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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 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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Objective:To investigate the phylogenetic and antigenic characteristics of hemagglutinin (HA) gene of influenza B/Victoria lineage (BV) viruses in Beijing during the 2021-2022 influenza surveillance season, and to analyze whether the circulating BV viruses match the vaccine strain.Methods:Pharyngeal swab specimens from influenza like-illness (ILI) cases in the 2021-2022 influenza surveillance season were collected from surveillance network labs in Beijing and cultured in MDCK cells and chicken embryo to isolate BV viruses. Nucleic acids of the viruses were extracted, and the HA gene was amplified and sequenced. The nucleotide and amino acid sequence identity of the HA gene was analyzed using MEGA5.0 software. A phylogenetic tree of HA gene was constructed using the maximum likelihood method. The N-glycosylation sites in HA were predicted online. Three-dimensional structure of HA was constructed using SWISS-MODEL homologous modeling. Hemagglutination inhibition (HI) test was performed to analyze the antigenicity of BV viruses.Results:A total of 402 BV viruses were collected and 58 strains with full-length HA gene sequences were chosen for further analysis. Compared with the HA gene of this year′s vaccine strain (B/Washington/02/2019), there were 27 amino acid mutations, 11 of which were located in four different antigenic determinants. The phylogenetic analysis revealed that three subgroups of 1A.3, 1A.3a1, and 1A.3a2 co-circulated in Beijing with 54 strains (54/58, 93.10%) clustered to the Clade 1A.3a2, two strains (2/58, 3.45%) clustered to the Clade 1A.3a1, and two strains (2/58, 3.45%) in the same subgroup (Clade 1A.3) as the vaccine component BV strain in 2021-2022. Compared with the vaccine strain (B/Washington/02/2019), two BV strains had an additional N-glycosylation site at residue 197, while the other 56 strains showed no change in N-glycosylation sites. Antigenic analysis showed that 35 BV strains (35/58, 60.34%) were antigenically similar to the vaccine strain and 23 strains (23/58, 39.66%) were low-response strains.Conclusions:Three subgroups of BV viruses co-circulated in Beijing during the 2021-2022 influenza surveillance season. The predominant subgroup was Clade 1A.3a2 (93.10%), showing a certain genetic distance with the vaccine strain (B/Washington/02/2019). Nearly 40% (39.66%) of the viruses were low-response strains. This study indicated that continuous monitoring of the variations of influenza epidemic strains and timely providing laboratory basis for screening vaccine component strains were the basic technical guarantee for coping with influenza pandemic.
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Objective:To clarify the M protein ( emm gene) types and drug susceptibility characteristic variations of Group A Streptococcus (GAS) in children in Beijing. Methods:The GAS strains isolated from throat swab samples of children diagnosed with scarlet fever and pharyngeal infection in scarlet fever etiology surveillance sentinel hospitals in 16 districts of Beijing in 2018, 2019 and 2021 were analyzed retrospectively.PCR amplification and sequencing were used for emm genotyping, and the minimum inhibitory concentrations (MIC) of 10 antibiotics were determined by the broth microdilution method.The data were analyzed using χ2 test and Fisher′ s exact method between groups. Results:A total of 557 GAS strains were collected, and 11 emm genotypes ( emm1, emm3, emm4, emm6, emm11, emm12, emm22, emm75, emm89, emm128, and emm212) were detected.Of 557 strains, 238 trains were of emm1 type (42.73%), 271 strains were of emm12 type (48.65%) and 48 strains were of other emm types (8.62%). The detection rates of emm1, emm12 and other emm type genes in 2018, 2019, and 2021 were [37.50% (105/280 strains), 57.14% (160/280 strains), 5.36% (15/280 strains)], [49.05% (129/263 strains), 39.54% (104/263 strains), 11.41% (30/263 strains)], and [28.57% (4/14 strains), 50.00% (7/14 strains), 21.43% (3/14 strains)], respectively.In children infected with emm12 in 2018 and 2019, there were more children under 6 years old than children over 6 years old (62.50% vs.46.88%, 46.36% vs.30.36%) (χ 2=7.182, 6.973; all P<0.05). Drug susceptibility testing results suggested that 225 randomly selected GAS strains were all 100.00% sensitive to 7 antibiotics including