Effect of Loulianwan on Gut Microbiota in db/db Mice with Type 2 Diabetes Mellitus Based on 16S rRNA Sequencing Technology / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Loulianwan on the gut microbiota of db/db mice with type 2 diabetes mellitus （T2DM）. MethodMale db/m+ mice aged 4-5 weeks were assigned to the normal group， and male db/db model mice of the same age were randomly divided into model group， metformin group （0.25 g·kg-1·d-1）， and Loulianwan group （13 g·kg-1·d-1）， with six mice in each group. Drug intervention lasted five weeks. The body weight， water intake， and fasting blood glucose （FBG） of the mice were recorded every week. After five weeks， the FBG， liver triglyceride （TG）， liver total cholesterol （TC）， glycated serum protein （GSP）， and fasting serum insulin （FINS） were detected， and the insulin resistance index （HOMA-IR） was calculated. The feces in the mouse intestines were collected， and the 16S rRNA sequencing technology was used to detect the structural changes in the fecal gut microbiota of mice in each group. ResultCompared with the normal group， the model group showed increased body weight， water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. Compared with the model group， the Loulianwan group showed reduced water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. The gut microbiota in the Loulian Lills group changed from phylum to genus level. The relative abundance of beneficial bacteria increased and the relative abundance of harmful bacteria decreased. Among them， the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides increased （P<0.01）. ConclusionLoulianwan can significantly improve glucose and lipid metabolism in db/db mice with T2DM， and its mechanism may be related to the increase in the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides in the intestine.

Construction of CeRNA network of differential LncRNAs in THP-1 cells infected by Dengue virus type Ⅲ and its antibody dependent enhancement / 中国生物制品学杂志

@#ObjectiveTo establish models of Dengue virus type Ⅲ（DENV-3,DV-3）infection and antibody dependent enhancement(ADE)infection at the acute monocytic leukemia cells(THP-1),investigate the differential expression of long non-coding RNAs(LncRNAs),map the competitive endogenous RNA(CeRNA)regulatory network and predict the translation function of LncRNAs.MethodsThe culture supernatant was harvested 6 d after C6/36 cells were infected with DENV-3,the virus titer was determined by CCID50,and the type and full-length genome amplification were identified by PCR;The DENV-3 standard plasmid was amplified,identified by PCR,and the standard curve was drawn;THP-1 cells were divided into negative control group(THP-1),direct infection group(DV-3),ADE group and blank control group[1640(-)]. After 48 h of infection,the total RNA was extracted and the copy number of intracellular virus nucleic acid was measured;Through the whole transcriptome sequencing technology,the CeRNA regulatory network was constructed for the top five up-regulated and down-regulated LncRNAs in THP-1 vs DENV3,THP-1 vs ADE,DENV3 vs ADE groups,and the functions of their coding proteins were analyzed.ResultsC6/36 cells infected with DENV-3 for 3 d showed obvious cell fusion,vacuoles and abscission;The virus had a titer of about 1. 0 × 104. 64PFU/mL and was identified as DENV-3 by PCR specific primers,of which the complete gene sequence was obtained;The number of viral nucleic acid copies in ADE group was significantly higher than those in DV-3 group and blank control group;In THP-1 vs DENV-3,the expression of cytohesin interacting protein(CYTIP)was predicted to be up-regulated;In THP-1 vs ADE,the expression of kinesin family5A(KIF5A)was predicted to be down-regulated;In DENV-3 vs ADE,the expression of cluster differentiation antigen 9(CD9)and insulin like growth factor 2(IGF2)was predicted to be up-regulated. All of these differential LncRNAs had open reading frames(ORFs). Except Lnc-SH3BP1 and Lnc-RPL41,all of the other LncRNAs had internal ribosome binding site(IRES).ConclusionIn DENV-3 infection of THP-1 cells and ADE infection mediated by DENV-3,the expression of LncRNAs has changed significantly,and may regulate the process of infection through a variety of biological functions,which is helpful for a deeper understanding of the mechanism of ADE infection.

