Objective:To develop and validate a liquid chromatography-tandem mass spectrometry method for determining levofloxacin in plasma sample from pediatric patients.Method:This is a prospective, observational study. The clinical residual plasma samples from healthy individuals for physical examination in Children's Hospital Affiliated to Zhengzhou University were collected as blank matrix. Plasma samples from five pediatric patients who did not receive levofloxacin or ciprofloxacin in the department of Respiration were collected for methodological evaluation. In addition, 34 clinical plasma samples from 22 pediatric patients (9 males and 13 females; mean age (8.1±3.7) years) using levofloxacin was collected, and their plasma concentrations were determined. Using ciprofloxacin as the internal standard, levofloxacin in plasma samples was quantified by liquid chromatography-tandem mass spectrometry following protein precipitation using acetonitrile. A C18 column (Shim-pac GIST-HP C18, 2.1 mm×100 mm, 3 μm) and mobile phase composed of water (containing 0.1% formic acid) and acetonitrile (containing 0.1% formic acid) with gradient elution at a flow rate of 0.4 ml/min were used to separate levofloxacin. The column temperature was 40 ℃, injection volume was 1 μl and the total analysis time was 9 min. Levofloxacin and ciprofloxacin were ionized with an ESI source in positive ion mode and detected in multiple reaction monitoring (MRM) mode. The detected ions of levofloxacin and ciprofloxacin were m/z 362.10→318.1 and 332.15→231.05, respectively. The method′s specificity, sensitivity, linearity, precision, accuracy, recovery rate, stability, matrix effect, and carry-over were validated. All statistical analyses were performed with SPSS statistical software (version 17.0). The normality of the data was detected by the K-S test. A P<0.05 was considered statistically significant for two tailed tests. Results:The LC-MS/MS method showed a good linearity within the range of 0.062 5-20 mg/L, with the lower detection limit of levofloxacin of 0.062 5 mg/L. The calibration curve for levofloxacin was Y=0.093X+0.010 ( R2>0.99). Under different quality control levels, the accuracy ranged from 92.57% to 104.39%, and the intra-day and inter-day imprecision ranged from 2.32% to 9.35%. These values were not affected by the normal matrix, 5% hemolysis matrix and 15% hyperlipidemia matrix. Furthermore, the levofloxacin plasma samples were stable in the short term. A total of 34 plasma samples from 22 patients were collected and analyzed. Only 2 plasma samples were below the lower limit of quantification, while the other plasma concentrations of levofloxacin were ranged from 0.091 to 6.755 mg/L. Cmax was (5.52 ± 1.09) mg/L. Conclusion:The LC-MS/MS method meets the requirements of the reference method and requires a small sample size (50 μl), making it suitable for the determination of levofloxacin in plasma from pediatric patients.

Objective:To clarify peripheral Th17 level in SSc patients and its correlation with disease.Methods:Chinese databases CNKI, CBM, Wanfang and VIP, and English databases PubMed, EMBASE, Web of Science, Cochrane Library and Science Direct were searched to collect a case-control study on the content of Th17 cells in peripheral blood of patients with SSc. The papers published when the database was first developed in 25 February 2021. Meta-analysis was conducted using Stata 12.0 software, and I2 and Egger tests were used to evaluate the heterogeneity and publication bias between studies. Results:A total of 26 case-controls were included in the study, including 1 160 patients with SSc and 778 healthy controls. Overall, the percentage of Th17 cells in SSc patients was higher than in healthy controls [SMD(95% CI)=1.85 (1.33, 2.38), P<0.001], which was most significant in IL-17 +Th17 concentration [SMD(95% CI)=1.88 (1.28, 2.48), P<0.001]. As for disease activity, the proportion of Th17 cells in active SSc patients was much higher than those of patients in remission [SMD(95% CI)=1.92 (1.12, 2.71), P<0.001]. SSc patients had a reduced Th17 level after receiving DMARDs treatment [SMD(95% CI)=-0.74 (-1.05, -0.42), P=0.029]. Conclusion:The number of Th17 cells increase significantly in the peripheral blood of patients with SSc, and is related to disease activity. DMARDs can be used to treat this disease by downregulating Th17 levels.