Penicillin, Levofloxacin, Meropenem, Linezolid, Cefotaxime, Cefepime and Vancomycin.The rates of resistance to Erythromycin, Tetracycline and Clindamycin were [88.57% (93/105 strains), 87.62% (92/105 strains), 86.67% (91/105 strains)], and [94.34% (100/106 strains), 94.34% (100/106 strains), 87.74% (93/106 strains)] in 2018 and 2019, respectively.The test strains were 100.00% (14/14 strains) resistant to the above 3 antibiotics in 2021.MIC 50 and MIC 90 values of Penicillin in 2018, 2019, and 2021 were (0.03 mg/L, 0.03 mg/L), (0.03 mg/L, 0.06 mg/L), and (0.06 mg/L, 0.06 mg/L), respectively.Among 225 GAS strains, 207 strains had drug resistance and were resistant to more than one drug.Specifically, 94.69% (196/207 strains) were resistant to Erythromycin, Tetracycline and Clindamycin.About 4.35% (9/207 strains) were resistant to both Erythromycin and Clindamycin.A total of 0.97% (2/207 strains) were resistant to Erythromycin and Tetracycline. Conclusions:The emm genotypes of GAS in children in Beijing are diverse in 2018, 2019 and 2021.The dominant genotypes are emm12 and emm1, and emm12 is the main epidemiological type.GAS strains maintain highly resistant to Erythromycin, Clindamycin and Tetracycline, and sensitive to Penicillin and other antibiotics.However, MIC 50 and MIC 90 of Penicillin shows an ascending trend.
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Objective To analyze the characteristics of super-antigen (SAg) of group A Streptococcus pyogenes (GAS),isolated from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017.Methods Throat swab specimens from patients with scarlet fever or pharyngeal infections were collected and tested for GAS.Eleven currently known SAg genes including SpeA,speC,speG,speH,speI,speJ,speK,speL,speM,smeZ and ssa were tested by real-time PCR while M protein genes (emm genes) were amplified and sequenced by PCR.Results A total of 377 GAS were isolated from 6 801 throat swab specimens,with the positive rate as 5.5％.There were obvious changes noticed among speC,speG,speH and speK in three years.A total of 45 SAg genes profiles were observed,according to the SAgs inclusion.There were significant differences appeared in the frequencies among two of the highest SAg genes profiles between emml and emml2 strains (x2=38.196,P＜0.001;x 2=72.310,P＜0.001).There also appeared significant differences in the frequencies of speA,speH,speI and speJ between emm 1 and emm 12 strains (x2 =146.154,P＜ 0.001;x2=52.31,P＜0.001;x2=58.43,P＜0.001;x2=144.70,P＜0.001).Conclusions Obvious changes were noticed among SAg genes including speC,speG,speH and speK from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017.SAg genes including speA,speH,speI and speJ appeared to be associated with the emm 1 and emm 12 strains.More kinds of SAg genes profiles were isolated form GAS but with no significant differences seen in the main SAg genes profiles,during the epidemic period.
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Objective To analyze the characteristics of super-antigen (SAg) of group A Streptococcus pyogenes (GAS),isolated from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017.Methods Throat swab specimens from patients with scarlet fever or pharyngeal infections were collected and tested for GAS.Eleven currently known SAg genes including SpeA,speC,speG,speH,speI,speJ,speK,speL,speM,smeZ and ssa were tested by real-time PCR while M protein genes (emm genes) were amplified and sequenced by PCR.Results A total of 377 GAS were isolated from 6 801 throat swab specimens,with the positive rate as 5.5％.There were obvious changes noticed among speC,speG,speH and speK in three years.A total of 45 SAg genes profiles were observed,according to the SAgs inclusion.There were significant differences appeared in the frequencies among two of the highest SAg genes profiles between emml and emml2 strains (x2=38.196,P＜0.001;x 2=72.310,P＜0.001).There also appeared significant differences in the frequencies of speA,speH,speI and speJ between emm 1 and emm 12 strains (x2 =146.154,P＜ 0.001;x2=52.31,P＜0.001;x2=58.43,P＜0.001;x2=144.70,P＜0.001).Conclusions Obvious changes were noticed among SAg genes including speC,speG,speH and speK from patients with scarlet fever or pharyngeal infections in Beijing between 2015-2017.SAg genes including speA,speH,speI and speJ appeared to be associated with the emm 1 and emm 12 strains.More kinds of SAg genes profiles were isolated form GAS but with no significant differences seen in the main SAg genes profiles,during the epidemic period.
