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BACKGROUND:Semen cuscutae has the effect of tonifying the liver and kidney system and benefiting the essence.The main pathogenesis of osteoarthritis is deficiency of the liver and kidneys.Therefore,it is hypothesized that there is a link between semen cuscutae and osteoarthritis. OBJECTIVE:To explore the potential relationship between osteoarthritis and semen cuscutae and validate the mechanism of semen cuscutae based on the network pharmacology and molecular docking analysis. METHODS:First,the active ingredients and targets of semen cuscutae were screened in TCMSP,and the genes related to osteoarthritis were collected in the disease databases GeneCard's,OMIM and TTD.The intersected genes were taken and then subjected to a series of analyses and screened for hub genes.Through the enrichment analysis of hub genes,the pathway of semen cuscutae acting on osteoarthritis was selected.The role of hub genes was verified by molecular docking.Therefore,the appropriate active ingredients of semen cuscutae were selected for experimental verification. RESULTS AND CONCLUSION:There were 11 active ingredients of semen cuscutae,66 intersection target genes of semen cuscutae and osteoarthritis,and 12 hub genes,including tumor necrosis factor,interleukin 1B,TP53,RAC-alpha serine/threonine protein kinase(AKT1),vascular endothelial growth factor A,matrix metalloproteinase 9,prostaglandin peroxidase 2,cystatinase 3,epidermal growth factor,peroxisome proliferator-activated receptor gamma,interleukin 10,vascular cell adhesion factor 1.After the enrichment analysis of the hub genes,the classical inflammatory pathway,nuclear factor-κB signaling pathway,was selected for subsequent validation of semen cuscutae to alleviate osteoarthritic inflammation.Through the results obtained after molecular docking of each active ingredient and the hub gene of the pathway prostaglandin peroxidase 2,sesamin with the highest affinity was selected for subsequent cell experiments,and the experimental results confirmed that sesamin,the active ingredient of semen cuscutae,could reduce the expression of cyclooxygenase 2 by inhibiting the nuclear factor-κB signaling pathway induced by interleukin-1β.To conclude,sesamin,the active ingredient of semen cuscutae,reduces the expression of cyclooxygenase 2 by inhibiting the nuclear factor-κB signaling pathway induced by interleukin-1β,thereby improving inflammation in osteoarthritis and expanding the therapeutic effect of semen cuscutae in osteoarthritis.
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Objective:To investigate the epidemiological characteristics, genotypes and genetic evolution of respiratory syncytial virus (RSV) isolated in Shanghai from April 2020 to December 2021, which was a period from the COVID-19 outbreak to the phase of regular epidemic prevention and control.Methods:This retrospective study collected the nasopharyngeal secretions or nasopharyngeal aspirates of children with acute respiratory tract infection (ARTI) admitted to the Shanghai Children′s Hospital from April 2020 to December 2021. PCR-capillary electrophoresis and RT-PCR were used for virus identification and the amplification of the gene fragment of the second hypervariable region of RSV G protein. Homology analysis and phylogenetic analysis were conducted using bioinformatics software. Chi-square test was used to compare the detection rates of RSV. Results:A total of 6 211 samples were collected and 13.62% (846/6 211) of them were positive for RSV. The positive rates of RSV in male and female patients were 14.07% (503/3 574) and 13.01% (343/2 637), respectively, with no significant gender difference (χ 2=1.467, P=0.226). The highest detection rate of RSV was found in children ≤6 months of age, and the rate of RSV infection decreased gradually with age (χ 2=352.942, P<0.001). No RSV-positive specimens were detected from April 2020 to August 2020, after which the detection rate of RSV gradually increased with two epidemic peaks occurring from December 2020 to February 2021 and from August to October 2021. The predominant epidemic subtype was RSV subtype B in 2020 and the first 9 months of 2021, which was gradually replaced by RSV subtype A in the last 3 months of 2021. The 176 strains of RSV subtype A obtained in this study were all ON1 genotype, and the nucleotide homology of the Shanghai epidemic strains was 90.20%-99.50%. All of the 250 strains of RSV subtype B were BA9 genotype, and the nucleotide homology of the Shanghai epidemic strains was 90.10%-100.00%. Conclusions:From April 2020 to December 2021, with the regular prevention and control of COVID-19, there is a change in the epidemic season of RSV. The prevalent genotypes of RSV subtypes A and B are ON1 and BA9, respectively, and the subtype A gradually replaces subtype B as the most prevalent subtype.
