AIM: To investigate the diagnostic and prognostic value of differential expression of Cyclin D1 and p53 in eyelid tumors.METHODS: This retrospective study enrolled patients who underwent surgical resection for eyelid tumors at our hospital between March 2018 and March 2023. Participants were categorized into benign and malignant groups based on tumor characteristics. Clinical data were collected. Genetic data for eyelid tumors were obtained from the GEO database, and differential gene analysis, including volcano plot visualization and KEGG pathway enrichment analysis, was performed using the Sangerbox 3.0 platform. Immunohistochemistry was used to detect the expression levels of Cyclin D1, p53, and BAX in tissue samples. Correlations with clinical features were analyzed using Spearman analysis, and prognostic factors were identified via Logistic regression analysis.RESULTS: This study included 69 patients with eyelid tumors(78 eyes), categorized into a benign group(37 patients, 41 eyes)and a malignant group(32 patients, 37 eyes)based on tumor characteristics. There were significant differences between the two groups in histological subtype, TNM staging, vascular invasion, differentiation status, and local infiltration(all P<0.05). Among benign tumors: pigmented nevi in 11 eyes(27%), hemangiomas in 9 eyes(22%), squamous cell papillomas in 5 eyes(12%), epidermoid cysts in 5 eyes(12%), seborrheic keratoses in 4 eyes(10%), neurofibromas in 3 eyes(7%), and both calcifying epithelioma and xanthelasma in 2 eyes each(5%); among malignant tumors: basal cell carcinoma in 18 eyes(49%), meibomian gland carcinoma in 8 eyes(22%), squamous cell carcinoma in 5 eyes(14%), sebaceous gland carcinoma in 4 eyes(11%), lymphoma and malignant melanoma each in 1 eye(3%). At the follow-up cutoff date of March 2025, the 2-year survival rate in the benign group(95%)was significantly higher than that in the malignant group(78%; P<0.05). Bioinformatics analysis identified 4 103 differentially expressed genes, including Cyclin D1, p53, and BAX, which were predominantly involved in pathways such as the p53 signaling pathway and calcium-related signaling. Spearman analysis revealed that local invasion(rs=0.71, P<0.05)and TNM stage(rs=0.73, P<0.05)correlated with Cyclin D1 expression; local invasion(rs=0.76, P<0.05)and histological subtype(rs=0.65, P<0.05)correlated with p53 expression. Logistic regression results indicated that Cyclin D1, p53, TNM staging, and local invasion were prognostic risk factors. ROC curve analysis demonstrated that the combined detection of these four indicators had the highest predictive value for prognosis(AUC=0.83).CONCLUSION: High expression of cyclin D1 and p53 serves as molecular markers for distinguishing benign from malignant eyelid tumors and assessing prognosis. Combined detection of these markers with TNM staging and local invasion demonstrates high predictive value for prognosis.

ObjectiveTaking Aurantii Fructus Immaturus juice（AFI）-processed Atractylodis Macrocephalae Rhizoma（AMR） as an example， this study aims to systematically compare the volatile and non-volatile components of AMR and its processed products， investigate the key differential components， evaluate their anti-inflammatory activities， and elucidate the synergistic mechanism of processing. MethodsThe chemical compositions of volatile and non-volatile components in AMR and AFI-processed AMR were systematically characterized using gas chromatography-mass spectrometry（GC-MS） and ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）， with relative mass fractions and response values determined separately. Volatile components were identified through searches in the National Institute of Standards and Technology（NIST）17 database， comparison with retention index（RI） and fragmentation pattern matching. Non-volatile components were identified by searching Waters Traditional Chinese Medicine （TCM） spectral library， in conjunction with PubChem and MassBank， characteristic fragmentation patterns and response values were also used to support identification. Differential components were screened using principal component analysis（PCA）， orthogonal partial least squares-discriminant analysis（OPLS-DA）， with variable importance in the projection（VIP） value >1. Components with high log₂fold change（FC） among major differential groups were selected as those exhibiting significant changes before and after processing. The anti-inflammatory activity of the differential compounds was evaluated by assessing their effects on nitric oxide（NO） production in a lipopolysaccharide（LPS）-induced RAW264.7 macrophage model. Enzyme-linked immunosorbent assay（ELISA） was used to detect the effects of the differential components on tumor necrosis factor（TNF）-α， interleukin（IL）-1β， IL-6， and monocyte chemotactic protein（MCP）-1 levels， and immunofluorescence（IF） was employed to assess their effects on nuclear transcription factor（NF）-κB p65 translocation， thereby elucidating the underlying molecular mechanisms. ResultsA total of 36 compounds were identified in the volatile components of AMR and AFI-processed AMR， among which， sesquiterpenes and monoterpenes were significantly increased after processing. In the non-volatile components， 36 compounds were identified， and the main differential components were flavonoids， sesquiterpenoids， and triterpenoids. Flavonoids were the primary differential components distinguishing AMR from its processed products， representing compounds directly introduced during processing. Five compounds， including atractylenolide Ⅲ， tangeritin， nobiletin， hesperidin and narirutin， were selected as representatives of three classes based on their most prominent differential expression among different compound types for subsequent anti-inflammatory activity studies. The results showed that 100 μmol·L^-1 tangerine and narirutin could significantly inhibit LPS-induced NO production（P<0.01） in a concentration-dependent manner. Tangeritin was able to significantly inhibit the levels of TNF-α and MCP-1 secreted by RAW264.7（P<0.05）， while narirutin significantly inhibited the levels of TNF-α， IL-1β， MCP-1 and IL-6（P<0.01）. IF revealed that both tangeritin and narirutin significantly blocked the translocation of NF-κB p65 from the cytoplasm to the nucleus. ConclusionAFI-processed AMR significantly alters the chemical composition profile of AMR， and the newly introduced flavonoid components during processing may be key to its enhanced anti-inflammatory effects.

