OBJECTIVE: To analyze the clinical phenotype and pathogenic mechanism of the ARSL gene variant in a fetus with nasal bone aplasia. METHODS: A 34-year-old pregnant woman who attended Quzhou Maternal and Child Health Care Hospital on January 3, 2023 was selected as the study subject. Whole exome sequencing (WES) was performed on the fetus. Bioinformatics analysis was carried out to identify and prioritize candidate gene variants, followed by Sanger sequencing for familial validation. A mutant plasmid expression vector was constructed and subsequently transfected into HEK293T cells to preliminarily investigate the pathogenetic mechanism of the identified variant. Additionally, a comprehensive review of literature was conducted to systematically summarize the associated clinical phenotypes. This study was approved by the Medical Ethics Committee of Quzhou Maternal and Child Health Care Hospital (Ethics No.: KY-2023-11). RESULTS: WES revealed that the fetus harbored a c.827del (p.L276Rfs*48) variant of the ARSL gene, for which its mother was heterozygous. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic(PVS1+PM2_Supporting). In vitro cellular function studies demonstrated that this variant can result in a substantial decrease in the expression of mutant mRNA, thereby preventing the production of normal ARSL protein. Clinical phenotypes resulting from ARSL gene variants exhibited considerable diversity, with nasal hypoplasia being the most common manifestation. CONCLUSION The c.827del (p.L276Rfs*48) variant of the ARSL gene can lead to degradation of mRNA via the nonsense-mediated mRNA decay pathway, resulting in reduced levels of ARSL protein. The pathogenetic mechanism underlying the ARSL gene variant may be associated with its haploinsufficiency effect.
Humans ; Female ; Pregnancy ; Adult ; Frameshift Mutation ; HEK293 Cells ; Nasal Bone/abnormalities* ; Fetus/abnormalities* ; Exome Sequencing

Objective:To assess the use of R-R interval (RRI) sequence data collected via wearable devices to evaluate emotional dynamic factors in adolescents with depressive disorders, and to analyze their impact on diagnosis and severity assessment.Methods:Clinical data were prospectively collected from 154 adolescent inpatients with depressive disorders (132 females, 22 males; age 12-18, mean: 13.5±1.6 years) treated at the Child Psychiatry Ward of Beijing Anding Hospital, Capital Medical University between January 2023 and January 2024.A control group of 152 healthy adolescents (62 females, 90 males; age 12-18, mean 14.5±1.3 years) was recruited during the same period. RRI data were obtained using the built-in photoplethysmography (PPG) sensor in the HUAWEI Band 7 wearable device. The device′s integrated emotion evaluation system extracted arousal and emotional valence (as indicators of emotional dynamics) from the collected RRIs. A Long Short-Term Memory (LSTM) network was employed to develop a model for depression diagnosis and severity prediction, while a random forest model was applied to generate receiver operating characteristic (ROC) curves to evaluate model performance. Binary Logistic regression was conducted to investigate the influence of arousal and emotional valence on depression diagnosis and severity.Results:A total of 429 records were collected and analyzed from 306 participants. The LSTM-based diagnosis and severity assessment models achieved area under the curve (AUC) of 0.896 9 and 0.715 3, respectively, indicating good model performance. Binary Logistic regression analysis showed that arousal and emotional valence had significant effects on diagnosis and severity. Specially, lower arousal in both the first 4 hours ( β=-8.906, 95%CI:-17.497 to -0.315) and second 4 hours ( β=-3.033, 95%CI:-5.109 to -0.957) significantly predicted positive depression diagnosis ( β=-1.219, 95%CI:-2.205 to -0.233), while emotional valence in the second 4 hours showed a trend toward a positive association ( β=0.675, 95%CI:-0.107-1.457). First 4-hour emotional valence: significantly positive association with severity ( β=0.322, 95%CI: 0.067-0.577), second 4-hour arousal level: negative association with severity ( β=-0.258, 95%CI:-0.527 to 0.011), whereas arousal in the second 4 hours had a marginal negative effect (β=-0.258, 95% CI:-0.527 to 0.011). Conclusion:RRI may serve as a useful auxiliary measure in diagnosing depressive disorders and predicting severity among adolescents. Wearable smart devices offer promising potential for screening emotional dynamic factors related to adolescent depression.

