BACKGROUNDOur previous research has suggested that platelet activating factor receptor was related to atherosclerosis. The present study investigated the effect of a platelet activating factor receptor antagonist-WEB2086 on angiogenesis in aortal plaque and ischemic hindlimb of apolipoprotein E-deficient mice.

METHODSEight-week-old apolipoprotein E-deficient mice were fed with a 0.15% cholesterol diet to develop advanced lesions. At age 32 weeks unilateral hindlimb ischemia was surgically induced and the mice were divided into two groups: with or without WEB2086 mixed with their drinking water (4.3 mg in 100 ml). At age 40 weeks blood was collected from the orbit for measurement of serum lipids and an enzyme linked immunosorbent assay was used to determine platelet activating factor and oxidized low density lipoprotein in the gastrocnemius and aorta. Whole-Mount CD31 stain and plaque-associated sprouting have been used to estimate angiogenesis in plaque from the aorta and laser Doppler perfusion imaging and immunohistochemical expression of von Willebrand factor have been used to estimate angiogenesis in ischemic hindlimb.

RESULTSThe lipid composition of serum was not different between the groups. However, the amount of platelet activating factor and oxidized low density lipoprotein detected in the aorta was significantly higher than that in the gastrocnemius of ischemic hindlimb. The ratio of lesion to aorta levels was significantly reduced by administration of WEB2086, (31.52 +/- 6.18)% vs (55.58 +/- 8.34)%, P < 0.01. The mean density of intimal capillaries in atherosclerotic plaque, (31.13 +/- 9.20)% vs (57.74 +/- 11.28)%, P < 0.01, and the mean number of sprouts per aorta were significantly reduced, 183.92 +/- 34.17 vs 392.54 +/- 76.79, P < 0.01, in the WEB2086 group. Blood flow (0.85 +/- 0.12 vs 0.45 +/- 0.06, P < 0.01) and capillary density of ischemic hindlimb (1.18 +/- 0.17 vs 0.53 +/- 0.09, P < 0.01) were markedly increased in apolipoprotein E-deficient mice treated with WEB2086 versus controls.

CONCLUSIONThe study provides evidence that WEB2086 can inhibit angiogenesis in atherosclerotic plaque but promote it in ischemic hindlimb.

Animals ; Apolipoproteins E ; deficiency ; Atherosclerosis ; physiopathology ; Azepines ; pharmacology ; Capillaries ; Cholesterol ; blood ; Hindlimb ; blood supply ; Ischemia ; physiopathology ; Lipoproteins, LDL ; blood ; physiology ; Male ; Mice ; Neovascularization, Physiologic ; drug effects ; Platelet Activating Factor ; analysis ; Platelet Membrane Glycoproteins ; antagonists & inhibitors ; Receptors, G-Protein-Coupled ; antagonists & inhibitors ; Triazoles ; pharmacology

OBJECTIVETo explore the influence of oxidized high-density lipoprotein (oxHDL) on the maturation and migration of bone marrow-derived dendritic cells (BMDCs) from C57BL/6J mice.

METHODSThe C57BL/6J mice bone marrow cell suspension was prepared and purified. Recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4) were used to promote monocytes to differentiate and suppress lymphocytes. Then 50 microg/mL oxHDL was added to stimulate BMDCs, using 50 microg/mL high-density lipoprotein (HDL) as homologous protein control, PBS as negative control, and 1 microg/mL lipopolysaccharide (LPS) as positive control. The CD86 and MHCII expression rates were detected with fluorescence-activated cell sorting (FACS). Liquid scintillation counting (LSC) was used in mixed lymphocyte reactions (MLRs) to reflect the ability of BMDCs in stimulating the proliferation of homologous T cells. Levels of cytokines IL-12 and IL-10 were detected by ELISA. The cell migration was evaluated with the transwell system.

RESULTSCompared with PBS group, the expressions of CD86 and MHCII, counts per minute of MLRs, secretion of IL-12 and IL-10, and number of migrated cells in oxHDL group and LPS group significantly increased (all P<0.05), while the increment was less in oxHDL group than LPS group. The number of migrated cells in oxHDL group was about twice of that in HDL group.

