Active components and potential mechanism of Taohong Siwu Decoction in regulating ischemic stroke based on target cell trapping combined with network pharmacology, molecular docking, and experimental validation / 中国中药杂志

The potential anti-stroke active components in Taohong Siwu Decoction(THSWD) were identified by target cell trapping coupled with ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS). The underlying mechanism of active components in THSWD in the treatment of ischemic stroke(IS) was explored by network pharmacology, molecular docking, and experimental validation. The UPLC-Q-TOF-MS technology combined with the UNIFI data analysis platform was used to analyze the composition of the cellular fragmentation fluid after co-incubation of THSWD with target cells. The targets of potential active components and IS were collected by network pharmacology, and the common targets underwent protein-protein interaction(PPI), Gene Ontology(GO), and Kyoto Encyclopedia of Genes and Genomes(KEGG) signaling pathway enrichment analyses. The target cell trapping component-core target-signaling pathway network was constructed, and the active components were molecularly docked to the top targets in the PPI network, followed by pharmacodynamic validation in vitro. Fifteen active components were identified in the target cellular fragmentation fluid, including bicyclic monoterpenes, cyanoglycosides, flavonols, quinoid chalcones, phenylpropanoids, and tannins. As revealed by the analysis of network pharmacology, THSWD presumably regulated PI3K-AKT, FoxO, MAPK, Jak-STAT, VEGF, HIF-1, and other signaling pathways to affect inflammatory cascade reaction, angiogenesis, oxidative stress, pyroptosis, apoptosis, and other pathological processes via paeoniflorin, butylphthalide, dehydrated safflower yellow B, 3,4-dicaffeoylquinic acid, amygdalin, paeoniflorin, and ligusticolactone. Molecular docking and in vitro pharmacodynamic validation revealed that the target cell trapping active components could promote neovascularization in rat brain microvascular endothelial cells(rBMECs) in the oxygen-glucose deprivation/reoxygenation(OGD/R) model. The application of target cell trapping coupled with UPLC-Q-TOF-MS technology can rapidly screen out the potential active components in THSWD. The active components of THSWD can be predicted to intervene in the pathogenesis of IS through network pharmacology, and molecular docking combined with experimental validation can further clarify the efficacy, thus providing a theoretical basis for research ideas on the pharmacodynamic substance basis of traditional Chinese medicine compounds.

Pharmacodynamic Mechanism of Kuanxiong Aerosol for Vasodilation and Improvement of Myocardial Ischemia / 中国结合医学杂志

OBJECTIVE@#To explore the effect of Kuanxiong Aerosol (KXA) on isoproterenol (ISO)-induced myocardial injury in rat models.@*METHODS@#Totally 24 rats were radomly divided into control, ISO, KXA low-dose and high-dose groups according to the randomized block design method, and were administered by intragastric administration for 10 consecutive days, and on the 9th and 10th days, rats were injected with ISO for 2 consecutive days to construct an acute myocardial ischemia model to evaluate the improvement of myocardial ischemia by KXA. In addition, the diastolic effect of KXA on rat thoracic aorta and its regulation of ion channels were tested by in vitro vascular tension test. The influence of KXA on the expression of calcium-CaM-dependent protein kinase II (CaMK II)/extracellular regulated protein kinases (ERK) signaling pathway has also been tested.@*RESULTS@#KXA significantly reduced the ISO-induced increase in ST-segment, interventricular septal thickness, cardiac mass index and cardiac tissue pathological changes in rats. Moreover, the relaxation of isolated thoracic arterial rings that had been precontracted using norepinephrine (NE) or potassium chloride (KCl) was increased after KXA treatment in an endothelium-independent manner, and was attenuated by preincubation with verapamil, but not with tetraethylammonium chloride, 4-aminopyridine, glibenclamide, or barium chloride. KXA pretreatment attenuated vasoconstriction induced by CaCl2 in Ca2+-free solutions containing K+ or NE. In addition, KXA pretreatment inhibited accumulation of Ca2+ in A7r5 cells mediated by KCl and NE and significantly decreased p-CaMK II and p-ERK levels.@*CONCLUSION@#KXA may inhibit influx and release of calcium and activate the CaMK II/ERK signaling pathway to produce vasodilatory effects, thereby improving myocardial injury.