Objective:To analyze the application effect of the information platform based on the Internet of Things (IoT) in the screening and management of patients with chronic obstructive pulmonary disease (COPD).Methods:In this cross-sectional study, 151 062 residents who participated in the screening in the districts covered by 33 general hospitals and 289 primary medical institutions within the framework of Henan Provincial Center for COPD Prevention and Treatment from November 2021 to October 2022 were included as the subjects. Spirometer based on the IoT was used to conduct pulmonary function tests for subjects who scored 16 points or more with COPD Screening Questionnaire (COPD-SQ), and the subjects were evaluated and managed through the structured electronic data in the information platform. The distribution characteristics and follow-up of the subjects and COPD patients were described and the application effect of this strategy in patients screening and management was analyzed.Results:A total of 151 062 residents with complete basic information in the information platform completed the questionnaire. High-risk population of COPD accounted for 26.5% (40 042/151 062) of the population who received the questionnaire screening, and more than 50% had respiratory symptoms, such as chronic cough (59.4%), or shortness of breath (77.6%). The proportion of high-risk population increased with age, especially after 50 years old. Compared with non-smokers, the proportion of high-risk group was significantly higher (77.1% vs 16.4%) in the group with smoking index over 600. Biofuel exposure (61.3% vs 22.1%) and family history of respiratory diseases (64.2% vs 22.6%) were associated with an increased proportion of high-risk groups, with statistically significant differences ( P<0.001). 5 268 patients were diagnosed with COPD by pulmonary function tests, and the prevalence of COPD in high-risk groups was 27.8% (5 268/18 965), the prevalence rate of male was 34.0% (3 942/11 588), which was higher than that of female 18.0% (1 326/7 377). 2 950 patients (56.0%) completed at least one follow-up of symptom questionnaire and 510 patients (9.7%) completed more than one follow-up of pulmonary function test. Conclusion:The screening and management strategy of COPD based on the IoT and information technology can improve the efficiency of COPD screening, and improve the status quo of under-diagnosis and discontinuous follow-up of COPD.

Humans ; Multiple Pulmonary Nodules ; Lung ; Lung Neoplasms/surgery* ; Solitary Pulmonary Nodule/surgery* ; Bronchoscopy

ObjectiveTo compare the current development of physical therapists in China, Japan and South Korea. MethodsThe condition of configuration, education and practice of physical therapists in China, Japan and South Korea were browsed from the official websites and existing data. ResultsThe allocation, education and training mechanism, and the professional accreditation system needs to be optimized in China. ConclusionThe physical therapists should be allocated based on rehabilitation service settings and needing, trained based on the World Health Organization's Rehabilitation Competency Framework and the International Classification of Functioning, Disability and Health, and the professional system should be constructed based on the World Health Organization's Rehabilitation Competency Framework and national professional admission systems.

Objective: To investigate the role of long non-coding RNA double homeobox A pseudogene 9 (DUXAP9) in head and neck squamous cell carcinoma (HNSCC) and to evaluate the expression level, molecular function and mechanism of DUXAP9 in HNSCC cells. Methods: Differential expression of lncRNAs between normal and tumor tissues in HNSCC tissues were screened using lncRNA microarray, the expression level of DUXAP9 in HNSCC tissues and its relationship with prognosis were analyzed in the TCGA database. The expression levels of DUXAP9 in HNSCC tissues and cell lines were detected using qRT-PCR. The function in HNSCC cells after DUXAP9 silencing was evaluated using the CCK-8 assay, wound healing assay, Transwell migration assay and subcutaneous xenograft assay in nude mice. Changes in the transcription and translation of epithelial-mesenchymal transition (EMT)-related proteins in head and neck squamous cell carcinoma cells after DUXAP9 silencing were detected using qRT-PCR and Western blot. Results: lncRNA microarray results showed that, compared to adjacent normal tissues, DUXAP9 was abnormally upregulated in HNSCC tissues. Analysis from TCGA database showed that, compared to HNSCC patients with low DUXAP9 expression, HNSCC patients with high DUXAP9 expression had poorer survival. The relative expression of DUXAP9 in HNSCC tissues and 4 HNSCC cell lines increased compared to paired adjacent normal tissues as detected using qRT-PCR. Silencing DUXAP9 significantly inhibited the proliferation, migration and expression of EMT-related genes in HNSCC cells. The silencing of DUXAP9 significantly inhibited subcutaneous tumorigenesis of the HNSCC cell line CAL27 in nude mice. Conclusion Silencing DUXAP9 significantly inhibited the proliferation of HNSCC cells and subcutaneous xenografts in nude mice. DUXAP9 may mediate the migration of head and neck squamous cell carcinoma cells via the EMT pathway.

Sedentariness is the most common bad habit of the elderly,which seriously affects their health and quality of life. This article summarized the research status,evaluation methods and influencing factors of sedentary behavior of the elderly,in order to provide reference for the intervention research of sedentary behavior of the elderly in China.