Application of single cell sequencing technology in liver regeneration / 中华消化外科杂志

The liver is a highly proliferative organ. As the liver injured, the hepatocytes can quickly enter the cell cycle to restore the volume and function of liver. Liver regeneration involves complex processes that depend on the interaction of many different cell types. As limited by the average cell change level in tissues, traditional sequencing methods can only acquire the average genetic information reflecting dominant cell subpopulations, but ignore the secondary cell subpopu-lations, which leads to the loss of cellular heterogeneity information. Single-cell sequencing tech-nology can analyze the biological behavior of single cell, which helps to better understand the distri-bution, interaction and cell heterogeneity of different cells during liver regeneration. The authors review the application of single cell sequencing technology in liver regeneration.

Advances in international newborn screening / 中华实用儿科临床杂志

Newborn screening is an important national public health policy and measure to reduce birth defects and improve the quality of China′s birth population.In the early 1960s, Dr.Robert Guthrie of the United States invented the first newborn screening test, namely, semi-quantitative determination of phenylalanine in dry blood filter paper for screening phenylketonuria.In the 1990s, tandem mass spectrometry (MS/MS) began to be applied to the screening of genetic metabolic diseases in newborns.This technology enabled the detection of multiple diseases by one test, and increased the types of diseases detected.In the past 10 years, with the development of screening technology, the invention of new drugs, the improvement of treatment methods, and especially the application of new technologies such as newborn genetic screening, the source of mutations can be identified at the molecular level.Moreover, newborn screening is extended to patients who are not candidates for MS/MS.Many genetic diseases are able to be screened and diagnosed early.Effective management and quality control of newborn disease screening are prerequisites for improving the quality and accuracy of results.Secondary and multi-level detection strategies, different biochemical or biochemical genetic testing methods, and the integration of targeted and non-targeted multi-omics data have a wide range of applications and great clinical value.

Study on RHD and RHCE gene mRNA based on three generation sequencing technology / 中国输血杂志

【Objective】 To establish RH gene mRNA sequencing method based on nanopores sequencing and to explore the RHD and RHCE mRNA transcripts in D positive and Del individuals. 【Methods】 From March 2021 to May 2022, 5 RhD positive samples and 5 Del samples screened out by hospitals in Chengdu were sent to our laboratory for futher examination. The erythrocytes and buff coat were isolated, then DNA and RNA were extracted.All 10 samples were genotyped by PCR-SSP. After the mRNA was reversely transcribed into cDNA, the full-length mRNA of RHD and RHCE genes were simultaneously amplified by a pair of primers. Sanger sequencing and third-generation sequencing technology based on Nanopore were used to sequence the amplified products, and the types and expressions of different splices of RHD and RHCE gene mRNA transcripts were analyzed. 【Results】 The method established in this study can simultaneously amplify the full length transcripts of RHD and RHCE. Ten different RHD gene mRNA transcripts and nine RHCE gene mRNA transcripts were detected in 10 samples. RHD full-length transcript (RHD-201) can be detected in RhD Del type, but the expression amount was significantly lower than that in RhD positive samples. The expression amount of transcript RHD-207 (Del789) in Del samples was significantly higher than that in RhD positive samples. The transcript RHD-208 (Del8910+ 213) was only detected in RhD Del type individuals, and no significant difference was found between other RHD transcripts and all RHCE transcripts in the two phenotypes. 【Conclusion】 In this study, an analytical method for sequencing full-length transcript isomers of RHD and RHCE mRNA via the third generation was successfully established, and complex alternative splicing patterns were found in RHD and RHCE genes, providing a new method for the study of alternative splicing of blood group gene variants mRNA.