Visualization Analysis of Henoch-schonlein Purpura Treated by Traditional Chinese Medicine Based on Citespace / 中国实验方剂学杂志

Objective::To finding the main research contents, research frontier, author and institutional cooperation of traditional Chinese medicine(TCM) for treating henoch-schonlein purpura(HSP). Providing reference for the research and development of TCM for treating the disease. Method::Using Citespace to analyze 2 878 TCM articles related to HSP retrieved from CNKI, cluster analysis and burst analysis of literature keywords, co-occurring authors and institutional cooperation analysis. Result::Since 1995, the number of related literature was growing rapidly and had been stable at more than 100 per year after 2005.Cluster analysis showed 32 clusters, consisting of 396 nodes and 638 lines. The main clustering results include Children with allergic purpura, blood-activating and stasis-resolving drug, Henoch-Schonlein purpura nephritis, blood-cooling drugs, clinical observation, etc. Break analysis yielded 52 emergent words. It can be seen that TCM treatment of HSP is mainly based on cooling blood, followed by activating blood to eliminate stagnation and clearing heat. Commonly used drugs are Moutan Cortex, Paeoniae Radix Rubra, and Rehmanniae Radix, etc. Clinically, it pays more attention to the experience of famous doctors, research on Children with allergic purpura, etc.The author's cooperation network has obtained the maps of the three main cooperation teams with DING Ying, SUN Yi-qiu and HE Ping as the core. The Density of institutional cooperation network is 0.007 1. Conclusion::The main research contents of TCM for treating HSP include Children with allergic purpura, blood-activating and stasis-resolving drug, HSP nephritis, blood-cooling drugs, clinical observation, etc. Children with allergic purpura, experience from famous doctor, HSP nephritis and clinical efficacy is the foremost current research hotspot. A number of research teams have been formed that are relatively stable, but the institutional cooperation is scattered.

Linc-OIP5 in the breast cancer cells regulates angiogenesis of human umbilical vein endothelial cells through YAP1/Notch/NRP1 signaling circuit at a tumor microenvironment

BACKGROUND: LincRNAs have been revealed to be tightly associated with various tumorigeneses and cancer development, but the roles of specific lincRNA on tumor-related angiogenesis was hardly studied. Here, we aimed to investigate whether linc-OIP5 in breast cancer cells affects the angiogenesis of HUVECs and whether the linc-OIP5 regulations are involved in angiogenesis-related Notch and Hippo signaling pathways. METHODS: A trans-well system co-cultured HUVECs with linc-OIP5 knockdown breast cancer cell MDA-MB-231 was utilized to study the proliferation, migration and tube formation abilities of HUVECs and alterations of related signaling indicators in breast cancer cells and their conditioned medium through a series of cell and molecular experiments. RESULTS: Overexpressed linc-OIP5, YAP1, and JAG1 were found in breast cancer cell lines MCF7 and MDA-MB-231 and the expression levels of YAP1 and JAG1 were proportional to the breast cancer tissue grades. MDA-MB-231 cells with linc-OIP5 knockdown led to weakened proliferation, migration, and tube formation capacity of co-cultured HUVECs. Besides, linc-OIP5 knockdown in co-cultured MDA-MB-231 cells showed downregulated YAP1 and JAG1 expression, combined with a reduced JAG1 level in conditioned medium. Furthermore, a disrupted DLL4/Notch/NRP1 signaling in co-cultured HUVECs were also discovered under this condition. CONCLUSION: Hence, linc-OIP5 in MDA-MB-231 breast cancer cells may act on the upstream of the YAP1/Notch/NRP1 signaling circuit to affect proliferation, migration, and tube formation of co-cultured HUVECs in a non-cellular direct contact way through JAG1 in conditioned medium. These findings at least partially provide a new angiogenic signaling circuit in breast cancers and suggest linc-OIP5 could be considered as a therapeutic target in angiogenesis of breast cancers.