ObjectiveTo evaluate the clinical efficacy of Maxing Loushi decoction in the treatment of acute exacerbation of chronic obstructive pulmonary disease （AECOPD） with phlegm turbidity obstructing lung syndrome， and to investigate its effects on inflammatory factors， immune function， and the programmed death-1（PD-1）/programmed death-ligand 1 （PD-L1） signaling pathway. MethodsA randomized controlled study was conducted， enrolling 90 hospitalized patients with AECOPD and phlegm turbidity obstructing lung syndrome in the Respiratory and Emergency Departments of Dongzhimen Hospital， Beijing University of Chinese Medicine， from April 2024 to December 2024. Patients were randomly assigned to a control group and an observation group using a random number table， with 45 patients in each group. The control group received conventional Western medical treatment， while the observation group received additional Maxing Loushi decoction for 14 days. Clinical efficacy， COPD Assessment Test （CAT） score， modified Medical Research Council Dyspnea Scale （mMRC）， 6-minute walk test （6MWT）， serum inflammatory factors， T lymphocyte subsets， and serum PD-1/PD-L1 levels were compared between the two groups before and after treatment. ResultsThe total clinical effective rate was 78.57% （33/42） in the control group and 95.35% （41/43） in the observation group， with the observation group showing significantly higher efficacy than that of the control group. The difference was statistically significant （χ² = 5.136， P<0.05）. After treatment， both groups showed significant reductions in CAT and mMRC scores （P<0.05， P<0.01） and significant increases in 6MWT compared to baseline （P<0.01）. The observation group demonstrated significantly greater improvements than the control group in this regard. Levels of inflammatory markers including C-reactive protein （CRP）， procalcitonin （PCT）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， monocyte chemoattractant protein-1（MCP-1）， and macrophage inflammatory protein-1α （MIP-1α） were significantly reduced in both groups （P<0.05， P<0.01）， with greater reductions in the observation group （P<0.05， P<0.01）. CD8⁺ levels were significantly reduced （P<0.01）， while CD3⁺， CD4⁺， and CD4⁺/CD8⁺ levels were significantly increased in both groups after treatment （P<0.05， P<0.01）， with more significant improvements observed in the observation group （P<0.05， P<0.01）. Serum PD-1 levels were reduced （P<0.05， P<0.01）， and PD-L1 levels were increased significantly in both groups after treatment （P<0.05， P<0.01）， with more pronounced changes in the observation group （P<0.05）. ConclusionMaxing Loushi decoction demonstrates definite therapeutic efficacy as an adjunctive treatment for patients with AECOPD and phlegm turbidity obstructing lung syndrome. It contributes to reducing serum inflammatory factors， improving immune function， and regulating the PD-1/PD-L1 signaling pathway.

ObjectiveTo evaluate the clinical efficacy of Maxing Loushi decoction in the treatment of acute exacerbation of chronic obstructive pulmonary disease （AECOPD） with phlegm turbidity obstructing lung syndrome， and to investigate its effects on inflammatory factors， immune function， and the programmed death-1（PD-1）/programmed death-ligand 1 （PD-L1） signaling pathway. MethodsA randomized controlled study was conducted， enrolling 90 hospitalized patients with AECOPD and phlegm turbidity obstructing lung syndrome in the Respiratory and Emergency Departments of Dongzhimen Hospital， Beijing University of Chinese Medicine， from April 2024 to December 2024. Patients were randomly assigned to a control group and an observation group using a random number table， with 45 patients in each group. The control group received conventional Western medical treatment， while the observation group received additional Maxing Loushi decoction for 14 days. Clinical efficacy， COPD Assessment Test （CAT） score， modified Medical Research Council Dyspnea Scale （mMRC）， 6-minute walk test （6MWT）， serum inflammatory factors， T lymphocyte subsets， and serum PD-1/PD-L1 levels were compared between the two groups before and after treatment. ResultsThe total clinical effective rate was 78.57% （33/42） in the control group and 95.35% （41/43） in the observation group， with the observation group showing significantly higher efficacy than that of the control group. The difference was statistically significant （χ² = 5.136， P<0.05）. After treatment， both groups showed significant reductions in CAT and mMRC scores （P<0.05， P<0.01） and significant increases in 6MWT compared to baseline （P<0.01）. The observation group demonstrated significantly greater improvements than the control group in this regard. Levels of inflammatory markers including C-reactive protein （CRP）， procalcitonin （PCT）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α）， monocyte chemoattractant protein-1（MCP-1）， and macrophage inflammatory protein-1α （MIP-1α） were significantly reduced in both groups （P<0.05， P<0.01）， with greater reductions in the observation group （P<0.05， P<0.01）. CD8⁺ levels were significantly reduced （P<0.01）， while CD3⁺， CD4⁺， and CD4⁺/CD8⁺ levels were significantly increased in both groups after treatment （P<0.05， P<0.01）， with more significant improvements observed in the observation group （P<0.05， P<0.01）. Serum PD-1 levels were reduced （P<0.05， P<0.01）， and PD-L1 levels were increased significantly in both groups after treatment （P<0.05， P<0.01）， with more pronounced changes in the observation group （P<0.05）. ConclusionMaxing Loushi decoction demonstrates definite therapeutic efficacy as an adjunctive treatment for patients with AECOPD and phlegm turbidity obstructing lung syndrome. It contributes to reducing serum inflammatory factors， improving immune function， and regulating the PD-1/PD-L1 signaling pathway.

