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This paper compared the differences between two kinds of Bufonis Venenum produced by Bufo gargarizans gargarizans and B. gararizans andrewsi, and verified the rationality of the market value orientation of Bufonis Venenum based on the zebrafish mo-del. Twenty batches of Bufonis Venenum from Jiangsu province, Hebei province, Liaoning province, Jilin province, and Liangshan, Sichuan province, including B. gargarizans gargarizans and B. gararizans andrewsi, were collected. The UHPLC-LTQ-Orbitrap-MS combined with principal component analysis was used to compare the differences between two kinds of Bufonis Venenum. According to the limiting conditions of VIP>1, FC<0.5 or FC>2.0, and peak total area ratio>1%, 9 differential markers were determined, which were cinobufagin, cinobufotalin, arenobufagin, resibufogenin, scillaredin A, resibufagin, 3-(N-suberoylargininyl)-arenobufagin, 3-(N-suberoylargininyl)-marinobufagin, and 3-(N-suberoylargininyl)-resibufogenin. The content of 20 batches of Bufonis Venenum was determined according to the Chinese Pharmacopoeia(2020 edition) by high-performance liquid chromatography, and the 2 batches of Bufonis Venenum, CS7(8.99% of total content) and CS9(5.03% of total content), with the largest difference in the total content of the three quality control indexes of the Chinese Pharmacopoeia(bufalin, cinobufagin, and resibufogenin) were selected to evaluate their anti-liver tumor activity based on the zebrafish model. The tumor inhibition rates of the 2 batches were 38.06% and 45.29%, respectively, proving that only using the quality control indexes of the Chinese Pharmacopoeia as the value orientation of Bufonis Venenum market circulation was unreasonable. This research provides data support for the effective utilization of Bufonis Venenum resources and the establishment of a rational quality evaluation system of Bufonis Venenum.
Subject(s)
Animals , Zebrafish , Bufanolides/analysis , Bufonidae , Chromatography, High Pressure Liquid , Quality Control , Cell Line, TumorABSTRACT
ObjectiveTo establish blood stasis models in zebrafish using three inducers and select the optimal model for evaluating the activity of Notoginseng Radix et Rhizoma in promoting blood circulation. MethodArachidonic acid （AA）， ponatinib， and isoprenaline （ISO） were used to induce blood stasis models in zebrafish. A normal group， a model group， a positive drug group， and Notoginseng Radix et Rhizoma water extract freeze-dried powder groups at different concentrations were set up. The staining intensity of cardiac erythrocytes and the fluorescence intensity of cardiac apoptotic cells were calculated， the anti-thrombotic effect and anti-myocardial hypoxia activity of Notoginseng Radix et Rhizoma were evaluated. The activities of water extract and 70% methanol extract of Notoginseng Radix et Rhizoma were compared based on the preferred AA- and ISO-induced blood stasis models in zebrafish and the difference in the chemical composition was analyzed by UHPLC LTQ-Orbitrap MS/MS. ResultAfter induction by AA and ponatinib， the staining intensity of cardiac erythrocytes was reduced （P<0.01）， and the fluorescence intensity of cardiac apoptotic cells increased after the induction by ISO （P<0.01）. The freeze-dried powder of the water extract of Notoginseng Radix et Rhizoma could antagonize the thrombosis in the AA-induced model （P<0.01） and the myocardial apoptosis in the ISO-induced model （P<0.05）， while no significant improvement in the thrombosis was observed in the ponatinib-induced model. The freeze-dried powder of 70% methanol extract of Notoginseng Radix et Rhizoma could inhibit myocardial apoptosis in the ISO-induced blood stasis model （P<0.01）， and the effect was stronger than that of the freeze-dried powder of Notoginseng Radix et Rhizoma water extract. The difference in chemical composition lay in some saponins （such as ginsenoside Re）， amino acids， and acetylenic alcohols. ConclusionAA， ponatinib， and ISO all can serve as inducers for the blood stasis model in zebrafish. AA- and ISO-induced models can be used to evaluate the activity of freeze-dried powder of Notoginseng Radix et Rhizoma water extract in promoting blood circulation. The chemical compositions of the freeze-dried powders of Notoginseng Radix et Rhizoma extracted with water and 70% methanol are quite different. For the ISO-induced blood stasis model， the freeze-dried powder of Notoginseng Radix et Rhizoma extracted with 70% methanol has a stronger ability against myocardial hypoxia. Saponins and acetylenic alcohols may be closely related to the effects of promoting blood circulation and resolving blood stasis.
