BACKGROUND:Ankylosing spondylitis is closely related to the occurrence and development of psoriasis,but the key genes and regulatory mechanisms are still unclear.OBJECTIVE:To establish an artificial neural network model of genes shared by ankylosing spondylitis and psoriasis based on the GEO database and evaluate its effect,and also to determine whether there is a causal relationship between the expression of key genes and the two diseases using Mendelian randomization.METHODS:Datasets GSE25101(816 ankylosing spondylitis samples and 816 healthy control samples),GSE30999(85 psoriasis samples and 85 healthy control samples),GSE73754(52 ankylosing spondylitis samples and 20 healthy control samples),and GSE14905(33 psoriasis samples and 49 healthy control samples)were downloaded from the GEO database.GSE25101 and GSE30999 were used as the training datasets of ankylosing spondylitis and psoriasis,respectively,and their respective differentially expressed genes were identified through difference analysis to obtain the common driver genes of the two diseases,and the key core genes were further screened out based on Mendelian randomization.The key core genes were further screened out,and artificial neural network models were constructed based on the key core genes and validated in external datasets GSE73754 and GSE14905,followed by the construction of the corresponding nomogram to predict the incidence rates of the diseases.Also,the results of immune infiltration in ankylosing spondylitis and psoriasis were analyzed.Finally,Mendelian randomization was used to assess causal relationships between key genes and diseases,and drug-gene interactions were analyzed using the Dgidb database to predict drug targets.RESULTS AND CONCLUSION:(1)A total of 61 differential genes were obtained in ankylosing spondylitis and 4 309 differential genes were obtained in psoriasis.Eight shared differential genes were obtained after intersection,and five key genes(DNMT1,GNG11,CDC25B,S100A8,and S100A12)were further screened by machine learning.The key genes were utilized to build artificial neural network models of ankylosing spondylitis and psoriasis,with the area under curve values of 0.979 and 0.989 in the training sets GSE25101 and GSE30999,respectively,and 0.818 and 0.874 in the external validation datasets GSE73754 and GSE14905,respectively.(2)Nomogram was constructed based on the five core genes,and the calibration curves showed that the predicted probabilities of the nomogram models were almost the same as that of the ideal model.Immune cell infiltration showed that the key genes were associated with activated B cells,natural killer cells,γδ T cells,follicular helper T cells,monocytes,plasma cell-like dendritic cells,and neutrophils.Mendelian randomization showed that S100A8 was a risk factor for the occurrence of ankylosing spondylitis and psoriasis.Finally,DGldb screening was utilized to obtain 81 targeted drugs,only 16 of which,including methotrexate,atogepant,ubrogepant,rimegepant,eptinezumab,azacitidine,selenium,hydroxyurea,ifosfamide,floxuridine,curcumin,mitoxantrone,cisplatin,arsenic trioxide,diethylstilbestrol,and decitabine,were approved by the U.S.Food and Drug Administration.(3)A large number of successful cases have been accumulated in international databases,research results and data analysis of European groups,especially in genomics and disease phenotyping studies.These experiences provide valuable references for the epidemiological characterization of diseases in China,genetic diversity and their response to the environment and lifestyle.(4)An artificial neural network model of the common driver genes of ankylosing spondylitis and psoriasis was constructed and validated,the causal relationship between the key genes and the pathogenesis of the two diseases was discovered,and the targeted drugs for potential treatments were predicted,which hopefully provides a new perspective for exploring their pathogenesis and therapeutic directions.