Clinical application of tibial composite saphenous nervetissue flap for the treatment of traumatic bone-skin defect of leg / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the effect of tibial composite saphenous nerve tissue flap in repairing traumatic bone-skin defect of leg.</p><p><b>METHODS</b>From February 2009 to May 2015, 23 patients with focile skin defect caused by trauma were treated, including 18 males and 5 females, aged from 8 to 59 years old with an average age of 35.6 years old. Length of tibia bone cortex defect ranged from 4 to 12 cm, width ranged from 2 to 2.5cm, skin defect ranged from 8 cm× 3 cm to 18 cm× 11 cm, all the wound surfaces had purulent exudation. Cross shift and ipsilateral shift methods of tibial composite saphenous nerve tissue flap were used to reconstruct continuity of the injured limb tibial brace and repair wound surface. Cutting flap ranged from 9 cm× 4 cm to 20 cm × 12 cm, bone flap length was 6 to 15 cm, width was 1.5 to 2.5 cm. Fracture healing time and recovery of limb function were observed.</p><p><b>RESULTS</b>At 3 to 5 months after operation, bone flap and area of affected area were healed with an average of 4.7 months. Twenty-three cases were followed up for 6 months to 5 years with an average of 30.3 months. Thirteen patients were able to walk without abnormal gait and could be engaged into original work. According to Enneking evaluation systems, 13 patients obtained excellent results, 8 moderate and 2 poor.</p><p><b>CONCLUSIONS</b>With tibial composite saphenous nerve tissue flap transplantation at stage one to repair tibia and skin defects according to specific condition of affected limbs has a high rate of success and creates less damage to region of limb injury, which is an ideal method to repair skin defect of focile.</p>

Effects of zinc on biological behavior of hepatocellular carcinoma cell line BEL-7404 / 中国病理生理杂志

AIM:To observe the effects of exogenous zinc on the biological behavior of hepatocellular carcino -ma ( HCC) cell line BEL-7404.METHODS: BEL-7404 cells were cultured with zinc sulfate at various concentrations . The intracellular concentration of zinc , cell viability , cell cycle , cell apoptosis and migration and invasion abilities were measured by TSQ fluorescent probe , MTT assay, DNA ploid analysis, acridine orange/ethidium bromide fluorescence stai-ning and Transwell assay , respectively .The mRNA and protein expression levels of albumin in the BEL-7404 cells were determined by real-time PCR and Western blot , respectively .RESULTS: With the elevated concentration of zinc in cul-ture condition, the concentration of zinc in the BEL-7404 cells was increased (P<0.05).The cell viability and migration and invasion abilities were decreased , while the apoptotic rate was increased (P<0.05).The cells in G0/G1 phase were decreased, while the cells in G2/M phase were increased.Additionally, the mRNA and protein expression of albumin also increased (P<0.05).CONCLUSION:The zinc ion inhibits the cell viability as well as migration and invasion abilities , blocks the cells in G 2/M phase , and may reduce cell malignant phenotype .

Mental health is the most important factor influencing quality of life in elderly left behind when families migrate out of rural China / A saúde mental é o fator mais importante que influencia a qualidade de vida de idosos deixados para trás quando as famílias emigram da China rural / La salud mental es el factor más importante que influye en la calidad de vida en los adultos mayores que se quedan cuando las familias emigran fuera de las zonas rurales de China

OBJECTIVES: to investigate the quality of life and the associated factors on left behind elderly in rural China. METHOD: the research was conducted cluster sampling to select 456 elderly left behind when family members migrated out of rural China to participate in a cross-sectional study by completing a general data questionnaire and Quality of Life questionnaire. RESULTS: 91.5% of the elderly requested psychological counseling and education. For the elderly, scores for mental health (39.56±13.73) were significantly lower compared with Chinese standard data (61.6±13.7) (P<0.001). Age, chronic disease type, gender, residence pattern and economic support from children were the main psychological factors influencing this population. CONCLUSION: mental health is the most important factor influencing quality of life of elderly left behind when family members migrated out of rural China. Our study suggested a need to widely establish a general practitioner team for psychological intervention for improving rural elderly people's Quality of Life level, especially in the psychological domain. .

