ObjectiveTo investigate the effects of aconitine, mesaconitine and hypaconitine on calcium release in isolated adult rat cardiac myocytes.MethodsThe left ventricular cardiac myocytes isolated from adult Sprague-Dawley rats were perfused withacnitine, mesaconitine and hypaconitine at 0.3 μmol/L, 1μmol/L, 3 μmol/L for 12 min. The spontaneous calcium release (SCR) rate, the end-diastolic[Ca2+](F0) and the calcium transient amplitude (ΔF) were detected 4 min, 8 min and 12 min after the perfusion. 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 0.3 μmol/L, the changes of systolic dynamics and calcium transient were detected for the positive inotropic effect. Results Any of aconitine, mesaconitine and hypaconitine induced SCR, mesconitine-induced SCR rate was highest at low concentration (0.3 μmol/L), and aconitine-induced SCR rate highest at high concentration (3 μmol/L). Compared with the control, 12 min after the perfusion with acnitine, mesaconitine and hypaconitine at 3 μmol/L elevated F0 (1.459 ± 0.379, 1.585 ± 0.493, 1.213 ± 0.254vs.1.079 ± 0.108, allP<0.05) and ΔF(1.615 ± 0.455, 2.210 ± 0.756, 1.528 ± 0.422vs. 1.036 ± 0.125, allP<0.05), mesaconitine with ΔF higher than aconitine and hypaconitine. At low concentration (0.3 μmol/L), compared with control, aconitine, mesaconitine and hypaconitine increased ΔF (0.409 ± 0.127, 0.423 ± 0.107, 0.414 ± 0.118vs.0.260 ± 0.065;P<0.05 orP<0.01) and contraction amplitudes (5.464% ± 2.239%, 7.449% ± 2.548%, 5.524% ± 1.645%vs.3.428% ± 0.911%;P<0.05 orP<0.01), prolonged the time to peak of calcium transient (0.041 ± 0.016 s, 0.039 ± 0.009 s, 0.038 ± 0.011 svs.0.032 ± 0.007 s;P<0.05 or P<0.01); compared with aconitine, mesaconitine and hypaconitine decreased calcium transient time constant (0.301 ± 0.054 s, 0.324 ± 0.064 svs.0.361 ± 0.076 s;P<0.05 orP<0.01) and diastolic t50 (0.124 ± 0.035 s, 0.126 ± 0.040 svs.0.157 ± 0.056 s;P<0.05 orP<0.01).ConclusionsAconitine, mesaconitine and hypaconitine reveal the positive inotropic effects couple with the toxic effects. Increased[Ca2+]in cardiac myocytes is the key factor for the positive inotropic effects, but also the risk factor for SCR.

ISG15, the first ubiquitin-like molecule identified two decades ago, is encoded by interferon stimulated gene 15 ( ISG 15), where its robust expression can be induced by viral infections or interferon treatments. ISG 15 conjugate to other proteins as the ubiquitin and was found to be involved in innate immune response. However, the functions of ISG15 modification remained unclear. We cloned bovine ISG15(bisg15) into a prokaryotic expression vector pET28a( + ) with a His-tag to generate a soluble form of bISG15 fusion protein, and purified with Ni-NTA Sepharose chromatography. The purified protein was concentrated and used to immune Balb/c mice to raise the antiserum, which could specifically recognize bISG15 expressed in eukaryotic cells by Western blot analysis. The concentrated bISG15 protein and its antiserum were then used to establish an in vitro bISG15 modification system. Our studies have demonstrated that cellular proteins could be conjugated to bISG15 with this system.

OBJECTIVETo explore evaluation strategies for middle ear dysfunction in cleft palate patients, to optimize the diagnosis and treatment of this dysfunction, and ultimately to improve the comprehensive treatment of cleft palate.

METHODSThe relationship among abnormal tympanic types (B, C, and Anomaly), effusion rate, tympanic pressure, and hearing loss were analyzed. We collected relevant information on 469 ears of cleft palate patients and traced one-year longitudinal changes in the tympana of 124 ears from 62 patients with both cleft lip and cleft palate.

RESULTSThe effusion rates of cleft palate patients with type B, type C, and type Anomaly were 50.3% (97/193), 34.8% (8/23), and 20.9% (53/253), respectively. The tympanic pressure of the ears with and without effusion showed no significant difference (P>0.05). The hearing loss in type B cleft palate patients with middle ear effusion was worse than that in patients without effusion (P=0.001). However, the hearing loss in type Anomaly showed no difference (P>0.05). The constituent ratio of each tympanic type remained constant during the period between cheiloplasty and palatoplasty for cleft lip and palate patients (P>0.05).

CONCLUSIONCleft palate patients of all tympanic types may all suffer from middle ear effusion at different rates. Examination by centesis is suggested for ears with abnormal tympanic types. Early aggressive therapy is essential for type B cleft palate patients with middle ear effusion to avoid hearing loss. However, catheterization may be not necessary for type Anomaly patients, and conservative observation should be performed instead. Myringotomy with grommet insertion during palatoplasty does not delay treatment timing for patients with both cleft lip and cleft palateg.

