ObjectiveTo explore the clinical features and causative genes of short stature children with unknown etiology， providing evidence for precise clinical diagnosis and treatment. MethodsThe study recruited children with suspected but undiagnosed short stature from the pediatric endocrinology department in our hospital between January 2018 and August 2022. A retrospective analysis was performed on the clinical manifestations， laboratory test and whole exome sequencing （WES） results. Causative genes were classified and analyzed according to different pathogenic mechanisms. ResultsA total of 48 children （30 boys and 18 girls） were enrolled， aged 7.73 ± 3.97 years， with a height standard deviation score （ HtSDS） of -3.63 ± 1.67. Of the patients， 33 （68.8%） suffered from facial anomalies， 31 （64.6%） from skeletal abnormalities， 26 ［54.2%， 61.5% of whom born small for gestational age （SGA）］ from perinatal abnormalities， 24 ［50.0%， 87.5% of whom with growth hormone （GH） peak concentration below normal］ from endocrine disorders and 21（43.8%） had a family history of short stature. Laboratory tests showed that GH peak concentration following stimulation test was （9.72 ± 7.25） ng/mL， IGF-1 standard deviation score was -0.82 ± 1.42， the difference between bone age and chronological age was -0.93 ± 1.39 years. Of the 25 cases with mutant genes found by WES， 14 （56.0%） had pathogenic mutation， 6 （24.0%） likely pathogenic mutation， and 5 （20.0%） mutation of uncertain significance. Pathogenic and likely pathogenic variants were identified in 14 genes， including 10 affecting intracellular signaling pathways （PTPN11， RAF1， RIT1， ARID1B， ANKRD11， CSNK2A1， SRCAP， CUL7， SMAD4 and FAM111A） and 4 affecting extracellular matrix （ECM） components or functions （ACAN， FBN1， COL10A1 and COMP）. ConclusionsA rare monogenic disease should be considered as the possible etiology for children with severe short stature accompanied by facial anomalies， disproportionate body types， skeletal abnormalities， SGA， GH peak concentration below normal and a family history of short stature. WES played an important role in identifying the monogenic causes of short stature. This study indicated that affecting growth plate cartilage formation through intracellular signaling pathways and ECM components or functions was the main mechanism of causative genes leading to severe short stature in children. Further research may help discover and study new pathogenic variants and gene functions.

ObjectiveTo evaluate the dynamic disinfection effect of the upper-room 222 nm ultraviolet radiation on the air in different areas of the emergency department, and to provide references for a new solution of air disinfection that man-machine coexisted in the medical and healthcare institutions. MethodsThe upper-room 222 nm ultraviolet radiation air sterilizers were installed at a height of 2.3‒2.6 m from the ground in the observation room, computed tomography (CT) scanning room, rescue room and consulting room of the emergency department in Beijing Tiantan Hospital, Capital Medical University . The test area was divided into a 222 nm ultraviolet group and a control group according to whether the 222 nm ultraviolet germicidal air disinfection was conducted or not. The experimental group started air disinfection at8:00 a.m., and the air sampling was conducted from 9:00 a.m. to 16:00 p.m., with a 10 min sampling interval of every 1 hour. While the control group only collected air sample with the same air sampling method used in the experimental group, without air disinfection. The air microbial sampler with six-level sieve impingement was used for the air sampling, and the differences in the total number of airborne bacterial colonies were compared between the two groups. ResultsA total of 128 air samples were collected in the trial, of which 64 were from the experimental group and 64 from the control group. The total number of airborne bacterial colonies in the experimental group was lower than that in the control group (all P<0.001), and was maintained at a lower level throughout the entire period. The killing rate of 222 nm ultraviolet radiation to airborne microorganisms was approximately 55.76% to 76.33% in different sampling areas. The qualified rates of the total number of dynamic airborne bacterial colonies in the observation room, rescue room and consulting room were improved from 12.50%, 37.50% and 25.00% to 81.25%, 100.00% and 100.00%, respectively (all P<0.001). Over 62.50% of the time, the air quality in the consulting room and CT room in the 222 nm ultraviolet group met the environment standards for airborne bacterial colony criteria of class Ⅰ or class Ⅱ. ConclusionThe upper-room222 nm ultraviolet radiation germicidal disinfection can effectively reduce the total number of airborne bacterial colonies and improve the environment for emergency department, and the continuous using of it is helpful for keeping the air safe and clean.

