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ObjectiveIn order to understand the quality differences between wild and cultivated Bupleurum chinense(BC), modern analytical techniques were used to systematically compare the quality of wild and cultivated BC in terms of appearance characteristics, primary and secondary metabolites. MethodSamples of wild and cultivated BC were collected from the main production areas of Shanxi, Shaanxi and Hebei, and images of BC were collected and their length and diameter were measured using vernier caliper to compare and analyze the characteristics of the two. Referring to the method under extract of CP in the 2020 edition of Chinese Pharmacopoeia, the extract contents of the two species were determined. The cellulose, hemicellulose and lignin compositions of both were determined using fiber analyzer. Quantitative determination of representative saikosaponins, flavonoids and saccharides in BC by ultra performance liquid chromatography(UPLC), headspace gas chromatography-mass spectrometry(HS-GC-MS) was used to determine the types and relative contents of volatile components, and UPLC-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) coupled with multivariate statistical analysis was used to screen and identify the differential compounds between wild and cultivated BC. ResultThere were significant differences in the appearance characteristics between wild and cultivated BC, the wild BC had a large root head, twisted and thick axial root, rough epidermis, and often had a stem base and lateral root with dark color and strong odor. However, the cultivated BC has long and straight taproots, delicate epidermis, few lateral roots, light root color and light smell. In terms of primary and secondary metabolites, the contents of alcohol-soluble extract and lignin of wild BC was significantly higher than those of cultivated BC, while the contents of water soluble extract and quercitrin was higher than those of cultivated BC, but the difference was not significant. The contents of cellulose, five saikosaponins, rutin, narcissoside and isorhamnetin-3-O-glucoside in cultivated BC were significantly higher than those of wild BC, and the total water-soluble polysaccharides, sucrose, hemicellulose and starch of cultivated BC were higher than those of wild BC, but the difference was not significant. The results of HS-GC-MS identification showed that a total of 67 volatile components were identified in wild and cultivated BC, 59 in wild BC and 51 in cultivated BC, with a total of 43 compounds in both, and the screening based on variable importance in the projection(VIP) value>1 revealed that the differential components were mainly concentrated in the aromatic and fatty acid compounds. The results of UPLC-Q-TOF-MS-based non-targeted metabolomics combined with multivariate statistical analysis showed that the two were significantly different in saikosaponins and the differential compounds had higher response values in cultivated BC. ConclusionThere are significant differences in the appearance, primary and secondary metabolite contents between wild and cultivated BC. At present, the quality evaluation system of cultivated BC is not perfect, and this study provides theoretical references for updating and revising the quality evaluation standard of cultivated BC and guiding the production of high-quality BC.
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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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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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By reviewing ancient materia medica, prescription and medical books, combined with modern literature, the paper made textual research on the name, origin, producing area, quality evaluation, harvesting and processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix, so as to provide a basis for the selection and use of these two herbs in the development of famous classical formulas. Through textual research, it can be found that Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix were mixed together in the early history of China, but the distinction was first made during the Southern and Northern dynasties, and since then there have been constant controversies, and it is not until contemporary times that they are distinguished clearly. In the past dynasties, Duhuo and Qianghuo were used as the rectification of names, some aliases and trade names were also seen. Angelica biserrata is the mainstream origin of Angelicae Pubescentis Radix in the past dynasties, and there are many plants belonging to Angelica, Heracleum and Aralia, which are also used as this medicine. However, the origin of Notopterygii Rhizoma et Radix used in the past dynasties is mostly Notopterygium incisum or N. franchetii, which is relatively uniform. The producing areas of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly concentrated in the western and northwestern regions of China, among which Angelicae Pubescentis Radix is mainly produced in Hubei, Chongqing, Sichuan, Shaanxi and other places, and the border area between Hubei and Chongqing is the geo-authentic area. Notopterygii Rhizoma et Radix is mainly produced in Sichuan, Gansu, Qinghai, Shaanxi and others with the western and northern Sichuan and southern Gansu as the geo-authentic areas. Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix in the past dynasties were harvested in spring and autumn, especially in February and August of the lunar calendar. Angelicae Pubescentis Radix with strong main roots, few branches, firm texture and strong aroma is superior, and Notopterygii Rhizoma et Radix with strong rhizomes, tightly raised knots, purple-brown skin, tight cross-section, strong aroma and silkworm-like shape is superior. The processing methods of Angelicae Pubescentis Radix and Notopterygii Rhizoma et Radix are mostly cut after cutting the reeds, and the raw product is used as medicine. Based on the above research results, it is recommended that the roots of A. biserrata should be used for Angelicae Pubescentis Radix and the roots of N. incisum should be used for Notopterygii Rhizoma et Radix in the development of famous classical formulas, and raw products should be used in the formulas that do not specify processing requirements.