OBJETIVOS: investigar a qualidade de vida e fatores associados de idosos deixados para trás na China rural. MÉTODO: foi realizada amostragem por conglomerado para selecionar 456 idosos deixados para trás quando os membros da família emigram da China rural. Este é um estudo transversal com preenchimento de um questionário de dados gerais e de qualidade de vida. RESULTADOS: 91,5% dos idosos convidados solicitaram aconselhamento e educação psicológicos. Para os idosos, os escores de saúde mental (39,56±13,73) foram significativamente menores em comparação aos dados padrões chineses (61,6±13,7) (p<0,001). A idade, tipo de doença crônica, gênero, tipo de moradia e suporte econômico dos filhos foram os principais fatores psicológicos que influenciaram essa população. CONCLUSÃO: a saúde mental é o fator mais importante que influencia a qualidade de vida de idosos deixados para trás quando os familiares emigraram da China rural. Este estudo sugere a necessidade de criação de ampla equipe de generalistas para intervenção psicológica, com o intuito de melhorar o nível de qualidade de vida de idosos de áreas rurais, especialmente no domínio psicológico. .

OBJETIVOS: investigar la calidad de vida y los factores asociados a los adultos mayores que se quedan en las zonas rurales de China. MÉTODO: la investigación se realizó por medio de muestreo por conglomerados para seleccionar 456 adultos mayores que se quedaron cuando los miembros de la familia emigraron de zonas rurales de China, para participar en un estudio de corte transversal, completando un cuestionario de datos generales y cuestionario de calidad de vida. RESULTADOS: el 91.5% de los adultos mayores solicitó asistencia psicológica y educación. Para los adultos mayores, las puntuaciones de salud mental (39.56±13.73) fueron significativamente más bajos en comparación con los datos estándar de China (61.6±13.7) (p<0.001). La edad, el tipo de enfermedad crónica, el género, el patrón de residencia y el apoyo económico de los hijos fueron los principales factores psicológicos que influencian esta población. CONCLUSIÓN: la salud mental es el factor más importante que influye la calidad de vida de los adultos mayores que se quedan cuando los miembros de la familia emigran de las zonas rurales de China. Nuestro estudio sugiere la necesidad de establecer ampliamente un equipo médico general para la intervención psicológica para mejorar el nivel de la Calidad de Vida de la población rural de edad avanzada, sobre todo en el dominio psicológico. .

The role of arecoline on hepatic insulin resistance in type 2 diabetes rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effects of arecoline on hepatic insulin resistance in type 2 diabetes rats and to elucidate its possible mechanism.</p><p><b>METHODS</b>Forty five Wistar rats were fed with high fructose diet for 12 weeks to induce type 2 diabetic rat model. rats were randomly divided into 5 groups (n = 8): control group, model group and model group were treated with different dose (0, 0.5, 1, 5 mg/kg) of arecoline. After 4 weeks, the fasting blood glucose, blood lipid and insulin level measured , mRNA expression of liver constitutive androstane receptor (CAR), pregnane X receptor (PXR), glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were detected by reverse transcription polymerase chain reaction (RT-PCR), the protein expression of p-AKT and glucose transporter4 (GLUT4) were detected by Western blot.</p><p><b>RESULTS</b>1.5 mg/kg arecoline could significantly decrease the level of fasting blood glucose, blood lipid, blood insulin level and liver G6Pase, PEPCK, IL-6, TNF-alpha mRNA level in type 2 diabetes rats. 1.5 mg/kg arecoline also could significantly increase CAR, PXR mRNA level and p-AKT and GLUT4 protein expression.</p><p><b>CONCLUSION</b>Arecoline improved hepatic insulin resistance in type 2 diabetes rats by increasing the mRNA levels of CAR and PXR leading to the creased glucose metabolism and inflammation related genes expression.</p>

Protective effect of medicated serum prepared with taohong siwu tang on hydrogen peroxide-induced human umbilical vein endothelial cells / 中国中药杂志