Objective To study the mechanism of Wenyang Jieyu Granules via cAMP-MAPK signaling pathway in hippocampus of chronic unpredictable mild stress(CUMS)model rats.Methods CUMS was used to establish depression model in rats.The depression state of rats was reflected by sucrose preference test,forced swimming test and open field test.The expressions of cAMP,phosphorylated extracellular signal-regulated kinase(p-ERK)and Raf protein kinase-1(Raf-1)in rat hippocampus were detected by enzyme-linked immunoassay(ELISA).Western blot was used to detect the expressions of cyclic adenosine phosphate response element binding protein(CREB),phosphorylated cyclic adenosine phosphate response element binding protein(p-CREB),extracellular signal-regulated kinase(ERK)and Mitogen-activatsion ofed Extracellular signal-regulated Kinas(MEK)in rat hippocampus.The mRNA expres cAMP,CREB,cyclic adenosine phosphate dependent protein kinase A(PKA),brain-derived neurotrophic factor(BDNF),and rat sarcoma(Ras)in rat hippocampus was detected by RT-PCR.The expression of caspase-3(caspase-3)in rath hippocampus was observed by immunohistochemistry.Results Compared with the model group,the body weight,sucrose preference rate and open field distance of model rats were increased in the positive control group and Wenyang Jieyu Granules group(P<0.05,P<0.01),and reduce the immobility time of forced swimming(P<0.05,P<0.01).It also increased the levels of cAMP,p-ERK,Raf-1,CREB,p-CREB,p-CREB/CREB,ERK and MEK in hippocampus of CUMS model rats(P<0.05,P<0.01).At the same time,the expression of cAMP,CREB,PKA,BDNF and Ras mRNA in the hippocampus of model rats were significantly improved(P<0.05,P<0.01),and he expression of caspase-3 in the hippocampus was reduced(P<0.05).Conclusion Wenyang Jieyu granules may exert antidepressant effect on model rats through cAMP and MAPK signaling pathways.

Objective:To study the complete genome characterization of Human Astrovirus (HAstV) in Shandong Province.Methods:Stool samples from acute flaccid paralysis (AFP) surveillance in Shandong Province from 2020 to 2022 were collected, and HAstV nucleic acid was examined by real-time quantitative PCR (qPCR). Next-generation sequencing (NGS) was conducted for the positive samples to obtain complete genome sequences and identify the genotype. Homology comparison and phylogenetic analysis were performed by using BioEdit and Mega software.Results:A total of 667 samples were examined by qPCR, of which 14 were HAstV-positive (2.1%), including HAstV-1 ( n=6), MLB1 ( n=6), MLB2 ( n=1), and VA2 ( n=1). The complete genome sequences were obtained from 11 samples. The six HAstV-1 sequences of this study had 98.2% to 99.9% nt similarities with each other and 87.6% to 98.6% with those from other regions. The four MLB1 sequences of this study had 99.1% to 99.9% nt similarities with each other and 92.2% to 99.4% with those from other regions. The VA2 sequence of this study had 96.0% to 96.3% nt similarities with those from other regions. Phylogenetic analysis based on ORF2 region showed that the local HAstV-1 sequences were most closely related to Japanese strains, and had distinct topology with phylogenies based on ORF1a and ORF1b regions. Conclusion:The complete genome sequences of 11 HAstV strains are obtained, and the VA2 complete genome is found.