CONCLUSIONOxHDL may promote the maturation and migration of BMDCs in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; physiology ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; physiology ; Humans ; Lipoproteins, HDL ; metabolism ; pharmacology ; Lipoproteins, LDL ; metabolism ; pharmacology ; Mice ; Mice, Inbred C57BL

OBJECTIVE To explore the effect of water extract of notopterygium incisum (WN) on neuropathic pain and its molecular biological mechanism.METHODS Pain behavior test to detect the effects of WN on thermal hyperalgesia and me-chanical hyperalgesia in acute pain and chronic constriction injury(CCI)induced neuropathic pain model.Immunohistochemis-try and qPCR were used to detect the expression of TRPV1 .Calcium imaging was used to detect calcium influx of capsaicin (TRPV1 agonist) in CCI mice after oral administration of WN.RESULTS WN significantly reduced the thermal hyperalgesia and mechanical hyperalgesia in acute pain model and CCI model (P<0.001).The expression of TRPV1 in protein level and mRNA level in DRG neurons were significantly inhibited by administration of WN(P<0.05).The response of DRG neurons to capsaicin was significantly inhibited by the treatment of WN(P<0.001).CONCLUSION WN alleviated CCI-induced me-chanical allodynia and thermal hypersensitivity via modulating TRPV1 .Our results revealed a new molecular biological mecha-nism of analgesia effect to traditional Chinese medicine.

BACKGROUNDStudy of the relationship between mast cells and atherosclerosis is mostly dependent on pathological observation and cytology experiments. To investigate the effects of mast cells degranulation on plaque and their possible mechanisms we used apolipoprotein E knockout mice which had been placed perivascular common carotid collar with mast cells degranulator compound 48-80.

METHODSForty apolipoprotein E knockout mice were fed a western-type diet and operated on with placement of perivascular right common carotid collar. Four weeks after surgery, the mice were intraperitoneally injected with compound 48-80 (0.5 mg/kg) or D-Hanks every other day for 4 times. The serum lipids and activity of tryptase were measured. Tissue sections were stained with hematoxylin and eosin. Corresponding sections were stained with toluidine blue and immunohistochemically with antibodies against macrophage-specific antigen, alpha-smooth muscle actin, interleukin-1beta and von Willebrand factor. Simultaneously, basic fibroblast growth factor was detected by in situ hybridization and immunofluorescence.

RESULTSNo pathological change was observed in common carotid non-collar placement but atherogenesis in common carotid collar placement of both groups. There was a significant increase in plaque area ((5.85+/-0.75) x 10(4) vs (0.86+/-0.28) x 10(4) microm(2), P<0.05), the degree of lumen stenosis ((81+/-15)% vs (41+/-12)%, P<0.05), the activity of tryptase in serum ((0.57+/-0.13) U/L vs (0.36+/-0.10) U/L, P<0.05), and the percentage of degranulated mast cells ((80.6+/-17.8)% vs (13.5+/-4.1)%, P<0.05). The expressions of macrophage-specific antigen, alpha-smooth muscle actin, interleukin-1beta, basic fibroblast growth factor and the density of neovessel in plaque were more in the compound 48-80 group than in the control group.

CONCLUSIONSPerivascular common carotid collar placement can promote atherosclerotic plaque formation in apolipoprotein E knockout mice. Compound 48-80 increases plaque area and the degree of lumen stenosis by the mechanism that compound 48-80 promotes proliferation of smooth muscle cells and aggregation of macrophages. Compound 48-80 promotes angiogenesis in plaque. The mechanism is potentially that compound 48-80 increases the expressions of basic fibroblast growth factor mRNA and protein in plaque. Compound 48-80 enhances the expression of interleukin-1beta in plaque.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; chemically induced ; genetics ; metabolism ; pathology ; Carotid Arteries ; drug effects ; pathology ; Fluorescent Antibody Technique ; Immunohistochemistry ; In Situ Hybridization ; In Vitro Techniques ; Male ; Mast Cells ; drug effects ; metabolism ; Mice ; Mice, Knockout ; p-Methoxy-N-methylphenethylamine ; pharmacology

Objective:To compare the consistency and detection capability of seven 2019-nCoV nucleic acid detection kits, and provide reference for detection method selection of clinical laboratory and diagnosis of new coronavirus pneumonia.Methods:Two batches of pharyngeal swab samples were collected from tenpatients with confirmed infection of 2019-nCoV and 10 suspected patients with negative 2019-nCoV test results during January 29 to February 5, 2020 in Shenzhen Luohu People′s Hospital. Seven kinds of kits were labeled as ato g and used for nucleic acid detection respectively to evaluate the consistency of the test results of the clinical samples. A 2019-nCoV positive specimen was selected and diluted to 5-concentration gradient plates (Level-1 to 5) with RNase-free water. The positive detection rate and intra-batch repeatability of different brands of kits were compared.Results:The negative and positive coincidence rates of twenty clinical samples tested by six kinds of kits were 100%, and the positive and negative coincidence rate was 8/10 and 10/10 for the other kit, respectively. The results of intra-batch repeatability showed the CVs of viral loads tested by these seven kits were all less than 5%. In the concentration range of Level-1 to 3, the detection capability for open reading frame (ORF)1ab gene of Kit b,d and f was lower than Kit a,c,e and g, and the detection capability of kit e and g was the highest (14/15). The detection capability for N gene of Kit a (15/15) was higher than the other 5 kits. The comprehensive analysis of the detection capability for ORF1ab and N gene showedthat Kit d had the lowest detection capability (ORF1ab:40%,N:53%), and there was no significant difference in the detection capability of Kit a, b, c, e, and f.Conclusions:There was no significant difference in the accuracy and repeatability of the seven kits for positive samples with high viral loads, and the detection performance was good; but some kits had poor detection capability for weak positive samples. It is suggested that the weak positive samples should be rechecked by at least two manufacturers′ kits to ensure the accuracy of the results.