BACKGROUND:Ankylosing spondylitis is closely related to the occurrence and development of psoriasis,but the key genes and regulatory mechanisms are still unclear.OBJECTIVE:To establish an artificial neural network model of genes shared by ankylosing spondylitis and psoriasis based on the GEO database and evaluate its effect,and also to determine whether there is a causal relationship between the expression of key genes and the two diseases using Mendelian randomization.METHODS:Datasets GSE25101(816 ankylosing spondylitis samples and 816 healthy control samples),GSE30999(85 psoriasis samples and 85 healthy control samples),GSE73754(52 ankylosing spondylitis samples and 20 healthy control samples),and GSE14905(33 psoriasis samples and 49 healthy control samples)were downloaded from the GEO database.GSE25101 and GSE30999 were used as the training datasets of ankylosing spondylitis and psoriasis,respectively,and their respective differentially expressed genes were identified through difference analysis to obtain the common driver genes of the two diseases,and the key core genes were further screened out based on Mendelian randomization.The key core genes were further screened out,and artificial neural network models were constructed based on the key core genes and validated in external datasets GSE73754 and GSE14905,followed by the construction of the corresponding nomogram to predict the incidence rates of the diseases.Also,the results of immune infiltration in ankylosing spondylitis and psoriasis were analyzed.Finally,Mendelian randomization was used to assess causal relationships between key genes and diseases,and drug-gene interactions were analyzed using the Dgidb database to predict drug targets.RESULTS AND CONCLUSION:(1)A total of 61 differential genes were obtained in ankylosing spondylitis and 4 309 differential genes were obtained in psoriasis.Eight shared differential genes were obtained after intersection,and five key genes(DNMT1,GNG11,CDC25B,S100A8,and S100A12)were further screened by machine learning.The key genes were utilized to build artificial neural network models of ankylosing spondylitis and psoriasis,with the area under curve values of 0.979 and 0.989 in the training sets GSE25101 and GSE30999,respectively,and 0.818 and 0.874 in the external validation datasets GSE73754 and GSE14905,respectively.(2)Nomogram was constructed based on the five core genes,and the calibration curves showed that the predicted probabilities of the nomogram models were almost the same as that of the ideal model.Immune cell infiltration showed that the key genes were associated with activated B cells,natural killer cells,γδ T cells,follicular helper T cells,monocytes,plasma cell-like dendritic cells,and neutrophils.Mendelian randomization showed that S100A8 was a risk factor for the occurrence of ankylosing spondylitis and psoriasis.Finally,DGldb screening was utilized to obtain 81 targeted drugs,only 16 of which,including methotrexate,atogepant,ubrogepant,rimegepant,eptinezumab,azacitidine,selenium,hydroxyurea,ifosfamide,floxuridine,curcumin,mitoxantrone,cisplatin,arsenic trioxide,diethylstilbestrol,and decitabine,were approved by the U.S.Food and Drug Administration.(3)A large number of successful cases have been accumulated in international databases,research results and data analysis of European groups,especially in genomics and disease phenotyping studies.These experiences provide valuable references for the epidemiological characterization of diseases in China,genetic diversity and their response to the environment and lifestyle.(4)An artificial neural network model of the common driver genes of ankylosing spondylitis and psoriasis was constructed and validated,the causal relationship between the key genes and the pathogenesis of the two diseases was discovered,and the targeted drugs for potential treatments were predicted,which hopefully provides a new perspective for exploring their pathogenesis and therapeutic directions.

The study aims to investigate the impacts of prolyl oligopeptidase (POP) on the replication of foot-and-mouth disease virus (FMDV) in BHK-21 cells. Firstly, the effects of FMDV replication on POP expression in BHK-21 cells were analyzed by Western blotting and Real-time reverse transcription polymerase chain reaction (RT-qPCR). Secondly, a eukaryotic expression plasmid for POP was constructed, and the effects of POP overexpression on the replication of two different serotypes of FMDV were assessed by Western blotting, RT-qPCR, and virus titer assays. Thirdly, specific small interfering RNAs (siRNAs) targeting POP were synthesized, and their efficiency in interfering with endogenous POP expression was identified by RT-qPCR. The impacts of downregulating endogenous POP expression on FMDV replication were further evaluated by Western blotting, RT-qPCR, and virus titer assays. The results indicated that FMDV infection did not significantly affect POP expression in BHK-21 cells. Overexpression of POP dose-dependently enhanced the replication of both FMDV/O and FMDV/A serotypes. Conversely, siRNA-mediated downregulation of endogenous POP expression markedly suppressed FMDV/O replication. This study is the first to demonstrated that the role of the host POP protein in promoting FMDV replication in BHK-21 cells, thereby providing a critical theoretical foundation and potential molecular targets for developing efficient candidate cell strains for foot-and-mouth disease inactivated vaccines.
