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ObjectiveBased on the experience of traditional quality evaluation， the quality of Atractylodis Macrocephalae Rhizoma（AMR） with different production methods such as direct seeding， transplanting after seedling raising， topping and non-topping， and difference in growth years was compared. MethodVernier caliper was used to measure the trait data of AMR in different production methods. Paraffin sections of AMR with different production methods were made by saffron solid green staining， and the microstructure was observed. The contents of water-soluble and alcohol-soluble extracts in AMR with different production methods were determined according to the 2020 edition of Chinese Pharmacopoeia. The content of water-soluble total polysaccharides in AMR with different production methods was detected by sulfuric acid-anthrone method. Fiber analyzer was used to detect the content of fiber components in AMR with different production methods. The contents of monosaccharides， oligosaccharides and some secondary metabolites in AMR with different production methods were detected by ultra performance liquid chromatography（UPLC）， and the differences of chemical components were compared by multivariate statistical analysis methods such as principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA）. ResultIn terms of traits， the 3-year-old AMR with direct seeding and without topping was close to the high-quality AMR with "phoenix-head and crane-neck， strong sweetness and clear aroma" recorded in ancient materia medica， followed by the 3-year-old AMR with topping after transplanting， while the 2-year-old AMR with topping after transplanting with high market circulation rate was generally fat and strong with mild odor. In the microscopic aspect， the arrangement of xylem vessels and fiber bundles in the 3-year-old samples formed two obvious rings. Compared with the 2-year-old samples cultivated in Bozhou and Zhejiang， the 3-year-old samples without topping after transplanting had more wood fibers. In terms of chemical composition， the contents of 70% ethanol extract， fructose， glucose， sucrose， 1-kestose， atractylenolide Ⅰ， chlorogenic acid， neochlorogenic acid， cryptochlorogenic acid and other components in 3-year-old AMR with direct seeding and without topping were significantly higher than those in the other three samples（P<0.05）. The contents of cellulose， 70% ethanol extract， sucrose， atractylenolide Ⅰ， atractylone and other components in 3-year-old AMR with topping after transplanting were significantly higher than those in the 2-year-old AMR with high market circulation rate（P<0.05）， while the contents of water-soluble extract and water-soluble total polysaccharides in 2-year-old samples with topping after transplanting were significantly higher than those in the 3-year-old AMR with topping after transplanting， direct seeding and without topping（P<0.05）. ConclusionUnder the current mainstream production mode， too much manual intervention makes AMR heavily enriched in polysaccharides and increased the yield， but the accumulation of sweet substances， fragrant substances and fiber substances is insufficient， which affects its quality. The current quality standard of AMR has some shortcomings in guiding the high quality production of it， it is suggested to revise the quality standard of AMR， supplement the quantitative analysis of secondary metabolites， and strengthen the production of imitation wild AMR.
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ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties， the chemical components of Sojae Semen Nigrum（SSN） and Sojae Semen Praeparatum（SSP） were systematically compared and analyzed， and the main fermentation products in different fermentation time were quantitatively analyzed， so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used for the structural identification of the chemical constituents of SSN and SSP， and with the aid of Progenesis QI v2.3 software， the negative ion mode was employed for principal component analysis（PCA） pattern recognition， and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis（OPLS-DA） for two-dimensional data to obtain S-plot， and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC， and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period， and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate（PITC） pre-column derivatization and high performance liquid chromatography（HPLC） gradient elution， and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value（TAV）. ResultA total of 19 kinds of differential components were screened out， mainly soybean saponins and isoflavones， and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP， glycoside bond hydrolysis mainly occurred， and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones， the content of flavor substances such as amino acids increased gradually. In the post-fermentation process， protein degradation mainly occurred， after 8 d of post-fermentation， the content of isoflavones was basically stable， while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP， such as the TAV of glutamate is always ahead of other flavor substances， and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation， combined with the traditional evaluation index of "fragrant and strong"， it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant， delicious and slightly sweet"， and at the same time， the post-fermentation index compounds such as glutamic acid， alanine and valine should be added as the quality control indicators of SSP， so as to standardize the production process and improve the quality of SSP.