Objective To discover the host factor interacting with severe acute respiratory syndrome coronavirus-2(SARS-CoV-2)papain-like protease(PLpro)and explore the potential mechanism.Methods The second-generation proximity-dependent biotin identification(BioID2)approach combined with mass spectrometry analysis was used to search for the potential host factors.Immunofluorescence and co-immunoprecipitation(Co-IP)assay were used to verify the interactions between DEAD-box helicase 3(DDX3)and PLpro.The influence of PLpro on DDX3-inhibitor of kappa B kinase ε(IKKε)-TANK-binding kinase 1(TBK1)and DDX3-mitochondrial antiviral signaling protein(MAVS)complexes was also investigated by Co-IP.The effect of PLpro on interferon-β(IFN-β)immune pathway and the protease activity on substrates were studied via luciferase activity assay.Results DDX3 could co-locate and interact with PLpro intracellularly.PLpro might possibly inhibit both the formation of DDX3-MAVS complex and the interactions between DDX3-IKK-ε-TBK1.PLpro could negatively regulate type Ⅰ interferon pathway.Overexpression of DDX3 could lead to a significant increase in the cleavage activity of PLpro/PLP-TM that might be significantly decreased in case of inventions with DDX3 expressions.Conclusion DDX3 may be one of the host factors that interact with SARS-CoV-2 PLpro.PLpro negatively regulates IFN-β immune pathway induced by DDX3,which may provide a favorable immune environment for virus replication.

Purpose/Significance To explore the feasibility of applying blockchain technology to the current healthcare system of hos-pitals,and to achieve the purpose of protecting patients'privacy to the greatest extent possible at a lower cost.Method/Process 505 questionnaires are randomly distributed and collected from people of different age groups in Beijing,Tianjin,Shanghai and Shenzhen who have a certain degree of understanding of blockchain technology,and the results are analyzed.Result/Conclusion Different age groups are highly concerned about personal privacy and privacy protection,and are willing to accept blockchain as an emerging technology.There is a greater demand and acceptance for the application of blockchain technology in the primary health care systems.

Objective To analyze the distribution characteristics and clinical significance of allergens in children with allergic diseases in Yangzhou area. Methods A total of 1, 025 children who were treated in the Affiliated Hospital of Yangzhou University from December 2022 to December 2023 and tested for allergens were selected as study objects. The detection results of allergens were collected and the distribution characteristics of allergens were analyzed. Results Among the 1, 025 children with allergy-related diseases, 83.41% (855 cases) tested positive for allergens, with a positive rate of 66.24% (679 cases) for inhalant allergens and 61.17% (627 cases) for food allergens. The most common allergens were milk (36.20%), molds (33.27%), and house dust mites (30.34%). There was no statistically significant difference in the positive rate of allergens among different age groups (P>0.05). The positive rate of allergen detection in boys was higher than that in girls, the difference was statistically significant (P < 0.05). The positive rate of inhalation allergens in children with respiratory symptoms was higher than that of food allergens, and the positive rate of food allergens in children with digestive tract symptoms was higher than that of inhalation allergens (P < 0.05). For children with immune system diseases such as skin symptoms and allergic purpura, there was no statistically significant difference in the positive rates of inhalant and food allergens (P>0.05). Conclusion The most common allergens in children with allergic diseases in Yangzhou area are milk, molds, and dust mites. Allergen testing can provide reference opinions for the prevention, diagnosis, and treatment of allergic diseases in children in Yangzhou area.

