further development and utilization of A. stolonifera. The wild and cultivated A. stolonifera from different altitudes were collected, and the agronomic characters, moxa yield, volatile components, flavonoids, and phenolic acids were determined. The results showed that the cultivated species were taller and stronger, with more leaves and branches, than the wild species. The moxa yield and combustion quality of wild products were higher than those of cultivated products. The content of main volatile components in cultivated products was higher than that in wild products. The content of flavonoids and phenolic acids in wild products was higher than that in cultivated products. At high altitude, the ignition performance, combustion persistence, comprehensive combustion performance, and heat release during combustion of the wild and cultivated A. stolonifera. were optimal. At middle altitude, the content of main characteristic volatile components and flavone phenolic acids in the leaves of the cultivated and wild A. stolonifera were the highest. At low altitude, the combustion quality and the content of the above components of the cultivated A. stolonifera decrease significantly. Considering the combustion quality and the content of the internal components of the leaf lint, the middle and high altitude areas are suitable for the artificial cultivation of A. stolonifera.


Subject(s)
Artemisia , Agriculture , Flavonoids , Plant Leaves , Drugs, Chinese Herbal
2.
China Journal of Chinese Materia Medica ; (24): 3715-3721, 2023.
Article in Chinese | WPRIM | ID: wpr-981503

ABSTRACT

The purpose of this study was to analyze the effects of shading intensity on the growth, yield, and quality of Artemisia stolonifera so as to provide references for the artificial cultivation of A. stolonifera. The seedlings of A. stolonifera with consistent growth underwent shading treatment at four shading intensity levels(0, 55%, 85%, and 95%) with different layers of black shading nets. The agronomic indexes, yield, moxa yield, total ash, quality characteristics of moxa during combustion and pyrolysis, main volatile components, flavonoids, and phenolic acids were measured. The results showed that under shading conditions, the stem diameter, leaf width, 5-leaf spacing, branch number, and yield of A. stolonifera decreased significantly, while the plant height, leaf length, leaf number, chlorophyll content, and moxa yield increased first and then decreased with the increase in shading intensity. The burning performance of moxa under natural light was better than that under moderate and severe shading conditions. The content of eucalyptol first increased and then decreased with the increase in shading intensity. The humulene content was negatively correlated with shading intensity. Other major volatile components showed no significant difference under various shading conditions. The content of neochlorogenic acid, cryptochlorogenic acid, isoschaftoside, and isochlorogenic acid B was positively correlated with shading intensity, while the content of chlorogenic acid, isochlorogenic acid A, and isochlorogenic acid C decreased first and then increased with the increase in shading intensity. To sum up, A. stolonifera is a light-loving plant, and shading can greatly reduce the yield, the content of internal components, and the burning performance of moxa. It is the main reason why A. stolonifera is mainly distributed in the forest edge, open forest, roadside, and wasteland grass in the middle and high mountains in the wild. For artificial domestication and cultivation of A. stolonifera, it is better to select plots with sufficient light.

3.
China Journal of Chinese Materia Medica ; (24): 3693-3700, 2023.
Article in Chinese | WPRIM | ID: wpr-981501

ABSTRACT

The quality of moxa is an important factor affecting moxibustion therapy, and traditionally, 3-year moxa is considered optimal, although scientific data are lacking. This study focused on 1-year and 3-year moxa from Artemisia stolonifera and A. argyi(leaf-to-moxa ratio of 10∶1) as research objects. Scanning electron microscopy(SEM), Van Soest method, and simultaneous thermal analysis were used to investigate the differences in the combustion heat quality of 1-year and 3-year moxa and their influencing factors. The results showed that the combustion of A. stolonifera moxa exhibited a balanced heat release pattern. The 3-year moxa released a concentrated heat of 9 998.84 mJ·mg~(-1)(accounting for 54% of the total heat release) in the temperature range of 140-302 ℃, with a heat production efficiency of 122 mW·mg~(-1). It further released 7 512.51 mJ·mg~(-1)(accounting for 41% of the total heat release) in the temperature range of 302-519 ℃. The combustion of A. argyi moxa showed a rapid heat release pattern. The 3-year moxa released a heat of 16 695.28 mJ·mg~(-1)(accounting for 70% of the total heat release) in the temperature range of 140-311 ℃, with an instantaneous power output of 218 mW·mg~(-1). It further released 5 996.95 mJ·mg~(-1)(accounting for 25% of the total heat release) in the temperature range of 311-483 ℃. Combustion parameters such as-R_p,-R_v, D_i, C, and D_b indicated that the combustion heat quality of 3-year moxa was superior to that of 1-year moxa. It exhibited greater combustion heat, heat production efficiency, flammability, mild and sustained burning, and higher instantaneous combustion efficiency. This study utilized scientific data to demonstrate that A. stolonifera could be used as excellent moxa, and the quality of 3-year moxa surpassed that of 1-year moxa. The research results provide a scientific basis for the in-depth development of A. stolonifera moxa and the improvement of moxa quality standards.