BACKGROUND:Multiple studies have confirmed that mitogen-activated protein kinase(MAPK)signaling pathway is involved in cell proliferation,and microRNA(miR)is involved in the occurrence and development of hypertrophic scars.Therefore,the role of miR-27a-3p and MAPK signaling pathways in pathological scar formation has been further explored. OBJECTIVE:To explore the effect of miR-27a-3p on the proliferation of human hypertrophic scar fibroblasts through the MAPK signaling pathway. METHODS:The primary fibroblasts were isolated and collected from the skin samples.The primary fibroblasts were observed by inverted microscope and verified by immunofluorescence.The relative expression level of miR-27a-3p in tissues was detected by qRT-PCR.The target genes of hsa-miR-27a-3p were predicted using the database,and then the predicted target genes were enriched by gene ontology function analysis and biological pathway enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes.There were seven groups:blank control,negative control,miR-27a-3p mimic,miR-27a-3p inhibitor,miR-27a-3p mimic+p38 MAPK inhibitor,miR-27a-3p mimic+extracellular regulated protein kinase inhibitor,miR-27a-3p mimic+c-Jun N-terminal kinase inhibitor.Western blot was used to detect the levels of extracellular regulated protein kinase,c-Jun N-terminal kinase inhibitor.and p38 kinase and their phosphorylation levels.Cell counting kit-8 and EdU were used to detect cell proliferation. RESULTS AND CONCLUSION:Compared with normal skin fibroblasts,hypertrophic scar fibroblasts had stronger proliferative activity(P<0.05)and faster proliferation level(P<0.001).Compared with normal skin,miR-27a-3p was highly expressed in hypertrophic scars(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p could promote cell proliferation activity(P<0.001)and proliferation levels(P<0.001).Compared with the negative control group,knockdown of miR-27a-3p could inhibit the proliferation activity(P<0.05)and proliferation levels(P<0.001).Compared with the negative control group,overexpression of miR-27a-3p promoted the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 mitogen-activated protein kinase(P<0.05).Compared with the negative control group,knockdown of miR-27a-3p inhibited the phosphorylated levels of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK(P<0.05).Compared with the miR-27a-3p mimic group,specific inhibitors of extracellular regulated protein kinase,c-Jun N-terminal kinase,and p38 MAPK reversed the effects of miR-27a-3p on the proliferative activity(P<0.01)and proliferation level(P<0.001)of fibroblasts.To conclude,these results suggest that miR-27a-3p promotes the proliferation of human hypertrophic scar fibroblasts by activating the MAPK signaling pathway.

