Objective: To investigate the therapeutic effect of patchouli alcohol on mice with lung-heat syndrome based on the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-kappa B(NF-κB) signaling pathway. Methods: First, network pharmacology was used to predict the potential targets of patchouli alcohol in the treatment of lung-heat syndrome, and a "component-disease-key target" network was constructed for pathway analysis. Then, 40 BALB/c mice were assigned to the normal, lung-heat model, honeysuckle, and low-dose and high-dose patchouli alcohol groups. All groups, except the blank group, were intranasally infected with 50 μL (103 TCID50) of influenza virus solution. After two hours of infection, mice were treated once a day for seven consecutive days. The therapeutic mechanism of patchouli alcohol was explored by measuring pulmonary inflammatory factors, the PI3K/Akt/NF-κB pathway, hypothalamic fever markers (PGE2, cAMP, cGMP levels), rectal temperature, and tissue energy metabolism. Results: Network pharmacology identified 135 target genes related to patchouli alcohol and lung-heat syndrome, with the key targets being STAT3, H1F1A, and NF-κB1. In animal experiments, patchouli alcohol significantly alleviated influenza virus-induced lung inflammatory damage in mice with lung-heat syndrome, inhibited the expression of TNF-α and IL-6 in lung tissues(P<0.01), and suppressed the activation of the PI3K/Akt/NF-κB pathway. It also reduced hypothalamic levels of PGE2 and cAMP(P<0.01), suppressed the increase in rectal temperature, significantly decreased liver glycogen and pyruvate levels(P<0.01), and increased the activities of SDH, LDH, and Na+ -K+ -ATPase in the liver(P<0.01) Conclusion Patchouli alcohol improves the symptoms of lung-heat syndrome in mice by inhibiting the activation of the PI3K/Akt/NF-κB pathway, reducing proinflammatory cytokines and inflammatory damage, and regulating hypothalamic fever markers and energy metabolism.

Objective To explore the clinical application value of MRI fat quantification parameter[verte-bral bone marrow fat fraction(FF)]combined with 25-hydroxyvitamin D[25(OH)D]in predicting fracture risk in patients with osteoporosis.Methods A total of 90 patients with osteoporosis who were admitted to the hospital from January 2023 to April 2024 were selected as the subjects.Among them,50 patients with osteoporotic vertebral com-pression fractures were included in the fracture group,and 40 patients without fractures were included in the control group.All patients underwent the iterative decomposition of water and fat with echo asymmetry and least-squares estimation(IDEAL-IQ)method of MRI to measure the FF of each vertebra among L1-5 and the average FF of L1-5.Serum 25(OH)D level was detected by electrochemiluminescence method.FF and serum 25(OH)D levels of the two groups were compared.The correlation of FF,25(OH)D and bone mineral density(BMD)was analyzed.Multi-variate logistic regression analysis was conducted to screen the risk factors for fracture in patients with osteoporosis.Receiver operating characteristic(ROC)curves were used to evaluate the predictive value of FF,25(OH)D,and their combination for fracture in patients with osteoporosis.Results Patients in the fracture group were older than those in the control group.BMD and serum 25(OH)D level were lower than those of the control group(P<0.05).The FF of L2 and average FF of L1-5 in the fracture group were higher than those in the control group(P<0.05).Correlation analysis results showed that the FF of L2 and the average FF of L1-5 were negatively correlated with BMD(P<0.05),while serum 25(OH)D level was positively correlated with BMD(P<0.05).Multivariate logistic regression analysis showed that age and FF of L2 were independent risk factors for fracture in patients with osteoporo-sis,while BMD and 25(OH)D were protective factors(P<0.05).ROC curves indicated that the AUC values of FF of L2 and 25(OH)D for predicting fracture were 0.714(95%CI:0.606～0.822)and 0.774(95%CI:0.672～0.876).The AUC of joint prediction was 0.923(95%CI:0.867～0.978),which was significantly larger than that of separate prediction(P<0.05).Conclusions FF of L2 and serum 25(OH)D are related to fracture in patients with osteopo-rosis.Age and BMD are factors influencing the occurrence of fracture in patients with osteoporosis.FF of L2 and 25(OH)D have certain predictive value for fracture risk in patients with osteoporosis,and combined detection of the two can improve predictive efficiency.