The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4％.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.

Codon optimization was performed for the M and HN genes of bovine parainfluenza virus type 3(BPIV3),and the recombinant shuttle plasmid Dual-M+HN was constructed.BPIV3 VLPs was prepared using the baculovirus expression system,and verified by Western blot,IFA and elec-tron microscopy.The successfully verified virus-like particle(VLPs)were mixed with MF59 adjuvant and CpG-ODN immunoenhancer to immunize mice by intramuscular-injection,and BPIV3 inactivated vaccine group and adjuvant control group were set up.The immune effect of BPIV3 VLPs was evaluated by monitoring mouse serum specific antibodies,neutralizing antibodies and hemagglutination inhibition antibodies.The results showed that the optimized codon adaptation in-dex(CAI)of the M and HN protein genes were 0.96 and 0.95,respectively,and the CG content reached 54.1％and 53.1％,respectively.The constructed recombinant plasmid was transformed in-to DHI0Bac for blue and white spot screening.The validated recombinant rod particles were trans-fected into Sf9 cells to obtain the rod-shaped virus pFastBac-M+HN.Under electron microscopy,BPIV3 VLPs with a diameter of approximately 180 nm were observed.IFA and Western blot ex-periments confirmed the successful expression and biological activity of the target protein.Through protein optimization,it was found that the protein expression was highest at an infection dose of MOI=5.After mixing 50 μg VLPs with MF59 adjuvant and CpG-ODN,mice were immunized by intramuscular injection.The results showed that the antibodies in the VLPs immunized group be-gan to rise at 2 weeks of the first immunization and reached their peak at 21 days of the second im-munization,with an average IgG antibody titer of 1∶40 228;The average titer of neutralizing anti-bodies is 1∶298;The titer of hemagglutination inhibition antibody is 1∶549,reaching the level of inactivated vaccine(P≥0.05),indicating that the VLPs prepared in this experiment can induce hu-moral immune response in the body.In summary,this study successfully prepared VLPs capable of self-assembly expression of BPIV3 HN and M proteins,and induced humoral immune response in mice,providing research basis for subsequent BPIV3 VLPs vaccine research.

The codon was optimized for the bovine nebovirus(BNeV)VP1 gene and the recombi-nant plasmid pFastBac-Dual-VP1 was constructed,and BNeV-VP1 virus-like particles(VLPs)were prepared using a baculovirus expression system,and identified by Western blot,indirect im-munofluorescence and electron microscopy.Successfully validated VLPs were mixed with MF59 adjuvant and CpG-ODG,and mice were immunised by intramuscular injection and evaluated for immunity effects.The results showed that the optimized CAI(codon adaptation index)of VP1 gene was 0.93 and the GC content was 60.4％.The constructed recombinant plasmid was trans-formed into DH10Bac for blue-white spot screening,and after successful verification,it was trans-fected into SF9 cells to obtain recombinant baculovirus Baculo-BNeV-VP1.BNeV virus-like parti-cles with diameters ranging from 35 to 40 nm were observed under the electron microscope,and both IFA and Western blot experiments proved that the target proteins were successfully ex-pressed and biologically active,and protein optimisation revealed that the highest protein expres-sion was found at the infectious dose MOI=0.5.Mice were immunized by intramuscular injection after 50 μg of VLPs were mixed with MF59 adjuvant and CpG-ODN.The results showed that the VLPs immunization group produced IgG antibodies 7 days after the first dose,and the antibody ti-ter increased gradually,reaching a maximum of 1∶102 400,and declined at 35 d,but still main-tained a high level;The blocking titer BT50 is up to 640,which can induce the production of BNeV VP1-specific blocking antibody in mice.In this study,the baculovirus expression system was used to express the VP1 protein of BNeV,and BNeV VLPs were successfully constructed,which could induce humoral immune response in mice,which provided a reference for the follow-up research of BNeV vaccine.