Subject(s)
Artemisia , Hot Temperature , Moxibustion , Plant Leaves
4.
Chinese Journal of Experimental Traditional Medical Formulae ; (24): 187-194, 2019.
Article in Chinese | WPRIM | ID: wpr-802253

ABSTRACT

Objective: To study the origin of Nanyang Chrysanthemi Flos materia medica and its historical origin, in order to evaluate the quality of Nanyang Chrysanthemi Flos by HPLC method, and define the advantages of Nanyang Chrysanthemi Flos with the origin of Nanyang. Method: Records of Nanyang Chrysanthemi Flos in the "Chinese Medical Code" and related ancient documents were studied to explain the origin and application. The genetic relationship between Nanyang Chrysanthemi Flos and other pieces was revealed in ancient literatures. Then 8 chemical constituents in 14 batches of Chrysanthemi Flos were determined by HPLC multi-components quantitation,and the comprehensive weighted score analysis was performed based on the results. The HPLC fingerprints were established,and the similarity analysis and clustering analysis were made to comprehensive evaluation the quality of Nanyang Chrysanthemi Flos and define the genetic relationship between Nanyang and other pieces at the chemical composition level. Result: The results of the herbal textual research show that Nanyang Chrysanthemi Flos spread from place to place since the Han dynasty,and impact many medicinal chrysanthemums of later generations. HPLC fingerprints similarity and cluster analysis also indicated the genetic relationship between Nanyang Chrysanthemi Flos and other species at the chemical level. The comprehensive score analysis results show that Nanyang Chrysanthemi Flos is of good quality and very suitable for medicinal purposes. Conclusion: Nanyang has been a quality production area of Chrysanthemi Flos since ancient times to present. Nanyang Chrysanthemi Flos is very suitable for medicinal purposes and worthy of promotion.

5.
China Journal of Chinese Materia Medica ; (24): 3504-3508, 2017.
Article in Chinese | WPRIM | ID: wpr-335827

ABSTRACT

In order to evaluate the quality of Artemisia argyi from Qichun, Ningbo, Anguo and Nanyang, the contents of eupatilin and jaceosidin were determined by RP-HPLC. The determination was performed on Agilent Eclipse XDB-C₁₈ (4.6 mm×250 mm, 5 μm) with mobile phase consisted of acetonitrile-0.2% phosphoric acid(35∶65) at the flow rate 1.0 mL•min ⁻¹. The detection wavelength was 350 nm and the column temperature was 25 ℃. The results showed that the amount of eupatilin and jaceosidin had a clear linear relationship in the range of 0.003-0.126 g•L ⁻¹ (r=0.999 9) and 0.005-0.200 g•L ⁻¹ (r=0.999 9), and the average recovery rates for them were 99.14% (n=6, RSD 1.2%) and 99.40% (n=6, RSD=0.73%), respectively. The results showed that RP-HPLC can be used for the quantification of eupatilin and jaceosidin in the folium of A. argyi. With this method, we found there was no significant difference of jaceosidin content within all the samples collected, but the content of eupatilin was significantly higher in samples from Qichun, Ningbo, Xiangyang and Nanyang, located in the south of Huaihe River compared with these from other areas.

6.
China Journal of Chinese Materia Medica ; (24): 793-798, 2015.
Article in Chinese | WPRIM | ID: wpr-330231

ABSTRACT

To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.


