OBJECTIVE To analyze the microbial community structure in Bronchoalveolar Lavage Fluid(BALF)u-sing high-throughput sequencing technology,with a particular focus on the abundance and distribution of Staphy-lococcus aureus.METHODS A total of 1,650 bronchoalveolar lavage fluid(BALF)samples were collected from the First People's Hospital of Yunnan Province between 2022 and 2023.S.aureus gene detection was performed using BALF culture,isothermal amplification and microfluidic technology.Representative 21 S.aureus-positive samples were grouped into S.aureus pneumonia group,SARS-CoV-2 co-infection group and chronic obstructive pulmona-ry disease(COPD)group,followed by 16S rDNA amplicon high-throughput sequencing.RESULTS The microbial community compositions among the three groups exhibited significant differences.Species-level hierarchical cluste-ring analysis revealed that S.aureus was more frequently detected and accounted for a higher proportion in the S.aureus pneumonia group,while the COPD group was dominated by Achromobacter.In contrast,the SARS-CoV-2 co-infection group was mainly composed of Methylobacterium aquatique and Achromobacter.Only in the S.au-reus pneumonia group was S.aureus the dominant bacterium.CONCLUSION The composition of the BALF micro-biota is closely related to different disease states,providing an important reference for a deeper understanding of the microbiological basis of pulmonary infectious diseases.

Objective:To investigate the effects of decellularised extracellular matrix material(dECM)extract on macrophage survival,migration,phagocytosis,pro-inflammatory factor expression and ROS production.Methods:CCK-8 method was used to detect whether the extract of dECM material had cytotoxicity on macrophages;effects of dECM material extracts on macrophage recruit-ment and chemotaxis were examined by Transwell migration assay;effects of dECM material extract on macrophage phagocytosis were detected by pHrodo staining;effects of dECM material extract on expression of pro-inflammatory genes and pro-inflammatory specific functional molecules were detected by RT-qPCR and immunofluorescence staining;detection of the effect of dECM material extracts on macrophage ROS production by the DCFH-DA fluorescent probe approach.Results:Compared with DMEM complete medium,①CCK-8 assay showed that dECM material extract had no toxic effect on macrophages,and could promote the formation of macrophage colonies;②pHrodo staining assay showed that dECM material extract did not affect phagocytosis of macrophages;③Transwell migra-tion assay showed that dECM material extract did not promote macrophage migration;④RT-qPCR results showed that dECM material extracts were able to down-regulate gene expressions of pro-inflammatory cytokines and specific functional molecules;⑤Flow cytome-try results showed that dECM material extract could reduce the production of ROS by macrophages.dECM material extract had excel-lent biocompatibility for macrophages.Conclusion:dECM material extract is non-toxic to macrophages and is able to reduce macro-phage pro-inflammatory gene expression and ROS production.

Objective:To investigate the effects of decellularised extracellular matrix material(dECM)extract on macrophage survival,migration,phagocytosis,pro-inflammatory factor expression and ROS production.Methods:CCK-8 method was used to detect whether the extract of dECM material had cytotoxicity on macrophages;effects of dECM material extracts on macrophage recruit-ment and chemotaxis were examined by Transwell migration assay;effects of dECM material extract on macrophage phagocytosis were detected by pHrodo staining;effects of dECM material extract on expression of pro-inflammatory genes and pro-inflammatory specific functional molecules were detected by RT-qPCR and immunofluorescence staining;detection of the effect of dECM material extracts on macrophage ROS production by the DCFH-DA fluorescent probe approach.Results:Compared with DMEM complete medium,①CCK-8 assay showed that dECM material extract had no toxic effect on macrophages,and could promote the formation of macrophage colonies;②pHrodo staining assay showed that dECM material extract did not affect phagocytosis of macrophages;③Transwell migra-tion assay showed that dECM material extract did not promote macrophage migration;④RT-qPCR results showed that dECM material extracts were able to down-regulate gene expressions of pro-inflammatory cytokines and specific functional molecules;⑤Flow cytome-try results showed that dECM material extract could reduce the production of ROS by macrophages.dECM material extract had excel-lent biocompatibility for macrophages.Conclusion:dECM material extract is non-toxic to macrophages and is able to reduce macro-phage pro-inflammatory gene expression and ROS production.

OBJECTIVE To analyze the microbial community structure in Bronchoalveolar Lavage Fluid(BALF)u-sing high-throughput sequencing technology,with a particular focus on the abundance and distribution of Staphy-lococcus aureus.METHODS A total of 1,650 bronchoalveolar lavage fluid(BALF)samples were collected from the First People's Hospital of Yunnan Province between 2022 and 2023.S.aureus gene detection was performed using BALF culture,isothermal amplification and microfluidic technology.Representative 21 S.aureus-positive samples were grouped into S.aureus pneumonia group,SARS-CoV-2 co-infection group and chronic obstructive pulmona-ry disease(COPD)group,followed by 16S rDNA amplicon high-throughput sequencing.RESULTS The microbial community compositions among the three groups exhibited significant differences.Species-level hierarchical cluste-ring analysis revealed that S.aureus was more frequently detected and accounted for a higher proportion in the S.aureus pneumonia group,while the COPD group was dominated by Achromobacter.In contrast,the SARS-CoV-2 co-infection group was mainly composed of Methylobacterium aquatique and Achromobacter.Only in the S.au-reus pneumonia group was S.aureus the dominant bacterium.CONCLUSION The composition of the BALF micro-biota is closely related to different disease states,providing an important reference for a deeper understanding of the microbiological basis of pulmonary infectious diseases.