Cleft Lip ; Cleft Palate ; Ear, Middle ; physiology ; Humans ; Middle Ear Ventilation ; Otitis Media with Effusion ; diagnosis ; epidemiology

Hand Deformities ; pathology ; Humans ; Klippel-Feil Syndrome ; pathology ; Thumb

OBJECTIVEThe purpose of this study was to establish a palatal organ culture method and to investigate the palatogenesis in vitro.

METHODS20 pregnant 14-day mice were killed, embryos were separated ascetically, and palatal shelves were dissected and placed on a modified Trowell's system. All explants were cultured 24 h and 48 h respectively. Finally, all explants were embedded and stained by Hematoxylin and Eosin.

RESULTSAll explants grew healthy. After incubation for 24 h, medial edge epithelium maintained, whereas after 48 h, medial edge epithelium disappeared, bilateral mesenchymal cells contacted, palates fused.

CONCLUSIONThis method provides an effective way for investigating the etiology of cleft palate in vitro.

Animals ; Cleft Palate ; Epithelium ; Female ; In Vitro Techniques ; Mice ; Organ Culture Techniques ; Palate ; cytology ; Pregnancy

Objective:To investigate the risk factors affecting the delay of recovery in patients under general anesthesia.Methods:Patients who underwent radical mastectomy for breast cancer in our hospital from Jul. 2020 to Aug. 2022 were selected as the research objects, and the effective data of 80 patients were obtained after screening. The patients were divided into the non-delayed recovery group (54 cases) and the delayed recovery group (26 cases). The general conditions and perioperative data of the two groups were compared, and the high-risk factors affecting delayed recovery from anesthesia were analyzed using the Logistic hazard proportional regression model. Results:In general, there were no significant differences in body mass index, hypertension, diabetes, tumor size, tumor subtype, tumor location, or tumor stage between the non-delayed awakening group and the delayed awakening group (all P>0.05). The average age of patients in the wake-up delay group was (53.28±11.01), the proportion of anemia was 42.30% (11/26), and the proportion of ASA Ⅱ patients was 76.92% (20/26) compared with the average age of the non-wake-up delay group (46.89±6.91) ( t=3.17, P=0.002), the proportion of anemia was 20.37% (11/54) ( χ2=3.17, P=0.040), the proportion of ASA Ⅱ patients was 27.78% (15/54) ( χ2=17.22, P<0.001), which was significantly increased. In the perioperative data, there was no statistical significance in the intraoperative combined epidural anesthesia between the non-delayed recovery group and the delayed recovery group (all P>0.05). The data of the patients in the delayed recovery group, including average intraoperative blood loss (234.14±32.28), operation time (229.47±29.84), anesthesia time (246.14±35.64) and intraoperative compound sevoflurane inhalation accounted for 69.23% (18/26) ,which were significantly increased compared with the data of non-delayed recovery group following, including the average intraoperative blood loss (215.48±29.54) ( t=2.57, P=0.012), operation time (206.35±27.41) ( t=3.43, P=0.001), anesthesia time (215.61±28.54) ( t=4.13, P<0.001), intraoperative compound sevoflurane inhalation (44.44%, 24/54). Through Logistic hazard ratio regression analysis, it was found that age ( OR=1.15, 95% CI: 1.05-1.30, P=0.008), ASA Ⅱ grade ( OR=9.49, 95% CI: 2.05-60.94, P=0.008), intraoperative bleeding volume ( OR=1.04, 95% CI: 1.01-1.08, P=0.012), operation time ( OR=1.05, 95% CI: 1.01-1.08, P=0.009), and anesthesia time ( OR=1.04, 95% CI: 1.02-1.07, P=0.004) were high-risk factors affecting the delayed recovery from anesthesia. Conclusion:Increasing age, high grade of ASA, heavy intraoperative blood loss, long operation time and anesthesia time are independent risk factors affecting delayed recovery from anesthesia.

Purpose/Significance The scoping review summarizes the application of large language models in clinical practice,and provides references for their promotion.Method/Process PubMed,Embase,Wanfang and CNKI databases are searched to screen the lit-erature related to the application of large language models in clinical practice,and the content of the included literature is extracted,sum-marized and analyzed.Result/Conclusion Large language models have application value in providing treatment suggestions,assisting disease diagnosis,health education,analyzing text image data,etc.However,their performance in answer accuracy and individualization is not satisfactory.In general,large language models show significant potential in clinical practice,but necessary measures must be taken to control the application risks and confirm the scope of application.