Subject(s)
Alleles , DNA Primers , Genetics , DNA, Plant , Genetics , Medicine, Mongolian Traditional , Molecular Sequence Data , Plants, Medicinal , Classification , Genetics , Polymerase Chain Reaction , Methods
7.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 1054-1059, 2015.
Article in Chinese | WPRIM | ID: wpr-237901

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of Wenyang Huazhuo Tongluo Recipe (WYHZTLR) containing serum on transforming growth factor β1 (TGF-β1)/Smad signaling pathway of skin fibroblasts in systemic sclerosis (SSc).</p><p><b>METHODS</b>Totally 36 SSc patients were randomly assigned to Chinese medicine (CM) group, Western medicine (WM) group, and integrative medicine (IM) group according to random digit table, 12 in each group. Patients in the CM group took WYHZTLR decoction (one dose per day). Patients in the WM group took penicillamine tablet (0. 125 g each time, bid) and Prednisone Acetate Tablet (PAT 20 mg, qd). Patients in the IM group took penicillamine, PAT, and WYHZTLR decoction (in the same dosage of corresponding drugs as aforesaid). All patients were treated for one month to get drug containing serum. Besides, 10 untreated SSc patients' serum was taken as the control group. Healthy subjects' skin fibroblasts were originated from healthy skin tissue of the upper arms of 2 female patients undergoing plastic surgery. Corresponding serum of each group was added in the culture system of SSc patients' and healthy subjects' dermal fibroblasts respectively. Expression levels of TGF-β1 receptor type I (TGF-β1 RI), TGF-β1 receptor II (TGF-β1 RII), p-Smad2/3 and Smad7 protein were examined by Western blot. Expression levels of collagen type I and collagen type III (Col-I, Col-III) mRNA were examined by reverse transcription PCR. Contents of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in the supernatant of SSc, skin fibroblasts were examined by ELISA.</p><p><b>RESULTS</b>Compared with the control group, expression levels of TGF-β1 R I and p-Smad2/3 protein significantly decreased, but expression levels of Smad7 protein significantly increased in skin fibroblasts of SSc patients and healthy subjects of WM, CM, and IM groups (P <0.05, P <0. 01). Meanwhile, the expression level of TGF-β1 RII decreased in the IM group (P <0. 05, P <0. 01). Compared with the WM group, expression levels of TGF-β1 RI and p-Smad2/ 3 protein significantly decreased, but that of Srnad7 protein significantly increased in IM groups (P <0. 01). mRNA expression levels of Col-I and Col-II in SSc skin fibroblasts significantly decreased more in WM, CM, and IM groups than in the control group (P <0. 05, P <0. 01). Besides, the expression level of Col-III mRNA was significantly lower in the IM group than in the WM group (P <0.01). Compared with the control group, serum levels of MMP-9 and MMP-9/TIMP-1 ratios increased more obviously in WM, CM, and IM groups (P <0.05, P <0.01). But expression levels of TIMP-1 decreased significantly in CM and IM groups (P <0.01). Expression levels of MMP-9 and MMP-9/TIMP-1 ratios increased more in the IM group than in the WM group (P <0. 01). Expression levels of TIMP-1 decreased more in CM and IM groups than in the WM group (P <0.01).</p><p><b>CONCLUSION</b>WYHZTLR containing serum could reduce expression levels of Col-I and Col-III possibly through regulating key signal molecules, such as TGF-β1 RI, p-Smad2/3, and Smad7 in TGF-β1/Smad signaling pathway of SSc skin fibroblasts, and inhibiting transduction of TGF-β1/Smad signaling pathway.</p>


Subject(s)
Female , Humans , Collagen Type I , Collagen Type III , Drugs, Chinese Herbal , Pharmacology , Therapeutic Uses , Fibroblasts , Matrix Metalloproteinase 9 , Scleroderma, Systemic , Drug Therapy , Metabolism , Signal Transduction , Smad2 Protein , Smad7 Protein , Transforming Growth Factor beta1 , Metabolism
8.
Journal of Experimental Hematology ; (6): 1015-1020, 2013.
Article in Chinese | WPRIM | ID: wpr-283990