<p><b>OBJECTIVE</b>To study the protective effect of medicated serum prepared with Taohong Siwu Tang on hydrogen peroxide-injured human umbilical vein endothelial cells (HUVECs).</p><p><b>METHOD</b>Sprague Dawley rats were orally administered with 10 mL x kg(-1) extracts from Taohong Siwu Tang (1.75 g crude drug), twice a day for three days, in order to prepare medicated serum of Taohong Siwu Tang. The effect of medicated serum pre-treated with Taohong Siwu Tang (with concentrations of 5%, 10%, 15%) on reduction of H2O2-induced cell activity was detected MTT. Cell morphological changes were observed under microscope. The effect of medicated serum prepared with Taohong Siwu Tang on the apoptosis and antioxidant capacity of HUVECs was detected with the content of malondialdehyde (MDA) and dehydrogenase (LDH), the activity of superoxide dismutase (SOD) and AO/EB staining. The expression of Caspase-3 was determined by western blot.</p><p><b>RESULT</b>Compared with the blank serum control group, cell were significantly less active after being damaged by H2O2 (400 micromol x L(-1)). Medicated serum could significantly improve the SOD activity, reduce the levels of LDH and MDA, and inhibite the expression of Caspase-3. AO/EB staining and inverted microscope also showed that medicated serum prepared with Taohong Siwu Tang could reduce cell apoptosis induced by H2O2.</p><p><b>CONCLUSION</b>Medicated serum prepared with Taohong Siwu Tang has significant protective effect on HUVECs injured by H2O2.</p>

Effect of Taohong Siwu decoction on angiogenesis of medicine-induced incomplete-abortion in early pregnancy rats and expressions of Ang-1, Ang-2 and Tie-2 / 中国中药杂志

<p><b>OBJECTIVE</b>To observe the effect of Taohong Siwu decoction (THSWD) on micro-vascular density (MVD) in rat uterus, the content of angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) in serum, and the expression of tyrosine kinasa receptor (Tie-2) in uterus.</p><p><b>METHOD</b>Early pregnancy rats were intragastrically administrated with misoprostol (100 microg x kg(-1)) and mifepristong (8.3 mg x kg(-1)) to established the incomplete-abortion model. The incomplete-abortion rats were randomly divided into the model group (the same volume of distilled water), the positive control group (at the daily dose of 4.3 g x kg(-1) Motherwort Particles), and THSWD-treated groups (at the daily dose of 18.0, 9.0 and 4.5 g x kg(-1)). Pregnant rats were taken as the control group (the same volume of distilled water). After the successive oral administration for 7 days, blood was collected from aorta abdominalis, and rat uterine tissues were collected. The content of serum Ang-1 and Ang-2 were detected by ELISA; And the levels of Tie-2 and MVD in uterine tissues were detected by SP immunohistochemistry.</p><p><b>RESULT</b>THSWD remarkably increased the levels of MVD in uterus of medicine-induced abortion rats, the content of Ang-1 and Ang-2 in serum, and the expression of Tie-2 in uterine tissues.</p><p><b>CONCLUSION</b>THSWD has the effect in markedly promoting angiogenesis in incomplete-abortion rats. Its mechanism may be related to the regulation of concentrations of Ang-1 and Ang-2 in serum and Tie-2 in uterine tissues.</p>

Thinking on the design scheme of clinical trial with the example of migraine treated by acupuncture / 中国针灸

Through the examples of explanatory and pragmatic trials on acupuncture for migraine and the comparison between research objectives, experiment environment and trial designs, the design of current clinical trial should focus on: (1) Making a point of continuum research on efficacy-effectiveness at trial design type; (2) Thinking highly of therapeutic effect criteria that could have a better show of trial purpose; (3) More use of qualitative research embedded in the randomized controlled trials. With these improvements, the effects of acupuncture could be evaluated more scientifically, objectively and comprehensively.

Impact of lesion size on the detection rate of non-palpable breast malignant lesions / 中国医学科学院学报

