Immunotherapy has emerged as a revolutionary approach to treat immune-related diseases. Dendritic cells (DCs) play a pivotal role in orchestrating immune responses, making them an attractive target for immunotherapeutic interventions. Modulation of gene expression in DCs using genome editing techniques, such as the CRISPR-Cas system, is important for regulating DC functions. However, the precise delivery of CRISPR-based therapies to DCs has posed a significant challenge. While lipid nanoparticles (LNPs) have been extensively studied for gene editing in tumor cells, their potential application in DCs has remained relatively unexplored. This study investigates the important role of cholesterol in regulating the efficiency of BAMEA-O16B lipid-assisted nanoparticles (BLANs) as carriers of CRISPR/Cas9 for gene editing in DCs. Remarkably, BLANs with low cholesterol density exhibit exceptional mRNA uptake, improved endosomal escape, and efficient single-guide RNA release capabilities. Administration of BLAN_mCas9/gPD-L1 results in substantial PD-L1 gene knockout in conventional dendritic cells (cDCs), accompanied by heightened cDC1 activation, T cell stimulation, and significant suppression of tumor growth. The study underscores the pivotal role of cholesterol density within LNPs, revealing potent influence on gene editing efficacy within DCs. This strategy holds immense promise for the field of cancer immunotherapy, offering a novel avenue for treating immune-related diseases.

OBJECTIVES: To compare the application value of the likelihood ratio (LR) method and identity by state (IBS) method in the identification involving half sibling relationships, and to provide a reference for the setting of relevant standards for identification of half sibling relationship. METHODS: (1) Based on the same genetic marker combinations, the reliability of computer simulation method was verified by comparing the distributions of cumulated identity by state score (CIBS) and combined full sibling index in actual cases with the distributions in simulated cases. (2) In different numbers of three genetic marker combinations, the simulation of full sibling, half sibling and unrelated individual pairs, each 1 million pairs, was obtained; the CIBS, as well as the corresponding types of cumulative LR parameters, were calculated. (3) The application value of LR method was compared with that of IBS method, by comparing the best system efficiency provided by LR method and IBS method when genetic markers in different amounts and of different types and accuracy were applied to distinguish the above three relational individual pairs. (4) According to the existing simulation data, the minimum number of genetic markers required to distinguish half siblings from the other two relationships using different types of genetic markers was estimated by curve fitting. RESULTS: (1) After the rank sum test, under the premise that the real relationship and the genetic marker combination tested were the same, there was no significant difference between the simulation method and the results obtained in the actual case. (2) In most cases, under the same conditions, the system effectiveness obtained by LR method was greater than that by IBS method. (3) According to the existing data, the number of genetic markers required for full-half siblings and half sibling identification could be obtained by curve fitting when the system effectiveness reached 0.95 or 0.99. CONCLUSIONS When distinguishing half sibling from full sibling pairs or unrelated pairs, it is recommended to give preference to the LR method, and estimate the required number of markers according to the identification types and the population data, to ensure the identification effect.
Humans ; Siblings ; Genetic Markers ; Computer Simulation ; Irritable Bowel Syndrome/genetics* ; Reproducibility of Results ; Genotype

OBJECTIVES: To investigate the risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture and establish a nomogram model for predicting the risk of neonatal asphyxia. METHODS: A retrospective study was conducted with 613 cases of neonatal asphyxia treated in 20 cooperative hospitals in Enshi Tujia and Miao Autonomous Prefecture from January to December 2019 as the asphyxia group, and 988 randomly selected non-asphyxia neonates born and admitted to the neonatology department of these hospitals during the same period as the control group. Univariate and multivariate analyses were used to identify risk factors for neonatal asphyxia. R software (4.2.2) was used to establish a nomogram model. Receiver operator characteristic curve, calibration curve, and decision curve analysis were used to assess the discrimination, calibration, and clinical usefulness of the model for predicting the risk of neonatal asphyxia, respectively. RESULTS: Multivariate logistic regression analysis showed that minority (Tujia), male sex, premature birth, congenital malformations, abnormal fetal position, intrauterine distress, maternal occupation as a farmer, education level below high school, fewer than 9 prenatal check-ups, threatened abortion, abnormal umbilical cord, abnormal amniotic fluid, placenta previa, abruptio placentae, emergency caesarean section, and assisted delivery were independent risk factors for neonatal asphyxia (P<0.05). The area under the curve of the model for predicting the risk of neonatal asphyxia based on these risk factors was 0.748 (95%CI: 0.723-0.772). The calibration curve indicated high accuracy of the model for predicting the risk of neonatal asphyxia. The decision curve analysis showed that the model could provide a higher net benefit for neonates at risk of asphyxia. CONCLUSIONS The risk factors for neonatal asphyxia in Hubei Enshi Tujia and Miao Autonomous Prefecture are multifactorial, and the nomogram model based on these factors has good value in predicting the risk of neonatal asphyxia, which can help clinicians identify neonates at high risk of asphyxia early, and reduce the incidence of neonatal asphyxia.
Infant, Newborn ; Humans ; Male ; Pregnancy ; Female ; Nomograms ; Retrospective Studies ; Cesarean Section ; Risk Factors ; Asphyxia Neonatorum/etiology*