ABSTRACT

There are many evidences that dendritic cells (DC) can establish and maintain immunological tolerance through inducing the differentiation of regulatory T cells Treg. This study was purposed to explore the possibility to gene-rate Treg from bone marrow-derived DC (BM-DC) or spleen-derived DC (spDC) generated CD4(+) CD25(+) FOXP3(+) Treg by induction. Bone marrow immature DC (imDC) induced from bone marrow precursor cells of C57BL/6 mice by GM-CSF and IL-4; after culture for 6 day, imDC were stimulated by LPS for additional 16 hours and the mature DC (mDC) have been got; the spDC were collected from spleen of C57BL/6 mice by MACS. Co-culturing fresh BALB/c mouse CD4(+) T cells with these three sorts of DC above mentioned respectively was performed to generate CD4(+) CD25(+) FOXP3(+) Treg. The expression of FOXP3 in CD4(+) T cells was detected by flow cytometry, and the capacity of different DC generated CD4(+) CD25(+) FOXP3(+) Treg was evaluated. The results showed that stimulated by C57BL/6 immature or mature DC, the positive rate of FOXP3 in BALB/c CD4(+) T cells increased from (8.57 ± 1.14)% to (15.80 ± 1.35)%, (17.93 ± 1.45)% respectively (P < 0.01); while stimulated by spDC, the positive rate of FOXP3 in BALB/c CD4(+) T cells decreased from (8.57 ± 1.14)% to (3.95 ± 0.79)% (P < 0.05). It is concluded that the BM-DC but not spDC can generate Treg from CD4(+) T cells, BM-DC may mediate immune tolerance rather than the immune response.


Subject(s)
Animals , Female , Mice , Bone Marrow Cells , Cell Biology , Cell Differentiation , Cells, Cultured , Coculture Techniques , Dendritic Cells , Cell Biology , Forkhead Transcription Factors , Metabolism , Mice, Inbred BALB C , Mice, Inbred C57BL , Spleen , Cell Biology , T-Lymphocytes, Regulatory , Cell Biology
9.
Journal of Southern Medical University ; (12): 1914-1917, 2011.
Article in Chinese | WPRIM | ID: wpr-265753

ABSTRACT

<p><b>OBJEVTIVE</b>To construct a eukaryotic expression vector of short-hairpin RNA (shRNA) targeting human Akt gene and assess the effect of Akt gene silencing on the growth of colon cancer Lovo cells.</p><p><b>METHODS</b>Two shRNAs targeting human Akt gene were cloned into pENTRTM/U6 plasmid to obtain the entry clones, and the positive clones were verified by sequencing. After recombination of the pENTRTM/U6 entry constructs and Plenti6/Block-iT DEST vector, the positive clones were confirmed by sequencing. Lovo cells were transfected by the entry vector and DEST Vector, and RT-PCR and Western blotting were performed to detect the interference of Akt gene expressions.</p><p><b>RESULTS</b>The pENTRTM/U6 entry clones carrying Akt shRNA and pLenti6/DEST-pENTRTM/U6-Akt shRNA were successfully constructed. Both of the vectors were transfected into Lovo cells and resulted in obvious knockdown of the mRNA and protein expressions of Akt.</p><p><b>CONCLUSION</b>The Akt siRNA expression vector constructed can significantly inhibit Akt gene expression in Lovo cells, which facilitates further studies of Akt function and tumor gene therapy.</p>


Subject(s)
Humans , Cell Line, Tumor , Colonic Neoplasms , Pathology , Genetic Vectors , Genetics , Proto-Oncogene Proteins c-akt , Genetics , RNA Interference , RNA, Small Interfering , Genetics , Transfection
10.
Journal of Southern Medical University ; (12): 1841-1843, 2010.
Article in Chinese | WPRIM | ID: wpr-330827

ABSTRACT

<p><b>OBJECTIVE</b>To construct pPICZalphaA-soluble interleukin-1 receptor type I (sIL-1RI) recombinant expression vector containing the gene fragment encoding the extracellular domain of sIL-1RI for its expression in Pichia pastoris.</p><p><b>METHODS</b>sIL-1RI gene was amplified by RT-PCR and inserted into the yeast expression vector pPICZalphaA by digestion ligation. The recombinant plasmid pPICZalphaA-sIL1RI was transformed into E.coli Stb13, and the positive clones were analyzed by PCR and DNA sequencing. The pPICZalphaA-sIL1RI recombinant plasmid was electroporated into GS115 cells and the transformants were analyzed by PCR. After phenotype identification, the recombinant strains were induced by methanol to express the target protein, which was analyzed by Western blotting of the cell extract and supernatant.</p><p><b>RESULTS</b>The recombinant plasmid pPICZalphaA-sIL-1RI was constructed successfully, and the results of Western blotting showed that yeast induced by methanol expressed a protein of about 39 kD.</p><p><b>CONCLUSION</b>sIL-1RI protein has been successfully expressed in P.pastoris expression system, which provides the basis for further study of sIL-1RI.</p>