Foot-and-Mouth Disease Virus/genetics* ; Virus Replication/genetics* ; Prolyl Oligopeptidases ; Serine Endopeptidases/physiology* ; Animals ; Cell Line ; RNA, Small Interfering/genetics* ; Foot-and-Mouth Disease/virology* ; Cricetinae

Objective:To explore clinical validation standard for intelligent ventilation mode of ventilator and promote ventilator's clinical application.Methods:By real-time quantification of physiological parameters related to patient's ventilation and oxygenation,the change of respiratory function was analyzed,and the settings of ventilator's parameters were adjusted synchronously by using feedback signals to realize personalized support of ventilation.Combining with artificial intelligence(AI)technique,the clinical validation standard for intelligent ventilation mode was investigated from three aspects:requirements of clinical application and function of intelligent ventilation mode of ventilator,validation for platform of simulation experiment,and early clinical evaluation and interventional trials.Results:The clinical validation standards for intelligent ventilation mode of ventilator should meet the requirements of clinical application and function,and should complete preclinical evaluation of intelligent ventilation mode of ventilator,and assess safety and usability of that by the platform of simulation trials,and should conduct early clinical evaluations and interventional clinical trials so as to further assess safety and effectiveness of that before it was used formally in clinical application.Conclusion:The clinical validation standard of intelligent ventilation mode of ventilator can form standard validation process for intelligent ventilation mode,and promote the construction of standard validation platform,and facilitate clinical application and iterative optimization of intelligent ventilation mode,and improve medical quality.

Objective To observe the value of ultrasound,CT and the combination for evaluating thyroid cancer(TC)involved peripheral structures.Methods Totally 102 patients with advanced TC were enrolled,and the value of ultrasound and CT for evaluating TC involved peripheral structures were analyzed taken post resection pathology as gold standards.Results The ultrasonic detection rate of TC involved laryngeal recurrent nerve and strap muscle was 47.37％(18/38)and 72.73％(24/33),while the CT detection rate was 15.79％(6/38)and 39.39％(13/33),respectively,the formers were better than the latters(both P＜0.05).Ultrasound had high sensitivity(88.24％),specificity(97.06％),and accuracy(94.12％)for diagnosing TC involved trachea,also had fair sensitivity(72.73％,72.73％),high specificity(96.70％,97.10％)and accuracy(94.12％,89.22％)for diagnosing TC involved thyroid cartilage and strap muscle.CT had high sensitivity(90.91％),specificity(92.31％)and accuracy(92.16％)for diagnosing TC involved thyroid cartilage,while had fair sensitivity(50.00％—76.47％)and high specificity(82.35％—88.46％)for diagnosing TC involved trachea,esophagus and blood vessels.The sensitivity and accuracy of combination of ultrasound and CT for evaluating TC involved peripheral structures were both higher than of each single method alone.Conclusion Combination of ultrasound and CT was valuable for evaluating TC involved peripheral structures.

Objective To observe the value of ultrasound,CT and the combination for evaluating thyroid cancer(TC)involved peripheral structures.Methods Totally 102 patients with advanced TC were enrolled,and the value of ultrasound and CT for evaluating TC involved peripheral structures were analyzed taken post resection pathology as gold standards.Results The ultrasonic detection rate of TC involved laryngeal recurrent nerve and strap muscle was 47.37％(18/38)and 72.73％(24/33),while the CT detection rate was 15.79％(6/38)and 39.39％(13/33),respectively,the formers were better than the latters(both P＜0.05).Ultrasound had high sensitivity(88.24％),specificity(97.06％),and accuracy(94.12％)for diagnosing TC involved trachea,also had fair sensitivity(72.73％,72.73％),high specificity(96.70％,97.10％)and accuracy(94.12％,89.22％)for diagnosing TC involved thyroid cartilage and strap muscle.CT had high sensitivity(90.91％),specificity(92.31％)and accuracy(92.16％)for diagnosing TC involved thyroid cartilage,while had fair sensitivity(50.00％—76.47％)and high specificity(82.35％—88.46％)for diagnosing TC involved trachea,esophagus and blood vessels.The sensitivity and accuracy of combination of ultrasound and CT for evaluating TC involved peripheral structures were both higher than of each single method alone.Conclusion Combination of ultrasound and CT was valuable for evaluating TC involved peripheral structures.