Objective:To investigate the characteristics of visual and auditory sustained attention and sex differences in pupils of different grades.Method:Totally 231 students from grades 1 to 6 participated in this stud-y.The development and sex differences of sustained attention were measured by average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates in visual and auditory two-choice re-action time(CRT)tasks.Result:The students in grade 5 had significantly slower visual average reaction times and coefficients of variation for reaction times(P＜0.001 or P＜0.05)than those in grade 3.The students in grade 5 had significantly lower auditory average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates(P＜0.001 or P＜0.01)than those in grade 1.There was no significant sex differ-ences in visual or auditory average reaction times,coefficients of variation for reaction times or omission error rates(Ps＞0.05).Conclusion:There would be age differences in characteristics of both visual and auditory sustained at-tention in pupils,and may be no sex difference in visual or auditory sustained attention.

Objective:To evaluate the influence of different positive end-expiratory pressure on the ventilation of laryngeal mask airway in pediatric patients undergoing laparoscopic surgery through ultrasound assessment.Methods:In this randomized controlled trial, 90 pediatric patients of both sexes, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, aged 2-10 yr, with a body mass index of 12-22 kg/m 2, scheduled for elective laparoscopic surgery, were divided into 3 groups ( n=30 each) by a random number table method: group P0, group P3, and group P5. Each group adopted the pressure-controlled mode with an inspiration/expiration ratio of 1∶2, a flow rate of 2 L/min, and a respiratory rate of 16-26 breaths/min. The positive end-expiratory pressure was set at 0, 3 and 5 cmH 2O(1 cmH 2O=0.098 kPa) in P0, P3 and P5 groups respectively. The ultrasonic scores and misalignment rate of the laryngeal mask airway were recorded every 5 min following the pneumoperitoneum, and the occurrence of intraoperative hypoxemia (SpO 2 < 92%) and P ETCO 2 ≥ 45 mmHg(1 mmHg=0.133 kPa) was recorded. The development of hypoxemia, blood staining on the laryngeal mask airway and hoarseness after laryngeal mask airway removal were also recorded. Results:Compared with P0 group, the ultrasound scores were significantly increased at 15 min of pneumoperitoneum in P3 group ( P<0.05), and the ultrasound scores and the laryngeal mask airway misplacement rate were significantly increased, and no significant change was found in the other parameters in P5 group ( P<0.05). There were no statistically significant differences in each parameter between P5 group and P3 group ( P>0.05). There was no statistically significant difference in the incidence of postoperative complications among the three groups ( P>0.05). Conclusions:The laryngeal mask airway provides better ventilatory effect in pediatric patients undergoing laparoscopic surgery when the PEEP is set at 3 cmH 2O.

Objective:To analyze the effect of continuous positive airway pressure (CPAP) on maternal and neonatal outcomes in pregnant women with obstructive sleep apnea syndrome (OSAS), especially on the incidence of hypertensive disorder in pregnancy (HDP) in women with moderate to severe OSAS.Methods:A total of 180 pregnant women with OSAS who were diagnosed through sleep monitoring during pregnancy due to high-risk factors of OSAS and registered in Peking University People′s Hospital from January 2021 to May 2024 were selected as the study subjects. Clinical data were collected from medical records for retrospective analysis. According to whether they received standardized treatment with CPAP, they were divided into the CPAP treatment group (42 cases) and the control group (138 cases). The CPAP treatment group consisted of 9 pregnant women with moderate to severe OSAS, while the control group consisted of 34 pregnant women with moderate to severe OSAS. The maternal and neonatal outcomes, the incidence of HDP, placental weight after delivery and placental weight/neonatal birth weight ratio were compared between the two groups.Results:(1) The average gestational age of pregnant women in the CPAP treatment group was higher than that in the control group [(38.7±1.0) vs (38.0±1.4) weeks], the proportion of infants small for gestational age (SGA) in the CPAP treatment group was lower [0 (0/42) vs 12.3% (17/138)], and the birth weight of infants in the CPAP treatment group was bigger [(3 396±475) vs (3 082±710) g); the differences between the two groups were statistically significant (all P<0.05). There were no significant differences between the CPAP treatment group and the control group in terms of delivery mode, rates of postpartum hemorrhage and preterm birth, umbilical artery blood gas analysis pH<7.1, lactate≥6.0 mmol/L, base excess<-12.0 mmol/L and the incidence of gestational diabetes mellitus and HDP (all P>0.05). (2) The placental weight of the CPAP treatment group was significantly lower than that of the control group [(554.0±70.6) vs (615.7±119.1) g], the placental weight/newborn birth weight ratio of the CPAP treatment group was significantly lower than that of the control group (median: 0.17 vs 0.19), and the differences were statistically significant (all P<0.05). (3) The incidence of HDP in pregnant women with moderate to severe OSAS in the CPAP treatment group was lower than that in the control group [1/9 vs 61.8% (21/34)], and the difference was statistically significant ( P<0.05). Conclusions:CPAP treatment could prolong the gestational age in pregnant women with OSAS, reduce the incidence of SGA, increase the birth weight of infants, and reduce the incidence of HDP in pregnant women with moderate to severe OSAS, and is worth promoting in clinical practice. The improvement of neonatal outcomes by CPAP treatment is closely related to the placenta, which is worthy of further exploration.