OBJECTIVE To investigate the effects of Wenyang jieyu decoction on phosphoinositide 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway and neurotransmitters in rats with kidney-yang deficiency depression. METHODS The SD rats were divided into blank group， model group， fluoxetine group （positive control of western medicine， 4.17 mg/kg）， Xiaoyao powder group （positive control of TCM， 1.88 g/kg） and Wenyang jieyu decoction low-dose， medium-dose and high-dose groups （1.25， 2.50， 5.00 g/kg）， with 15 rats in each group. Except for blank group， the other groups were treated with corticosterone 20 mg/kg subcutaneously to induce kidney-yang deficiency depressed model， meanwhile the mice were given relevant medicine intragastrically， once a day， for 28 consecutive days. The general conditions of rats were observed. The sucrose preference rate and the static time of forced swimming were detected， and organ indexes of rats were calculated. The levels/contents of neurotransmitters in serum were detected， the expressions of PI3K/Akt pathway-related proteins in hippocampus were detected， and the number of dendritic spines was determined. RESULTS Compared with blank group， model group suffered from the symptoms such as hair loss， fear of cold， curling up； sucrose preference rate， indexes of adrenal gland， thymus gland and spleen，serum levels of cyclic adenosine phosphate （cAMP）， brain-derived neurotrophic factor and gamma-aminobutyric acid， the ratio of cAMP to cyclic guanosine monophosphate， the contents of norepinephrine， dopamine and 5-hydroxy-tryptamine， the expressions of PI3K， Akt， mammalian target of rapamycin， and the number of dendritic spines in the hippocampus were significantly decreased （P＜0.01）. The time of immobility， level of glutamic acid and protein expression of glycogen synthetase kinase-3β were prolonged and increased （P＜0.01）. Compared with model group， depression symptoms of rats in each administration group were improved， and the above indexes were mostly reversed （P＜0.05 or P＜0.01）. CONCLUSIONS Wenyang jieyu decoction can improve depression-like behavior and the deficiency of kidney-yang， regulate the secretion of neurotransmitters， and activate PI3K/Akt signaling pathway， thus playing a role in protecting hippocampal neurons.

Objective To investigate a convenient and quantitative solution to activation levels and functional characterization of CAR-T cells by inserting T cell activity-responsive promoter (TARP) nanoluciferase reporter gene system into a lentiviral plasmid containing the gene encoding the chimeric antigen receptor (CAR). Methods The recombinant plasmid was constructed by using whole gene synthesis and molecular cloning techniques. The lentivirus was packaged and was infected with human primary T lymphocytes. Flow cytometry was used to detected the positive rate of lentivirus-infected T cells. The functional characterization of CAR-T cells was identified by luciferase reporter gene system, Western blot, flow cytometry, and small animal live imaging techniques. Results The results of enzyme digestion identification and the plasmid sequencing showed that the recombinant plasmids were constructed, and flow cytometry displayed the normal preparation of CAR-T cells. This system could dynamically respond to the activation of CAR-T cells by luciferase reporter gene system. The functional assay in vitro confirmed that the system could reflect the exhaustion of CAR-T cells, and the small animal live imaging results demonstrated that the system can be used as a tracer of CAR-T cells in mice. Conclusion TARP nanoluciferase reporter gene system provides a more convenient, sensitive and quantitative method for evaluating CAR-T cells activation level, exhaustion phenotype and tracing.
Humans ; Animals ; Mice ; T-Lymphocytes ; Cell Line, Tumor ; Receptors, Chimeric Antigen/genetics* ; Promoter Regions, Genetic ; Immunotherapy, Adoptive/methods*

Background/Aims: Existing hepatocellular carcinoma (HCC) prediction models are derived mainly from pretreatment or early on-treatment parameters. We reassessed the dynamic changes in the performance of 17 HCC models in patients with chronic hepatitis B (CHB) during long-term antiviral therapy (AVT). Methods: Among 987 CHB patients administered long-term entecavir therapy, 660 patients had 8 years of follow-up data. Model scores were calculated using on-treatment values at 2.5, 3, 3.5, 4, 4.5, and 5 years of AVT to predict threeyear HCC occurrence. Model performance was assessed with the area under the receiver operating curve (AUROC). The original model cutoffs to distinguish different levels of HCC risk were evaluated by the log-rank test. Results: The AUROCs of the 17 HCC models varied from 0.51 to 0.78 when using on-treatment scores from years 2.5 to 5. Models with a cirrhosis variable showed numerically higher AUROCs (pooled at 0.65–0.73 for treated, untreated, or mixed treatment models) than models without (treated or mixed models: 0.61–0.68; untreated models: 0.51–0.59). Stratification into low, intermediate, and high-risk levels using the original cutoff values could no longer reflect the true HCC incidence using scores after 3.5 years of AVT for models without cirrhosis and after 4 years of AVT for models with cirrhosis. Conclusions The performance of existing HCC prediction models, especially models without the cirrhosis variable, decreased in CHB patients on long-term AVT. The optimization of existing models or the development of novel models for better HCC prediction during long-term AVT is warranted.