Objective:To explore clinical validation standard for intelligent ventilation mode of ventilator and promote ventilator's clinical application.Methods:By real-time quantification of physiological parameters related to patient's ventilation and oxygenation,the change of respiratory function was analyzed,and the settings of ventilator's parameters were adjusted synchronously by using feedback signals to realize personalized support of ventilation.Combining with artificial intelligence(AI)technique,the clinical validation standard for intelligent ventilation mode was investigated from three aspects:requirements of clinical application and function of intelligent ventilation mode of ventilator,validation for platform of simulation experiment,and early clinical evaluation and interventional trials.Results:The clinical validation standards for intelligent ventilation mode of ventilator should meet the requirements of clinical application and function,and should complete preclinical evaluation of intelligent ventilation mode of ventilator,and assess safety and usability of that by the platform of simulation trials,and should conduct early clinical evaluations and interventional clinical trials so as to further assess safety and effectiveness of that before it was used formally in clinical application.Conclusion:The clinical validation standard of intelligent ventilation mode of ventilator can form standard validation process for intelligent ventilation mode,and promote the construction of standard validation platform,and facilitate clinical application and iterative optimization of intelligent ventilation mode,and improve medical quality.

To investigate the role of chebulagic acid(CA)in regulating lipopolysaccharide(LPS)-in-duced inflammatory response in endometrial tissue of dairy cows,and to provide new ideas for the treatment and new drug development of endometritis in dairy cows.The endometrial tissues of dairy cows were isolated and cultured in vitro,stimulated with 1 mg/L LPS for 1 h and then co-cultured with CA(12.5,25.0,50.0,100.0 mg/L).Then the endometrial tissues of different treat-ment groups were collected for experiments.The protein secretion and gene expression levels of TNF-α,IL-6 and IL-1β were detected by ELISA and real-time fluorescence quantitative PCR.HE staining was used to observe the degree of endometrial tissue damage.The expressions of high mobility protein 1(HMGB-1)and hyaluronidase binding protein 2(HABP-2)were detected by immunofluorescence.The phosphorylation levels of ERK,p65 and IκBα were detected by Western blot.The results showed that the protein secretion levels of TNF-α at 6,24 h and IL-6 at 6,12 and 24 h,and the gene expression levels of IL-1β and IL-6 at 6,9,and 12 h and TNF-α at 6 h were sig-nificantly down-regulated after CA treatment with the LPS-induced endometrial tissue inflammation response model of dairy cows.HE staining showed that compared with the LPS group,the LPS+CA group had some improvements,the degree of epithelial cell exfoliation was reduced,the struc-ture of glands and blood vessels was relatively complete,the degree of inflammatory cell infiltra-tion was reduced,and there was no obvious necrosis or hemorrhage.The expression of HMGB-1 and HABP-2 in LPS+CA group was also significantly down-regulated.The phosphorylation levels of ERK,IκBα and p65 in the LPS+CA group were significantly decreased.In conclusion,CA can reduce LPS-induced inflammation in the endometrial tissue of dairy cows by inhibiting the activa-tion of MAPK and NF-κB pathways and down-regulating the expression of inflammatory factors in the uterine tissue.It is concluded that CA may be a potential therapeutic agent for endometritis in dairy cows and deserves further research and development.