<p><b>OBJECTIVE</b>To evaluate the impact of lesion size on the detection rate of non-palpable breast malignant lesions and determine whether lesion size should prompt biopsy of non-palpable breast lesions.</p><p><b>METHODS</b>The study included 816 ultrasonographically detected non-palpable breast lesions. We divided the lesions into five groups based on their largest diameters: ≤0.5cm, 0.6-1.0cm, 1.1-1.5cm, 1.6-2.0 cm, and >2.0 cm. The detection rate of malignancies of different sizes were compared among these lesions, Breast Imaging Reporting and Data System (BI-RADS) category 2-3 lesions, and BI-RADS grades 4-5 lesions. The feasibility of using lesion size as biopsy indicator for BI-RADS category 2-3 non-palpable breast lesion was analyzed using ROC curve.</p><p><b>RESULTS</b>Of these 816 lesions, 100 (12.3%) were found to be malignant lesions. The detection rate of malignancy significantly increased along with the increase of lesion size (P<0.05). When the BI-RADS category was not considered, the frequency of malignancy in the >2.0 cm group was significantly higher than in other groups (P<0.05) The frequencies of malignancy in the 0.6-1.0 cm group, 1.1-1.5 cm group, and 1.6-2.0 cm group were higher than that in ≤0.5 cm group, but the difference was not significant (P>0.05) For BI RADS category 4 and 5 lesions, the frequency of malignancy in >2.0 cm group was higher than in other groups, but significant difference was only seen between >2.0 cm group and ≤0.5 cm group (P<0.05).</p><p><b>CONCLUSIONS</b>Lesion size may influence the detection rate of malignancy of non palpable breast lesions, and can be used as biopsy indicator of non palpable breast lesions in BI-RADS 2,3 category When we use 1.25cm as threshold,the sensitivity and specificity may be satisfying.</p>

Clinical study of safflower injection in treating and preventing the vascular crisis after free flap transplantation / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the clinical efficacy of Safflower Injection (Al) in treating and preventing the vascular crisis after free flap transplantation.</p><p><b>METHODS</b>Sixty patients undergoing free flap transplantation were randomly assigned to the treatment group and control group according to the visiting sequence, thirty in each. Free flap transplantation was performed on all patients, and medication was given 0. 5 h before flap vascular anastomosis, 1-7 days after surgery. Twenty mL Al was intravenously dripped to patients in the treatment group after adding in 250 mL 5% glucose injection, while Dextran-40 was intravenously dripped to patients in the control group. The medication was conducted once per day. The hemorheology and four indices of blood coagulation [prothrombin time (PT), international normalized ratio (INR), activated partial thromboplastin time (APTT), fibrinogen (FIB)] were compared between the two groups before operation (TO), during operation (T1), 24 h after operation (T2), three days after operation (T3), and seven days after operation (T4). Meanwhile, flaps were observed and adverse reaction recorded. The clinical efficacy and safety were compared.</p><p><b>RESULTS</b>Better result was obtained in the treatment group when compared their clinical efficacy (86. 67% vs 60. 00%, P<0.05). The whole blood high and low viscosity, plasma viscosity, red blood cell (RBC) volume, RBC aggregation index all decreased, and RBC deformed index increased in the two groups at T4, showing statistical difference when compared with those at T3 (P<0.05, P<0.01). There was no statistical significance in the four indices of blood coagulation when compared with any time point in the same group (P>0.05). There was no statistical significance in hemorheology and the four indices of blood coagulation between the two groups at the same time point (P>0.05). The adverse reaction rate in the treatment group was lower than that in the control group, showing statistical difference (13.33% vs 30.00%, P<0.05).</p><p><b>CONCLUSIONS</b>AI could effectively prevent and treat the vascular crisis after free flap transplantation. It had less adverse reaction and good safety. It was better than Dextran-40. It was a safe and effective drug to prevent the vascular crisis.</p>

Proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood under the culture conditions of hypoxia and serum deprivation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood (PB-MSCs) were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair.</p><p><b>METHODS</b>MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor (G-CSF) and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension (20% O2 and 2% O2), with or without 10% fetal bovine serum (FBS) conditions: 20% O2 and 10% FBS complete medium (normal medium, N), 20% O2 and serum deprivation medium (D), 2% O2 and 10% FBS complete medium (hypoxia, H), 2% O2 and serum deprivation (HD). Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling (TUNEL) staining.</p><p><b>RESULTS</b>Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow (BM-MSCs), and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II (MHC II). They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia.</p><p><b>CONCLUSIONS</b>PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.</p>

Effect of sufentanil on calcium-activated potassium currents in rat abdominal aortic smooth muscle cells / 第二军医大学学报

Objective: To investigate the influence of sufentanil on calcium-activated K+ channels (IKCa) in rat abdominal aortic smooth muscle cells, and to investigate its role in dilation of blood vessels. Methods: Rat abdominal aortic smooth muscle cells (AASMCs) were freshly obtained by enzymatic digestion. Whole-cell voltage-clamp technique was used to assess the effect of sufentanil (1 × 10-8, 3 × 10-8, 1 × 10-7 mol/L) on IKCa. Results: Sufentanil significantly increased the amplitude of IKc compared with the control group (P < 0.05 or P < 0.01). The effect of sufentanil was reversible and in a concentration-dependent manner. Conclusion: Sufentanil can promote the activation of KCa channel in rat AASMCs, which might be related to the vasodilatory effect of sufentanil observed in clinical practice.

