BACKGROUND:Early diagnosis and treatment of intervertebral disc degeneration is particularly important.Pyroptosis of nucleus pulposus cells plays an important role in the early process of intervertebral disc degeneration,but the role of pyroptosis-related molecules of nucleus pulposus cells in early intervertebral disc degeneration and related molecular markers are still unclear. OBJECTIVE:To explore the diagnostic and therapeutic value of differentially expressed genes related to pyroptosis during intervertebral disc degeneration. METHODS:Public datasets of the GEO database were integrated for differential analysis,and intersected with 33 pyroptosis-related genes previously reported.Using Gene Ontology analysis and Kyoto Encyclopedia of Genes and Genomes,pathway enrichment analysis was performed in genes related to pyroptosis during intervertebral disc degeneration.Enrichment analysis of the intervertebral disc degeneration dataset was conducted using gene set enrichment analysis.To construct the immune cell spectrum in intervertebral disc degeneration samples,the correlation between the differentially expressed pyroptosis-related genes and immune cells was analyzed.Protein interaction networks were built.Screening of the Hub gene to identify key genes associated with intervertebral disc degeneration in protein interaction networks.The receiver operating characteristic curve of the differentially expressed pyroptosis-related genes was plotted,and the area under the curve was calculated.The clinical diagnostic value of target genes was explored.qRT-PCR was applied to verify the difference in the expression of pyroptosis-related genes between normal human nucleus pulposus cells and degenerated human nucleus pulposus cells with intervertebral discs. RESULTS AND CONCLUSION:(1)4426 differentially expressed genes associated with intervertebral disc degeneration were obtained,and 14 differentially expressed pyroptosis-related genes were obtained after the intersection.(2)Gene Ontology and Kyoto Encyclopedia of Genes and Genomes revealed important enrichment pathways,mainly related to inflammation,cell cycle,infection and NOD-like receptor pathway.Gene set enrichment analysis showed that pathways such as amino acid metabolism and P53 transcription regulation were significantly enriched in patients with intervertebral disc degeneration.Immune infiltration analysis suggested that the occurrence and development of intervertebral disc degeneration were closely related to immune cells.(3)A total of five Hub genes were screened,namely IL1-β,Caspase-1,AIM2,Caspase-5,and NLRC4.(4)Acquisition and identification of key biomarkers for intervertebral disc degeneration:After intersecting the two datasets of GEO with the pyroptosis-related gene,it was found that NLRP3 had significant expression differences,and the receiver operating characteristic curve showed that NLRP3 had clinical diagnostic significance.(5)qRT-PCR showed that the expression of Hub genes such as IL1-β,Caspase-5 and NLRC4 was significantly increased in the nucleus pulposus cells of intervertebral disc degeneration(P<0.05),and there was no difference in Caspase-1,AIM2 and NLRP3 expression(P>0.05).(6)It is suggested that cell pyroptosis plays an important mechanism in the occurrence and development of intervertebral disc degeneration,among which pyroptosis-related molecules NLRP3,IL1-β,Caspase-5 and NLRC4 have early diagnosis and treatment value.

