Angiogenesis opens a way for Chinese medicine to treat stroke / 中国结合医学杂志

Based on the pathophysiology of the brain, advance in angiogenesis induced by stroke, and evidences of Chinese-medicine-mediated angiogenesis, the possibility to study the stroke-treating mechanism of Chinese medicine in angiogenesis was discussed. And regarding our previous work on angiogenesis modulated by qi-tonifying and stasis-eliminating therapy following intracerebral hemorrhage, we proposed some questions, which should be taken into account in the further work.

Electroacupuncture at Zusanli (ST36) accelerates intracerebral hemorrhage-induced angiogenesis in rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of electro-acupuncture on intracerebral hemorrhage (ICH)-induced angiogenesis and hypoxia-inducible factor-1α (HIF-1α) expression in rats.</p><p><b>METHODS</b>Adult male Sprague-Dawley rats were randomly divided into 4 groups of 24 rats each. ICH was induced in 3 groups by stereotactic injection of collagenase type VII into the right globus pallidus; of these, one group was not further treated, the second group underwent Zusanli (ST36)-acupuncture, and the third group underwent non-acupoint acupuncture. The fourth group underwent sham operations. Acupuncture was performed by stimulation with electrical needles at frequencies of 2-20 Hz for 30 min per day. Angiogenesis on days 3, 7 and 14 was assessed by double immunolabeling, and expression of HIF-1α was evaluated by immunohistochemistry, quantitative real time reverse transcription-polymerase chain reaction and Western blotting.</p><p><b>RESULTS</b>5-Bromo-2-deoxyuridine (BrdU)-labeled nuclei in cerebral endothelial cells (ECs) resided around the hematoma and the labeling peaked from 7 to 14 days (P<0.01). HIF-1α positive microvessels with a dilated outline were detected in perihematomal tissues after ICH, with the vessels extending into the clot from the surrounding area beginning on day 7. Following ICH, HIF-1α protein levels increased (P<0.05), but HIF-1α mRNA levels did not change. Electro-acupuncture at the Zusanli (ST36) acupoint increased BrdU-labeled nuclei in cerebral ECs (P<0.05) and up-regulated the expression of HIF-1α protein (P<0.05), but had little effect on the spatial distribution of HIF-1α or on HIF-1α mRNA levels.</p><p><b>CONCLUSIONS</b>Electro-acupuncture treatment at the Zusanli (ST36) acupoint may accelerate ICH-induced angiogenesis by up-regulating HIF-1α protein, and may enhance recovery following hemorrhagic cerebral injury.</p>

A strategy for detecting absorbed bioactive compounds for quality control in the water extract of rhubarb by ultra performance liquid chromatography with photodiode array detector / 中国结合医学杂志

<p><b>OBJECTIVE</b>To detect absorbed bioactive compounds of the water extract whose pharmacodynamic effect was craniocerebral protection for quality control assessment.</p><p><b>METHODS</b>Anthraquinones in water extract of rhubarb (WER), in cerebrospinal fluid (CSF) of patients with traumatic brain injury (TBI) and in ipsilateral cortex of TBI rats following oral WER were respectively explored by ultra performance liquid chromatography with photodiode array detector (UPLC-PDA) method developed in the present study. The effects of anthraquinones absorbed into injured cortex on superoxidase dismutase (SOD) activity in TBI rats were detected. The antioxidative anthraquinones absorbed into target organ were evaluated for quality control of WER.</p><p><b>RESULTS</b>Anthraquinones in WER were aloe-emodin, rhein, emodin, chrysophanol, and physcion. Only the last anthraquinone was found in CSF and in ipsilateral cortex under this chromatographic condition. Physcion increased SOD activity in TBI rats significantly.</p><p><b>CONCLUSIONS</b>Physcion was the main active compound of rhubarb against craniocerebral injury via antioxidant pathway. According to our strategy, the exploration of physcion suggested the possibility of a novel quality control of WER in treating TBI injury.</p>

Ursolic acid inhibits T-cell activation through modulating nuclear factor-κ B signaling / 中国结合医学杂志

<p><b>OBJECTIVE</b>To investigate the effects of ursolic acid (UA) on T-cell proliferation and activation, as well as to examine its effect on nuclear factor-κB (NF-κB) signaling pathway in T cells.</p><p><b>METHODS</b>T-cells isolated from BALB/c mice were incubated with UA at concentrations ranging from 5-30 μmol/L in the presence of phorbol 12-myristate 13-acetate (PMA) or PMA plus ionomycin. The proliferation of T cells was measured by the MTT assay. The expressions of CD69, CD25, and CD71 on T-cell surface were analyzed using flow cytometry. The level of interleukin-2 (IL-2) in the culture supernatant of activated T cells was quantified by enzyme-linked immunosorbent assay (ELISA). The level of phosphorylated IκB-α (p-IκB-α) in total protein and p65, a subunit of NF-κB, nuclear translocation were measured by Western blot analysis.</p><p><b>RESULTS</b>UA in a dose-dependent manner significantly decreased the proliferation and inhibited the surface expressions of CD69, CD25, and CD71 in murine T lymphocytes upon in vitro activation (P<0.01). Significant reduction of IL-2 production was found in activated T cells treated with UA (P<0.01). The PMA-induced increase in p-IκB-α protein was inhibited, and nuclear translocation of p65 from the cytoplasm was blocked by UA.</p><p><b>CONCLUSION</b>UA is a potent inhibitor for T cell activation and proliferation; these effects are associated with the inhibition of NF-κB signaling pathway.</p>