Survey on host animal and molecular epidemiology of hantavirus in Chuxiong prefecture, Yunnan province / 中华流行病学杂志

Objective To determine the hosts of hantavirus (HV) and its molecular epidemiological characteristics, to provide evidence for prevention and control on hemorrhagic fever with renal syndrome (HFRS). Methods Rodents were captured by a special trap within the residential area. The antigens of HV in lung tissues were detected by direct immuno-fluorescence assay (DFA). Nucleotide sequences of HV were amplified by RT-PCR with HV genotype-specific primer. The amplified genes were then sequenced. Phylogenetic tree were built on nucleotide sequence with Clusta1X 1.83 software. Results 1421 rodents were captured and classified into 8 species of 4 Genera in the epidemic area within 10 counties of Chuxiong prefecture, Yunnan province, between 2005 and 2006. Out of the 1421 rodents, 1056 (74.31%) of them were Rattus norvegicas and 280 (19.70%) belonged to Rattus flavipectus. The antigens of HV were detected by DFA in lung tissues and the total positive rate of HV was 5.15% (53/ 1029). After applying the sequencing nucleotide method to the 53 positive specimens, data showed that 21 specimens were positive and all of them belonged to Seoul type ( 15 samples were from Rattus norvegicus, 4 samples Rattasflavipectas, 2 samples Rattus nitidas). The partial S segments from 12 specimens were sequenced which appeared homologic with R22, L99 and HLD65 from GenBank in relatively high level (87.1%-99.7%). When compared to 76-118 strain of Hantaan type, their homologic degree was only 64.4%-69.1%. Results from Phylogenetic analysis showed that 12 specimens belonged to Seoul type. As for their homology, they were significantly similar to Seoul type and could be tentatively divided into two subtypes S1 and S3. Conclusion It was confirmed that the Seoul type virus, as HFRS' s pathogenetic agent mainly carried by rats, prevailed widely in Chuxiong prefecture. Owing to the local ecological environment, we also noticed the characteristics of different HV subtypes among Seoul type.

The proapoptotic activity of C-terminal domain of apoptosis-inducing factor (AIF) is separated from its N-terminal

Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein that mediates both NADH-oxidizing and caspase-independent apoptosis. Further, the proapoptotic activity of AIF is located in the C-terminus of AIF, although the precise minimum sequence responsible for apoptosis induction remains to be investigated. In the present study, we generated two truncated AIFs, AIFΔ1-480-FLAG, which is a FLAG-tagged C-terminal peptide comprising amino acids from 481 to 613, and AIF360-480 containing amino acids from 360 to 480 of AIF. We used confocal microscopy to demonstrate that both the truncated proteins are expressed and located in the cytoplasm of transfected cells. AIFΔ1-480 but not AIF360-480 induces apoptosis in transfected cells. We also found that the expression of AIFΔ1-480 could initiate the release of cytochrome c from the mitochondria. The suppression of caspase-9 via siRNA blocked the proapoptotic activity of AIFΔ1-480. Therefore, AIFΔ 1-480 is sufficient for inducing caspase-9-dependent apoptotic signaling, probably by promoting the release of cytochrome c. At last, we generated a chimeric immuno-AIFΔ 1-480 protein, which comprised an HER2 antibody, a Pseudomonas exotoxin A translocation domain and AIFΔ 1-480. Human Jurkat cells transfected with the immuno-AIFΔl-480 gene could express and secrete the chimeric protein, which selectively recognize and kill HER2-overexpressing tumor cells. Our study demonstrates the feasibility of the immuno-AIFΔl-480 gene as a novel approach to treating HER2-overexpressing cancers.

