Understanding microbial-host interactions in the oral cavity is essential for elucidating oral disease pathogenesis and its systemic implications. In vitro bacteria-host cell coculture models have enabled fundamental studies to characterize bacterial infection and host responses in a reductionist yet reproducible manner. However, existing in vitro coculture models fail to establish conditions that are suitable for the growth of both mammalian cells and anaerobes, thereby hindering a comprehensive understanding of their interactions. Here, we present an asymmetric gas coculture system that simulates the oral microenvironment by maintaining distinct normoxic and anaerobic conditions for gingival epithelial cells and anaerobic bacteria, respectively. Using a key oral pathobiont, Fusobacterium nucleatum, as the primary test bed, we demonstrate that the system preserves bacterial viability and supports the integrity of telomerase-immortalized gingival keratinocytes. Compared to conventional models, this system enhanced bacterial invasion, elevated intracellular bacterial loads, and elicited more robust host pro-inflammatory responses, including increased secretion of CXCL10, IL-6, and IL-8. In addition, the model enabled precise evaluation of antibiotic efficacy against intracellular pathogens. Finally, we validate the ability of the asymmetric system to support the proliferation of a more oxygen-sensitive oral pathobiont, Porphyromonas gingivalis. These results underscore the utility of this coculture platform for studying oral microbial pathogenesis and screening therapeutics, offering a physiologically relevant approach to advance oral and systemic health research.
Coculture Techniques/methods* ; Humans ; Fusobacterium nucleatum/physiology* ; Gingiva/microbiology* ; Keratinocytes/microbiology* ; Host Microbial Interactions ; Mouth/microbiology* ; Host-Pathogen Interactions ; Epithelial Cells/microbiology* ; Cells, Cultured ; Porphyromonas gingivalis

Host-derived small RNAs are emerging as critical regulators in the dynamic interactions between host tissues and the microbiome, with implications for microbial pathogenesis and host defense. Among these, transfer RNA-derived small RNAs (tsRNAs) have garnered attention for their roles in modulating microbial behavior. However, the bacterial factors mediating tsRNA interaction and functionality remain poorly understood. In this study, using RNA affinity pull-down assay in combination with mass spectrometry, we identified a putative membrane-bound protein, annotated as P-type ATPase transporter (PtaT) in Fusobacterium nucleatum (Fn), which binds Fn-targeting tsRNAs in a sequence-specific manner. Through targeted mutagenesis and phenotypic characterization, we showed that in both the Fn type strain and a clinical tumor isolate, deletion of ptaT led to reduced tsRNA intake and enhanced resistance to tsRNA-induced growth inhibition. Global RNA sequencing and label-free Raman spectroscopy revealed the phenotypic differences between Fn wild type and PtaT-deficient mutant, highlighting the functional significance of PtaT in purine and pyrimidine metabolism. Furthermore, AlphaFold 3 prediction provides evidence supporting the specific binding between PtaT and Fn-targeting tsRNA. By uncovering the first RNA-binding protein in Fn implicated in growth modulation through interactions with host-derived small RNAs (sRNAs), our study offers new insights into sRNA-mediated host-pathogen interplay within the context of microbiome-host interactions.
Fusobacterium nucleatum/growth & development* ; RNA-Binding Proteins/genetics* ; Bacterial Proteins/genetics* ; RNA, Bacterial/metabolism* ; Humans ; RNA, Transfer/metabolism*

Objective:To evaluate the clinical effect of orbital fat releasing and repositioning through supra-suborbicularis oculi fat (SOOF) in lower eyelid blepharoplasty.Methods:The authors retrospectively analyzed the clinical data of patients who received lower eyelid blepharoplasty in the Department of Plastic Surgery, Beijing Tsinghua Changgung Hospital from January 2017 to January 2023. Lower eyelid blepharoplasty with orbital fat releasing and repositioning through supra-SOOF was performed. The tear trough ligament and orbicularis retaining ligament were released between the orbicularis oculi muscle and SOOF via skin approach. Orbital septal fat was released to this space, lateral fat flap was fixed to the SOOF capsule, medial fat flap was fixed to the superficial fascia of the levator labii superioris muscle, orbital septum was repositioned, and lateral canthus was suspended. After the operation, lower eyelid morphology and complications were evaluated.Results:A total of 58 patients were enrolled, including 8 males and 50 females with the age of (49.8 ± 10.4) years (32 to 70 years). All patients had obvious lower eyelid bags on both sides, combining the tear trough deformity and (or) palabromalar groove. Fifty-seven patients were followed up for (29.1 ± 16.9) months (6-67 months). All patients were satisfied with the operation. The appearance of lower eyelid was greatly improved, with elimination of fat prominence and tear trough deformity. Mild lower eyelid ectropion was observed two cases 1 week after the operation, and were relieved by moderate massage 2 weeks postoperatively. In one case, mild lower eyelid retraction was observed 1 month postoperatively, with no complaint of discomfort or further treatments. Bulbar conjunctival hemorrhage occurred in 1 case and recovered without further interventions.Conclusion:Lower eyelid blepharoplasty with orbital fat releasing and reposting through supra-SOOF can effectively release the tear trough ligament and the orbicularis retaining ligament, with stronger fat fixation and less surgical difficulty. After the operation, eyelid contour was flat and tight with fewer complications and higher patient satisfaction.