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Objective To investigate the genetic characteristic of whole-genome of influenza A/H3N2 viruses in severe acute respiratory infection cases in Beijing area.Methods From 2014 to 2016,the viral RNA was extracted from 32 strains isolated from SARI cases,then sequenced by Ion Torrent PGM Sequencer.The phylogeny and molecular features of whole-genome were analyzed by Mega and Consurf software.Results The HA gene of tested strains isolated in 2014-2015 influenza season belonged to lineage 3C.3a and 3C.2a,while those isolated in 2015-2016 influenza season belonged to cluster 3C.2a.Moreover,compared with the vaccine strains,7 variant amino acids of protein of HA1 were identified,and two of them were located in antigenic sites.All isolates were sensitive to neuraminidase inhibitors while showed resistance to blockers for M2 ion channel.Conclusion The phylogenetic features of isolates studied in this study are similar with that of current circulating strains.However,the difference between isolates and vaccines should not be overlooked.
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Objective To explore the relationship between superantigen and M protein gene (emm)-types genes of Group A Streptococcus pyogenes (GAS) isolated from patients with scarlet fever in Beijing from May 2012 to July 2013 .Methods GAS was isolated from specimens of patients with scarlet fever . Superantigen genes (speA ,speB ,speC ,speF ,speG ,speH ,speI ,speJ ,speL ,speK ,speM ,ssa ,and smeZ) ,and emm gene were amplified by polymerase chain reaction .Rate and proportion were compared by chi-square test .Results Of the 423 GAS strains isolated from patients with scarlet fever from 2012 to 2013 ,most of the isolates possessed speB (97 .6% ) ,speC (99 .8% ) ,speF (98 .3% ) ,speG (99 .8% ) , smeZ (94 .1% ) and ssa (88 .4% ) ,and some of them possessed speH (54 .6% ) ,speI (53 .4% ) ,speA (45 .2% ) and speJ (43 .5% ) ,but very few isolates possessed speK (2 .4% ) ,speL (1 .4% ) and speM (1 .7% ) .Type emm12 (59 .5% ) and type emm1 (37 .4% ) were the main types of GAS .Most of the emm12-type isolates possessed speH (84 .8% ) and speI (84 .0% ) compared with only 4 .0% of speH and 3 .4% of speI in type emm1 .Most of type emm1 possessed speA (95 .3% ) and speJ gene (94 .6% ) compared with only 17 .3% of speA and 14 .8% of speJ in type emm12 .The superantigen genes profiles were significant different between emm 1-type and emm 12-type isolates (P< 0 .05) .Conclusion Type emm1 and type emm12 are epidemic strains in patients with scarlet fever from 2012 to 2013 in Beijing ,and emm gene-types are associated with superantigen genes profiles .
