Comparison of growth and quality of wild and cultivated Artemisia stolonifera / 中国中药杂志

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.

Effects of shading intensity on growth and quality of Artemisia stolonifera / 中国中药杂志

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.

Quality of moxa from Artemisia argyi and A. stolonifera in different storage years based on simultaneous thermal analysis / 中国中药杂志

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.

Quality of moxa with different leaf-to-moxa ratios based on correlation analysis of thermogravimetric properties, cellulose content, and microscopic characteristics of non-secretory trichomes / 中国中药杂志

The quality of moxa is a key factor affecting the efficacy of moxibustion. Traditional moxa grades are evaluated by the leaf-to-moxa ratio, but there is a lack of support from scientific data. Scanning electron microscopy(SEM), Image Pro Plus, Van Soest method, and stimultaneous thermal analysis(TGA/DSC) were used to characterize the scientific implication of the combustion differences between moxa with different leaf-to-moxa ratios(processed by crusher). The results showed that the median lengths from non-secretory trichomes(NSTs) of natural NSTs and moxa with leaf-to-moxa ratios of 3∶1, 5∶1, 10∶1, and 15∶1 were 542.46, 303.24, 291.18, 220.69, and 170.61 μm, respectively. The cellulose content of moxa increased significantly(P<0.05) with the increase in leaf-to-moxa ratio and the combustion parameters(T_i, t_i, D_i, C,-R_p,-R_v, S, D_b, and J_(total)) all showed an increasing trend. The correlation results showed that the burning properties of moxa(T_i,-R_v, t_i, and J_2) were significantly and positively correlated with cellulose content. NSTs with a length of 1-200 μm were significantly and positively correlated with J_2. NSTs with a length of 200-600 μm were significantly and positively correlated with J_1, T_(peak2), T_(peak1), and-R_v, and negatively correlated with J_(total), T_b, and t_b. As the leaf-to-moxa ratio increases, the NSTs in the moxa become shorter and the cellulose content increases, which is more conducive to ignition performance, heat release, and a milder, longer-lasting burn. The "NSTs-cellulose-TGA/DSC" quantitative evaluation method scientifically reveals the scientific connotation of the combustion of moxa with different leaf-to-moxa ratios and provides a scientific basis for the establishment of quality evaluation methods for moxa with different leaf-to-moxa ratios.

[Research progress on chemical constituents from Artemisiae Argyi Folium and their pharmacological activities and quality control].

Artemisiae Argyi Folium is a traditional medicine commonly used in traditional Chinese medicine(TCM). It contains volatile oil, flavones, phenylpropanoids, terpenes and other chemical constituents. It has the functions of antibacterial, antiviral, hemostasis, anti-tumor, liver protection, analgesic, anti-inflammatory, anti-oxidation, relieving cough and asthma and other pharmacological activities. At present, many useful researches on the quality of moxa floss and Artemisiae Argyi Folium have been carried out on the contents of volatile oil, flavones, phenylpropanoids, the storage time of Artemisiae Argyi Folium, the processing of moxa, the genuineness of Artemisiae Argyi Folium, and their heat release properties in combustion. This paper summarized the literature on the chemical composition, pharmacological activities and quality control of Artemisiae Argyi Folium and provided the basis for the further development and utilization of Artemisiae Argyi Folium.

[Comparison of chemical components between Artemisia stolonifera and Artemisia argyi using UPLC-Q-TOF-MS].

The chemical component information of samples was analyzed by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). The leaves and flosses of Artemisia argyi and A. stolonifera from different places, were distinguished by principal component analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). Then, compounds with significant differences(P<0.01) in abundance were screened out according to their VIP values and t-test. Eighteen components in all samples have been filtered and identified, including flavonoids and chlorogenic acids, the content of the 12 of which were measured by UPLC-UV, which are different in presence and content. Hispidulin in A. argyi is not detected in A. stolonifera. Schaftaside, isochlorogenic acid B, and isochlorogenic acid C are differential compounds of A. argyi and A. stolonifera leaves. Isochlorogenic acid A, isochlorogenic acid C and jaceosidin are differential compounds of A. argyi and A. stolonifera floss. There are significant differences in the contents of jaceosidin and schaftoside in the four famous A. argyi. In addition, the content of isochlorogenic acid A in wild A. stolonifera is higher than that in cultivated A. stolonifera. The results of the study successfully clarified the differences between A. stolonifera and A. argyi, and provided theoretical and data references for the further development and utilization of A. stolonifera.

[Textual research on traditional Chinese medicine Jiuniucao].

Jiuniucao in Qizhou, known as "Qiai", was precious and expensive in the Ming and Qing Dynasties. But the authentic plant of Jiuniucao is not mentioned in the Ben Cao Tu Jing and other medical books in the Song, Ming and Qing Dynasties. In history, mugwort leaf originates from many species of plants, Jiuniucao may be one of it. So this paper is to identify the original plant of Jiuniucao and clarify the historical origin of Jiuniucao and mugwort leaf. The textual research and geographical origin analysis of Jiuniucao in ancient literature was conducted. Then field investigation and sampling of Jiuniucao in Qizhou was proceeded, combing with feature comparison of Jiuniucao and Artemisia specimen in the Herbarium of Institute of Botany, Chinese Academy of Sciences. The study concluded that A. stolonifera is the authentic Jiuniucao for medical use. Jiuniucao was also an important original plant of mugwort leaf and it is worth further development and utilization.