Objective: To isolate a bacteriophage against pan-drug resistant Klebsiella pneumoniae in a burn patient, and to study its biological characteristics, genomic information, and effects on bacterial biofilm. Methods: (1) In 2018, pan-drug resistant Klebsiella pneumoniae UA168 (hereinafter referred to as the host bacteria) solution isolated from the blood of a burn patient in Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (hereinafter referred to as Ruijin Hospital) was used to isolate and purify the bacteriophage against pan-drug resistant Klebsiella pneumoniae from the sewage of Ruijin Hospital with sewage co-culture method, drip plate method, and double-agar plate method. The bacteriophage was named as phage KP168 and the plaque morphology was observed. (2) The phage KP168 solution was taken for cesium chloride density gradient centrifugation and dialysis, and then the morphology of phage KP168 was observed through transmission electron microscope after phosphotungstic acid negative staining. (3) The phage KP168 solution was taken to determine the lytic ability of the phage KP168 against 20 strains of pan-drug resistant Klebsiella pneumoniae isolated from the burned patients′ blood in Ruijin Hospital by the drip plate method, and then the lysis rate was calculated. (4) The phage KP168 solution at a initial titer of 9.3×10¹¹ plaque-forming unit (PFU)/mL (400 μL per tube) and the host bacteria solution at a concentration of 1×10⁹ colony-forming unit (CFU)/mL (4 mL per tube) were conventionally shaking cultured together for 4 hours at multiplicity of infection (MOI) of 10.000, 1.000, 0.100, 0.010, or 0.001, respectively (1 tube per MOI). The titer of phage KP168 was measured by the double-agar plate method (the measurement method was the same below) to select the optimal MOI. The experiment was repeated three times. (5) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (4 mL per tube) and the phage KP168 solution at an adjusted titer of 5×10⁷ PFU/mL (400 μL per tube) were mixed at the MOI of 0.005. The plaques were counted 0 (immediately), 1, 2, 3, 4, 5, 15, and 30 minutes (1 tube at each time point) after mixing by the double-agar plate method (the counting method was the same below), and the percentage of adsorbed phages was calculated to screen for the optimal adsorption time. The experiment was repeated three times. (6) The host bacteria solution at a concentration of 1×10⁹ CFU/mL (300 μL per tube) and the phage KP168 solution at a titer of 5×10⁸ PFU/mL (60 μL per tube) were mixed at MOI of 0.005 and conventionally shaking cultured after standing for the optimal adsorption time. The phage KP168 titer was measured 0 (immediately), 10, 20, 30, 40, 50, 60, 70, 80, 90, and 100 minutes after culture, and a one-step growth curve was drawn. The experiment was repeated three times. (7) The phage KP168 solution at a titer of 2.5×10¹⁰ PFU/mL was left to stand for 1 hour at 37, 40, 50, 60, or 70 ℃ (3 tubes at each time point, 1 mL per tube) for counting the plaques, and then the thermal stability curve was drawn. SM buffer at a pH values of 5.0, 6.0, 7.0, 7.4, 8.0, 9.0, or 10.0 were added to the phage KP168 solution at a titer of 3.0×10¹⁰ PFU/mL, respectively. The mixed solution was left to stand for 1 hour at 37 ℃ (3 tubes of each pH, each tube containing 100 μL phage KP168 solution and 900 μL SM buffer), and then the plaques were counted, and an acid-base stability curve was drawn. (8) The phage KP168 solution was taken for DNA extraction and sequencing after dialysis as in experiment (2). The whole genome was annotated with Prokka to obtain the coding sequence of phage KP168. Nucleotide′s BLAST function was used to proceed nucleic acid sequence alignment for finding a known phage with the highest similarity to the phage KP168 nucleic acid sequence, and Blastx function was used to translate the coding sequence into protein for its function prediction. The comparison with Antibiotic Resistance Genes Database and Virulence Factors Database was proceeded. (9) In a 96-well plate, at a MOI of 1.000, 0.100, 0.010 or 0.001 (3 wells per MOI), 20 μL phage KP168 solution at a initial titer of 5.8×10¹⁰ PFU/mL was added to 200 μL host bacteria solution at a concentration of 1.5×10⁸ CFU/mL (the same concentration below) for co-cultivation for 48 hours. After 200 μL host bacteria solution was left to stand for 48 hours, 20 μL phage KP168 solution at