Objective To carry out carrier screening for spinal muscular atrophy(SMA)in 3 302 preg-nant women from Xinjiang region and preliminarily determine the SMA carrier frequency among pregnant women in the region.Methods A total of 29 089 pregnant women who underwent prenatal visits at this hos-pital from April 2020 to February 2023 were educated,of whom 3 302 were received SMA carrier screening.SMA carriers were screened by detecting the copy numbers of SMN1 exons 7(E7)and 8(E8)using quantita-tive fluorescence PCR.Multiplex ligation-dependent probe amplification(MLPA)was used for prenatal diag-nosis of high-risk fetuses in couples who were both SMA carriers.Results The acceptance rate of SMA carri-ers screening was 11.35％.Among the 3 302 pregnant women,58 were found to be SMA carriers,and the total carrier frequency was 1.76％(1/57).Among them,there were 45 cases of Han ethnicity,with a carrying fre-quency of 1.63％(1/61),and there were 13 cases of ethnic minorities,with a carrying frequency of 2.39％(1/42).Among the 58 carriers,46 spouses were received SMA screening,and the results showed that two couples were both SMA carriers.Further prenatal diagnosis was performed,and the results of MLPA indicated that the fetuses were all heterozygous deletions of SMN1 E7 and E8,suggesting continued pregnancy.Conclusion This study has preliminarily determined the SMA carrier frequency among pregnant women in Xinjiang region.SMA carrier screening in pregnant women and prenatal diagnosis of high-risk fetuses are of great significance for the prevention and control of birth defects.

Objective To investigate the value of contrast-enhanced ultrasound(CEUS)combined with shear wave elastography(SWE)in the diagnosis of liver tumors.Methods This study was conducted according to the PRISMA guideline,with a PROSPERO registration number of CRD42023491288.PubMed,Embase,the Cochrane Library,CNKI,VIP,and Wanfang Data were searched for articles on CEUS combined with SWE in the diagnosis of liver tumors published from January 2000 to October 2023,and a total of 12 articles were included,with 1 328 patients in total.The QUADAS-2 tool was used to assess the quality of the articles included.Stata 15.0 software was used to calculate pooled sensitivity,specificity,positive likelihood ratio,negative likelihood ratio,diagnostic odds ratio,and heterogeneity.The summary receiver operating characteristic(SROC)curve was plotted,and the area under the SROC curve(AUC)was calculated.Results There were 1 457 lesions for the patients included,among whom there were 764 malignant lesions and 693 benign lesions,with a positive rate of 52.44%and a negative rate of 47.56%.Calculations obtained a pooled sensitivity of 0.94(95%confidence interval[CI]:0.91—0.96),a specificity of 0.92(95%CI:0.87—0.95),a positive likelihood ratio of 12.00(95%CI:7.40—19.40),a negative likelihood ratio of 0.06(95%CI:0.04—0.10),and a diagnostic odds ratio of 191(95%CI:87—417).The tests for heterogeneity showed Q=54.78,df=11.00,P<0.001,and I2=79.92%(95%CI:69.18%—90.66%),with an AUC of 0.98.Conclusion CEUS combined with SWE has a relatively high diagnostic value for benign and malignant liver tumors and thus holds promise for clinical application.