Objectives:QT interval prolongation during treadmill test exercise is one of the clinical feature of patients with long QT syndrome(LQTS).This study aimed to explore the feasibility and efficacy of treadmill exercise testing as an auxiliary diagnosis tool for LQTS in clinical practice.Methods:We enrolled normal healthy individuals,common cardiovascular disease patients,and clinically diagnosed or suspected LQTS patients,who underwent treadmill exercise test from July 2023 to July 2024 at Fuwai Hospital.A special treadmill exercise testing procedure was designed to record the QT interval correction(QTc)intervals of the twelve lead electrocardiogram at 6 time points when performing the exercise tablet,including supine,sitting,standing,peak exercise,and recovery at 1-minute and 4-minute.The differences in QTc intervals among healthy group,cardiovascular diseases group,and suspected LQTS group were compared.Results:A total of 80 cases were consecutively enrolled,including 37 normal healthy controls,25 patients with common cardiovascular disease,and 18 patients with suspected LQTS.The QTc intervals at 6 points did not differ significantly between normal healthy controls and patients with cardiovascular disease,with QTc intervals less than 480 ms at all measurement.For patients with suspected LQTS,67.7%(12/18)of these patients presented a QTc interval≥480 ms at the 4-minute during recovery period.Among them,5 cases were confirmed to have pathogenic gene mutations of LQTS by genetic testing(including 1 case with a lying electrocardiogram QTc interval of 489 ms diagnosed with LQTS 1 type and a QTc interval of 636 ms during the 4-minute recovery period after exercise);5 clinically diagnosed patients(negative or undetectable in genetic testing)with a Schwartz score≥4,and the remaining 2 patients had a Schwartz score of 3.The remaining 5/18 patients,include 2 patients with clinical Schwartz scores≥4 and 3 patients with clinical suspicion(Schwartz scores 2-3)had a 4 min QTc interval of 445-480 ms during exercise recovery.Another patient with clinical suspicion(Schwartz score 3)had a 4 min QTc interval of<445 ms during exercise recovery and a negative genetic test at a later stage.Receiver operating characteristic curve analysis showed a sensitivity of 83.3%and specificity of 98.4%for QTc interval≥482 ms during the 4-minute recovery period of exercise as the LQTS diagnostic cutoff.Conclusions:This study results suggest that this special treadmill exercise testing protocol is effective in identifying LQTS and has strong feasibility and generalizability for clinical practice.

Objective To study how lipid bilayer fluidity modulates the interaction between β1 integrin and CD40L,as well as the formation of CD40L-mediated tumor cell contact interfaces.Methods Supported lipid bilayers(SLB)with different fluidities were prepared through adjusting the 1,2-dioleoyl-sn-glycero-3-[N-(5-amino-l-carboxypentyl)iminodiacetic acid]succinyl nickel salt(DGS-NTA)content.The functionalization of lipid bilayers was achieved by anchoring fluorescently labeled CD40L molecules onto the membrane surface.The contact interface formation of PC9 cells on the functionalized lipid bilayers was observed through confocal fluorescence imaging and fluorescence recovery after photobleaching(FRAP)experiments,and data of two dimensional(2D)reaction kinetics of β1 integrin and CD40L were extracted from Zhu-Golan plots.Results The diffusion coefficient of molecules in lipid bilayer was negatively correlated with DGS-NTA content.High fluidity of lipid bilayer promoted CD40L accumulation at cell contact interface and expanded the cell contact area.The 2D dissociation constants(2D Kd)of β1 integrin-CD40L complexes were approximately 13,31 and 65 molecules/μm2 for the three lipid bilayers with high,moderate and low fluidities,respectively.Conclusions High fluidity of lipid bilayers significantly facilitates diffusion and aggregation of CD40L to the cell contact interface,thus enhancing β1 integrin-CD40L interaction and the stability of cell contact interfaces.

