Background: The treatment landscape for melanoma, a particularly malignant skin cancer, is constrained by notable drug resistance and toxicity. β-Asarone, a natural compound from Acorus tatarinowii, has shown anticancer potential. Disruption of calcium homeostasis and mitochondrial dysfunction are key regulators of tumor cell survival and death. Objective: This research was conducted to investigate the impact of β-Asarone on B16F10 melanoma cells, focusing on its potential to induce apoptosis by modulating calcium signaling and mitochondrial function. Methods: Cell proliferation and apoptosis were evaluated using CCK-8, colony formation, EdU, and TUNEL assays. Intracellular calcium levels and mitochondrial membrane potential were measured using Fluo-4 AM, Rhod-2 AM, and JC-1 staining. Reactive oxygen species (ROS) generation and adenosine triphosphate (ATP) levels were assessed by fluorescent probes and ATP assay. Western blotting was utilized to detect apoptosis-related proteins, AMP-activated protein kinase (AMPK) pathway activation, and mitochondrial dynamics (OPA1, DRP1, FIS1). Results: Treatment with β-Asarone notably inhibited the proliferation of B16F10 cells while simultaneously inducing apoptosis. Fluorescent probe analysis revealed that β-Asarone triggered cytosolic and mitochondrial Ca 2+ overloaded in both the cytosol and mitochondria, accompanied by decreased mitochondrial membrane potential, elevated ROS levels, and reduced ATP production. Western blot analysis showed increased expression of DRP1 and FIS1, decreased OPA1, and enhanced AMPK phosphorylation, indicating that β-Asarone promotes mitochondrial fission through AMPK activation, likely driven by intracellular calcium imbalance. Conclusion This study demonstrates that β-Asarone induces apoptosis in B16F10 melanoma cells by triggering Ca 2+ overload and mitochondrial dysfunction.

Background/Aims: The accuracy of endosonographers in diagnosing gastric subepithelial lesions (SELs) using endoscopic ultrasonography (EUS) is influenced by experience and subjectivity. Artificial intelligence (AI) has achieved remarkable development in this field. This study aimed to develop an AI-based EUS diagnostic model for the diagnosis of SELs, and evaluated its efficacy with external validation. Methods: We developed the EUS-AI model with ResNeSt50 using EUS images from two hospitals to predict the histopathology of the gastric SELs originating from muscularis propria. The diagnostic performance of the model was also validated using EUS images obtained from four other hospitals. Results: A total of 2,057 images from 367 patients (375 SELs) were chosen to build the models, and 914 images from 106 patients (108 SELs) were chosen for external validation. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of the model for differentiating gastrointestinal stromal tumors (GISTs) and non-GISTs in the external validation sets by images were 82.01%, 68.22%, 86.77%, 59.86%, and 78.12%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in the external validation set by tumors were 83.75%, 71.43%, 89.33%, 60.61%, and 80.56%, respectively. The EUS-AI model showed better performance (especially specificity) than some endosonographers.The model helped improve the sensitivity, specificity, and accuracy of certain endosonographers. Conclusions We developed an EUS-AI model to classify gastric SELs originating from muscularis propria into GISTs and non-GISTs with good accuracy. The model may help improve the diagnostic performance of endosonographers. Further work is required to develop a multi-modal EUS-AI system.

This study explores the effects of enhancing the definitive hematopoiesis(DH)signals during the differentiation of human embryonic stem cells(hESCs)into hematopoietic stem/progenitor cells on the generation efficiency and effector function of natural killer(NK)cells generated from hESCs(also known as hESC-NK cells).The hESC(H1)were transformed into embryoid bodies(Ebs)by centrifugation,and during the induction of K562,were used to analyze the efficiency of hematopoietic differentiation,the efficiency of NK cell generation from hESC,the in vitro effector functions,and the expression of effector function related surface receptors.Compared to the control group,the DH group had a significant increase in the number of arterial hematopoietic endothelial cells(CD34+DLL4+)and a significant decrease in primitive hematopoietic related cells(CD34-CD43+)on day 8 of hematopoietic differentiation(P<0.05).On day 28 of NK cell differentiation,the DH group demonstrated a significant increase in the number of NK cells(CD45+CD56+),while a slight increase in the expression of effector function-related molecules such as IFN-γ,Granzyme B,Perforin and CD107a without statistical significance.Furthermore,the activation receptors CD16a and CD69 were significantly increased,NKP46 was significantly decreased,the inhibitory receptor NKG2A was significantly increased,while CD96 was significantly decreased on hESC-NK cells of DH groups(P<0.05).Conclusively,enhancing the signals for definitive hematopoiesis during hESC differentiation into hematopoietic stem/progenitor cells significantly improves the yield of NK cells and the expression of CD16a without affecting their in vitro effector functions.Our study provides a new approach to improving the efficiency of hESC-NK cell or iPSC-NK cell generation.