Proteomics study on the essence of wind syndrome caused by gan-yang hyperactivity in Chinese medicine / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To preliminarily study the essence of wind syndrome caused by Gan-yang hyperactivity (WSGH) in Chinese medicine at the protein expression level.</p><p><b>METHODS</b>WSGH was strictly differentiated from wind stirring due to yin deficiency syndrome in patients with intracerebral hemorrhage (ICH) and those with cerebral infarction (CI); from Gan-yang hyperactivity syndrome in patients with cervical spondylosis (CS); from wind syndrome induced by blood deficiency in patients with Parkinson's disease (PD) according to Chinese medicine syndrome typing standard. Control studies were performed. Peripheral blood mononuclear cells (PBMCs) were isolated from all patients of the aforesaid syndromes and healthy subjects. The total proteins were extracted, two-dimensional gel electrophoresis (2-DE) conducted and analyzed by PDQuest software. The peptide mass fingerprint (PMF) was determined using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). The SwissProt database was inquired using Mascot reference system. Proteins of different and same expressions in PBMCs of patients suffering from the same disease of different syndromes, different diseases of the same syndrome, and syndromes of the same kind were compared.</p><p><b>RESULTS</b>The 2-DE map of PBMCs' total proteins in the aforesaid syndrome groups and healthy subjects was established. Through comparison, analysis, and appraisement, there was 1 protein dot of the same expression and 22 protein dots of different expressions between ICH patients of WSGH and ICH patients of wind stirring due to yin deficiency syndrome. There were 6 protein dots of the same expression and 21 protein dots of different expressions between CI patients of WSGH and CI patients of wind stirring due to yin deficiency syndrome. There were 3 protein dots of the same expression and 12 protein dots of different expressions between CS patients of WSGH and CS patients of Gan-yang hyperactivity syndrome. There was no protein dot of the same expression and 12 protein dots of different expressions between PD patients of WSGH and PD patients of wind syndrome induced by blood deficiency. There were 13 protein dots of the same expression in different diseases of the same syndrome. There was 1 protein dot (Thioredoxin-dependent peroxide reductase, TPx) of the same expression in the four diseases of the same kind syndrome.</p><p><b>CONCLUSIONS</b>Different connotations of the essence existed (having multiple different protein expressions) in patients with the same disease of different syndromes. Syndromes of the same kind share the same material bases (having the same protein expression). These suggested that Chinese medicine syndrome has its own material bases and essence findable.</p>

Effect of Bizhongxiao decoction on proteomics of peripheral blood mononuclear cells in patients with rheumatoid arthritis / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To probe in the possible acting mechanism of Bizhongxiao Decoction (BZXD) for treatment of early active rheumatoid arthritis (RA) by way of observing the two-dimensional gel electrophoresis map of proteins in peripheral blood mononuclear cells (PBMCs) of healthy persons and RA patients (intervened or un-intervened with BZXD), analyzing the differential proteins and seeking out the RA associated proteins.</p><p><b>METHODS</b>Eighteen patients with early active RA were randomized into the BZXD group and the methotrexate (MTX) group, nine in each group, they were treated with BZXD (contained 15 Chinese herbs, as Herba Hedyotis diffusae, Herba Sarcandrae glabrae, Radix Salviae miltiorrhizae, Caulis Trachelosperi, Rhizoma Drynariae, Semen Coicis, etc.) and MTX combined with nimesulide Tablets respectively, three months as a treatment course, and their blood samples were collected for observation. Besides, blood samples from 9 healthy persons were taken as normal controls. PBMCs were isolated from blood using lymphozytes separation medium, and total protein in the cells was extracted through immobilized pH gradient two-dimensional gel electrophoresis. After Coomassie brilliant blue G250 staining, gel-image analysis was performed using PDQuest software. The differentially expressed proteins were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS). Then partial proteins were validated by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The 2-DE protein profile of PBMCs from healthy persons and RA patients before and 3 months after treatment were obtained, and 23 differential protein spots were found, 14 from 18 differential protein spots were successfully identified, of which 8 proteins were up-regulated and 6 proteins were down-regulated in RA patients as compared with control. After 3-month treatment, 5 differentially expressed proteins showed more obvious in the BZXD group than in the MTX group. RT-PCR verified that the expression of ApoA-I in all the three groups was consistent with the outcomes of 2-DE.</p><p><b>CONCLUSIONS</b>Some differentially expressed proteins exist in the PBMCs of RA patients, which may play a potential role in the pathogenesis of RA; BZXD may treat RA by way of regulating the expression of some differential proteins in patients.</p>