a titer of 1×10^6, 1×10^7, 1×10^8, 1×10^9, or 1×10¹⁰ PFU/mL (3 wells per titer) was added respectively for action for 4 hours. In both experiments, 200 μL host bacteria solution added with 20 μL SM buffer (3 wells) acted as a negative control, and 220 μL LB culture medium (3 wells) acted as a blank control. Absorbance values were measured by a microplate reader, and inhibition/destruction rates of biofilm were calculated. The experiments were both repeated three times. Results: (1) The plaques of phage KP168 successfully isolated and purified were transparent and round, and its diameter was approximately 1.5 mm. (2) The phage KP168 has a regular polyhedron structure with a diameter of about 50 nm and without a tail. (3) The phage KP168 could lyse 13 of 20 strains of Klebsiella pneumoniae from burned patients, with a lysis rate of 65.0%. (4) When MOI was 1.000, the titer was the highest after co-culturing the phage KP168 with the host bacteria for 4 hours, which was the optimal MOI. (5) After the mixing of the phage KP168 with the host bacteria for 4 minutes, the percentage of the adsorbed phage reached the highest, which was the optimal adsorption time. (6) The one-step growth curve showed that during the lysis of the host bacteria by phage KP168, the incubation period was about 10 minutes, and the lysis period was about 40 minutes. (7) With the condition of 40 ℃ or pH 7.4, the number of plaques and the activity of phage KP168 reached the highest. (8) The genome of phage KP168 was a linear double-stranded DNA with a length of 40 114 bp. There were 48 possible coding sequences. It had the highest similarity to Klebsiella phage_vB_Kp1. The most similar known proteins corresponding to the translated proteins of coding sequences contained 23 hypothetical proteins and 25 proteins with known functions. No resistance genes or virulence factor genes were found. The GeneBank accession number was KT367885. (9) After 48 hours of co-cultivation of the phage KP168 and the host bacteria at each MOI, the inhibition rates of biofilm were similar, with an average of about 45%. After the phage KP168 with a titer of 1×10⁹ PFU/mL acted on the biofilm formed by the host bacteria for 4 h, the destruction rate of biofilm was the highest, reaching an average of 42%. Conclusions In this study, a bacteriophage against pan-drug resistant Klebsiella pneumoniae from a burn patient, phage KP168, is isolated from sewage, which belongs to the tailless phage. It has a wide host spectrum, short adsorption time, and short incubation period, with certain thermal and acid-base stability. Its genomic information is clear, and it does not contain resistance genes or virulence factor genes. It also has an inhibitory effect on the formation of bacterial biofilm and a destructive effect on the formed bacterial biofilm.

Objective To investigate the treatment outcome,laryngeal preservation and side-effect in locally advanced hypopharyngeal carcinoma treated with combined Hilical tomotherapy (HT) or intensitymodulated radiotherapy (IMRT) and chemotherapy and/or EGFR inhibitor (Cetuximab or Nimotuzumab).Methods A total of 68 patients (20 cases with T1-2N1-3M0 and 48 cases with T3-4N1-3M0) with locally advanced hypopharyngeal cancer were treated individualy with non-surgical combined modality treatments including induction chemotherapy followed by concurrent chemoradiotherapy,induction chemotherapy followed by concurrent radiotherapy and EGFR inhibitor,concurrent chemoradiotherapy and EGFR inhibitor,and concurrent radiotherapy and EGFR inhibitor.HT was used in 40 patients and IMRT in 28 patients.Side-effects were evaluated with the established Common Terminology Criteria for Adverse Events (CTCAE) version 3.0 criteria.Results The average follow-up time was 25.7 months (range 3-69 months).All patients completed the planned radiotherapy without treatment breaks,and 66 (97.0％) of 68 patients completed the planned chemotherapy.The 2-year and 3-year overall survival rates were 78.8％ and 64.7％ respectively,with an organ preservation rate of 84.2％.The most common side-effect greater than or equal to grade 3 was oropharyngeal mucositis.No patient dependent on a percutaneous gastrostomy and tracheostomy tube.Conclusion Hypopharyngeal carcinoma can be treated with non-surgical combined modality treatment including HT or IMRT,with a high laryngeal organ preservation rate and minimal toxicities.