Objective: To compare the irradiated dose to unprotected lymph node stations (LNS) between volume-modulated arc therapy (VMAT) and 5-field intensity-modulated radiotherapy (5F-IMRT) in the treatment of patients with upper thoracic esophageal cancer. Methods: A total of 20 patients were selected for re-planning. LNS were not included in the GTV and CTV, instead, LNS were contoured as normal tissues. However, LNS were not constrained in the VMAT and 5F-IMRT inverse optimization for protection. Dosimetric parameters of conformal index (CI), homogeneity index (HI) of targets, V₉₅, V₁₁₀ of planning target volume (PTV), D_mean, V₅, V₂₀, V₃₀ of lung, D_mean, V₂₅ of heart, D_max of spinal cord, MU, as well as the equivalent uniform dose (EUD) and V₄₀ of LNS were compared between the two plans. Results: 5F-IMRT was superior in PTV_V_95% (t=-9.4, P<0.05), but worse in terms of CI (t=-5.3, P<0.05) compared with VMAT. 5F-IMRT reduced the V₅ of lung by 10.9% (t=-7.8, P<0.05) and the D_max of spinal cord by 9% (t=-10.2, P<0.05), but increased the MU (t=-6.2, P<0.05) compared with VMAT. The average EUD and V₄₀ of LNS in upper thoracic were significantly increased by 4.7% and 2.4% in 5F-IMRT compared with VMAT, respectively. The irradiated doses to LNS were significantly associated with the volume of PTV (R=0.716-0.933, P<0.05) expect for 106tbL. Conclusions The irradiated doses to unprotected LNS were less for IMRT plans and were highly associated with PTV volume in patients with upper thoracic esophageal cancer.

Objective To investigate the effects of cryptotanshinone and tanshinone Ⅱ A,two major monomer components of tanshinone,on the proliferation,differentiation and lipid synthesis of rat meibomian gland epithelial cells in vitro.Methods The eyelid meibomian gland tissue was isolated from 2-month-old SD rats and co-cultured with 3T3 trophoblasts for 5 days.Cryptotanshinone and tanshinone Ⅱ A were prepared with DMSO into different concentrations.The cells were grouped to 0.125 μmol/L,0.250 μmol/L,0.500 μmol/L,1.250 μmol/L and 2.500 μmol/L drug groups and treated for 48 hours,respectively.Only dimethyl sulfoxide (DMSO) was added in medium in the DMSO control group.The expressions of keratin 14 (K14) and p63 in frozen sections of meibomian gland tissue and clones of meibomian gland cells were detected by immunofluorescence.Real-time fluorescence quantitative PCR was used to assay the relative expression of K16,cell proliferation related antigen 67 (Ki67) and CCAAT enhancer binding protein α (C/EBPcα) in meibomian gland cell clones.Crystal violet staining and oil red staining were used to evaluate the colony formation rate and lipid synthesis of meibomian gland epithelial cells.Results Primary cultured meibomian gland cells were cloned on day 4-5 in vivo,and the cloning area was increased on day 7 after culture,p63 and K14 were positively expressed in clones.Compared with the DMSO control group,the relative expression levels of Ki67 mRNA were significantly elevated in the 0.500 μmol/L,1.250 μmol/L and 2.500 μmol/L cryptotanshinone groups (all at P ＜ 0.05).The relative expressions of Ki67 mRNA in the 0.500 μmol/L and 1.250 μmol/L tanshinone Ⅱ A groups were significantly higher than those in the DMSO control group (all at P＜0.05).No significant difference was found in the relative expression of K16 and C/EBPα mRNA among different concentrations of cryptotanshinone or tanshinone Ⅱ A group (all at P＞0.05).No lipid drop was found in the tarsal gland cell clones;however,the accumulation of lipid was seen in the cell clusters at the margin of the clones by oil red O staining.The average clone formation rate of tarsal gland cells in the 1.250 μ mol/L cryptotanshinone group was (2.55±0.20)％,which was significantly higher than (2.05±0.13)％ in the DMSO control group (t =4.379,P＜0.05).The average clone formation rate of tarsal gland cells in the 1.250 μmol/L tanshinone Ⅱ A group was (2.25±0.20)％,there was no significant difference between 1.250 μmol/L tanshinone Ⅱ A group and DMSO control group (t=1.616,P＞0.05).Conclusions Cryptotanshinone and tanshinone Ⅱ A promote the proliferation of meibomian gland epithelia cells,but play less impacts to lipid synthesis of meibomian gland epithelia cells in vitro.cryptotanshinone promote the clone tormation of meibomian gland epithelia cells.