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Objective:To explore the effect of Ershiwuwei Zhenzhu tablets (EZT) on microvascular function. Method:The zebrafish models of thrombosis, microvascular defect and vascular endothelial injury were induced by using arachidonic acid, simvastatin and ponatinib respectively, and treated with EZT, astragaloside or asprin. To evaluate the protective effect of EZT on vascular endothelium and its effect on thrombus formation, zebrafish heart output and blood flow velocity were counted, and the vascular area of the zebrafish intestine and the intervascular diameter were calculated. The thrombus in the tail vein was observed under microscope. Result:Compared with model group,EZT improved the cardiac output (PP-1, and promoted angiogenesis in zebrafish at concentrations of 0.11, 0.33, 1 mg·L-1. Compared with the model group, the vascular diameter of the zebrafish internode was significantly increased at the concentrations of 33 mg·L-1 (P-1 (P-1(PConclusion:EZT could improve microvascular dysfunction, and its mechanism may be related to the reduction of vascular endothelial damage to promote its angiogenesis and the improvement of microvascular hemodynamics to reduce thrombus formation.
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Objective @# A titanium personalized orthodontic bracket is designed and manufactured using the technology of computed tomography (CT), computer aided design (CAD), finite element analysis (FEA), and selective laser melting (SLM) to well match the patient tooth so as to realize mold-free manufacturing. @*Methods@#A model of a titanium personalized orthodontic bracket which is built by CT scanning machine and Pro/E software, is imported into Ansys software to carry out finite element simulation analysis with nonlinear contact method. Then, the titanium personalized orthodontic bracket is proposed after the model data above is imported to SLM molding equipment via titanium powder melted with layer by layer using a high-speed scanning galvanometer. @*Results @#The maximum equivalent stress of the titanium personalized orthodontic bracket is distributed uniformly and reasonably, the titanium personalized orthodontic bracket formed by SLM molding equipment can provide high accuracy and there is a high similarity between the bottom of the orthodontic bracket and the tooth surface shape.@*Conclusion @#The combination of CT, CAD, FEA, SLM technology can fulfill model-free manufacturing of the personalized orthodontic bracket and thus shorten the manufacturing cycle.
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A method for the simultaneous determination of oxygen and nitrogen in synthetic diamonds by inert gas high temperature extraction-impulse heating method was proposed. The sample weight, the selection of analysis power and the calibration curves of oxygen and nitrogen were discussed. Oxygen and nitrogen in analytical samples are determined. Values of RSDs (n=7) for oxygen and nitrogen were less than 4. 5% and 4. 0% respectively. The analytical results of oxygen and nitrogen obtained by the proposed method were in good agreement with those by inert gas fusion-impulse heating method.
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A method for the determination of Fe, Co, Mn and Ni in synthetic diamonds by inductively coupled plasma atomic emission spectrometry ( ICP-AES) was proposed. The synthetic diamond sample was decomposed completely, while the sample was burned in air at 1000 ℃ for 10 h, and then a mixed acid of H2 SO4 , aqua regia and HClO4 was used for the dissolving the residue of the sample. In this method, the limits of detection of Fe, Co, Mn and Ni were 0. 0147, 0. 0018, 0. 0006 and 0. 0027 mg/L, respectively. Under the optimum condition, Fe, Co, Mn and Ni in synthetic diamond sample were determined. The values of RSDs (n=7) were less than 0. 5%. The recoveries of added standard were 94. 0%-105. 0%.
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Studies demonstrated that natural killer T (NKT) cells are involved in the diverse immunomodulatory responses.But the mechanism that NKT cells regulate pathogenic T helper cells (Th) are still unclear.Recent researches found that Th17 cells are associated with a variety of autoimmune disease and play a crucial role in autoimmune diseases,which is very different from previous thesis that Th1 and Th2 are the key factors in the immune response in the patients with autoimmune disease.Studies both in vitro and in vivo also documented that invariant NKT cells (iNKT) inhibit the differentiation of CD4+ T cells toward Th1 and Th17 cells at the different mechanisms.The concept of NKT cells,the regulation of NKT cells to Th1 and Th17 cells in the autoimmune diseases were reviewed.