The application and significance in prenatal diagnosis using G-banding, fluorescence in situ hybridization and comparative genomic hybridization / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the procedure and the value of G-banding, fluorescence in sit hybridization (FISH) and comparative genomic hybridization (CGH) techniques in prenatal diagnosis.</p><p><b>METHODS</b>Karyotype analyses with three diagnostic procedures, G-banding, G-banding and FISH, G-banding, FISH and CGH, were performed in the amniotic fluid samples taken from 102 fetuses at gestational ages 16-24 weeks. And the significance was valued in prenatal diagnosis.</p><p><b>RESULTS</b>In the first procedure of karyotype analysis, 98 cases were diagnosed, 2 cases were not conformed while 2 cases were failed in all 102 cases. In the second procedure, 2 cases were determined, 1 case was not conformed and 1 case was still failed. In the third step, 2 cases were diagnosed. The diagnostic rate of the karyotype reached to 100% (102/102 cases) using all the three procedures. In total, seven cases with chromosomal abnormality were diagnosed. Four cases, 1 case and 2 cases were identified in the first step (4/7, 57.1%), the second (1/7, 14.3%) and the third (2/7, 28.5%), respectively.</p><p><b>CONCLUSION</b>It can help improve the diagnostic rate of chromosomal aberrations and standardize diagnostic procedure to perform the three detecting steps in prenatal diagnosis.</p>

Scanning of drug targets related to uterus contraction from the uterine smooth muscles by cDNA microarray / 中南大学学报(医学版)

OBJECTIVE@#To screen the differentially expressed gene profile from the smooth muscles in the fundus uterus at the active stage of labor, and to provide candidate genes for picking out the drug targets related to uterine contraction.@*METHODS@#Differentially expressed genes of uterine smooth muscles in the corpus from pro and post spontaneous parturition and those induced by oxytocin,as well as those from the corpus and the lower portion spontaneous parturition,were scanned respectively by human full-length genetic cDNA microarray with 8064 probe sets. Semi-quantitative RT-PCR was applied to testify the expression of voltage dependent calcium channel-L subtype (CACNA). The differentially expressed genes in the structure and function of the drug targets were picked out by bio-informatics to serve as candidate drug targets related to uterine contraction.@*RESULTS@#The expressions of 29 genes were upregulated in fundus smooth muscles from the pro and post natural parturition, the pro and post inductive parturition of oxytocin, and the natural parturition. The expression of CACNA gene in RT-PCR was in accordance with that in the microarray. Among the 29 genes, neuromedin B receptor (NMBR) gene and neuropeptide Y (NPY) gene were the genes which not only had the targets of uterine contracted medicine, but also could contract the uterine. The differential expression ratios of NMBR in the above 3 types of uterine myometrium were 6.9,11.3, and 9.0, respectively while those of NPY were 6.0,29.8, and 2.9 respectively.@*CONCLUSION@#NMBR, whose expression in the uterine smooth muscles is always up-regulated at different parturition conditions, is likely to be an ideal candidate target of uterotonic drugs.

Effect of bizhongxiao decoction on the expression of 2-dimensional polyacrylamide gel electrophoresis protein in the synovial tissue with collagen-induced arthritis in rats / 中南大学学报(医学版)

OBJECTIVE@#To explore the effect of bizhongxiao decoction (BZXD) on the protein maps of BZXD-treated synovitis of collagen-induced arthritis (CIA) in rats in 2-dimensional gel electrophoresis (2-DE), and to provide new clues for illuminating the active mechanism of BZXD in treating the rheumatoid arthritis (RA).@*METHODS@#Seventy SD rats were randomly divided into nor- mal group, model group and BZXD group. The experimental arthritis rat model was established by subcutaneouly injecting Type II collagen and complete Freunds adjuvant. The total proteins of synovial tissue of rat joints in the normal group, model group and BZXD group were seperated by 2-DE respectively. The gels of the 3 groups were stained by Coomassie brilliant blue. Electron pictures were obtained by scanning the gels, and then the differential proteins among the normal group, model group and BZXD group were examined by comparing the spots density volume in the gels. The electrophoregrams of the gels were analyzed in Pdquest software.@*RESULTS@#The incidence of arthritis in the rats was approximately 88%. The 2-DE maps of rat synovial tissue in the normal group, model group and BZXD group were well duplicated. The average protein spots in the normal group, model group and BZXD group were 947 +/- 39, 994 +/- 41, and 1031 +/- 52, and the match rates were 92%, 91%, and 94.2% respectively. The average deviations of spot position were (0.896 +/- 0.217) mm in isoelectric focusing (IEF) and (1.102 +/- 0.104) mm in sodiumdo-decylsufate-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Three hundred twenty-eight differential proteins were observed between the model group and BZXD group, of which 174 were up-regulated, 147 were down-regulated in the BZXD group, and 7 proteins were expressed only in the model group. One hundred ninty-three differential proteins were displayed between the model group and the normal group, of which 123 proteins were up-regulated and 70 were down-regulated in the model group.@*CONCLUSION@#2-DE protein expression profiles of synovial tissue in CIA rats are established, and many differential proteins are discovered. Further analysis on the differential proteins may serve as a new method to study the moleculer mechanism of BZXD in treating the rheumatoid arthritis.