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Objective To understand the mutations of macrolide resistance gene locus (23S rRNA) of Mycoplasma pneumoniae (MP) and its correlation with clinical features .Methods A total of 354 respiratory tract samples were collected from children pa-tients with pneumonia .MP and its mutations in 23S rRNA gene locus were detected by real-time PCR .The children cases of MP positive were divided into the mutation group and non-mutation group .Then the clinical data were compared between the two groups .Results Among 354 respiratory tract samples ,166 cases(46 .9% ) were MP positive ,moreover the mutation of 23S rRNA gene locus existed in 135 MP positive samples with the positive detection rate of 81 .3% ,while no 23S rRNA gene locus mutations were detected in 31 samples .Analyzing the clinical data of the mutation group and non-mutation group found that there was no sta-tistical difference in the aspects of age and gender between the two groups .The occurrence rates of severe pneumonia and extrapul-monary complications in the mutation group were higher than those in the non-mutation group (P<0 .05) ,moreover the average hospitalization time and fever duration in the mutation group were longer than those in the non-mutation group (P<0 .05) .Conclu-sion 23S rRNA gene locus mutation has higher detection rate ,prompting that MP shows high resistant rate to macrolides ,which could provide a certain basis for treatment of M P infections .
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Objective To investigate the factors affecting the isolation of fastidious bacteria such as Streptococcus pneumoniae and Haemophilus influenzae from lower respiratory tract specimens in children.Methods A total of 210 lower respiratory tract specimens from children were collected and inoculated on both blood agar plate and chocolate agar plate 0.5 h,2 h,6 h,12 h,and 24 h after collection.The effect of specimen turnaround time,transport medium,and inoculation medium on the isolation of fastidious bacteria was studied.Results The results of 200 qualified specimens showed that the isolation of S.pneumoniae and H.influenzae significantly decreased with the increasing of specimen turnaround time (P＜0.05),but no significant difference was observed between 0.5 h and 2 h timepoints.The isolation of fastidious bacteria in semi-solid medium was non-significantly higher than the other two transport medium (P＞0.05).No significant difference was found in the isolation and growth ofS.pneumoniae and H.influenzae on different inoculation media (P＞0.05).Conclusions The appropriate specimen turnaround time,transport medium,and inoculation medium are important for improving the isolation of fastidious bacteria from lower respiratory tract in children.
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Objective To understand the drug resistance and antibiotic resistance mechanism ofβ-lactam antibiotics of invasive Streptococcus pneumoniae isolated from Shanghai Children′s Hospital, provides the reference for the rational use of antimicrobial agents.Methods This study is based on the research of the mechanism of drug resistance.62 isolates of invasive Streptococcus pneumoniae were collected from Shanghai Children′s Hospital from January 2005 to December 2011.Minimum inhibitory concentrations ( MIC) of strains to 9 antimicrobial agents were determined by E-test method.The penicillin binding protein coding genes pbp2x, pbp2b, and pbp1a of Streptococcus pneumoniae were amplified by PCR.Then, the correlation between the gene mutation andβ-lactam antibiotics resistant level were analyzed.The murM gene of Streptococcus pneumoniae was amplified by PCR and the correlation of mutation and β-lactam antibiotics resistant level was analyzed.Results Out of 62 strains of invasive Streptococcus pneumoniae from children, the detection rate of penicillin resistant Streptococcus pneumoniae was 43.6% (27/62).Between penicillin intermediate Streptococcus pneumoniae ( PISP ) ( 100%, 25/25 ) and penicillin sensitive Streptococcus pneumoniae (PSSP) (3/10), the difference of gene mutation rate near the pbp2b conserved sequence was statistically significant (χ2 =21.875, P<0.01).The same situation occurred between penicillin resistant Streptococcus pneumoniae (PRSP)(100%, 27/27)and PSSP (3/10) (χ2 =23.310, P<0.01).Also the difference of gene mutation rate of PISP (84%, 21/25) vs PSSP (0) and PSSP (0) vs PRSP (85.2%, 23/27) near or in the pbp2x conserved sequence were statistically significant (χ2 =21.000, P <0.01;χ2 =22.513,P<0.01).The difference of gene mutation rate near the pbp1a conserved sequence and Insertion sequence, which were statistically significant, occurred between PISP and PSSP (χ2 =13.22,P<0.01), between PRSP and PSSP (χ2 =37.000,P<0.01), between PISP and PRSP (χ2 =10.211,P=0.001). MurM gene mutation rate was statistically significant different between the 2 group penicillin MIC≥8 mg/L or ceftriaxone MIC≥2 mg/L group (95.8%, 23/24) and penicillin MIC<8 mg/L or ceftriaxone MIC<2 mg/L group (0) (χ2 =56.2,P =0.002 6).Conclusions The resistance phenomenon of invasive Streptococcus pneumoniae in Shanghai Children′s Hospital is serious.The gene mutations of pbps and murM play a role in amide in the beta of antibiotic resistance, and there is a certain correlation with the antibiotic resistance level.