Survey on human rabies cases and its viral molecular biological features in Baoshan city, Yunnan province / 中华流行病学杂志

Objective To understand the epidemiological features of two rabies cases in Baoshan city year 2006 and 2007 and to analyze its source of infection.Methods Questionnaires were used to do the epidemiologieal survey on each of the rabies cases.Brain timue samples of rabies patients were collet to detect the rabies virus by direct immunofluoreseence assay(DFA)and RT-PCR assay.Homology and phylogenetic tree were analyzed.based on the whole nucleotide and deduced amino acid sequence of P,M and N gene of rabies virus followed by molecular epidemiological analysis.Results In July 2006,one human rabies case was identified in Longyang district,and another one in Tengchong county in Baoshan city in 2007.The degrees of exposure of these two patients was all at degreeⅢ.Two brain tissue samples among the dead patients(No.CYN0601H and CYN0701H)were confirmed positive by both DFA and RTPCR assay.The homology analysis of P,M and N gene sequences among CYN0601H,CYN0701H and other rabias strains isolated from other provinces and other counties.showed that the samples in Baoshan city shared the highest homology with the strains in Thailand.Phylogenetic analysis indicated that the two samples were very dose and all belonged to genetype 1 Lyssavirus,with the closest relationship between samples in Baoshan city and strains in Thailand.Conclusion It Was confirmed on the virus molecular level that the two patients in Baoshan city were both suffered from rabies.The prevalent strains in Baoshan city WaS probably imported from foreign country,suggesting that prevention and control measures on rabies virus in the boarder areas of Yunnan should be strengthened.

Aging impairs insulin-stimulated glucose uptake in rat skeletal muscle via suppressing AMPKalpha

Insufficient intracellular fat oxidation is an important contributor to aging-related insulin resistance, while the precise mechanism underlying is unclear. AMP-activated protein kinase (AMPK) is an important regulator of intracellular fat oxidation and was evidenced to play a key role in high-glucose and high-fat induced glucose intolerance. In the present study, we investigated whether altered AMPK expression or activity was also involved in aging-related insulin resistance. Insulin sensitivity of rats' skeletal muscles was evaluated using in-vitro glucose uptake assay. Activity of alpha subunit of AMPK (AMPKalpha) was evaluated by measuring the phosphorylation of both AMPKalpha (P-AMPKalpha) and acetyl-CoA carboxylase (P-ACC), while expression of AMPKalpha was assessed by determining the mRNA levels of AMPKalpha1 and AMPKalpha2, and protein contents of AMPKalpha. Compared with 4-month old rats, 24-month old rats exhibited obviously impaired insulin sensitivity. At the same time, AMPKalpha activity significantly decreased, while AMPKalpha expression did not alter during aging. Glucose transporter 4 expression also decreased in old rats. Compared with 24-month old rats, administration of the specific activator of AMPK, 5-aminoimidazole-4-carboxamide riboside (AICAR), significantly elevated AMPKalpha activity and GluT4 expression. Also, aging-related insulin resistance was significantly ameliorated by AICAR treatment. In conclusion, aging-related insulin resistance is associated with impaired AMPKalpha activity and could be ameliorated by AICAR, thus indicating a possible role of AMPK in aging-induced insulin resistance.

Isolation of Japanese Encephalitis Virus from Brain Tissues of Bat in Yunnan Province / 中国病毒学

In July 1997, a strain (GB30) of virus was isolated from 60 samples of brain tissues of Murina aurata (Chiroptera: Vespertilionidae) co llec ted in Gengma county, Yunnan province, China. Isolation of virus was negative fr om 4 samples of brain tissues of Rousettus leschenaulti (Chiroptera: Pteropo did ae) collected in Gengma. GB30 virus strain could regularly cause illness and dea th in suckling mice, produced evident CPE in BHK21 cells. It agglutinated red b lood cells of dove at pH5.75～7.4. This virus has been identified serologically by hemagglutination inhibition and immunofluorescent tests using Japanese enceph alitis (JE), dengue (DEN) type 1,2,3,4, and chikungunya (CHIK) viruses monoclona l antibodies, and JE and sindbis (SIN) viruses immune sera. It showed specific r eaction to JE virus only and no reaction with DEN 1～4, CHIK and SIN viruses. Th erefore it can be identified as JE virus. This is the first report on the isolat ion of JE virus from Murina aurata. The results showed that bats are conside red as the reservoir and amplifier host of JE virus transmission in nature.