Objective To investigate the effect of intravitreal injection of fibrillin-2(FBN2)recombinant protein on FBN2-deficient retinopathy.Methods Thirty-two SPF-grade C57BL/6J mice were randomly divided into 4 groups:nor-mal control group,negative control group,FBN2 knockdown group,and FBN2 recombinant protein group,with 8 mice in each group.The right eyes were taken as the experimental eyes.Mice in the normal control group did not receive any inter-vention,mice in the negative control group were intravitreally injected with 3 μL empty vector(1 mg·L-1),and mice in the FBN2 knockdown group and FBN2 recombinant protein group were intravitreally injected with 3 μL adeno-associated vi-rus(1 mg·L-1).After 4 weeks,mice in the FBN2 recombinant protein group were intravitreally injected with 3 μL FBN2 recombinant protein(1 mg·L-1).Then,electroretinogram(ERG)and optical coherence tomography(OCT)were used to measure the amplitude of Rod-b and Max-a waves and the changes in the retinal structure.Real-time quantitative poly-merase chain reaction(RT-PCR)and Western blot were used to detect changes in FBN2,microfibril-associated glycopro-tein 2(MAGP-2),collagen I(COL1)mRNA and protein expression in the mouse retina.Results The ERG findings showed that compared with the negative control group and normal control group,the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 knockdown group and FBN2 recombinant protein group decreased(all P＜0.05);com-pared with the FBN2 knockdown group,the amplitude of Rod-b and Max-a waves in the retina of mice in the FBN2 recom-binant protein group significantly increased(both P＜0.05).The OCT findings showed that compared with the FBN2 knock-down group,the structure of the retinal pigment epithelium and the light reflex in the FBN2 recombinant protein group be-came more regular.The RT-PCR detection results showed that compared with the FBN2 knockdown group,the expression of FBN2 mRNA in the retinal tissue of mice in the FBN2 recombinant protein group significantly increased,while the ex-pression of COL1 and MAGP-2 mRNA significantly decreased(all P＜0.05).Western blot assay results showed that com-pared with the FBN2 knockdown group,the expression of FBN2 protein in the retinal tissue of mice in the FBN2 recombi-nant protein group increased significantly,while the expression of COL1 and MAGP-2 proteins decreased significantly(all P＜0.05).Conclusion Intravitreal injection of FBN2 recombinant protein can compensate for the endogenous deficiency of FBN2 in mice with FBN2-deficient retinopathy and achieve therapeutic effects by regulating COL1 and MAGP-2 expres-sion.

Objective:To investigate the expression of latent transforming growth factor-β-binding protein (LTBP), transforming growth factor-β (TGF-β), cyclin-dependent kinase 2 (CDK2) and cyclin D2 (CCND2) in fibrillin-2 ( FBN2) interfering induced mouse retinopathy. Methods:Twenty-seven 8-week-old C57BL/6J mice were randomly divided into normal control group, empty vector group and FBN2 interference group according to the random number table method, with 9 mice in each group.The normal control group was not treated.The empty vector group and FBN2 interference group were intravitreally injected with 3 μl empty vector and 3 μl adeno-associated virus (AAV) carrying the sh-FBN2 interference plasmid in the right eye, respectively.The structural and functional changes of the retina were detected at 4 weeks after injection by optical coherence tomography (OCT) and full-field electroretinography (ERG).The expression and distribution of FBN2 protein in the retina were detected by immunofluorescence staining.The mRNA and protein expression levels of FBN2, LTBP-1, TGF-β2, CDK2 and CCND2 in mouse retina were detected by real-time fluorescence quantitative PCR and Western blot.All experiments complied with the ARVO statement.The research scheme was approved by the Experimental Animal Ethics Committee of Shandong University of Traditional Chinese Medicine (No.2019036).Results:Four weeks after injection, the results of OCT examination showed that compared with normal control and empty vector groups, the retinal pigment cortex of the FBN2 interference group was irregular with high density reflection areas.Full-field ERG results showed that compared with normal control and empty vector groups, the amplitude of Rod-a, Rod-b, Max-a and Max-b waveforms in FBN2 interference group decreased, and the differences were statistically significant (all at P<0.05).The results of immunofluorescence staining showed that FBN2 was expressed in the whole retina, and the fluorescence intensity of FBN2 was weaker in FBN2 interference group than that in normal control and empty vector groups.The fluorescence intensity of FBN2 in normal control group, empty vector group and FBN2 interference group was 16.21±2.21, 15.57±3.63 and 5.32±1.06, respectively, with a statistically significant overall difference ( F=66.03, P<0.05).The fluorescence intensity of FBN2 protein in FBN2 interference group was significantly lower than that in empty carrier group and normal control group (both at P<0.05).Compared with normal control and empty vector groups, the relative expression levels of LTBP-1 and TGF-β2 mRNA and protein were significantly increased in FBN2 interference group, while the relative expression levels of FBN2, CDK2 and CCND2 mRNA and protein were significantly decreased, and the differences were statistically significant (all at P<0.05). Conclusions:The increase of LTBP-1 and TGF-β2 and the decrease of G1/S phase related proteins CDK2 and CCND2 are involved in the development of FBN2-deficient retinopathy.