ABSTRACT:OBJECTIVE To study the effects of Qingluo Tongbi Compound(QLT)on pain behavior and COX-2 mRNA ex-pression in dorsal root ganglion(DRG)and blood PGE2 concentration,and to explore the mechanisms of pain in rheumatoid ar-thritis.METHODS The mice were randomly divided into control group,celecoxib 30 mg∕kg,QLT 4.35 g∕kg,QLT 8.70 g∕kg and QLT 1 7.4 g∕kg groups.The mice pain threshold change were measured by hot plate method,and body torsion times were tested by acetic acid twisting method.The collagen-induced arthritis(CIA) was induced by collagenⅡ in DBA∕1 mice. The mice were randomly divided into control group,CIA group,celecoxib 30 mg∕kg group,QLT 8.70 g∕kg.The paws swell-ing,mechanical withdrawal threshold(MWT)and thermal withdrawal latency(TWL)were recorded.After treated with corre-sponding drugs by intragastric administration for four weeks,COX-2 mRNA in DRG and PGE2 in mice were quantified.RE-SULTS QLT reduced pain threshold and body torsion times in pain model mice(P <0.01);compared with the CIA group,the QLT group decreased the paws swelling(P <0.05),up-regulated MWT(P <0.01) and PWL(P <0.05),down-regulated the expression level of COX-2 mRNA in DRG and PGE2 concentrations in blood.CONCLUSION QLT shows certain analgesic ac-tion,which might be related to the inhibitory effect of COX-2 mRNA expression in DRG and PGE2 in blood.

OBJECTIVETo investigate the protective effect of retarded removal of the unilateral necrotic testis after long-time (> 24 h) spermatic cord torsion on the contralateral testis in rats.

METHODSThirty-three male SD rats aged 21 -42 days were divided into a sham-operation group (n = 11), a torsion-reservation group (n = 12) and a torsion-orchiectomy group (n = 10). The rats of the sham-operation group received dartos pouch orchidopexy on the left testis, while those of the latter two groups underwent 720 degrees unilateral spermatic cord torsion on the left side. Ninety-six hours later, the rats of the torsion-reservation group received detorsion with the ipsilateral testis preserved, while those of the torsion-orchiectomy group underwent orchiectomy. Three months after operation, blood samples were obtained from the rats for measurement of serum testosterone and antisperm antibodies by ELISA, and meanwhile testes and epididymides were harvested for determination of the volumes of various structures and the diameter of seminiferous tubules with stereological methods.

RESULTSThere were no significant differences in the level of serum testosterone among the three groups. Anti-sperm antibody positive was found in only 1 animal in the torsion-reservation group. The Leydig cell nuclei in the contralateral testis appeared larger in the torsion groups than in the sham-operation group. Marked morphological changes were observed in 1, 3 and 0 of the animals in the sham-operation, torsion-reservation and torsion-orchiectomy group, respectively, mainly including atrophy of seminiferous tubules and reduced number of spermatogenic cells. The volume of the contralateral testis was increased by 19% and 21% in the torsion-reservation and torsion-orchiectomy group, respectively, in comparison with that in the sham-operation group (P < 0.05). No significant differences were observed in the volume of seminiferous tubules of the contralateral testis among the sham-operation, torsion-reservation and torsion-orchiectomy groups ([1.15 +/- 0.07], [1.30 +/- 0.04] and [1.35 +/- 0.05] cm3). The volume of the interstitial tissue was significantly increased in the latter two groups ([0.36 +/- 0.02 and 0.34 +/- 0.03] cm3) as compared with the former ([0.25 +/- 0.02] cm3) (P < 0.05). The diameters of the seminiferous tubules exhibited no significant differences among the three groups ([226.00 +/- 7.00], [223.00 +/- 6.00] and [221.00 +/- 3.0] microm).