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ObjectiveBased on the traditional quality evaluation methods summarized in previous dynasties， this paper systematically contrasted cultivated Astragali Radix（CA） and wild-simulated Astragali Radix（WA） from the aspects of character， microstructure and chemical composition by modern technological means. MethodThe collected CA and WA were compared in characters and microscopic characteristics in cross section， and comparative analysis were performed on the contents of cellulose， extracts， carbohydrate， total flavonoids， total saponins， etc. Then ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometer（UPLC-Q-TOF-MS） and desorption electrospray ionization mass spectrometry imaging（DESI-MSI） were used to comparatively analyze the secondary metabolites and their spatial distributions in the xylem and phloem of CA and WA. ResultIn terms of characters， the characters and sectional features of WA was consistent with the characteristics of high-quality Astragali Radix， while the CA was quite different from the traditional high-quality Astragali Radix. In terms of microscopy， the phellem layer of CA was thin， and the section fissures were mostly distributed through the cambium in a long strip shape without obvious growth ring characteristics. The cork layer of WA was thick， and the cracks in the section were distributed in the center of the xylem and the outer edge of the phloem in an irregular cavity shape. The cambium was tight without cracks， and had obvious characteristics of a growth ring. In terms of chemical composition， the contents of water-soluble extract， 80% ethanol extract and sucrose of CA was significantly higher than those of WA， while the contents of total saponins， lignin and hemicellulose were significantly lower than those of WA. And the contents of 100% ethanol extract， total polysaccharides and total flavonoids in both of them were generally similar， but slightly higher in WA. The contents of 2 kinds of monoacyl-substituted flavonoid glycosides in the xylem of WA was significantly higher than those of CA， while the contents of 2 kinds of flavonoid aglycones and one flavonoid glycoside were on the contrary. The contents of 7 saponins in phloem of WA were significantly higher than those of CA. ConclusionThere are significant differences between CA and WA in characters， microstructure and chemical components， in which CA has a fast growth rate and a short planting period， and the primary metabolites such as water-soluble extracts and sucrose are more enriched， which is the reason for its firm texture and sweetness being significantly higher than those of WA. However， the contents of lignin， hemicellulose and some secondary metabolites in WA are significantly higher than those in the CA， which are close to the traditional description of characters and quality. Based on the results of this study， it is suggested to strengthen the production of WA， improve the supply capacity of WA， and gradually upgrade the current standard. It is recommended to increase the contents of monoacyl-substituted flavonoid glycosides， total saponins and other indicators that can characterize different production methods， so as to guide the high-quality production of Astragali Radix.
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By reviewing the research history on quality comparison between wild and cultivated Chinese crude drugs， this paper systematically combed the relevant research reports since the 1950s， and summarized and analyzed the results of existing comparative studies， and found that the existing comparative research on the quality of wild and cultivated Chinese crude drugs were mainly focused on several aspects， including characteristics， microstructures， chemical compositions， pharmacodynamic effects， and genetic diversity. Among these， comparative studies of chemical compositions have been the dominant approach， with a particular emphasis on comparing the contents of index components. However， research on pharmacodynamic effects remained relatively limited. Due to various factors such as sample quantity， sample origin， growth period and cultivation methods， the differences in quality between wild and cultivated Chinese crude drugs vary significantly. In general， most wild Chinese crude drugs exhibited higher quality than cultivated products， with significant differences in their characteristics. The contents and proportions of some chemical components underwent noticeable changes， particularly with a marked increase in the proportion of primary metabolites after cultivation. The quality of cultivated Chinese crude drugs is closely related to the cultivation practices employed. Chinese crude drugs produced through wild nurturing， simulated wild planting， ecological cultivation， and other similar methods demonstrate quality levels comparable to those of wild Chinese crude drugs. Based on the analysis results， it is recommended to explicitly specify the cultivation practices and cultivation period of cultivated Chinese crude drugs in comparative studies of the quality between wild and cultivated Chinese crude drugs. Multiple technical approaches， including characteristics， microscopy， non-targeted metabolomics combined with quantitative analysis of differential components， and bioefficacy evaluation， should be employed to comprehensively assess the quality disparities between wild and cultivated Chinese crude drugs. Moreover， research efforts should be intensified to investigate the changes in pharmacodynamic effects resulting from differences in plant cell wall composition， primary metabolites， and secondary metabolites， in order to guide the production of high-quality Chinese crude drugs.