ObjectiveTo explore the impact of subjective vertical perception impairment after stroke on visuospatial cognition, balance, walking and activities of daily living, to investigate the mediating role of visuospatial cognition and lateropulsion. MethodsFrom February to December, 2023, 96 stroke patients were selected from the Rehabilitation Medicine Center of the First Affiliated Hospital of Nanjing Medical University. They were divided into vertical perception impairment group (n = 53) and non-vertical perception impairment group (n = 43). They were assessed with National Institutes of Health Stroke Scale (NIHSS), the Scale for Contraversive Pushing (SCP), Burke Lateropulsion Scale (BLS), Line Bisection Test (LBT), Line Cancellation Test (LCT), Star Cancellation Test (SCT), Subjective Visual Vertical (SVV), Berg Balance Scale (BBS), Holden Functional Ambulation Categories (FAC) and Barthel Index (BI). ResultsScores of SVV orientation, SVV uncertainty, NIHSS, SCT, SCP, BLS, BBS, FAC and BI were worse in the vertical perception impairment group than in the non-vertical perception impairment group (|t| > 2.414, Z = -3.481, P < 0.05). Scores of SVV were correlated with SCT, BLS and BBS (|r| ≥ 0.273, P < 0.05). After controlling for age and gender, SVV orientation score did not directly impact BBS score (β = -0.011, P = 0.920). However, it exerted a partial mediating effect through BLS (effect = -0.173, 95%CI -0.278 to -0.076) and a chain-mediated effect through SCT and BLS (effect = -0.073, 95%CI -0.137 to -0.027), impacting BBS score. ConclusionSubjective vertical perception impairment results in poorer visuospatial cognition, balance, walking and activities of daily living in stroke patients. This influence on balance function is mediated through the mediating effects of visuospatial cognition and lateropulsion.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

Objective To investigate the mechanism of 2,6-dimethoxy-1,4-benzoquinone(DMQ),an active ingredients in fermented wheat germ extract,for inhibiting NLRP3 inflammasome activation and alleviating septic shock in mice.Methods Cultured murine bone marrow-derived macrophages(BMDM)stimulated with lipopolysaccharide(LPS)were treated with DMQ,followed by treatment with Nigericin,ATP,and MSU for activating the canonical NLRP3 inflammasome;the non-canonical NLRP3 inflammasome was activated by intracellular transfection of LPS,and AIM2 inflammasome was activated using Poly A:T.In human monocytic THP-1 cells,the effect of Nigericin on inflammasome activation products was examined using Western blotting and ELISA.Co-immunoprecipitation was performed to explore the mechanism of DMQ-induced blocking of NLRP3 inflammasome activation.In a male C57BL/6J mouse model of LPS-induced septic shock treated with 20 and 40 mg/kg DMQ,the levels of IL-1β and TNF-α in the serum and peritoneal lavage fluid were determined using ELISA,and the survival time of the mice within 36 h was observed.Results Treatment with DMQ effectively inhibited LPS-induced activation of canonical NLRP3 inflammasome in mouse BMDM and human THP-1 cells and also inhibited non-canonical NLRP3 inflammasome activation in mouse BMDM,but produced no significant effect on AIM2 inflammasome activation.DMQ significantly blocked the binding between ASC and NLRP3.In the mouse models of septic shock,DMQ treatment significantly reduced the levels of IL-1β in the serum and peritoneal fluid and obviously prolonged survival time of the mice.Conclusion DMQ can effectively block ASC-NLRP3 interaction to inhibit NLRP3 inflammasome activation and alleviate LPS-induced septic shock in mice.

Objective To investigate the effect of serum containing Huangqi decoction on the proliferation, migration, and tubulogenesis of rat liver sinusoidal endothelial cells (LSECs) induced by vascular endothelial growth factor (VEGF) and its mechanism of action based on the protein kinase B (AKT)/mammalian target of rapamycin (mTOR) signaling pathway. Methods The serum containing Huangqi decoction was prepared, and rat LSECs were isolated and cultured in vitro . Then rat LSECs were randomly divided into blank group, VEGF group, serum control group, and low-, middle-, and high-dose serum containing Huangqi decoction groups. MTT colorimetry, Transwell assay, and tubulogenesis assay were used to measure the proliferation, migration, and tubulogenesis abilities of LSECs in each group, and Western Blot was used to measure the protein expression levels of platelet endothelial cell adhesion molecule-1 (CD31), endothelin-1 (ET-1), and endothelial nitric oxide synthase (eNOS), as well as AKT, phosphorylated-AKT (p-AKT), mTOR, and phosphorylated mTOR (p-mTOR) in the AKT/mTOR signaling pathway. A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the Tukey's test was used for further comparison between two groups. Results Compared with the blank group, the VEGF group and the serum control group had significantly promoted proliferation, migration, and angiogenesis of rat LSECs (all P < 0.05), and Western Blot showed significant increases in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.05). There were no significant differences in the above indices between the VEGF group and the serum control group (all P > 0.05). Compared with the serum group, the middle- and high-dose serum containing Huangqi decoction groups had significantly inhibited proliferation, migration, and angiogenesis of rat LSECs induced by VEGF (all P < 0.05), and Western Blot showed significant reductions in the expression levels of CD31, ET-1, eNOS, and AKT/mTOR signaling pathway-related proteins (all P < 0.01). Conclusion A relatively high dose of serum containing Huangqi decoction can significantly inhibit the proliferation, migration, and tubulogenesis of rat LSECs induced by VEGF, possibly by regulating the AKT/mTOR signaling pathway.