Genetics and muscle pathology in the diagnosis of muscular dystrophies: An update

Muscular dystrophies are a clinically and genetically heterogeneous group of disorders involving the skeletal muscles. They have a progressive clinical course and are characterized by muscle fiber degeneration. Congenital muscular dystrophies (CMD) include dystroglycanopathies, merosin-deficient CMD, collagen VI-deficient CMD, SELENON-related rigid spine muscular dystrophy, and LMNA-related CMD. Childhood and adult-onset muscular dystrophies include dystrophinopathies, limb-girdle muscular dystrophies, Emery-Dreifuss muscular dystrophy, facioscapulohumeral muscular dystrophy, and myotonic dystrophy. Traditionally, muscle biopsy and histopathology along with special pathology techniques such as immunohistochemistry or immunoblotting were used for the diagnosis of muscular dystrophies. However, recent advances in molecular genetic testing, especially the next-generation sequencing technology, have revolutionized the diagnosis of muscular dystrophies. Identification of the underlying genetic basis helps in appropriate management and prognostication of the affected individual and genetic counseling of the family. In addition, identification of the exact disease-causing mutations is necessary for accurate prenatal genetic testing and carrier testing, to prevent recurrence in the family. Mutation identification is also essential for initiating mutation-specific therapies (which have been developed recently, especially for Duchenne muscular dystrophy) and for enrolment of patients into ongoing therapeutic clinical trials. The 'genetic testing first' approach has now become the norm in most centers. Nonetheless, muscle biopsy-based testing still has an important role to play, especially for cases where genetic testing is negative or inconclusive for the etiology.

Clinical phenotype and genetic features of 16p11.2 microdeletion-related epilepsy in children / 中国当代儿科杂志

OBJECTIVES@#To study the clinical phenotype and genetic features of 16p11.2 microdeletion-related epilepsy in children.@*METHODS@#The medical data of 200 children with epilepsy who underwent a genetic analysis of epilepsy by the whole exon sequencing technology were collected retrospectively, of whom 9 children with epilepsy had 16p11.2 microdeletion. The clinical phenotype and genetic features of the 9 children with 16p11.2 microdeletion were analyzed.@*RESULTS@#The detection rate of 16p11.2 microdeletion was 4.5% (9/200). The 9 children with 16p11.2 microdeletion were 3-10 months old. They experienced focal motor seizures with consciousness disturbance, and some of the seizures developed into generalized tonic-clonic seizures. The interictal electroencephalogram showed focal or multifocal epileptiform discharge, and all 9 children responded well to antiepileptic drugs. The 9 children had a 16p11.2 deletion fragment size of 398-906 kb, and the number of deleted genes was 23-33 which were all pathogenic mutations. The mutation was of maternal origin in 2 children, of paternal origin in 1 child, and de novo in the other children.@*CONCLUSIONS@#16p11.2 microdeletion can be detected in some children with epilepsy. Most of the 16p11.2 microdeletion is de novo mutation and large gene fragment deletion. The onset of 16p11.2 microdeletion-related epilepsy in children is mostly within 1 year of life, and the epilepsy is drug-responsive.

Application of reactive soft tissue preservation in extraction sockets of periodontitis: recent progress and perspectives / 口腔疾病防治

@#Chronic periodontitis is a prevalent disease; if left untreated, it is a main indication for tooth extraction and can lead to tooth loss. The reactive soft tissue, formed as a result of the immune response to chronic inflammation, is left in the compromised socket. The major concern is how to deal with the residual reactive soft tissue. Conservative thought states that the reactive soft tissue should be completely debrided. In addition, novel practices concerning the reactive soft tissue were proposed in recent trials, which demonstrated that there might be merits for soft and hard tissue regeneration with preservation of the reactive soft tissue. Studies have shown that mesenchymal stem cells exist in inflammatory reactive soft tissue, stressing their potential in tissue regeneration. Although the therapeutic value is highly promising, the specific components of the reactive soft tissue and the standard on whether it should be preserved need further investigation.