OBJECTIVE: To derive the Chinese medicine (CM) syndrome classification and subgroup syndrome characteristics of ischemic stroke patients. METHODS: By extracting the CM clinical electronic medical records (EMRs) of 7,170 hospitalized patients with ischemic stroke from 2016 to 2018 at Weifang Hospital of Traditional Chinese Medicine, Shandong Province, China, a patient similarity network (PSN) was constructed based on the symptomatic phenotype of the patients. Thereafter the efficient community detection method BGLL was used to identify subgroups of patients. Finally, subgroups with a large number of cases were selected to analyze the specific manifestations of clinical symptoms and CM syndromes in each subgroup. RESULTS: Seven main subgroups of patients with specific symptom characteristics were identified, including M3, M2, M1, M5, M0, M29 and M4. M3 and M0 subgroups had prominent posterior circulatory symptoms, while M3 was associated with autonomic disorders, and M4 manifested as anxiety; M2 and M4 had motor and motor coordination disorders; M1 had sensory disorders; M5 had more obvious lung infections; M29 had a disorder of consciousness. The specificity of CM syndromes of each subgroup was as follows. M3, M2, M1, M0, M29 and M4 all had the same syndrome as wind phlegm pattern; M3 and M0 both showed hyperactivity of Gan (Liver) yang pattern; M2 and M29 had similar syndromes, which corresponded to intertwined phlegm and blood stasis pattern and phlegm-stasis obstructing meridians pattern, respectively. The manifestations of CM syndromes often appeared in a combination of 2 or more syndrome elements. The most common combination of these 7 subgroups was wind-phlegm. The 7 subgroups of CM syndrome elements were specifically manifested as pathogenic wind, pathogenic phlegm, and deficiency pathogens. CONCLUSIONS There were 7 main symptom similarity-based subgroups in ischemic stroke patients, and their specific characteristics were obvious. The main syndromes were wind phlegm pattern and hyperactivity of Gan yang pattern.
Humans ; Syndrome ; Ischemic Stroke ; Medicine, Chinese Traditional ; Liver ; Phenotype

Bronchoalveolar Lavage ; Humans ; Lung ; Pulmonary Alveolar Proteinosis/therapy*

Ambystoma mexicanum/immunology* ; Amputation ; Animals ; Biomarkers/metabolism* ; Blastomeres/immunology* ; Cell Lineage/immunology* ; Connective Tissue Cells/immunology* ; Epithelial Cells/immunology* ; Forelimb ; Gene Expression ; High-Throughput Nucleotide Sequencing ; Humans ; Immunity ; Peroxiredoxins/immunology* ; Regeneration/immunology* ; Regenerative Medicine/methods* ; Single-Cell Analysis/methods*

OBJECTIVE: To determine the environmental contamination degree of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in corona virus disease 2019 (COVID-19) wards, to offer gui-dance for the infection control and to improve safety practices for medical staff, by sampling and detecting SARS-CoV-2 nucleic acid from the air of hospital wards, the high-frequency contact surfaces in the contaminated area and the surfaces of medical staff's protective equipment in a COVID-19 designated hospital in Wuhan, China. METHODS: From March 11 to March 19, 2020, we collected air samples from the clean area, the buffer room and the contaminated area respectively in the COVID-19 wards using a portable bioaerosol concentrator WA-15. And sterile premoistened swabs were used to sample the high-frequency contacted surfaces in the contaminated area and the surfaces of medical staff's protective equipment including outermost gloves, tracheotomy operator's positive pressure respiratory protective hood and isolation clothing. The SARS-CoV-2 nucleic acid of the samples were detected by real-time fluorescence quantitative PCR. During the isolation medical observation period, those medical staff who worked in the COVID-19 wards were detected for SARS-CoV-2 nucleic acid with oropharyngeal swabs, IgM and IgG antibody in the sera, and chest CT scans to confirm the infection status of COVID-19. RESULTS: No SARS-CoV-2 nucleic acid was detected in the tested samples, including the 90 air samples from the COVID-19 wards including clean area, buffer room and contaminated area, the 38 high-frequency contact surfaces samples of the contaminated area and 16 surface samples of medical staff's protective equipment including outermost gloves and isolation clothing. Moreover, detection of SARS-CoV-2 nucleic acid by oropharyngeal swabs and IgM, IgG antibodies in the sera of all the health-care workers who participated in the treatment for COVID-19 were all negative. Besides, no chest CT scan images of medical staff exhibited COVID-19 lung presentations. CONCLUSION Good ventilation conditions, strict disinfection of environmental facilities in hospital wards, guidance for correct habits in patients, and strict hand hygiene during medical staff are important to reduce the formation of viral aerosols, cut down the aerosol load, and avoid cross-infection in isolation wards. In the face of infectious diseases that were not fully mastered but ma-naged as class A, it is safe for medical personnel to be equipped at a high level.
Betacoronavirus ; COVID-19 ; China ; Coronavirus Infections ; Humans ; Medical Staff ; Pandemics ; Pneumonia, Viral ; Protective Devices ; SARS-CoV-2 ; Severe Acute Respiratory Syndrome/prevention & control*