The fall armyworm (FAW), Spodoptera frugiperda, is a destructive pest native to America and has recently become an invasive insect pest in China. Because of its rapid spread and great risks in China, understanding of FAW genetic background and pesticide resistance is urgent and essential to develop effective management strategies. Here, we assembled a chromosome-level genome of a male FAW (SFynMstLFR) and compared re-sequencing results of the populations from America, Africa, and China. Strain identification of 163 individuals collected from America, Africa and China showed that both C and R strains were found in the American populations, while only C strain was found in the Chinese and African populations. Moreover, population genomics analysis showed that populations from Africa and China have close relationship with significantly genetic differentiation from American populations. Taken together, FAWs invaded into China were most likely originated from Africa. Comparative genomics analysis displayed that the cytochrome p450 gene family is extremely expanded to 425 members in FAW, of which 283 genes are specific to FAW. Treatments of Chinese populations with twenty-three pesticides showed the variant patterns of transcriptome profiles, and several detoxification genes such as AOX, UGT and GST specially responded to the pesticides. These findings will be useful in developing effective strategies for management of FAW in China and other invaded areas.
Animals ; China ; Genomics ; Humans ; Male ; Pesticides ; Spodoptera/genetics* ; Transcriptome

Trauma registration is an important tool to record the process and timeline in the treatment of trauma patients. The operation of trauma database is of great significance for reducing the mortality of patients, promoting the construction of trauma treatment system, and providing reference for policy-making. Trauma registration system has been established in the United States, United Kingdom, Germany and other developed countries for many years. However, the domestic system is still at an initial stage, and there are problems like data deficiencies, data incoherence, no item of complications, no treatment data after discharge and limits of human and financial resources. Therefore, there is room for improvements in terms of personnel fixation, financial support and continuous data monitoring should be further improved. In this study, the authors summarize the traum registration system from aspects of basic situation both at home and abroad, data analysis, clinical value, operation mechanism and challenges so as to provide important data for clinical researches.

Humans ; Air Pollutants ; Sleep Apnea, Obstructive ; Polysomnography ; Sleep ; Pulmonary Disease, Chronic Obstructive

Objective:To explore the clinical application and long-term safety of hydroxychloroquine sulfate (HCQ) in the treatment of rheumatic diseases.Methods:A multi-center cross-sectional study was conducted between August 2017 and August 2018 in a random sample of eleven medical institutions of rheumatology and immunology in China. Patients who took HCQ for more than 3 months were enrolled into this study. The cumulative dose and long-term side effects of HCQ were recorded. The changes of laboratory indexes before and after treatment with HCQ were analyzed. Categorical variables were presented with counts and proportions, and evaluated by Chi-square test. Continuous parametric data were presented as Mean±standard deviation, and evaluated by Student's t test or Mann-Whitney U test. P-values less than 0.05 were considered statistically significant. Results:A total of 886 patients with rheumatic diseases were enrolled into this study, including 505 cases with systemic lupus erythematosus (57.0%), 210 cases with rheumatoid arthritis (23.7%), 80 cases with Sj?gren's syndrome (9.0%), 57 cases with undifferentiated connective tissue disease (6.4%), 12 cases of systemic vasculitis (1.4%), 10 cases of mixed connective tissue disease (1.1%), 7 cases of myositis (0.8%) and 5 cases with systemic sclerosis (0.6%). The most common long-term side effects of HCQ was skin or mucous lesions (12.4%) and vision problems (8.0%). Other adverse reactions included problems of digestive system (3.0%), nervous system (2.1%), musculoskeletal system (1.1%) and cardiovascular system (0.9%). 140 cases (15.8%) had stopped taking HCQ during the treatment. More than half of them decided to stop taking medicine by themselves. Fifty-four patients (6.1%) stopped using HCQ due to side effects while 24 of them took it again, and another 12 patients (1.4%) stopped the drug due to remission of illness. Patients were divided into three groups according to the cumulative dose of HCQ: less than 500 g, 500-1 000 g and more than 1 000 g respectively. There was significant difference in the incidence of long-term side effects among the three groups ( χ2=6.382, P=0.041). The last group (more than 1 000 g) suffered the highest incidence of long-term adverse reactions (37.1%). No severe adverse drug reactions were observed in this study. Conclusion:Hydroxychloroquine is widely used in the treatment of rheumatic diseases. The incidence of long-term side effects is 20.4%, is 6.1% lead to drug withdrawal, which are especially related to the cumulative doses. It should be adjusted properly according to the clinical application.