Metagenomic study on the composition and function of oral microorganisms in healthy adults / 口腔疾病防治

Objective@#Metagenomic sequencing was used to explore the species composition and internal functional metabolic pathway of saliva and supragingival plaque microbial communities in healthy adults to provide a theoretical reference for the biological prevention and treatment of oral diseases.@*Methods@#Saliva and supragingival plaque samples were collected from healthy adults, total DNA was extracted, and a metagenomic library was constructed. The qualified library was sequenced via metagenomics, and the sequencing data were analyzed using bioinformatics and statistics. @*Results @#The main bacterial phyla in healthy oral samples were Proteobacteria (32.51%), Bacteroidetes (30.81%), and Actinobacteria (16.23%), and the main bacterial species were Corynebacterium matruchotii (3.84%), Haemophilus parainfluenzae (2.91%), and Prevotella melaninogenica (2.76%). The alpha diversity of the supragingival plaque group was higher than that of the saliva group, and there was a significant difference in the composition of the microbial community between the two groups (P<0.05). At the species level, Prevotella melaninogenica, Fusobacterium periodonticum, and Prevotella intermedia were more abundant in saliva samples than in supragingival plaque samples, while Corynebacterium matruchotii, Propionibacterium acidifaciens, and Rothia dentocariosa were more abundant in supragingival plaque samples than in saliva samples (P<0.05). High-quality gene sets of saliva and supragingival plaque in healthy adults were constructed based on metagenomic sequencing. The results of KEGG pathway functional metabolic differences showed that starch and sucrose metabolism, leucine and isoleucine degradation, and arginine biosynthesis in salivary microorganisms were more abundant than in supragingival plaque, while glycolysis/gluconeogenesis and carbon metabolism in supragingival plaque were more abundant than in saliva.@* Conclusion@#There are significant differences in the species composition and functional gene metabolic pathways of saliva and supragingival plaque microecology in healthy adults. The sensitivity of dominant species in different microecological regions to the identification of oral diseases may be different. In the microbiological study of oral diseases, appropriate samples should be selected according to different diseases.

Effects of nano titanium dioxide on gut microbiota based on human digestive tract microecology simulation system in vitro / 北京大学学报(医学版)

OBJECTIVE@#To explore the effects of oral exposure to titanium dioxide nanoparticles (TiO2 NPs) on the composition and structure of human gut microbiota.@*METHODS@#The particle size, shape, crystal shape and degree of agglomeration in ultrapure water of TiO2 NPs were characterized. The in vitro human digestive tract microecological simulation system was established by simulating the fluid environment and physical conditions of stomach, small intestine and colon, and the stability of the simulation system was evaluated. The bacterial communities were extracted from human feces and cultured stably in the simulated system. They were exposed to 0, 20, 100 and 500 mg/L TiO2 NPs, respectively, and the bacterial fluids were collected after 24 h of exposure. The effect of TiO2 NPs on the composition and structure of human gut microbiota was analyzed by 16S rRNA sequencing technology. Linear discriminant analysis effect size (LEfSe) was used to screen differential bacteria, and the Kyoto encyclopedia of genes and genomes (KEGG) database for functional prediction.@*RESULTS@#The spherical and anatase TiO2 NPs were (25.12±5.64) nm in particle size, while in ultra-pure water hydrated particle size was (609.43±60.35) nm and Zeta potential was (-8.33±0.22) mV. The in vitro digestive tract microecology simulation system reached a relatively stable state after 24 hours, and the counts of Enterococci, Enterobacte-rium, and Lactobacillus reached (1.6±0.85)×107, (5.6±0.82)×107 and (2.7±1.32)×107, respectively. 16S rRNA sequencing results showed that compared with the control group, the number and evenness of gut microbiota were not significantly affected at phylum, class, order, family and genus levels in TiO2 NPs groups (20, 100 and 500 mg/L). The relative abundance of some species was significantly changed, and a total of 42 different bacteria were screened between the TiO2 NPs groups (20, 100 and 500 mg/L) and the control group [linear discriminant analysis(LDA) score>3], represented by Enterobacter, Bacteroidaceae, Lactobacillaceae, Bifidobacteriaceae and Clostridium. Further predictive analysis of gut microbiota function showed that TiO2 NPs might affect oxidative phosphorylation, energy meta-bolism, phosphonate and phosphonate metabolism, and methane metabolism (P < 0.05).@*CONCLUSION@#In human digestive tract microecological simulation system, TiO2 NPs could significantly change the composition and structure of human gut microbiota, represented by Enterobacter and probiotics, and may further affect a variety of metabolism and function of the body.