Codon optimization was performed for the M and HN genes of bovine parainfluenza virus type 3(BPIV3),and the recombinant shuttle plasmid Dual-M+HN was constructed.BPIV3 VLPs was prepared using the baculovirus expression system,and verified by Western blot,IFA and elec-tron microscopy.The successfully verified virus-like particle(VLPs)were mixed with MF59 adjuvant and CpG-ODN immunoenhancer to immunize mice by intramuscular-injection,and BPIV3 inactivated vaccine group and adjuvant control group were set up.The immune effect of BPIV3 VLPs was evaluated by monitoring mouse serum specific antibodies,neutralizing antibodies and hemagglutination inhibition antibodies.The results showed that the optimized codon adaptation in-dex(CAI)of the M and HN protein genes were 0.96 and 0.95,respectively,and the CG content reached 54.1％and 53.1％,respectively.The constructed recombinant plasmid was transformed in-to DHI0Bac for blue and white spot screening.The validated recombinant rod particles were trans-fected into Sf9 cells to obtain the rod-shaped virus pFastBac-M+HN.Under electron microscopy,BPIV3 VLPs with a diameter of approximately 180 nm were observed.IFA and Western blot ex-periments confirmed the successful expression and biological activity of the target protein.Through protein optimization,it was found that the protein expression was highest at an infection dose of MOI=5.After mixing 50 μg VLPs with MF59 adjuvant and CpG-ODN,mice were immunized by intramuscular injection.The results showed that the antibodies in the VLPs immunized group be-gan to rise at 2 weeks of the first immunization and reached their peak at 21 days of the second im-munization,with an average IgG antibody titer of 1∶40 228;The average titer of neutralizing anti-bodies is 1∶298;The titer of hemagglutination inhibition antibody is 1∶549,reaching the level of inactivated vaccine(P≥0.05),indicating that the VLPs prepared in this experiment can induce hu-moral immune response in the body.In summary,this study successfully prepared VLPs capable of self-assembly expression of BPIV3 HN and M proteins,and induced humoral immune response in mice,providing research basis for subsequent BPIV3 VLPs vaccine research.

Objective:To observe the effects of Maxing Loushi decoction on the inflammatory response and inflammatory indicators in mice with chronic obstructive pulmonary disease (COPD) models.Methods:Thirty-six BALB/C mice were randomly divided into 4 groups by random number table method: 10 mice in the COPD model group (referred to as the model group), 10 mice in the Maxing Loushi decoction group (referred to as the traditional Chinese medicine group), 10 mice in the programmed death receptor-1 (PD-1) inhibitor group (referred to as the control group), and 6 mice in the normal group. The COPD models of mice in the model group, the traditional Chinese medicine group and the control group were prepared by cigarette smoking combined with lipopolysaccharide (LPS) induction method. During the modeling process, the model group and the traditional Chinese medicine group were respectively given normal saline and Maxing Loushi decoction by gavage. The control group was given intraperitoneal injection of PD-1 inhibitor, while the normal group was given intragastric administration of normal saline. Pathological changes of lung tissues in each group of mice were detected by HE staining. The levels of inflammatory factors [monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α)] in the plasma and alveolar lavage fluid (BALF) of mice in each group were determined by enzyme-linked immunosorbent assay (ELISA). The effects of Maxing Loushi decoction intervention on inflammatory responses and inflammatory factors were evaluated.Results:The lung tissue structure in the normal group was basically normal. There was no thickening of the alveolar walls, no infiltration of neutrophils in the tissues, and no obvious inflammatory infiltration. In the model group, the lung tissue structure was slightly abnormal. A small amount of alveolar atrophy could be observed, the alveolar walls were slightly thickened, and inflammatory infiltration could be seen. In the traditional Chinese medicine group, the lung tissue structure was slightly abnormal. A small amount of alveolar atrophy and collapse could be observed, the alveolar walls were not thickened, and individual neutrophil infiltration could be seen in the tissue. In the control group, the lung tissue structure was slightly abnormal, some alveoli atrophied, and a small amount of neutrophil infiltration could be seen in the tissue. The levels of MCP-1 and MIP-1α in plasma and lavage fluid of the model group were significantly higher than those of the normal group (all P<0.05), while the levels of MCP-1 and MIP-1α in plasma and lavage fluid of the traditional Chinese medicine group and the control group were significantly lower than those of the model group (all P<0.05). Moreover, the levels of plasma MCP-1 and MIP-1α in the traditional Chinese medicine group were significantly lower than those in the control group (all P<0.05), while there was no statistically significant difference in the levels of MCP-1 and MIP-1α in alveolar lavage fluid between the traditional Chinese medicine group and the control group (all P>0.05). The levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the model group were significantly higher than those of the normal group (all P<0.05), while the levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the traditional Chinese medicine group and the control group were significantly lower than those of the model group (all P<0.05). Moreover, the levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the traditional Chinese medicine group were significantly lower than those of the control group (all P<0.05). Conclusions:The intervention of Maxing Loushi decoction has a significant improvement effect on the inflammatory response of COPD model mice. Inflammatory factors such as MCP-1, MIP-1α, IL-6, and TNF-α can be used as indicators to determine the degree of COPD inflammation.