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Objective To explore the surgical technique and clinical effect of percutaneous lumbar endoscopic surgery for central disc herniation.Methods From February 2010 to April 2014,69 consecutive patients (36 males and 33 females) with central lumbar disc herniation underwent percutaneous endoscopic surgery were included in the study.The average age was 31.33±8.27 years,ranging from 16 to 56 years.All operations were performed by increasing the initial puncture angle.First enter the intervertebral space through the ventral nerve root,and then move the guide rod tip step by step to the top of the protrusion.Operative time,intraoperative blood loss and complications were recorded.Visual analogue scale (VAS), Japanese Orthopaedic Association (JOA) and Oswestry disability index (ODI) were followed up before surgery,immediately after surgery,3,12,24 months after surgery.The MacNab criteria were used to evaluate the efficacy at the last follow-up.During the follow-up period,X-ray,CT and MRI were performed to observe the instability and recurrence of the operative segment.Results All of the disc herniations were successfully removed without conversion to open surgery or revision.All patients were followed up for more than 24 months.The VAS scores of low back pain and leg pain were significantly lower than those before operation immediately,3 months,12 months and 24 months after operation (P<0.01).The JOA score was significantly higher in monthly follow-up (P<0.01),and ODI was significantly improved in 3 months and 24 months after operation (P< 0.01).There were 22 excellent cases (31.88%),44 good cases (63.77%),3 cases (4.35%) during the final follow-up,and the excellent and good rate was 95.65%.Dural sac injuries occurred in 1 patient and discharged from hospital in 2 weeks of symptomatic treatment.Lumbar instability and operative segment recurrence were not found during the follow-up period (2 years).Conclusion The improvement of the puncture angle and the technique innovation of the guide rod moving from intervertebral space to intraspinal canal step by step can effectively reduce the nerve root stimulation and injury during the puncture and the placement of the working passage,and significantly improve the clinical efficacy of percutaneous lumbar endoscopy in the treatment of central lumbar disc herniation.Microscopic management of the posterior longitudinal ligament can effectively reduce the recurrence rate and has no significant impact on spinal stability.
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Objective To explore the clinical application of antcrolateral thigh flap transplantation in repairing wound around the knee with descending genicular artery as the recipient vessel,when anterior or posterior tibial vessel could not be utilized.Methods From January,2015 to May,2017,free anterolateral thigh flaps obtained from anastomosis of descending genicular artery and great saphenous vein were transplanted to repair the skin soft tissue defect around the knee combined with tendon and bone exposure in 7 patients,after preoperative color Doppler sonography ultrasound (CDU) for precise positioning.There were 4 males and 3 females,with the flap area ranging from 18.0 cm×8.0 cm-38.0 cm×8.0 cm.All of the donor sites were sutured directly.Postoperative followedup was done termly.Results All the patients were followed-up for 6 to 14 months,with an average of 8.9 months.Typically,2 cases had large defect areas,with distal flap necrosis of 6.0 cm and 4.0 cm,respectively,which were resected and achieved secondary skin graft healing on the residual surface.Additionally,4 cases had completely survived flaps and achieved secondary skin graft healing on the residual surface.The remaining 1 case had completely survived flap,but the distal flap near the anteromedial tibia developed bone exposure as a result of the complicated osteomyelitis.As a result,the patient received gastrocnemius myocutaneous flap to repair the wound.Conclusion Anterolateral thigh flap transplantation in repairing skin soft tissue defect wound around the knee,with descending genicular artery as the recipient vessel,can achieve satisfactory clinical efficacy,which can serve as one choice for flap repair in wound around the knee.
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Objective To establish a simple and feasible method in analyzing the molecular weight distribution of peptides in thymosin.Methods Improved Tricine-SDS-PAGE electrophoresis was applied to detect the molecular weight of peptides and the content of thymosinα1 in thymosin.Results Tested the thymosin preparations on the domestic market using this im-proved method.It was demonstrated that the peptide molecular weight distribution in thymosin preparations was between 3.5~8.5 kD,also could detect the concentration of 1 μg thymosinα1 in thymosin by using this improved method.Conclusion This improved method is suitable for the analysis of peptides molecular weight distribution and the concentration of thy-mosinα1,so it can be used for control quality of thymosin preparations.