Objective:To construct a chronic obstructive pulmonary disease (COPD) assessment test (CAT) score prediction model based on a deep network fused with air data, and to explore its significance.Methods:From February 2015 to December 2017, the outdoor environmental monitoring air data near the residential area of the patients with COPD from the Respiratory Outpatient Clinics of Peking University Third Hospital, Peking University People′s Hospital and Beijing Jishuitan Hospital were collected and the daily air pollution exposure of patients was calculated. The daily CAT scores were recorded continuously. The CAT score of the patients in the next week was predicted by fusing the time series algorithm and neural network to establish a model, and the prediction accuracy of the model was compared with that of the long short-term memory model (LSTM), the LSTM-attention model and the autoregressive integrated moving average model (ARIMA).Results:A total of 47 patients with COPD were enrolled and followed up for an average of 381.60 days. The LSTM-convolutional neural networks (CNN)-autoregression (AR) model was constructed by using the collected air data and CAT score, and the root mean square error of the model was 0.85, and the mean absolute error was 0.71. Compared with LSTM, LSTM-attention and ARIMA, the average prediction accuracy was improved by 21.69%.Conclusion:Based on the air data in the environment of COPD patients, the fusion deep network model can predict the CAT score of COPD patients more accurately.

A new variant of concern for SARS-CoV-2,Omicron(B.1.1.529),was designated by the World Health Organization on November 26,2021.This study analyzed the viral genome sequenc-ing data of 108 samples collected from patients infected with Omicron.First,we found that the enrichment efficiency of viral nucleic acids was reduced due to mutations in the region where the primers anneal to.Second,the Omicron variant possesses an excessive number of mutations compared to other variants circulating at the same time(median:62 vs.45),especially in the Spike gene.Mutations in the Spike gene confer alterations in 32 amino acid residues,more than those observed in other SARS-CoV-2 variants.Moreover,a large number of nonsynonymous mutations occur in the codons for the amino acid residues located on the surface of the Spike protein,which could potentially affect the replication,infectivity,and antigenicity of SARS-CoV-2.Third,there are 53 mutations between the Omicron variant and its closest sequences available in public databases.Many of these mutations were rarely observed in public databases and had a low muta-tion rate.In addition,the linkage disequilibrium between these mutations was low,with a limited number of mutations concurrently observed in the same genome,suggesting that the Omicron vari-ant would be in a different evolutionary branch from the currently prevalent variants.To improve our ability to detect and track the source of new variants rapidly,it is imperative to further strengthen genomic surveillance and data sharing globally in a timely manner.

Animals ; CRISPR-Cas Systems ; Gene Editing ; Humans

Objective: To evaluate the effect of 3D-printed oval root canal preparation by using small field-of-view cone beam CT (CBCT) combined with computer-aided technologies. Methods: An extracted tooth with a suitable single oval canal was selected by small field-of-view CBCT scanning. Three-dimensional (3D)-printed resin teeth were obtained based on the CBCT data after data conversion and processing. 50 general dentists were selected to prepare the oval root canal of the resin teeth with X-Gold rotary Ni-Ti instruments. Small field-of-view CBCT was applied to scan the oval root canals before and after preparation. Then, computer-aided technologies were used to calculate and compare these CBCT data, analyzing the effects of oval root canal cleaning as well as the root canal deviation by 3D reconstruction and rendering of the images. Results : Among the 50 cases, the mean unprepared area of the oval root canal wall was（56.20 ± 11.91）% and the mean maximum deviation distance of the root canal was（0.140 ± 0.041）mm. There was no correlation between root canal cleanliness and deviation (r=0.18, P=0.212). Conclusion Combined with small field-of-view CBCT and computer-aided technology, we can effectively quantitatively evaluate the root canal preparation effect.