Pancreatic cancer is a common malignancy with the worst prognosis.Radical surgery has been the only curative treatment for pancreatic cancer.With the advancement of surgical techniques and the implementation of the concept of comprehensive treatment for cancer in recent years,neoadjuvant therapy for pancreatic cancer has received more attention.There are continuing controversies in the hotspots and difficulties,with opportunities and challenges coexisting.Four famous experts and their teams in pancreatic surgery discussed selection strategy of neoadjuvant therapy for pancreatic cancer based on clinical experiences.Professor Wang Chunyou proposed that surgery was prior for patients with a higher likelihood of achieving R0 resection for pancreatic cancer to avoid the possibility of tumor progression and loss the opportanity of radical resection during neoadjuvant therapy.For patients with less chance of radical resection for pancreatic cancer and unresectable pancreatic cancer,neoadjuvant therapy is worthy of a positive attempt.Professor Jin Gang and his team believed that neoadjuvant therapy played an important role in improving the survival time of patients with pancreatic head cancer,especially with borderline resectable pancreatic head cancer.After neoadjuvant therapy,pancreatic surgeons should pay attention to improvement of surgery safety and R0 resection rate.Professor Dai Menghua and his team suggested that patients with resectable pancreatic cancer and borderline resectable pancreatic cancer could benefit from neoadjuvant therapy,which required proof from clinical trials.Surgeons should choose the appropriate treatment strategy based on guidelines and individual conditions for patients with pancreatic cancer.Professor Shao Cheghao and his team suggested that surgical treatment after neoadjuvant therapy or translational therapy for locally advanced pancreatic head cancer is safe,effective and feasible,especially for pancreaticoduodenectomy with combined revascularization.For the treatment of patients with pancreatic head cancer after neoadjuvant chemotherapy,the choice of next treatment options,evaluation indicators,timing of surgery and surgical methods need to be further studied.

Objective In this study,folic acid(FA) was tested for antiteratogenic effects on Tetrachlorodibenzo-p-dioxin(TCDD)-induced cleft palate in fetal mice.Methods In the present study,pregnant mice were dosed with TCDD 24 μg/kg and with or without FA 5 mg/kg body weight on gestation day 10.Control group mice received sesame oil 50 ml/kg body weight on gestation day(GD)10.The mice were sacrificed on GD12.5,GD13.5,GD14.5,GD15.5 and GD16.5.From each pregnant mouse on GD16.5,embryos were obtained to examine under a dissecting microscope,and routine histology was performed for detection and classification of palatal clefts.The fetuses were prepared for histologic examination,scanning electron microscope and TdT-mediated X-dUTP nick and labeling(TUNEL).On GD12.5,GD13.5,GD14.5 and GD15.5.Meanwhile,real-time(RT)-PCR was employed to detect the mRNA expression levels about arylhydrocarbon receptor(AHR) and transforming growth factor(TGF)β3 in this animal model.Results Total frequencies of clefts were 70.2％ in TCDD group(group B) and 66.3％ in TCDD + FA group(group C) in relation to control fetuses(group A).Filopodia disappeared completely at the medial edge epithelia surface on GD15.5(group A),GD12.5(group B) and GD14.5(group C).the RT-PCR results showed that TGF-β3 expression was down-regulated on GD13.5 and GD14.5 compared to the control.Conclusions It is found that folic acid has no protects agaist 2.3.7.8-TCDD-indued cleft palate in the experiment.Meanwhile,TCDD repressed the TGF-β3 expression during the palatal development.Anormal apoptosis was induced by 2,3,7,8-TCDD at the medial edge epithelia(MEE) during the early development stage.

Objective:To investigate the effects of methyltransferases like 3(METTL3)mediated m6A modification of double homology cassette A pseudogene8(DUXAP8)on the proliferation,migration and invasion of salivary adenoid cystic carcinoma SACC-LM cells and its potential molecular mechanisms.Methods:Whole-transcriptome sequencing showed that DUXAP8 was highly ex-pressed in SACC than in para-cancerous tissues(P＜0.05).The m6A modification sites on DUXAP8 were predicted using the SRAMP website,and the mRNA and protein expression of m6A-modified genes and the genes associated with the epithelial-mesen-chymal transition(EMT)was measured by qRT-PCR and Western blot,respectively.METTL3 and DUXAP8 was knocked down or overexpressed in SACC-LM cells,and the proliferation,migration,and invasion of the cells were assessed by CCK-8,scratch and Transwell assays.The correlation between METTL3 and DUXAP8 was evaluated using MeRIP-qPCR.Results:The expression of DUXAP8 in SACC tumor was higher than that in para-cancerous tissues(P＜0.05).Knockdown of DUXAP8 reduced proliferation,migration and invasion of SACC-LM cells,as well as the expression of EMT-related genes(P＜0.05).Multiple m6A modification sites of high confidence were found on DUXAP8.METTL3 was highly expressed in tumor tissues,more than other related genes(P＜0.05)and enzyme-encoding genes in SACC-LM cells(P＜0.05).METTL3 was found to function as a methyltransferase to regulate the expression of DUX-AP8,and downregulation of METTL3 inhibited prolifera-tion,migration and invasion of SACC-LM cells and partially reversed the promotion of these activities induced by DUX-AP8 overexpression(P＜0.05).Conclusion:METTL3-me-diated m6A modification upregulated DUXAP8 expression,which promotes the proliferation,migration and invasion of SACC cells.