Objective: To investigate changes in the three⁃dimensional position of the maxillary canine during the distal movement of the maxillary first molar by a mini⁃implant combined with a CD appliance. Methods: Ten typodont models of class II malocclusion were selected, and one side was randomly chosen as the experimental group. The CD ap⁃ pliance was bonded to the maxillary canine and first molar of the experimental group, and 1.2 mm stainless steel wire was bent as the anchorage, which was fixed on the model to simulate mini⁃implants implanted in the zygomatic alveolar ridge. Then, 180 g orthodontic force was applied to the canine of the experimental group; the other side was recognized as the control group and was not used for strengthening. Six mini⁃implants were implanted in different parts of the mod⁃ el and used as a reference before and after the experiment. The models were placed in an incubator and heated at 56 ℃ in a water bath for 2 min. The models were scanned before and after thermostatic water bath treatment with a 3⁃shape scanner. Then, the digital models were overlapped through the reference points, and the positions of the canines and first molars were measured before and after the experiment. Results: The sagittal movement distance of the first molar in the experimental group was ( 0.25 ± 0.33) mm, and the vertical movement distance was (0.25 ± 0.28 )mm, which was significantly different from the control group (P < 0.05), while the transverse change was not significantly different (P > 0.05). There was no significant difference in the root position of the first molar in the experimental group (P > 0.05). The lateral and vertical displacement distances of the maxillary canine crown in the experimental group were (4.03 ± 2.11) mm and (1.86 ± 1.01) mm, respectively, which were significantly different from those in the control group (P < 0.001), while the sagittal changes showed no significant differences (P > 0.05). In the experimental group, there was no significant difference in the position of the apex of the maxillary cusp (P > 0.05). Conclusion Our in vitro study showed that the maxillary canines inclined buccally accompanied by a small amount of intrusion during molar distaliza⁃ tion by a mini⁃implant combined with a CD appliance.

OBJECTIVE: To explore the effect of electrical stimulation at auricular points (EAS) combined with sound masking on the expression of cAMP-response element binding protein (CREB), brain-derived neurotrophic factor (BDNF) and tyrosine receptor kinase B (TrkB) in the auditory cortex of tinnitus rats. METHODS: A total of 27 adult male SD rats were randomly divided into a control group, a model group and an EAS group. The rats in the model group and the EAS group were intervened with intraperitoneal injection of sodium salicylate to induce tinnitus model, while the rats in the control group were intervened with injection of 0.9% NaCl solution. After the model was successfully established, the rats in the EAS group were treated with electrical stimulation at "Shenmen" (TF) and "Yidan" (CO), combined with sound masking; the treatment was given once a day for 15 days. The gap prepulse inhibition of acoustic startle (GPIAS) and prepulse inhibition (PPI) testing were performed using the acoustic startle reflex starter package for rats. The expression of BDNF, TrkB, CREB and p-CREB in the auditory cortex of each group were measured with Western Blot analysis. RESULTS: ① Compared with the control group, the GPIAS values in 12 kHz, 16 kHz, 20 kHz and 28 kHz were significantly decreased in the model group (all <0.05); compared with the model group, GPIAS values in 12 kHz, 16 kHz, 20 kHz and 28 kHz were significantly increased in the EAS group (all <0.05). ② Compared with the control group, the expression of BDNF and p-CREB in the model group was significantly increased (<0.01), and the expression of TrkB in the model group was significantly increased (<0.05); the differences of expression of BDNF, TrkB, CREB and p-CREB between the model group and the EAS group had no statistics significance (all >0.05). CONCLUSION EAS could improve the GPIAS values of high-frequency background sound in tinnitus rats, which may be related with the upregulation of the BDNF/TrkB/CREB signaling pathway in the auditory cortex, leading to the reversion of the maladaptive plasticity.
Acupuncture Points ; Animals ; Auditory Cortex ; Brain-Derived Neurotrophic Factor ; metabolism ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Electric Stimulation ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, trkB ; metabolism ; Tinnitus ; metabolism ; therapy

We investigated the potential hepatoprotective effect of Radix Bupleuri (RB) by inducing acute liver injury (ALI) in an animal model using acetaminophen (APAP) after pretreatment with RB aqueous extract for three consecutive days. Compared to those of the APAP group, the biochemical and histological results of the RB pretreatment group showed lower serumaspartate transaminase (AST) and alanine transaminase (ALT) levels as well as less liver damage. Pharmacokinetic study of the toxicity related marker acetaminophen-cysteine (APC) revealed a lower exposure level in rats, suggesting that RB alleviated APAP-induced liver damage by preventing glutathione (GSH) depletion. The results of cocktail approach showed significant inhibition of CYP2E1 and CYP3A activity. Further investigation revealed the increasing of CYP2E1 and CYP3A protein was significantly inhibited in pretreatment group, while no obvious effect on gene expression was found. Therefore, this study clearly demonstrates that RB exhibited significant protective action against APAP-induced acute live injury via pretreatment, and which is partly through inhibiting the increase of activity and translation of cytochrome P450 enzymes, rather than gene transcription.