Objective: The purpose of this study was to explore the surgery-first approach in sequential combined orthodontic-orthognathic treatment to shorten total treatment duration and improve the clinical outcome. Methods: This study included 185 patients with Angle classⅢ malocclusion. The patients were divided into 3 different types according to cephalometry analyses and facial features. ①Type Ⅰ: mandibular prognathism or asymmetry mandibular prognathism; ②Type Ⅱ: mandibular prognathism and maxillary retrusion; ③ Type Ⅲ: mild Angle′s Class Ⅲ malocclusion, cross bite in anterior teeth, or normal overlap and overbite relation with midfacial hypoplasia. All of patients received surgery first approach therapy. The surgical procedures were chosen according to different malformation types. Type Ⅰ was treated with the sagittal split ramus osteotomy (SSRO). Type Ⅱ was treated by Le Fort Ⅰmaxillary osteotomy combined with SSRO. Type Ⅲ underwent anterior subapical osteotomy combined pyriform aperture augmentation with biomaterials as well as maxillary anterior orthodontics. All patients received postoperative rapid orthodontic treatment for 6-12 month after 2 weeks of operation. Using the straight arch wire techniques and the class Ⅲ intermaxillary traction, we removed the overcrowding upper and lower teeth, the compensatory axial tilt of teeth, and the deviation of the dental arch and maintained the neutral relationship of the molar. The mandible Hawley retaining devices were used during the maintaining stage. Results: The cases in study acquired satisfactory clinical outcome, which included the shortened overall treatment duration, the significantly improved facial features, the corrected occlusion relationship, and the restored function of mastication and temporomandibular joint. There were some complications as follows: intraoperative fracture (6 cases, 3.24%), the inferior alveolar nerve bundle injury (2 cases, 1.1%), and temporary open-bite that diminished by inter-maxillary elastic distraction one month after operation (19 cases, 10%). All cases in this study accepted postoperative orthodontic treatment. Follow-up time ranged from 6 months to 5 years. The cephalometric analysis results of 126 cases who had complete image data and over 6 months of follow-up showed that hard and soft tissue indexes were restored to normal range after combined orthognathic-orthodontic treatment. The stability of the maxillary and occlusive relationship of SFA(surgery-first approach) was similar to that of the COS(conventional orthodontics-first system) [relapse ratio=(T2-T1)/(T1-T0)×100%]. Over six months of follow-up , SNB and ANB showed that the average relapse ratio were 22% and 19.8%, whereas the relapse angle are less than 2°. Conclusions The Surgery-first approach could be used to treat most patients with Angle skeletal Class Ⅲ malocclusion, but the indications and the surgical procedures should be noticed and chosen.

Objective To explore the neuropsychological development in twin premature infants,and to analyze the risk factors for the retardation of development.Methods The premature infants hospitalized in the Neonatal Ward First Affiliated Hospital of Kunming Medical University from Jun.2010 to Jun.2012 were divided into study groups of twin premature infants and a control group of singleton premature infants.The neuropsychological development of 88 premature infants was evaluated at corrected gestational age of 1 year old by Bayley scales of infant development.According to normal lab findings of the Bayley scales of infant development,the premature infants were then divided into abnormal neuropsychological development group [mental development index(MDI) ≤79 scores] and normal neuropsychological development group (MDI ＞ 79 scores),and the data of 2 groups were statistically analyzed.Results The frequency distribution of MDI score in study group showed a skewed distribution,of which 59％ (27/46 cases) had abnormal neuropsychological development (MDI ≤ 79 scores); However,the frequency distribution of MDI score in control group showed an approximate normal distribution,of which only 12％ (5/42 cases) for abnormal neuropsychological development.Head circumference,body length,body weight,MDI,psycho-motor development index of twin premature infants of one-year old were lower than those of the singleton premature infants of the same age,and the differences were statistically significant (all P ＜0.05).The single factor analysis demonstrated that the risk factors for abnormal neuropsychological development (MDI ≤ 79 scores) of twin premature infants were gestational age,birth weight,mother' s gcstational age,the cultural degree of parents,mode of feeding,neonatal hyperbilirubinemia,neonatal hypoglycemia and neonatal sepsis.Multiple regression analysis showed that neonatal hyperbilirubinemia was the independent risk factor for abnormal neuropsychological development of twin premature infants.Conclusions At same gestational age,neuropsychological development of twin premature infants lagged behind singleton premature infants.Neonatal hyperbilirubinemia may be the risk factors for neuropsychological development of twin premature infants,so the earlier management of neonatal hyperbilirubinemia may be beneficial for the neuropsychological development of twin premature infants.