Analysis on Antidepressant Mechanism of Verbascoside Based on RNA-Seq Technology / 中国实验方剂学杂志

Objetive： To investigate the mechanism of antidepressant effect of verbascoside based on high-throughput sequencing technology （RNA-Seq），and to explore the possible targets and signaling pathways. MethodsForty C57BL/6 mice were randomly divided into control group，model group，fluoxetine group and verbascoside group，10 mice in each group. Except for the control group，all the other three groups were constructed with chronic unpredictable mild stimulation （CUMS） combined with solitary feeding for four weeks. Control group，fluoxetine group and verbascoside group were administered by gavage once daily for three weeks during the second week of modeling. The mice were assessed by sugar-water preference test，forced swimming test，open field test，elevated cross maze test，and water maze test. Enzyme linked immunosorbent assay（ELISA） was performed to detect the levels of major neurotransmitters and inflammatory factors in mice serum，and mRNA high-throughput sequencing was performed in the nucleus accumben and colon to screen differentially expressed genes and perform pathway enrichment analysis. Real-time polymerase chain reaction（Real-time PCR） was used to detect the mRNA expression of Gad，Slc32a1（VGAT） and brain-derived neurotrophic factor (BDNF） in nucleus accumbens. ResultCompared with the control group，the anxiety and depression-like behaviors in the model group increased，while the learning and memory ability decreased significantly. The content of neurotransmitter in serum decreased significantly. The content of pro-inflammatory factors increased significantly. The mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens decreased significantly，while that of BDNF increased significantly. Compared with the model group，the anxiety and depression-like behavior of mice in verbascoside group was significantly relieved. The neurotransmitter content increased significantly，and the pro-inflammatory factors decreased significantly. The mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens increased significantly，while that of BDNF decreased significantly.A total of 48 differentially expressed genes in nucleus accumbens and 43 differentially expressed genes in colon were screened by high-throughput sequencing. Differential genes in nucleus accumbens mainly focus on neuroactive ligand-receptor interaction， γ-aminobutyric acid（GABA）ergic synapse，synaptic vesicle cycle and other pathways. Colonic differential genes are mainly concentrated in GABAergic synapses，synaptic vesicle circulation，cyclic adenosine monophosphate（cAMP） signal pathway and other signal pathways. Compared with the control group，the mRNA expression of Gad and Slc32a1（VGAT） in nucleus accumbens of model group decreased significantly，while the mRNA expression of BDNF increased significantly. ConclusionVerbascoside has significant antidepressant effects. Its antidepressant effect may be related to the increase of monoamine neurotransmitters，the decrease of pro-inflammatory factors and the restoration of neurotransmitter homeostasis by increasing GABA，and it mainly acts through the signaling pathways such as neuroactive ligand-receptor interaction，GABAergic synapses，synaptic vesicle cycle and cAMP signaling pathway.

Application of single cell sequencing technology in atherosclerosis research / 中华神经科杂志

Atherosclerosis is one of the most common cardiovascular and cerebrovascular diseases, and it is also an important cause of stroke. However, the research on the pathogenesis of atherosclerosis is still incomplete. Single cell technology, as an emerging technology in the study of differences in cell biology, has become a new tool and provides a new way of exploring the etiology of atherosclerosis. This article reviewed the research progress of single cell sequencing technology in atherosclerosis in recent years.

Sequencing Analysis of MiRNA in the Exosomes from Normal and Oxygen and Glucose Deprivation Reperfusion Astrocytes / 中国生物化学与分子生物学报