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<p><b>OBJECTIVE</b>To investigate superantigen gene profiles of group A Streptococcus (GAS) isolated in Beijing pediatric patients in 2014, and to explore the the relationship between superantigen gene profiles with emm types, and GAS infections with diseases.</p><p><b>METHODS</b>A total of 259 GAS strains were isolated from pediatric patients clinically who diagnosed with scarlet fever and pharyngitis from 36 hospitals in Beijing from May to July, 2014.The Superantigens genes of strains were performed by Real-time PCR (speA, speB, speC, speF, speG, speH, speI, speJ, speK, speL, speM, smeZ, ssa). PCR amplification of GAS strain M protein N gene segments were carried ort; products after sequencing comparison were analyzed to determine the GAS types of emm. The differences in distributions of superantigen genes and emm types of GAS isolates were compared between subgroups.</p><p><b>RESULTS</b>Among the 259 GAS strains, the detection rates of 13 superantigens were as the following: speA 48.6% (126), speB 99.2% (257), speC 99.2% (257), speF 98.8% (256), speG 98.5% (255), speH 43.6% (113), speI 46.3% (120), speJ 49.0% (127), smeZ 99.2% (257) and ssa 98.5% (255), respectively, however, speK, speL, and speM were not found. Eleven superantigen gene profiles in all were observed (A-K). The percentage of emm1 strains harbored spe A and speJ were 94.2% (113/120), 95.0% (114/120), respectively, which were significantly higher than those of emm12 strains (5.6% (7/124), 5.6% (7/124), respectively; χ(2) = 191.20, 194.80, P < 0.001). The percentage of emm12 strains harbored speH and speI were 83.9% (104/124), 88.7% (110/124), respectively, which were significantly higher than those of emm1 strains (3.3% (4/120), 4.2% (5/120), respectively; χ(2) = 160.30, 174.90, P < 0.001).The superantigen genotypes of GAS strains and emm types, which were isolated from scarlet fever and pharyngitis cases, were not significant different (P > 0.05).</p><p><b>CONCLUSION</b>The GSA strains isolated in Beijing pediatric patients in 2014, the relevance ratio of speB, speC, speF, smeZ, speG, ssa were higher than others, while speK, speL, and speM were no detected in any GAS strains. The superantigen genes appeared to be associated with the emm type. Furthermore, emm type distribution and superantigen genes were not different between scalet fever and pharyngitis.</p>
Subject(s)
Child , Humans , Antigens, Bacterial , Genetics , Beijing , Genotype , Real-Time Polymerase Chain Reaction , Streptococcal Infections , Microbiology , Streptococcus pyogenes , Genetics , Allergy and Immunology , Superantigens , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To understand the distribution of emm gene types related to group A streptococcus-caused scarlet fever among children in Beijing and to analyze the relationship between the mutation of the emm types and scarlet fever.</p><p><b>METHODS</b>Nasopharyngeal swab samples were collected from the scarlet fever cases diagnosed in 36 hospitals in Beijing to isolate the GAS strains from May to July, betgween 2011 and 2014. Genotyping of emm gene was performed with PCR and N-terminal gene fragments of M protein were sequenced. Data of all the scarlet fever cases in Beijing that reported through the National Notifiable Infectious Disease Surveillance System (NNIDSS) , were gathered and analyzed.</p><p><b>RESULTS</b>Among the collected 2 161 nasopharyngeal swabs, 762 GAS strains were identified (35.3%). In addition, 7 emm types were detected, in which emm12 accounted for 69.4% (529/762) , emm1 accounted for 29.8% (227/762) , and other five types (emm 11, 22, 75, 89, and 128) accounted for 0.8% (6/762) , respctively. Compared with the emm types detected between 2011 and 2014, emm12, emm1 and other types accounted for 82.2% (295/359) , 16.7% (60/359) and 1.1% (4/359, including emm11, 22 and 89) in 2011 respectively.emm12, emm1 and emm75 accounted for 77.3% (123/163) , 23.9% (39/163) and 0.6% (1/163) respectively in 2012. emm12 and emm1 accounted for 50.7% (38/75) and 49.3% (37/75) in 2013 while emm12, emm1 and emm128 accounted for 44.2% (73/165) , 55.2% (91/165) and 0.6% (1/165) respectively in 2014. The differences of the constitution of emm types from 2011 to 2014 appeared statistically significant (P<0.001). In 2011 and 2012, major type appeared as emm12, but in 2014, emm1 became predominant. A total of 6 152 cases were reported in 2011, while 2 908, 2 048 and 3 918 cases were reported in 2012, 2013 and 2014 respectively. Age specific differences were noticed in the distribution of emm types GAS strains in 2011, with the number of emm12 strains detected higher in 1-5 year olds than in age group > 5 years (P<0.05). There were area specific differences in distribution of emm types of GAS strains seen in 2011 and 2013. In 2011, the number of emm1 strains detected in urban area was higher than in suburb area (P<0.05). However, in 2013, the number of emm1 strains detected in suburb area was seen higher than in urban area (P< 0.05).</p><p><b>CONCLUSION</b>GAS with emm12 and GAS emm1 appeared interchangeably predominant in Beijing from 2011 to 2014. Changes in predominant emm types seemed also related to the trends of incidence rates on scarlet fever.</p>