CONCLUSIONLong-time unilateral spermatic cord torsion may result in compensatory hypertrophy of the contralateral testis, and orchiectomy does not significantly affect the histology of the contralateral testis and epididymis.

Animals ; Epididymis ; pathology ; Male ; Necrosis ; Orchiectomy ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; pathology ; surgery ; Testis ; pathology ; surgery

OBJECTIVETo explore the infarct sites in patients with inferior wall acute myocardial infarction (AMI) concomitant with ST segment elevation in leads V1-V3 and leads V3R-V5R.

METHODSFive patients diagnosed as inferior, right ventricular, and anteroseptal walls AMI at admission were enrolled. Electrocardiographic data and results of isotope 99mTc-methoxyisobutylisonitrile (MIBI) myocardial perfusion imaging and coronary angiography (CAG) were analyzed.

RESULTSElectrocardiogram showed that ST segment significantly elevated in standard leads II, III, aVF, and leads V1-V3, V3R-V5R in all five patients. The magnitude of ST segment elevation was maximal in lead V1 and decreased gradually from lead V1 to V3 and from lead V1 to V3R-V5R. There was isotope 99mTc-MIBI myocardial perfusion imaging defect in inferior and basal inferior-septal walls. CAG showed that right coronary artery was infarct-related artery.

CONCLUSIONSThe diagnostic criteria for basal inferior-septal wall AMI can be formulated as follows: (1) ST segment elevates > or = 2 mm in lead V1 in the clinical setting of inferior wall AMI; (2) the magnitude of ST segment elevation is the tallest in lead V1 and decreases gradually from lead V1 to V3 and from lead V1 to V3R-V5R. With two conditions above, the basal inferior-septal wall AMI should be diagnosed.

Aged ; Coronary Angiography ; Diagnostic Errors ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; diagnostic imaging ; physiopathology ; Radionuclide Imaging

Mast cells are widely distributed in various parts of the body, especially in the mucosal surface between the body and the external environment. Mast cell is one of the important immune cells and plays important roles in innate immunity, adaptive immunity and immune regulation. Previous researches have shown that excessive activation of mast cells is closely related to the development of allergic and inflammatory diseases such as asthma, allergic rhinitis, food allergies, acute and chronic itching. Mast cells infiltrate into the inflammation site and release various allergic mediators during the occurrence and development of these diseases. Therefore, termination of mast cell activation can be one of the effective methods for the treatment of allergic and inflammatory diseases, and receptors related to mast cell activation are potential targets for the development of anti-allergic drugs. There are many receptors related to mast cell activation, and the effects mediated by different receptors varied from each other. In the recent years, new mast cell receptors are being discovered, but there are not many literatures discussing the possible functions of these newly discovered receptors. This review aims to summarize the receptors involved in mast cell activation and classify related receptors according to their effects.
Asthma ; immunology ; Humans ; Hypersensitivity ; immunology ; Immunity, Innate ; Inflammation ; immunology ; Mast Cells ; cytology ; immunology

Itching is an unpleasant sensation that can cause physical and psychological discomfort. It often occurs in a variety of diseases. In addition to the common chemical medicines, the application of traditional Chinese medicines(TCMs) have attracted much attention for itch. As one of the classic antipruritic formula, Xiaofengsan is widely used by the later generations of doctors based on the edition of Orthodox Manual of External Diseases(Wài Kē Zhèng Zōng). It can be used to remove wind, dehumidify, clear heat and nourish blood by taking into account the complicated pathogenesis of skin diseases. In modern clinics, Xiaofengsan is commonly used for the treatment of rubella, eczema, acute and chronic urticaria, contact dermatitis, allergic dermatitis, psoriasis and other types of dermatitis with remarkable antipruritic effect. Its mechanism is related to its anti-allergic reaction, anti-allergy, anti-inflammatory, anti-histamine,immune regulation etc. This article reviews the research literature of Xiaofengsan and its addition and subtraction in modern pharmacological experimental research and clinical treatment in the past 15 years, and analyzes the current research situation of Xiaofengsan in the topic of antipruritic effect. The focus is on the compatibility characteristics of formula and the component study of single herb after being decomposed of Xiaofengsan, and the main material basis of antipruritic effect is summarized. It was found that Schizonepetae Herba, Saposhnikoviae Radix, Arctii Fructus, Cicadae Periostracum and Sophorae Flavescentis Radix were the main medicines for its antipruritic effect,which was helpful to expand the clinical value of Xiaofengsan and deepen its mechanisms of anti-itching.