In this study, high-throughput sequencing technology was used to analyze the differentiallyexpressed microRNA (miRNA) of astrocyte-derived exosomes in control group and oxygen and glucosedeprivation/ reoxygenation (OGD/ R) group. Ultracentrifugation was used to extract exosomes from thesupernatant of astrocyte medium in the control group and OGD/ R group. Transmission electron microscopyshowed that exosomes had a typical vesicle shape with intact membrane and low electron content density. Nanoparticle tracking technology (NTA) detected astrocyte exosomes with a size of 100. 5 ± 31. 1 nm, accounting for 96. 8%. Western blot detection showed that the exosome contained exosome-specificproteins tumor-susceptibility protein (TSG101), heat shock protein 60 (Hsp60), ALG-2-interactingprotein X (ALIX). Compared with the control group, 41 miRNAs in the exosomes of the OGD / R groupwere significantly changed, of which 20 miRNAs were increased and 21 miRNAs were decreasedsignificantly (P < 0. 05). Gene ontology function (GO) analysis showed that significantly differentiallytarget genes were mainly involved in protein glycosylation, lipid metabolism, phosphorylation, Golgiapparatus, endoplasmic reticulum, endosome, cytoplasmic vesicles and cell protrusions, etc. KyotoEncyclopedia of Genes and Genomes (KEGG) pathway analysis found that differential miRNAs weremainly related to metabolic pathways and signaling pathways such as butyrate metabolism, ß-alaninemetabolism, fatty acid degradation, mitophagy and P53 signaling pathway. Sequencing analysis of theexosomal miRNAs derived from control and OGD / R astrocytes and target gene function enrichmentanalysis can be useful for the mechanism study of astrocyte exosomes in response to oxygen and glucosedeprivation reperfusion.

Research highlights of renal transplantation in 2020: voice from China / 器官移植

Renal transplantation is the optimal approach to improve the quality of life and restore normal life for patients with end-stage renal diseases.With the development of medical techniques and immunosuppressants, the shortterm survival of renal graft has been significantly prolonged, whereas the long-term survival remains to be urgently solved.Renal ischemia-reperfusion injury (IRI), acute rejection, chronic renal allograft dysfunction, renal fibrosis and other factors are still the major problems affecting the survival of renal graft.Relevant researches have always been hot spots in the field of renal transplantation.Meantime, 2020 is an extraordinary year.The novel coronavirus pneumonia (COVID-19) pandemic severely affects the development of all walks of life.Researches related to renal transplantation have also sprung up.In this article, the frontier hotspots of clinical and basic studies related to renal transplantation and the COVID-19 related researches in the field of renal transplantation in China were reviewed, aiming to provide novel therapeutic ideas and strategies.

Pathogeny research of pulmonary infection after renal transplantation based onmetagenomics nextgeneration sequencing / 中华器官移植杂志

Objective:Discuss the etiological characteristics of pulmonary infection after renal transplantation and the diagnostic value of metagenomics nextgeneration sequencing (mNGS) technique.Methods:A total of 40 patients with pulmonary infection who were admitted to the Department of Renal Transplantation of the First Affiliated Hospital of Medical College, Xi'an Jiaotong University from January, 2018 to January, 2021 were selected, and identification of pathogens through routine pathogen detection methods and mNGS. The routine pathogen detection methods included: blood culture, bronchoalveolar lavage fluid (BALF) and sputum culture and smear staining, lung histopathology, antigen detection and PCR, etc. BALF were used to search for pathogens by mNGS. Combined with the results of the two groups to give accurate anti-infection treatment, the clinical data were retrospectively analyzed.Results:Eventually 36 patients were cured and discharged, and 4 patients deaths. In 40 cases of pulmonary infection, the BALF mNGS pathogens detection of BALF was positive in 37 cases and negative in 3 patients, with a detection sensitivity of 92.5%. In addition, there were 15 cases of single pulmonary infection and 22 cases of mixed pulmonary infection, including 8 cases of bacterial infection, 9 cases of viral infection and 20 cases of fungal infection, among which pneumocystis (20/40, 50%) and cytomegalovirus (10/40, 25%) were the most common. In contrast, the positive rate of pathogens by routine detection were only 30% (12/40), and the difference between the two detection methods was statistically significant ( χ2=32.92, P<0.05). The diagnostic rates of mixed pulmonary infection were 55% and 10% respectively, the difference was statistically significant ( χ2=18.46, P<0.05), the single type pulmonary infection was 30% and 20% respectively, the difference was not statistically significant( χ2=2.99, P>0.05). Conclusions:mNGS has more advantages than routine pathogen detection methods in terms of pathogen species and distribution, detection time, sensitivity, mixed infection diagnosis rate and benefit. Using mNGS can be more efficient to find pathogens of pulmonary infection after renal transplantation, take accurate treatment, reduce costs, and improve cure rate, such as worth wide application..