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Aluminum is a light metal which is rich in the earth's crust and widely used. Recently, the adverse health effects of environmental and occupational aluminum exposure on human have attracted more and more attention. Aluminum exposure has toxic effects on the central nervous system and is believed to be closely related to the development and progression of Alzheimer's disease. The neurotoxic mechanism of aluminum is complex, especially the role of regulated cell death (RCD) in aluminum-induced neuronal death remains to be further studied. RCD refers to all modes of cell death regulated by multiple intracellular signal transduction pathways under physiological and pathological conditions, including apoptosis, necroptosis, autophagy, pyroptosis, and ferroptosis. This review summarized the morphological characteristics and mechanisms of each RCD mode in the process of aluminum-induced neuronal death, and discussed the relationship and transformation between different RCD modes, providing a new scientific basis for future studies on the treatment and intervention of neurotoxicity induced by aluminum exposure.
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By reviewing ancient materia medica， prescription books， medical books and modern literature， this article makes a systematic textual research on the name， origin， commercial specification， producing area， harvesting and processing of Scutellariae Radix used in famous classical formulas according to the historical development， in order to provide a reference for the development and utilization of famous classical formulas. Scutellariae Radix was first named as Qian， Qin and so on， while Huangqin was used as the official name in ancient literature， as well as many aliases such as Fuchang， Huangwen and Dufu. The main origin of Scutellariae Radix in the past dynasties was Scutellaria baicalensis， and other Scutellaria plants were also used for Scutellariae Radix at different times. The medicinal parts of Scutellariae Radix in all dynasties are roots， since the Northern and Southern dynasties， it has gradually differentiated into Ziqin and Kuqin with different efficacy according to their different growth stages and characteristics， and continued to this day. At present， most of Scutellariae Radix are Ziqin in the market， due to the influence of cultivation cost， index component content requirements and other factors. The production area of Scutellariae Radix recorded in ancient literature was wide and gradually expanded from the Yangtze River basin to the northward. Since modern times， Chengde city of Hebei province has been respected as a geo-authentic area， now S. baicalensis is cultivated in a wide area， spreading over north and northeast China. Scutellariae Radix was mostly used as raw products in the early period， and a variety of processed products appeared in the Ming dynasty. Development continues to this day， Scutellariae Radix mainly includes raw products and wine-processed products. According to the research conclusion， it is suggested that S. baicalensis should be used as Scutellariae Radix in famous classical formulas， and the selection of its commercial specifications and processed products can be comprehensively determined according to the requirements of original prescription and the clinical effect.
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Aiming at the current situation of performance testing of hemodialysis extracorporeal circulation tubing, which has slow efficiency, inaccurate measurement, and inconvenient testing, a portable detection system for testing the performance of hemodialysis extracorporeal circulation tubing is designed. The system mainly includes a hardware system and a software system. The hardware system uses STM32F407 single-chip microcomputer as the core to design the driving control of the roller pump; the software system uses the C++ real-time operating system, and the flow detection data is transmitted to the upper computer through RS485 communication and displayed. Experimental showed that the system detects the accuracy and the stability of the flow rate. It has the characteristics of stability and high precision. The relative error of the experimental measurement is within the range of ±10%. The weight of the whole machine is 2 kg, which improves the efficiency by 50% compared with the traditional detection method.
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Computers , Equipment Design , Extracorporeal Circulation , Microcomputers , Renal Dialysis , SoftwareABSTRACT
Tripartite motif (TRIM) family proteins are important effectors of innate immunity against viral infections. Here we identified TRIM35 as a regulator of TRAF3 activation. Deficiency in or inhibition of TRIM35 suppressed the production of type I interferon (IFN) in response to viral infection. Trim35-deficient mice were more susceptible to influenza A virus (IAV) infection than were wild-type mice. TRIM35 promoted the RIG-I-mediated signaling by catalyzing Lys63-linked polyubiquitination of TRAF3 and the subsequent formation of a signaling complex with VISA and TBK1. IAV PB2 polymerase countered the innate antiviral immune response by impeding the Lys63-linked polyubiquitination and activation of TRAF3. TRIM35 mediated Lys48-linked polyubiquitination and proteasomal degradation of IAV PB2, thereby antagonizing its suppression of TRAF3 activation. Our in vitro and in vivo findings thus reveal novel roles of TRIM35, through catalyzing Lys63- or Lys48-linked polyubiquitination, in RIG-I antiviral immunity and mechanism of defense against IAV infection.