[Thermodynamic characteristics of moxa floss and its biological effects and therapeutic mechanism on osteoarthritis rats].

Moxa floss is the main material of moxibustion, which plays a therapeutic role through the thermal effect of combustion. In this paper, TG/DSC thermal analysis method was used to study the characteristic parameters of combustion heat of moxa floss produced in Qichun, and the thermal therapeutic effect and mechanism of moxibustion were studied through moxibustion OA animal model. The results show that the combustion process of moxa floss can be divided into three stages: drying, combustion oxidation and carbonization, and ash burnout. The combustion properties of moxa floss are between herbaceous and woody, and tend to be woody, with flammable, slow and lasting combustion characteristics. Moxibustion can relieve the pathological state of knee joint synovium to a certain extent, reduce knee joint swelling and blood stasis in OA rats, reduce interstitial edema, and improve local inflammation. The mechanism and target point of moxibustion treatment for OA may be up-regulating TRPM3 gene to activate ion channels, affecting calcium metabolism and reducing OA swelling degree; down-regulation of GAPDH affects glucose metabolism of knee synovial cells and mediates anti-inflammatory effect. Down-regulation of pain-related gene MMP24 is helpful to relieve OA pain. Up-regulation of CTNNB1 activates Wnt/ß-catenin signaling pathway and affects differentiation and regeneration of OA chondrocytes. This study reveals the pyrolysis characteristics of moxa floss for the first time and discusses the biological effect and possible mechanism of moxibustion heat, providing new ideas for the quality evaluation of moxa floss and the mechanism of moxibustion therapy.

Herbal Textual Research on Genuine Medicinal Materials “Deng Ju” and Quality Evaluation of Nan Yang Chrysanthemi Flos / 中国实验方剂学杂志

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.

Quantative analysis of eupatilin and jaceosidin in folium of Artemisia argyi from different areas in China by RP-HPLC based on ancient medicine books / 中国中药杂志

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.

Investigation on reference intervals and regional differences of platelet indices in healthy Chinese Han adults.

BACKGROUND: Reference intervals are important for interpretation of clinical laboratory tests. The platelet (PLT) indices such as the mean platelet volume (MPV) and platelet distribution width (PDW) are newer hematological parameters, which have been recently reported as clinically valuable biomarkers. However, there are not many studies that have estimated the reference intervals for these parameters in healthy Chinese Han adults. OBJECTIVE: The objectives of this study were to establish reference values of PLT indices [including PLT count, MPV, PDW, platelet-large cell ratio (P-LCR), and plateletcrit (Pct)] for healthy Chinese Han adults. We also aimed to determine the region-based differences of PLT indices in China. METHODS: A total of 4,642 volunteers with a mean age of 43 were recruited from six regions of China. PLT indices were performed on Sysmex XE-2100 hematology analyzers, whose traceability was well verified. RESULTS: There were significant region-based differences for all PLT indices. Reference people in Chengdu had the lowest mean PLT count and Pct, but the highest MPV, PDW, and P-LCR among the six regions. Therefore, we derived the reference intervals in Chinese Han population excluding Chengdu reference people for PLT indices as PLT count: (127-341) × 10(9)/l; MPV: (9.20-13.30) fl; PDW: 9.90-19.00%; P-LCR: 18.10-52.00%; Pct: 16.00-41.00%. CONCLUSIONS: Region-specific reference intervals are essential as there were statistically significant region-related differences in the PLT parameters. The reference intervals established in this study differed from the existing reference values. Chengdu region may need proper specific reference ranges, which apply to their people, for all PLT parameters.

Study on identification of "Digeda" raw materials in Mongolian patent medicine by PCR amplification of specific alleles / 中国中药杂志

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.

Effects of Wenyang Huazhuo Tongluo Recipe Containing Serum on Transforming Growth Factor β1/ Smad Signaling Pathway of Skin Fibroblasts in Systemic Sclerosis / 中国中西医结合杂志

<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>

Bone marrow-derived dendritic cells rather than spleen-derived dendritic cells can generate regulatory T cells / 中国实验血液学杂志

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.

Construction of the expression vector for short-hairpin RNA-mediated Akt gene silencing in Lovo cells / 南方医科大学学报

<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>

Expression of recombinant extracellular region of human interleukin-1 receptor type I in Pichia pastoris / 南方医科大学学报

<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>

Assessment of the performance of an automated analysis system in detecting C3 and C4 / 南方医科大学学报

<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>

Determination of IgM-RF, IgA-RF, IgG-RF, IL-1RI, CDK2 and ROC evaluation in patients with rheumatoid arthritis / 南方医科大学学报

<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>

Isolation and characterization of human rheumatoid arthritis fibroblast-like synoviocytes / 南方医科大学学报

<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>

Inhibition of tissue factor expression in endothelial cells by lentivirus mediated shRNA / 中南大学学报(医学版)

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.