Objective:To evaluate the clinical effect of orbital fat releasing and repositioning through supra-suborbicularis oculi fat (SOOF) in lower eyelid blepharoplasty.Methods:The authors retrospectively analyzed the clinical data of patients who received lower eyelid blepharoplasty in the Department of Plastic Surgery, Beijing Tsinghua Changgung Hospital from January 2017 to January 2023. Lower eyelid blepharoplasty with orbital fat releasing and repositioning through supra-SOOF was performed. The tear trough ligament and orbicularis retaining ligament were released between the orbicularis oculi muscle and SOOF via skin approach. Orbital septal fat was released to this space, lateral fat flap was fixed to the SOOF capsule, medial fat flap was fixed to the superficial fascia of the levator labii superioris muscle, orbital septum was repositioned, and lateral canthus was suspended. After the operation, lower eyelid morphology and complications were evaluated.Results:A total of 58 patients were enrolled, including 8 males and 50 females with the age of (49.8 ± 10.4) years (32 to 70 years). All patients had obvious lower eyelid bags on both sides, combining the tear trough deformity and (or) palabromalar groove. Fifty-seven patients were followed up for (29.1 ± 16.9) months (6-67 months). All patients were satisfied with the operation. The appearance of lower eyelid was greatly improved, with elimination of fat prominence and tear trough deformity. Mild lower eyelid ectropion was observed two cases 1 week after the operation, and were relieved by moderate massage 2 weeks postoperatively. In one case, mild lower eyelid retraction was observed 1 month postoperatively, with no complaint of discomfort or further treatments. Bulbar conjunctival hemorrhage occurred in 1 case and recovered without further interventions.Conclusion:Lower eyelid blepharoplasty with orbital fat releasing and reposting through supra-SOOF can effectively release the tear trough ligament and the orbicularis retaining ligament, with stronger fat fixation and less surgical difficulty. After the operation, eyelid contour was flat and tight with fewer complications and higher patient satisfaction.

Sleep is an important activity of daily life for individuals,and sleep disorders can seriously affect their physical and mental health.This article summarizes the impact of vestibular stimulation on sleep,and reviews feasible and effective intervention methods from swing movement,galvanic vestibular stimulation,and weighted blankets stimulation,in order to provide new ideas and methods for the diagnosis and treatment of sleep disorders.

Objective:To explore the application experience of autologous fat transplantation in improving the facial contour of young cosmetic patients.Methods:From October 2017 to October 2020, the plastic surgery department of Beijing Tsinghua Changgung Hospital admitted 10 young cosmetic patients with poor facial contours, including 1 male and 9 females, aged 18-35 years, with an average of 28 years. Autologous subcutaneous fat was harvested by liposuction and static purification and then injected into the areas with poor facial contour with an amount of 10%-30% over.Results:The facial contour of 10 young cosmetic patients was well improved, and there were no postoperative complications such as facial asymmetry, local uneven skin, skin infection and necrosis or fat embolism. 8 cases were filled once and 2 cases were filled twice. The patients were followed up for 6-24 months and the postoperative effect was good. The excellent and good rate evaluated by patients, plastic surgeon and the third party doctor was more than 80%.Conclusions:The use of autologous fat to improve the facial contour of young cosmetic patients is easy to operate with less trauma and good effect, which is worthy of promotion.

Objective:To retrospectively review the efficacy of enhanced epidural anesthesia in the patients undergoing cesarean section.Methods:The data from patients who underwent cesarean section in our hospital from August 2008 to December 2021 were retrospectively collected. The method of epidural anesthesia, drug dosage, onset time, blocking level, anesthesia effect, remedial medication, and complications related to the method were recorded. The main points of enhanced epidural anesthesia included: ① test negative pressure by rotating the epidural puncture needle in situ to determine the position of the needle tip; ② catheterization after administering local anesthetic through epidural puncture needle to assess the clinical application of this technique. A questionnaire survey was conducted in 134 anesthesiologists with qualifications for epidural procedures. The questionnaire addressed whether to perform routine needle rotation during epidural procedures, direction of needle rotation, and the percentage of the dosage of local anesthetic administered through puncture needle. Results:A total of 10 054 patients received epidural anesthesia (10 016 cases) and switched to general anesthesia because of poor epidural effect or intraoperative emergencies (38 cases) were enrolled. The consumption of local anesthetics for epidural anesthesia was 18 (15, 20) ml.There were 7 287 cases with block plane records, of which the ratio of the block level reached T 6-8 was 86.84%. The time from the beginning of anesthesia to skin incision was 25 (20, 30) min. Sixteen patients (0.159%) switched to general anesthesia because of poor epidural effect.Among 10 016 patients underwent epidural anesthesia, 925 patients (9.235%) received intravenous adjunctive drugs.No patients experienced total spinal anesthesia, 25 patients (0.248%) had epidural perforation due to puncture or needle rotation, and 4 cases (0.039%) local anesthetic intoxication, 3 cases (0.030%) postoperative nerve injury, and 142 patients (1.412%) requiring intervention with vasoactive drugs due to hemodynamic instability within 30 min after epidural administration. Questionnaire survey: The proportion of anesthesiologists rotating epidural puncture needles was 85.1%, and the proportion of anesthesiologists using epidural puncture needles to inject local anesthesia drugs with a dose greater than 50% of the total dose before catheterization was 84.2%. Conclusions:Enhanced epidural anesthesia can provide effective anesthesia for the patients undergoing cesarean section without increasing the risk of complications related to epidural procedures.