Objective:To observe the effects of Maxing Loushi decoction on the inflammatory response and inflammatory indicators in mice with chronic obstructive pulmonary disease (COPD) models.Methods:Thirty-six BALB/C mice were randomly divided into 4 groups by random number table method: 10 mice in the COPD model group (referred to as the model group), 10 mice in the Maxing Loushi decoction group (referred to as the traditional Chinese medicine group), 10 mice in the programmed death receptor-1 (PD-1) inhibitor group (referred to as the control group), and 6 mice in the normal group. The COPD models of mice in the model group, the traditional Chinese medicine group and the control group were prepared by cigarette smoking combined with lipopolysaccharide (LPS) induction method. During the modeling process, the model group and the traditional Chinese medicine group were respectively given normal saline and Maxing Loushi decoction by gavage. The control group was given intraperitoneal injection of PD-1 inhibitor, while the normal group was given intragastric administration of normal saline. Pathological changes of lung tissues in each group of mice were detected by HE staining. The levels of inflammatory factors [monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α)] in the plasma and alveolar lavage fluid (BALF) of mice in each group were determined by enzyme-linked immunosorbent assay (ELISA). The effects of Maxing Loushi decoction intervention on inflammatory responses and inflammatory factors were evaluated.Results:The lung tissue structure in the normal group was basically normal. There was no thickening of the alveolar walls, no infiltration of neutrophils in the tissues, and no obvious inflammatory infiltration. In the model group, the lung tissue structure was slightly abnormal. A small amount of alveolar atrophy could be observed, the alveolar walls were slightly thickened, and inflammatory infiltration could be seen. In the traditional Chinese medicine group, the lung tissue structure was slightly abnormal. A small amount of alveolar atrophy and collapse could be observed, the alveolar walls were not thickened, and individual neutrophil infiltration could be seen in the tissue. In the control group, the lung tissue structure was slightly abnormal, some alveoli atrophied, and a small amount of neutrophil infiltration could be seen in the tissue. The levels of MCP-1 and MIP-1α in plasma and lavage fluid of the model group were significantly higher than those of the normal group (all P<0.05), while the levels of MCP-1 and MIP-1α in plasma and lavage fluid of the traditional Chinese medicine group and the control group were significantly lower than those of the model group (all P<0.05). Moreover, the levels of plasma MCP-1 and MIP-1α in the traditional Chinese medicine group were significantly lower than those in the control group (all P<0.05), while there was no statistically significant difference in the levels of MCP-1 and MIP-1α in alveolar lavage fluid between the traditional Chinese medicine group and the control group (all P>0.05). The levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the model group were significantly higher than those of the normal group (all P<0.05), while the levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the traditional Chinese medicine group and the control group were significantly lower than those of the model group (all P<0.05). Moreover, the levels of IL-6 and TNF-α in plasma and alveolar lavage fluid of the traditional Chinese medicine group were significantly lower than those of the control group (all P<0.05). Conclusions:The intervention of Maxing Loushi decoction has a significant improvement effect on the inflammatory response of COPD model mice. Inflammatory factors such as MCP-1, MIP-1α, IL-6, and TNF-α can be used as indicators to determine the degree of COPD inflammation.