Objective:To obtain the inter-fractional set-up errors of intensity-modulated radiotherapy (IMRT) of cervical cancer by cone-beam CT (CBCT), and to analyze the variations of the set-up errors on the cumulative dose deviation of the target volume.Methods:A total of 48 patients with cervical cancer who underwent IMRT were enrolled in this study, and the set-up errors of 696 CBCTs were obtained. The set-up errors were input into the treatment planning system, and the cumulative set-up error dose was obtained by superposing the set-up errors dose. The deviation percentage was calculated by the deviation formula and the standard planning dose.Results:The set-up errors caused the offset of isocenter distance by 0.58(0.36, 0.80) cm. Different statistical differences were noted between the cumulative set-up error dose and the standard planning dose by WilCoxon test. All the dose deviations in the target volume were reduced, and the differential dose volume histogram (DVH) appeared negatively skewed, and the peak value was declined. The DVH diagram shifted to the left with an inverse S-curve and the slope was increased. The HI deviation of the target volume from small to large were: CTV 1, CTV 2, GTV/CTV, CTV 3, CTV n, CTV all, and GTV nd; The HI deviation of the target volume were increased. Conclusions:The effect of set-up errors in IMRT of cervical cancer upon the cumulative doses of the target volume significantly differs. The cumulative dose of the target volume is reduced, and the uniformity of the target volume dose becomes worse. The uncertainty of the inter-fractional position leads to an increase or decrease in the the fractional doses of the target volume. The biological effect on tumor cells and the tumor recurrence remains to be investigated. In IMRT of cervical cancer, the CBCT position calibration is required before each treatment to ensure the dose accuracy of each structure in the target volume.

Different fragments of SARS-CoV-2 nucleocapsid (N) protein were expressed and purified, and a fluorescence immunochromatography method for detection of SARS-CoV-2 total antibody was established. The effect of different protein fragments on the performance of the method was evaluated. The N protein sequence was analyzed by bioinformatics technology, expressed in prokaryotic cell and purified by metal ion affinity chromatography column. Different N protein fragments were prepared for comparison. EDC reaction was used to label fluorescence microsphere on the synthesized antigen to construct sandwich fluorescence chromatography antibody detection assay, and the performance was systemically evaluated. Among the 4 prepared N protein fragments, the full-length N protein (N419) was selected as the optimized coating antigen, N412 with 0.5 mol/L NaCl was used as the optimal combination; deleting 91-120 amino acids from the N-terminal of N412 reduced non-specific signal by 87.5%. the linear range of detection was 0.312-80 U/L, the limit of detection was 0.165 U/L, and the accuracy was more than 95%. A fluorescence immunochromatographic detection method for analysis of SARS-CoV-2 total antibody was established by pairing N protein fragments. The detection result achieved 98% concordance with the commercially available Guangzhou Wanfu test strip, which is expected to be used as a supplementary approach for detection of SARS-CoV-2. The assay could also provide experimental reference for improving the performance of COVID-19 antibody detection reagents.
Antibodies, Viral ; COVID-19 ; Chromatography, Affinity ; Fluorescent Antibody Technique ; Humans ; Microspheres ; SARS-CoV-2 ; Sensitivity and Specificity

Hypervirulent Klebsiella pneumoniae, short for hvKP, is a hypervirulent variant of classical Klebsiella pneumoniae, which accounts for serious infection in healthy people, exhibits strong pathogenicity, high mortality and poor prognosis. At present, hvkp is of high prevalence all over the world, and the infection rate shows a continuous upward trend, which brings great challenges to public health security and clinical treatment. This paper summarized the research progress on virulence factors of hvkp, such as capsular polysaccharides, siderophore, lipopolysaccharide, adhesins and recently discovered Type Ⅵ secreting system, and aimed to deepen the understanding and recognition of hvKP.