Objective To analyze the biophysical dosimetric characteristics and clinical application ability of VMAT technology for breast cancer post-mastectomy.Methods 28 patients with breast cancer (10 at left side and the other at right side) were planned in different ways respectively.One was two 90 degree arc VMAT plan and the other were 5 beam IMRT plan.The dosimetric parameters of two different plans including tumor control probability (TCP),conformity index(CI),homogeneity index (HI),V95and V110 in target,normal tissue complication probability (NTCP),V5,V20,V30 for ipsilateral lung,NCTP,D V25 for heart,D for the contralateral breast in OARs,MU and times were compared.Results The average tumor control probability (TCP) in VMAT and IMRT group was(96 ±2)％ and (90 ±2)％ (t =-6.28,P ＜ 0.01),respectively.The PTV dose average homogeneity index (HI) of VMAT plans was better than that of IMRT plan (0.15 ±0.04 vs 0.22 ±0.02,t =13.29,P ＜0.01).For cancer position in left side,the mean dose of heart was decreased by 433.24 cGy in the VMAT plan.The NTCP of the hearts in VMAT plans had statistically significant difference compared with IMRT plans [(1.00±0.12)％ vs (1.70±0.13)％,t =2.14,P ＜0.05].For plans of right breast cancer,the average mean dose of hearts in two control group was (3.27 ± 0.26) Gy and (6.00 ± 0.47) Gy (t =9.21,P＜0.01).The total monitor unit (MU) was 530.7 in the VMAT arm and 693.9 in the IMRT arm (t =9.58,P ＜0.01).The treatment time was shorter in VMAT arm (t =8.40,P ＜0.05).Conclusions VMAT plans have better clinical value and more superior biophysical dosimetric characteristics for breast cancer post-mastectomy.

In this study, the current status for breast diseases in a region with high-incidence of cervical cancer were epidemiologically investigated. From March to August, 2009, 17618 women, from Wufeng area of Hubei province, China, were recruited to screen breast diseases by using breast infrared diagnostic apparatus. Other diagnostic methods, such as B-mode ultrasound, X-ray mammography, needle biopsy and pathological examination were, if necessary, used to further confirm the diagnosis. The screening showed that 5990 of 17618 cases (34.00%) had breast diseases, 5843 (33.16%) had mammary gland hyperplasia, 48 (0.27%) had breast fibroadenoma, 11 (0.06%) had breast carcinoma, and 88 (0.50%) had other breast diseases. The peak morbidity of breast cancer was found in the women aged 50-60 ages. The morbidity of breast cancer was significantly increased in women elder than or equal to 50 years old (n=8, 0.157%) in comparison with that in the subjects younger than 50 years old (n=3, 0.024%) (u=2.327, P<0.05). It was shown that the occurrence of breast diseases was concentrated in women aged 20-40 years, while the total morbidity reached its peak at the age of 30 years and then decreased sharply after age of 40. Compared with the patients elder than or equal to 40 years old (n=3289, 27.46%), the morbidity rate of breast diseases was significantly increased in women less than 40 years old (2648 cases, 47.18%; P<0.001). However, there was no significant difference in the morbidity of breast diseases between the age group of 20-29 years and that of 30-39 years (P=0.453), and both of them were high. There was no significant association between the morbidity of breast diseases and cervical cancer. Since the morbidity of breast diseases was higher among young women, more attention should be paid to the screening of breast diseases among young women for early diagnosis.

In this study,the current status for breast diseases in a region with high-incidence of cervical cancer were epidemiologically investigated.From March to August,2009,17618 women,from Wufeng area of Hubei province,China,were recruited to screen breast diseases by using breast infrared diagnostic apparatus.Other diagnostic methods,such as B-mode ultrasound,X-ray mammography,needle biopsy and pathological examination were,if necessary,used to further confirm the diagnosis.The screening showed that 5990 of 17618 cases (34.00％) had breast diseases,5843 (33.16％) had mammary gland hyperplasia,48 (0.27％) had breast fibroadenoma,ll (0.06％) had breast carcinoma,and 88 (0.50％) had other breast diseases.The peak morbidity of breast cancer was found in the women aged 50-0 ages.The morbidity of breast cancer was significantly increased in women elder than or equal to 50 years old (n=8,0.157％) in comparison with that in the subjects younger than 50 years old (n=3,0.024％) (u=2.327,P＜0.05).It was shown that the occurrence of breast diseases was concentrated in women aged 20-40 years,while the total morbidity reached its peak at the age of 30 years and then decreased sharply after age of 40.Compared with the patients elder than or equal to 40 years old (n=3289,27.46％),the morbidity rate of breast diseases was significantly increased in women less than 40 years old (2648 cases,47.18％; P＜0.001).However,there was no significant difference in the morbidity of breast diseases between the age group of 20-29 years and that of 30-39 years (P=0.453),and both of them were high.There was no significant association between the morbidity of breast diseases and cervical cancer.Since the morbidity of breast diseases was higher among young women,more attention should be paid to the screening of breast diseases among young women for early diagnosis.