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<p><b>OBJECTIVE</b>To observe and compare the clinical effect of Dingweiban and Xieban manipulation, and to compare the change of the deviation of spinous processes between two methods.</p><p><b>METHODS</b>One hundred and twenty-two cases were divided into two groups. Sixty-two cases were treated with Dingweiban manipulation method and 60 cases by Xieban manipulation. The changes of Fairbank scores, the clinical effects and the difference of the deviation of the spinous processes (L3, L4, L5) from the lumbar posterior-anterior X-ray were compared.</p><p><b>RESULTS</b>The scores before and after treatment and 3 months after treatment were compared. There were significant differences between two groups (P < 0.05) by nonparametric test. The result of Dingweiban manipulation group: 53 cases cured, 5 cases better, 3 cases effective and 1 case no effect. The result of clinical Xieban manipulation group: 43 cases cured, 6 cases better, 7 cases effective and 4 cases no effect. The clinical effects had significant differences after treatment and 3 months after treatment (P < 0.05, P < 0.0l) by nonparametric test. After the first treating, there was clear difference of the deviations' distance of the L4 spinous process compared with the Xieban manipulation group (P < 0.05). After the last treating, there were clear differences of the deviation distance of the L4 and L5 spinous processes compared with the Xieban manipulation group (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>Dingweiban manipulation is better than Xieban manipulation in effects and has influence on the deviation of spinous processes, especially for the L5 spinous process.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Intervertebral Disc Displacement , Therapeutics , Lumbar Vertebrae , Manipulation, Spinal , MethodsABSTRACT
<p><b>OBJECTIVE</b>To understand the evolution of HIV-1 CRF07_BC envelope, we performed a longitudinal study on two patients during their early HIV-1 infection.</p><p><b>METHODS</b>RNA was extracted from the plasma of the individuals and the C2-C5 fragments of the gp120 gene of HIV-1 were amplified by RT-PCR. Purified DNA segments were inserted into T easy vector and transformed into E. coli Top 10 competent cells. Positive clones were identified by blue-white screening, confirmed by PCR and sequenced by ABI 3700.</p><p><b>RESULTS</b>The samples were collected from the patients every 6 months from seroconversion time. The genetic diversity and divergence in env gene showed consistent increases over time. Our sequence analysis also revealed obvious non-synonymous change in env C1, C3 and V4 regions among these samples.</p><p><b>CONCLUSION</b>The results support the concept that the consistent pattern of viral evolution existed during early phase of HIV-1 infection. C1, C3 and V4 region of env gene may be mainly immunological target during AIDS progression.</p>
Subject(s)
Adult , Humans , Male , Genetic Variation , HIV Infections , Virology , HIV-1 , Genetics , Longitudinal Studies , env Gene Products, Human Immunodeficiency Virus , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To investigate the subtype distribution and the prevalence of sequence characteristics of HIV-1 strains in Beijing residents during 2006 and to analyze the relationship between distribution of HIV-1 subtypes and transmission routines.</p><p><b>METHODS</b>Blood samples from 32 new confirmed HIV-1 infected individuals from Beijing residents in 2006 and separated plasma specimens were collected. RNAs were extracted and the gag and env gene were amplified by RT-PCR and nest-PCR. PCR products were sequenced directly and phylogenetic analyses of gag and env gene were performed using the MEGA2 software.</p><p><b>RESULTS</b>Among 32 HIV-1 plasma samples, 22 gag and 4 env gene fragments were amplified and analyzed. Five HIV-1 subtypes or circulating recombinant forms(CRFs) of HIV-1 including Thai B (2 strains), B (9 strains), C (2 strains), CRF07_BC (5 strains), CRF01 AE (4 strains) were identified being circulated in Beijing. The gene divergences of gag gene inside the subtypes were 6.6%, 4.3%, 6.8%, 4.9% and 3.0% in subtype B, Thai B, C, CRF01_AE and CRF07_BC respectively. Subtypes B were predominant in Beijing, accounted for 40.9% among 22 samples.</p><p><b>CONCLUSION</b>Five HIV-1 subtypes were identified in Beijing and the surveillance of HIV-1 gene variation should be paid more attention to.</p>
Subject(s)
Humans , China , HIV-1 , Classification , Genetics , Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , env Gene Products, Human Immunodeficiency Virus , Genetics , gag Gene Products, Human Immunodeficiency Virus , GeneticsABSTRACT
OBJECTIVE@#To discuss forensic identification of floating shoulder injury (FSI).@*METHODS@#To analyze fifteen cases of FSI which were accepted from Jan. 1993 to Jan. 2006, including 15 shoulder neck fracture, 13 clavide stem fracture and 2 distal end clavide fracture, the function of shoulder joint was evaluated six months after injure considering the following three aspects: result of forensic examination such as X-ray photograph, CT and MRI, the injurers' symptom, objective sign and joint function, shoulder joint territory, degree of pain and local muscle power.@*RESULTS@#Basing on the curative effect standard of Herscovic, all cases were divided into good. Modest, worst, which included 2, 4, 9 cases respectively; referring the standard of GA35-92, GB18667-2002, all cases were divided into six, seven, eight, nine and ten degree, which included 2,9,2,1,1 cases respectively.@*CONCLUSION@#As a special powerful injure, FSI always companied with concurrent and multiple injure, and characterized by missed, incorrect and delayed diagnosis and infelicitous treatment, which lead to the high frequency and degree of injure. To prevent missed and incorrect forensic identification, we should have a full realization of the particularity of FSI, and evaluate the function of shoulder all-sidely, objectively and synseticaly.