Subject(s)
Humans , Escherichia coli , Metabolism , Gene Expression , Genetic Vectors , Pichia , Metabolism , Plasmids , Receptors, Interleukin-1 Type I , Genetics
11.
Journal of Southern Medical University ; (12): 462-465, 2009.
Article in Chinese | WPRIM | ID: wpr-233763

ABSTRACT

<p><b>OBJECTIVE</b>To isolate and characterize human rheumatoid arthritis (RA) fibroblast-like synoviocytes (FLSs).</p><p><b>METHODS</b>The synovial membrane tissues were obtained from 4 RA patients, 1 chondroma patient and 1 healthy subject and FLS were isolated by means of tissue culture. The cell morphology was observed by phase-contrast microscope and the cell surface markers were detected by flow cytometry.</p><p><b>RESULTS</b>The FLSs were successfully cultured from the synovial membrane tissues with good cell homogeneity after the third passage. The FLSs of the 3rd to 7th passages were stable and proliferated actively, followed by slow proliferation and aging since the 8th passage. Flow cytometry showed that the 4th-passage FLSs from the RA patients contained 99.04% CD90(+) cells, 2.73% CD3(+) cells, 0.29% CD3(-)CD19(+) cells, 2.81% CD3(-)CD16(+)CD56(+) cells, 5.89% CD14(+) cells, and 54.17% CD55(+) cells. The presence of interleukin-1 receptor type I (IL-1RI, 158.63-/+20.32 pg/ml) and IL-1beta (4.67-/+0.82 pg/ml) were detected in the cell culture supernatant of the 4th-passage FLSs from the RA patients by enzyme-linked immunosorbent assay ELISA.</p><p><b>CONCLUSION</b>FLSs from RA patients can be effectively culture by means of tissue culture, and the cultured FLSs show high expressions of CD90, IL-1RI and IL-1beta.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Arthritis, Rheumatoid , Pathology , Cell Proliferation , Cell Separation , Cells, Cultured , Fibroblasts , Pathology , Interleukin-1beta , Metabolism , Receptors, Interleukin-1 Type I , Metabolism , Synovial Membrane , Cell Biology , Pathology , Thy-1 Antigens , Metabolism
12.
Journal of Southern Medical University ; (12): 884-886, 2009.
Article in Chinese | WPRIM | ID: wpr-268820

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the performance of BNII auto-analyzer system in detecting C3 and C4.</p><p><b>METHODS</b>CLSI protocols (EP15-A, EP6-A, EP9-A2) and other relevant literatures were use to or evaluate the precision, accuracy, linearity of C3 and C4 detection by the auto-analyzer system, and the results were compared with the recognized standards.</p><p><b>RESULTS</b>The relative bias of C3 and C4 was less than one third of the CLIA'88 standard and the precision met the clinical requirement. The results tested by DADE BNII system were not compatible with those by Roche Modular System. C3 showed good linearity in the tests (R2>0.975, P<0.05) with a linearity range of 0.18-5.1 g/L. The linearity of C4 was not available because of lack of high-level samples.</p><p><b>CONCLUSION</b>The performances of DADE BNII System basically meet the recognized standards in clinical detection of C3 and C4, but the method comparison needs further validation.</p>


Subject(s)
Humans , Autoanalysis , Methods , Blood Chemical Analysis , Methods , Complement C3 , Complement C4 , Nephelometry and Turbidimetry , Methods , Proteins , Sensitivity and Specificity
13.
Journal of Southern Medical University ; (12): 1609-1611, 2009.
Article in Chinese | WPRIM | ID: wpr-282637