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Animals , Dogs , Humans , Mice , A549 Cells , Apoptosis Regulatory Proteins/immunology , DEAD Box Protein 58/immunology , HEK293 Cells , Influenza A Virus, H1N1 Subtype/immunology , Madin Darby Canine Kidney Cells , Mice, Knockout , Orthomyxoviridae Infections/pathology , Proteolysis , Signal Transduction/immunology , THP-1 Cells , TNF Receptor-Associated Factor 3/immunology , Ubiquitination/immunology , Viral Proteins/immunologyABSTRACT
Bone marrow stromal cells (BMSCs) are a kind of stem cells with multiple differentiation potential in bone marrow.BMSCs have been widely used in tissue engineering,cell transplantation,gene therapy and organ transplantation,due to their characteristics of wide range of sources,weak immunogenicity weak,easily transfected by exogenous gene,long survival time in the host,multi-directional differentiation,etc.Cerebral ischemia is caused by neurological impairment,which is the most common cause of death and quality-of-life impairments.The clinical manifestations of the patients with cerebral ischemia are motor function failure,sensory dysfunction and abnormal mental consciousness.A large number of studies have reported that BMSCs transplantation has the therapeutic effects of body sensory and motor function recovery,and can treat ischemic stroke.BMSCs transplantation has brought new hope for the clinical treatment of ischemic cerebrovascular disease.In this paper,the recent progress in the study of BMSCs transplantation for ischemic stroke was reviewed.The mechanism,pathways,influencing factors and clinical application of BMSCs transplantation were summarized.
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BACKGROUND: Hydroxyapatite/chitosan (HA/CS) complex may act as a drug carrier for drug release, but little is reported about the release amount and antibacterial effect of minocycline-HA/CS (Mino-HA/CS) complex. OBJECTIVE: To investigate the in vitro release and antibacterial property of Mino-HA/CS complex. METHODS: HA/CS and Mino-HA/CS were prepared using co-precipitation method. The surface and cross-section features of the complexes were observed under scanning electron microscopy. The porosities were measured according to Archimedes Principle. The release of minocycline hydrochloride was measured by high performance liquid chromatography with the simulated saliva as drug release media. In vitro antibacterial effect on Porphyromonas gingivalis and Staphylococcus aureus were measured by bacteria-inhibiting ring method. Biological toxicities were evaluated via cel counting kit-8cel proliferation assay. RESULTS AND CONCLUSION: The porosity of Mino-HA/CS was larger than that of HA/CS, with the average porosity of 53.99%. Single-day release amount of Mino-HA/CS could maintain at the level of 0.5-1 μg per day for a long-term. Bacteriostatic rings of Porphyromonas gingivalis and Staphylococcus aureus stil existed clearly after 7 days. Cel proliferation assays showed that Mino-HA/CS extract had the significant effect on promoting cel proliferation. These findings indicate that the Mino-HA/CS sustains the release of minocycline at a relatively stable level within a longer period, shows good inhibitory effect on Porphyromonas gingivalis and Staphylococcus aureus and promotes the proliferation of periodontal ligament cel s.
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Lysostaphin is highly effective on eliminating methicillin resistant Staphylococcus aureus (MRSA). In order to achieve controlled release of lysostaphin, a biocompatible drug carrier is needed. Hydroxyapatite/chitosan (HA/CS) composites were chosen to carry lysostaphin and sample composites with different weight ratios of HA to CS, including 80/20, 70/30, 60/40, and 40/60, were prepared. Multiple analyses were performed to determine the structural and physicochemical properties of the composites, including scanning electron microscopy, X-ray diffraction and Fourier transform infrared spectroscopy. We immersed HA/CS composites loaded with 1 wt% lysostaphin to test in vitro release activity and cultured MC3T3-E1 cells to carry out biocompatibility test. The result of the release behavior of the composites revealed that the controlled release of lysostaphin from 60/40 HA/CS composites was the highest release rate of (87.4 ± 2.8)%, which lasted for 120 hours. In biocompatibility testing, MC3T3-E1 cells were able to proliferate on the surface of these composites, and the extract liquid from the composites could increase the growth of the cells. These results demonstrate the controlled release of lysostaphin from HA/CS composites and their biocompatibility, suggesting the potential application of these composites to bone injury and infection applications.