Objective:To investigate the clinical features, gene mutation and follow-up outcome of children with paroxysmal kinesigenic dyskinesia(PKD).Methods:Clinical data was collected at Beijing Tiantan Hospital Affiliated to Capital Medical University from November 2018 to November 2019.In total, seven children with PKD were recruited, and peripheral blood samples for gene study were collected from six patients and their parents.Mutation analysis of PRRT2 gene was performed by PCR sequencing in children and by Sanger sequencing in patients.Results:Of the seven patients, four were male and three were female, and the median age of onset was 11 years and 6 months, ranging from 5 to 14 years.Among them, two patients were family cases and the other five patients were sporadic cases.The presentation were abnormal involuntary movements provoked by sudden movements, without loss of consciousness.Five patients exhibited dystonia and two patients had dystonia and choreoathetosis.The duration of the attacks lasted for a few seconds to 40 seconds.The frequency ranged from 5 to 15 times per day.PRRT2 mutations, c.649_650insC(P.R217PfsX8), were found in two patients with PKD families and three sporadic PKD cases.Conclusion:The onset age of PKD is pre-school or school age.The attacks manifest as dystonia or mixed with dystonia and choreoathetosis.PRRT2 is the main pathogenic gene of PKD and mutation c. 649_650insC is the hotspot mutation.Low-dose Carbamazepine has good effects.

Objective To study the effects of medicated serum with total saponins from Rhizoma Dioscreae Nipponicae (RDN) on VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α. To investigate the mechanism about total saponin from RDN inhibition of angiogenesis. Methods Medicated serum of total saponins from RDN and tripterygium (positive control) were prepared. Rat synovial cells RSC-364 were divided into four groups: the blank control,IL-17+TNF-α model,tripterygium medicated serum,and total saponins medicated serum groups. After one hour of incubation,all groups except for the blank control were incubated with both IL-17(10 μg·L-1 ) and TNF-α(10 μg·L-1 ) for 24 hours. VEGF mRNA expression in RSC-364 was detected by PrimeScriptTM real-time quantitative PCR (RT-PCR) detection kit,and the AP-1 DNA-binding activity was detected by electrophoretic mobility shift assay (EMSA). Results Compared with the control blank group,both of the VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α increased remarkably (P<0. 05,P<0.01). The VEGF mRNA expression and AP-1 activity in tripterygium medicated serum group and total saponins medicated serum group were remarkably lower than those of the model control group (P<0.05). There was no significant difference between the two medicated serum groups. Conclusion Serum medicated with total saponins from RDN can remarkably decrease VEGF mRNA expression and AP-1 activity,indicating that the total saponins from RDN could influence VEGF secretion by inhibiting the AP-1 signal transduction pathway,VEGF is the key factor of angiogenesis,thereby to restrain angiogenesis.

This paper was aimed to study effects of diosgenin on expressions of activator protein-1 (AP-1) and vascular endothelial growth factor (VEGF) in synovial tissues of collagen-induced arthritis (CIA) rats induced by bovine type II collagen, in order to investigate the possible mechanism of herbal medicine diosgenin in the treatment of rheumatoid arthritis (RA). After the CIA rats models were successfully established, rats were randomly divided into the blank control group, CIA model group, diosgenin group, and positive medicine control (tripterygium) group. The in situ hybridization was used to detect the expressions of AP-1 (c-fos and c-jun) in synovial tissues of the knee joint. The real-time PCR was used to detect the VEGF mRNA expression in synovial tissues of rats’ knee joints. The results showed that c-jun and c-fos, VEGF mRNA expressions in synovial tissues of rats’ knee joints were obviously higher than that of the blank control group (P<0.01). After treatment of diosgenin and tripterygium, the expressions of c-jun and c-fos, VEGF mRNA were significantly reduced (P<0.01). It was concluded that diosgenin may regulate the expression of VEGF in synovial tissues through c-jun and c-fos of AP-1 in order to inhibit synovial angiogenesis for the treatment of RA.

Objective To explore the susceptibility of epilepsy in rat with cerebral trauma. Methods An impact-acceleration head injury model was established with rats. After trauma, the electroencephalograph was recorded. Epileptic model wad established by injecting pentylenetetrazol (PTZ) intraperitoneally and the dosage of PTZ was recorded. Results The wave of delta and theta increased after trauma, alpha and beta decreased and there was significant difference among the power of delta, theta and alpha (P＜0.05). The dosage of rats with cerebral trauma was less than that in normal rats (P＜0.05). Conclusion The susceptibility of epilepsy in rat with cerebral trauma increases.