ABSTRACT

<p><b>OBJECTIVE</b>To assess the value of rheumatoid factors IgM-RF, IgA-RF, IgG-RF, interleukin-1 receptor type I (IL-1RI) and cyclin-dependent kinase 2 (CDK2) in the diagnosis of rheumatoid arthritis (RA).</p><p><b>METHODS</b>IgM-RF, IgA-RF, IgG-RF, IL-1RI and CDK2 were detected in 47 patients with active RA, 47 with inactive RA, 20 with active systemic lupus erythematosus (SLE), 20 with inactive SLE, 20 with acute upper respiratory tract infection (AURTI), and 20 healthy controls using enzyme-linked immunosorbent assay (IgM-RF, IgA-RF, IgG-RF, IL-1RI) and time-resolved fluoroimmunoassay (CDK2).</p><p><b>RESULTS</b>Patients with active and inactive RA showed significant differences in peripheral serum CDK2 and IgA-RF levels (P<0.05). Between active and inactive RA patients, RA patients and SLE patients, RA patients and AURTI patients, and RA patients and the control subjects, the area under curve (AUC) of the receiver operating characteristic curve (ROC) was 0.561, 0.814, 0.799, and 0.888 for IgM-RF, 0.596, 0.678, 0.729, and 0.850 for IgA-RF, 0.614, 0.718, 0.692, and 0.791 for IgG-RF, 0.646, 0.691, 0.762, and 0.835 for IL-1RI, 0.803, 0.753, 0.741, and 0.840 for CDK2, respectively.</p><p><b>CONCLUSIONS</b>IgM-RF may have high value in the diagnosis and differential diagnosis of RA, and CDK2 can be useful for differential diagnosis between active and inactive RA.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Area Under Curve , Arthritis, Rheumatoid , Blood , Diagnosis , Cyclin-Dependent Kinase 2 , Blood , Immunoglobulin A , Blood , Immunoglobulin G , Blood , Immunoglobulin M , Blood , ROC Curve , Receptors, Interleukin-1 Type I , Blood , Rheumatoid Factor , Blood
14.
Journal of Central South University(Medical Sciences) ; (12): 682-687, 2008.
Article in Chinese | WPRIM | ID: wpr-814015

ABSTRACT

OBJECTIVE@#To construct the recombinant lentivirus RNAi vector, and to determine whether the lentivirus mediated short hairpin RNA (shRNA) can inhibit the tissue factor (TF) expression in endothelial cells.@*METHODS@#Two short hairpin RNAs targeting to human TF were cloned into pENTRTM/U6 plasmid to obtain an entry clone, and the positive clones were verified by sequencing. A recombination reaction was performed between the pENTR/U6 entry construction and pLenti6/BLOCKiTTM-DEST vector, and then the positive clones were confirmed by sequencing. The 293FT cell line was transfected by the above recombined plasmid and lentivirus packing materials, the culture supernatant was harvested, and the virus titer was determined. RT-PCR and ELISA were used to observe the inhibition of TF gene expression after the lentivirus transduction in human umbilical vein endothelial cells.@*RESULTS@#The shRNA sequences targeting to human TF were cloned into the vectors, and an entry clone and an expression clone were constructed successfully, which were proved by sequence determination. Viral particles were packaged in the 293FT cell line, all virus stocks were collected, and the transfection titer was 5*10(5)/transduced unit. RT-PCR and enzyme linked immunosorbent assay demonstrated that the lentivirus stocks could suppress the TF expression in endothelial cells remarkably.@*CONCLUSION@#Lentivirus RNAi vectors containing human TF gene are successfully constructed, and lentivirus mediated shRNA can inhibit the TF expression in endothelial cells, which may provide a highly effective method for the prevention and treatment of thrombo-embolic diseases.


Subject(s)
Humans , Base Sequence , Down-Regulation , Endothelial Cells , Cell Biology , Metabolism , Genetic Vectors , Genetics , Lentivirus , Genetics , Molecular Sequence Data , RNA Interference , RNA, Messenger , Genetics , RNA, Small Interfering , Genetics , Recombinant Proteins , Genetics , Thromboplastin , Genetics , Umbilical Veins , Cell Biology
15.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 106-109, 2007.
Article in Chinese | WPRIM | ID: wpr-269125

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effects of dialysate contained Chinese herbs for replenishing qi and activating blood circulation on platelet membranous glycoprotein CD62P in patients with chronic renal failure (CRF) undergoing hemodialysis.</p><p><b>METHODS</b>Forty patients underwent maintaining hemodialysis were randomly assigned to two groups, the Western medicated group (WMG) and the Chinese herbs group (CHG). The content of CD62P in all patients was detected by ELISA before and after hemodialysis.</p><p><b>RESULTS</b>The levels of blood urea nitrogen, creatinine, potassium, hematocrit, platelet count and carbon dioxide combining power (CO2CP) as well as the expression of CD62P after treatment were significantly changed in both groups with significant difference as compared with those before treatment (both P < 0.05). And comparison between the two groups in expression of CD62P after treatment also showed significant difference (P < 0.05). But the improvement in TCM syndrome between the two groups was insignificantly different (P > 0.05).</p><p><b>CONCLUSION</b>Hemodialysis with dialysate containing Chinese herbs of replenishing qi and activating blood circulation can decrease the expression of platelet membranous glycoprotein CD62P, which may be associated with the mechanism of Chinese herbs in treating CRF.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Blood Circulation , Blood Platelets , Metabolism , Combined Modality Therapy , Dialysis Solutions , Drugs, Chinese Herbal , Therapeutic Uses , Enzyme-Linked Immunosorbent Assay , Kidney Failure, Chronic , Blood , Therapeutics , P-Selectin , Blood , Phytotherapy , Qi , Renal Dialysis , Methods
16.
Journal of Southern Medical University ; (12): 1661-1664, 2007.
Article in Chinese | WPRIM | ID: wpr-281568