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Objective To investigate the effect of dexmedetomidine on the expression of purinergic P2X4 receptor (P2X4R) mRNA and p38 mitogen-activated protein kinase (p38 MAPK) mRNA in spinal cord in a rat model of neuropathic pain (NP).Methods Seventy-two male SD rats weighing 180-220 g were randomly assigned into 3 groups ( n =24 each):sham operation group (group S),group NP and dexmedetomidine group ( group DEX).The animals were anesthetized with intraperitoneal 10％ choral hydrate 350 mg/kg.NP was induced by chronic constrictive injury (CCI).The right sciatic nerve was exposed and 4 ligatures were placed on sciatic nerve at 1 mm intervals with 4-0 silk thread in groups NP and DEX.In group S,the right sciatic nerve was only exposed but not ligated.Dexmedetomidine 50μg/kg was injected intraperitoneally daily staring from the end of operation,while the equal volume of normal saline was injected in groups S and NT.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured on day 1 before operation and on day 1,3,7 and 14 after operation.Six animals were sacrificed after MWT and TWL measurement on day 1,3,7 and 14 after operation in each group,the L4～6 segment of spinal cords was removed for determination of P2X4 R mRNA and p38 MAPK mRNA expression by RT-PCR.Results Compared with group S,MWT and TWL were significantly decreased and the expression of P2X4 R mRNA and p38 MAPK mRNA was significantly up-regulated after operation in groups NP and DEX ( P ＜0.05).Compared with group NP,TWL and MWT were significantly increased and the expression of P2X4 R mRNA and p38 MAPK mRNA was significantly down-regulated after operation in group DEX ( P ＜ 0.05 ).TWL and MWT were significantly higher and the expression of P2X4 R mRNA and p38 MAPK mRNA was significantly lower on day 1,3 and 14 after operation than on day 7 after operation in groups NiP and DEX ( P ＜ 0.05).Conclusion The mechanism by which dexmedetomidine attenuates NP is related to inhibition of the expression of P2X4 R mRNA and p38 MAPK in rats.
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Objective To investigate the effects of dexmedetomidine (Dex) on the neuronal apoptosis in spinal dorsal horn in a rat model of chronic neuropathic pain.Methods Seventy-two adult male SD rats weighing 180-220 g were randomly divided into 3 groups ( n =24 each):sham operation group (group S) ; chronic constrictive injury (CCI) group; Dex + CCI group (group D).Two ligatures were placed on right sciatic nerve at 1 mm intervals with 4-0 silk thread in groups CCI and D.In group D Dex 50 μg/kg was injected intraperitoneally once a day starting from the end of operation until the animals were sacrificed.Paw withdrawal threshold to mechanical stimulation with yon Frey filament (PWT) and paw withdrawal latency to thermal stimulation (PWL) were measured at one day before (T0,baseline) and on the 3rd,7th and 14th day after operation (T1,2,3).Six animals were sacrificed at each time points (T1,2,3) after the measurement of PWT and PWL.Their lumbar segments (L4,5)were removed for examination with transmission electron microscope and detection of Bcl-2 and caspase-3 expression (by immune-histochemistry).Results CCI significantly decreased PWT and PWL,increased Bcl-2 and caspase-3 expression at T1,2,3 and induced apoptosis in spinal dorsal horn neurons in group CCI as compared with group S.Intraperitoneal Dex significantly attenuated CCI-induced mechanical and thermal hyperalgesia and neuronal apoptosis in group D as compared with group CCI.Dex injected intraperitoneally further increased Bcl-2 expression but decreased caspase-3 expression in group D as compared with group CCI.Conclusion Reduction in neuronal apoptosis in spinal dorsal horn is involved in the attenuation of neuropathic pain by Dex.
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Objective To prepare totel glucosides in paeony(TGP) pellets by extrusion-spheronization and to optimize the formulation of TGP sustained-release pellets coated with Eudragit RS 100.Methods The TGP sustained-release pellets were prepared by mini Glatt fluidized bed coating method.The factors to influence the drug release behaviors and their properties were evaluated.Results The optimal TGP sustained-release pellet was shown significant sustained-releasing at the loading weight of copolymers of 6% and the plasticizer concentration of 10%.And the curves of cumulative drug release were fit for Peppas and Higuchi equation.Conclusion The pellets showed an obviously sustained-release effect.
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0.999 5), RSD were no more than 3.0%. The recovery rates were 98.0%-102.0%. There was no significant difference between the results of this method and EDTA-Na2 titration. Conclusion The method is rapid, accurate, simple, little disturbance and is acceptable in determination of total hardness in drinking water.