Distribution of bacteria infected by metagenomic sequencing technology in maxillofacial space / 华西口腔医学杂志

OBJECTIVES@#This study aimed to compare and analyze the consistency and difference between metageno-mic next-generation sequencing (mNGS) and conventional bacterial culture in the detection of pathogenic microorganisms in maxillofacial space infection, as well as to provide a new detection method for the early clinical identification of pathogenic bacteria in maxillofacial space infection.@*METHODS@#The clinical data of 16 patients with oral and maxillofacial space infections in the First Affiliated Hospital of Zhengzhou University from March 2020 to June 2020 were collected. mNGS and conventional bacterial culture methods were used to detect pus. We then analyzed and compared the test results of the two methods, including the test cycle, positive detection rate, anaerobic bacteria, facultative anaerobes and aerobic bacteria detection rates, distribution of pathogenic bacteria, relative species abundance, and resistance genes.@*RESULTS@#The average inspection period of mNGS was (18.81±3.73) h, and the average inspection period of bacterial culture was (83.25±11.64) h, the former was shorter than the latter (@*CONCLUSIONS@#Compared with conventional bacterial culture, mNGS has the characteristics of short test time, high sensitivity, and high accuracy. Thus, it is a new detection method for the early identification of pathogenic bacteria in maxillofacial space infection and is beneficial to the early clinical diagnosis and treatment of the disease.

Research strategies for endophytes in medicinal plants based on high-throughput sequencing and traditional culture and isolation methods / 中国中药杂志

The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.

Advance in study on 16S rRNA gene sequencing technology in oral microbial diversity / 中南大学学报(医学版)

The 16S rRNA gene is the most commonly used molecular marker for identifying microorganisms. It is used in sequencing technology, including the first-generation, the second-generation, and the third-generation sequencing technology. A large number of studies on the 16S rRNA gene have contributed to a deeper understanding of oral microbial diversity. In the healthy oral cavity, there is microbial diversity in time and space. With the occurrence or development of oral diseases such as caries, periodontal disease, or halitosis, the microbial diversity will be changed.

Research progress and application of nanopore sequencing technology / 生物工程学报

Sequencing technology has been greatly improved in terms of throughput and cost. The single-molecule nanopore DNA sequencing, one of the major branches of the third-generation sequencing technology, has made great contributions in the fields of medicine and life sciences due to its advantages of ultra-long reading length, real-time detection and direct detection of base methylation modification, etc. This article briefly describes the principle of nanopore sequencing technology, and discusses its application in clinical, animal, plant, bacterial and virus fields and its future development direction.

Effect of active peptides of Eupolyphaga on intestinal flora in rats with hyperlipemia / 中国药理学通报

Aim To investigate the effect of active peptide of Eupolyphaga (APE) on changes of intestinal flora and blood lipid in rats with hyperlipidemia. Methods SD rats fed with high-fat diet were induced to hyperlipemia model. Simvastatin being a positive drug, ELISA was used to investigate the effect of APE on blood lipids in hyperlipidemic rats. At the same time, the results of the microbial flora about APE model rats were measured using 16rS high-throughput assay technology (V4-V5). Results APE could significantly reduce the serum lipid index in model rats and improve the degree of liver pathological changes. Meanwhile, the intestinal flora results showed that when the relative abundance of hymenoscyphus, lactobacillus and bifidobacterium in firmicutes were enriched, the relative abundances of rumenococcus, plasmodium and prevotella in bacteroidetes and tenericutes were reduced. Conclusions APE could obviously reduce the level of blood lipid and repair the disordered intestinal flora of hyperlipidemia rats. It can be speculated that APE might play a role in lowering blood lipids through the intervention of lipid metabolism pathway by intestinal flora.