Objective:To investigate the risk factors of hypoparathyroidism (HPT) and hypocalcemia in patients undergoing total thyroidectomy (TT), and to explore the changes of parathyroid hormone and blood calcium after TT.Methods:The clinical data of 101 patients undergoing TT from November 2018 to September 2022 in the First Hospital of Hebei Medical University were retrospectively analyzed. The basic clinical data were recorded. The blood calcium and parathyroid hormone levels were measured before surgery and 1 d, 1 week after surgery. The occurrence of postoperative hypocalcemia was recorded. According to postoperative parathyroid hormone level, the patients were divided into control group (normal parathyroid function) and HPT group (reduced parathyroid hormone level). The patients with postoperative hypocalcemia symptoms were classified as the hypocalcemia symptoms group, and the patients without postoperative hypocalcemia symptoms were classified as the non-hypocalcemia symptoms group. Multivariate Logistic regression was used to analyze the independent risk factors of HPT and hypocalcemia in TT patients.Results:The postoperative parathyroid hormone level decreased in 41 cases (HPT group) and normal in 60 cases (control group). There were 24 patients with postoperative hypocalcemia symptoms (hypocalcemia symptoms group) and 77 patients without postoperative hypocalcemia symptoms (non-hypocalcemia symptoms group). The rate of using bipolar electric coagulation forceps in HPT group was significantly lower than that in control group: 31.71% (13/41) vs. 76.67% (46/60), while the rate of central lymph node dissection was significantly higher than that in control group: 82.93% (34/41) vs. 60.00% (36/60), and there were statistical differences ( P<0.01 and <0.05). Multivariate Logistic regression analysis result showed that TT combined with unilateral or bilateral central lymph node dissection was an independent risk factor for HPT in TT patients ( OR = 1.706 and 1.501, 95% CI 1.019 to 2.856 and 1.052 to 2.140, P<0.05). The preoperative serum calcium, postoperative serum calcium and postoperative parathyroid hormone in hypocalcemia symptoms group were significantly lower than those in hypocalcemia symptoms group: (2.32 ± 0.11) mmol/L vs. (2.37 ± 0.11) mmol/L, (2.16 ± 0.21) mmol/L vs. (2.25 ± 0.18) mmol/L and 3.00 (1.00, 5.45) ng/L vs. 19.90 (8.50, 33.80) ng/L, and there were statistical differences ( P<0.05 or <0.01). Multivariate Logistic regression analysis result showed that postoperative parathyroid hormone was an independent risk factor of hypocalcemia symptoms in TT patients ( OR = 0.927, 95% CI 0.883 to 0.974, P<0.01). In patients with HPT, the blood calcium at 1 week after surgery was significantly lower than that at 1 d after surgery: (2.07 ± 0.19) mmol/L vs. (2.17 ± 0.25) mmol/L, and there was statistical difference ( t = 2.05, P<0.05); the parathyroid hormone at 1 week after surgery was significantly higher than that at 1 d after surgery: 8.30 (3.55, 19.55) ng/L vs. 3.60 (1.00, 6.85) ng/L, and there was statistical difference ( Z = - 3.78, P<0.01). Conclusions:When performing TT, standardizing the surgical techniques, reducing unnecessary central lymph node dissection, and using bipolar electric coagulation forceps as much as possible can help to reduce the occurrence of postoperative HPT. The levels of postoperative parathyroid hormone and blood calcium should be promptly detected, the change of both should be given attention, and do a good job in preventing and treating hypocalcemia.

Objective:To investigate the characteristics of visual and auditory sustained attention and sex differences in pupils of different grades.Method:Totally 231 students from grades 1 to 6 participated in this stud-y.The development and sex differences of sustained attention were measured by average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates in visual and auditory two-choice re-action time(CRT)tasks.Result:The students in grade 5 had significantly slower visual average reaction times and coefficients of variation for reaction times(P＜0.001 or P＜0.05)than those in grade 3.The students in grade 5 had significantly lower auditory average reaction times,coefficients of variation for reaction times,omission error rates and commission error rates(P＜0.001 or P＜0.01)than those in grade 1.There was no significant sex differ-ences in visual or auditory average reaction times,coefficients of variation for reaction times or omission error rates(Ps＞0.05).Conclusion:There would be age differences in characteristics of both visual and auditory sustained at-tention in pupils,and may be no sex difference in visual or auditory sustained attention.