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the single-nucleotide polymorphisms (SNPs) at positions -572 and -174 in the promoter region of interleukin-6 (IL-6) gene and at -607 and -137 in the promoter region of interleukin-18 (IL-18) gene for their association with rheumatoid arthritis (RA) in the Chinese Han population in Guangdong Province.</p><p><b>METHODS</b>SNPs of IL-6 and IL-18 genes were detected in 120 patients with RA and 168 normal subjects using polymerase chain reaction with sequence-specific primers (PCR-SSP).</p><p><b>RESULTS</b>SNPs at -572 and -174 of IL-6 gene and at -607 and -137 of IL-18 gene were detected in this population. There was a significant difference in the genotype and allele frequency at -572 and -174 of IL-6 gene and -607 of IL-18 gene (P<0.001), but not in the distribution of genotype frequencies at -137 of IL-18 gene between the RA patients and healthy subjects (P=0.141). A significant difference was found, however, in the allele frequency at -137 of IL-18 (P=0.024). Logistic regression analysis revealed significant association of age, gender, IL-6 gene -572, -174 and IL-18 gene -137 SNPs with RA (P<0.05).</p><p><b>CONCLUSIONS</b>The polymorphisms of the promoter region of IL-6 gene at positions -572 and -174 is probably associated with RA, and further study is needed to understand the relation of the polymorphisms of IL-18 gene at positions -607C/A and-137G/C with RA.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Age Factors , Arthritis, Rheumatoid , Genetics , Asian People , Genetics , Case-Control Studies , China , Gene Frequency , Genetic Predisposition to Disease , Genotype , Interleukin-18 , Genetics , Interleukin-6 , Genetics , Logistic Models , Polymorphism, Single Nucleotide , Promoter Regions, Genetic
17.
Chinese Journal of Integrated Traditional and Western Medicine ; (12): 17-20, 2004.
Article in Chinese | WPRIM | ID: wpr-320261

ABSTRACT

<p><b>OBJECTIVE</b>To find the optimal dosage of Salvia injection in treating chronic hepatitis B caused liver fibrosis.</p><p><b>METHODS</b>Sixty-four patients, whose diagnosis was confirmed as chronic hepatitis B caused liver fibrosis and differentiated by TCM typing as blood stasis blocking Collaterals type, were selected and randomly divided by lottery method into the large, middle and small dose of SI treated groups and the control group. All the patients were treated with modified Gexia Zhuyu Decoction, to the patients in the SI groups, 24 ml, 16 ml and 8 ml of SI were additionally administered by intravenous dripping respectively. The therapeutic course was 45 days. The clinical symptoms and signs; liver functional indexes as alanine transaminase (ALT), aspartate aminotransferase (AST) and albumin (ALB); and liver fibrosis indexes as procollagen type III (PC-III), collagen type IV (C-IV) and hyaluronic acid (HA), were measured before and after treatment.</p><p><b>RESULTS</b>Different dosages of SI all could improve the clinical symptoms, and lower levels of ALT, AST, HA, PC-III and C-IV. Treatment of large dosage SI showed the best efficacy, superior to that of middle and small dosage SI, but no significant difference was found between the efficacy of the latter two.</p><p><b>CONCLUSION</b>Anti-liver fibrosis effect of large dosage SI is better than that of middle or small dosage SI.</p>


Subject(s)
Adult , Female , Humans , Male , Middle Aged , Collagen Type III , Blood , Collagen Type IV , Blood , Drugs, Chinese Herbal , Hepatitis B, Chronic , Blood , Hyaluronic Acid , Blood , Infusions, Intravenous , Liver Cirrhosis , Blood , Drug Therapy , Phytotherapy , Salvia miltiorrhiza
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