Background Conjunctivochalasis (CCh) is a common age-related ocular surface diseases.Researches showed that the inflammatory factors are upregulated in conjunctiva and tear of CCh patients,inferring inflammation participates in the pathogenesis of CCh,however,its mechanism is unclear now.Studies also showed that the expression of matrix metalloproteinases (MMPs) in CCh conjunctiva is elevated.However,the association between inflammatory factors and MMPs is unknown.Objective This research was to observe the effects of tumor necrosisfactor-α (TNF-α) and interleukin-1 β (IL-1 β) on the expression of MMP-1,MMP-3,tumor necrosis factor-stimulatedgene-6(TSG-6) and pentraxin-3 (PTX3) in cultured CCh fibroblasts.Methods Conjunctival fibroblasts wereisolated and cultured from CCh-derived and cataract-derived human conjunctiva explants.The cells were treated using20 ng/ml PBS,TNF-α or IL-1β for 4 hours,respectively.The expression levels of MMP-1,MMP-3,TSG-6 and PTX3genes and proteins were detected by quantitative real time PCR and Western blot assay,respectively.Results Thesecond-generation cells showed a long and fusiform shape with ovoid nucleus and radial agreement,and the cellprocessors connected each other.The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3mRNA in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-αand IL-1 β (MMP-1 mRNA:Fgroup =2.611,P =0.116;Fi =161.564,P =0.000;MMP-3 mRNA:Fgroup =5.201,P =0.029;Fintervene =211.021,P =0.000;TSG-6 mRNA:Fgroup =47.209,P =0.000;Fintervene =119.340,P =0.000;PTX3 mRNA:Fgroup =40.512,P =0.000;Fintervene =93.935,P =0.000).Compared with the cataract group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with PBS treatment or the cataract group with TNF-α and IL-1 β treatment (all at P＜0.05).Compared with the CCh group with PBS treatment,the expression levels of MMP-1,MMP-3,TSG-6 and PTX3 mRNA were significantly elevated in the CCh group with TNF-α and IL-1β treatment (all at P ＜ 0.05).The differences of the relative expression levels of MMP-1,MMP-3,TSG-6 and PTX3 proteins in the cells were significantly different between CCh group and cataract group after being treated by PBS,TNF-α and IL-1 β (MMP-1:Fgroup =84.702,P =0.000;Fint =48.900,P =0.000;MMP-3:Fgroup =112.818,P =0.000;Fint =194.980,P =0.000;TSG-6:Fgroup =56.867,P =0.000;Fint =70.356,P =0.000;PTX3:Fgroup =1.488,P =0.231;Fint =89.872,P =0.000),and the expression changes of MMP-1,MMP-3,TSG-6 and PTX3 proteins were coincident with the genes among the groups with various treatments.Conclusions The expressions of MMP-1,MMP-3,TSG-6 and PTX3 in conjunctival fibroblasts are upregulated in CCh eyes.The interaction of TNF-α and IL-1 β with MMPs is probably involved with the pathogenesis and development MMPs probably.

Objective To observe the changes of tear film stability,goblet cell and mucin 5AC expression in conjunctivochalasis patients,and explore the mechanism of conjunctivochalasis.Methods Conjunctivochalasis patients (30 cases) and single age-related cataract patients (15 cases) were collected as conjunctivochalasis group and normal control group.Eye symptom assessment (OSDI score),tear break-up time (BUT),Schirmer Ⅰ test,tear fern crystallization tests were performed for all the selected persons.Conjunctival rescent-shaped resections were made for all the conjunctivochalasis patients.Conjunctival tissue samples were stained by HE staining,AB staining,mucin 5AC immunohistochemical staining from the conjunctivochalasis group and norral control group respectively,and then statistical analysis was made.Results The OSDI score in the conjunctivochalasis group (37.80 ± 8.94) was significantly higher than that in the normal control group (11.40 ±4.08) (P ＜0.01).BUT in the conjunctivochalasis group (6.70 ± 2.76) s was significantly lower than that in the normal control group (13.67 ± 3.48) s (P ＜ 0.01).Schirmer Ⅰ test in the conjunctivochalasis group (6.23 ± 3.13) mum was significantly lower than the normal control group (13.40 ± 3.74)mm (P ＜ 0.01).Tear ferbing crystallization of the conjunctivochalasis group was decreased significantly compared with the normal control group (x2 =14.309,P =0.003).Light microscopic showed that conjunctival thickness was thinned,collagen fibers were less,elastic fiber was reduced,the lamina propria and interstitial were congestion and edema,the number of goblet cells was significantly reduced,and the positive staining of mucin 5AC staining was significantly lower in the conjunctivochalasis group than in normal control group (x2 =9.499,P =0.023).Conclusion For patients with conjunctivochalasis,the tear film function is affected,goblet cells are decreased,tear fern crystallization is decreased,mucin 5AC content is decreased,which finally leads the excessive conjunctival relaxation and abnormal ocular surface and tear.

AIM:To investigate the effect of growth differentiation factor 11 ( GDF11 ) on the expansion of CD8 +memory stem T cells ( Tscm) and to further improve the effect of adoptive immunotherapy.METHODS:Healthy human peripheral blood mononuclear cells ( PBMCs) were isolated by density gradient centrifugation at first.Among the i-solated PBMCs, CD8 +T cells were further purified with MACS microbeads.The CD8 +T cells were then randomly divided into experimental groups and control group.The same volume of different concentrations of GDF11 were added into the ex-perimental groups, and the same volume of PBS solution was added into the control group.Finally, the expansion of Tscm in experimental groups and control group was measured by flow cytometry at several time points.RESULTS:GDF11 sig-nificantly increased the number of Tscm in CD8 +T cells in vitro expansion and also dramatically increased the ratio of Tscm in CD8 +T cells.Furthermore, 400 μg/L GDF11 treatment for 3 weeks was the optimal condition to induce CD8 +Tscm. CONCLUSION:GDF11 effectively increases the number and ratio of Tscm in the CD8 +T cells in cell culture growth, thereby creating a new strategy to further improve the efficiency of adoptive immunotherapy.