BACKGROUND: Oral squamous cell carcinoma (OSCC) is a prevalent type of cancer with a high mortality rate in its late stages. One of the major challenges in OSCC treatment is the resistance to epidermal growth factor receptor (EGFR) inhibitors. Therefore, it is imperative to elucidate the mechanism underlying drug resistance and develop appropriate precision therapy strategies to enhance clinical efficacy. METHODS: To evaluate the efficacy of the combination of the Ca 2+ /calmodulin-dependent protein kinase II (CAMK2) inhibitor KN93 and EGFR inhibitors, we performed in vitro and in vivo experiments using two FAT atypical cadherin 1 ( FAT1 )-deficient (SCC9 and SCC25) and two FAT1 wild-type (SCC47 and HN12) OSCC cell lines. We assessed the effects of EGFR inhibitors (afatinib or cetuximab), KN93, or their combination on the malignant phenotype of OSCC in vivo and in vitro . The alterations in protein expression levels of members of the EGFR signaling pathway and SRY-box transcription factor 2 (SOX2) were analyzed. Changes in the yes-associated protein 1 (YAP1) protein were characterized. Moreover, we analyzed mitochondrial dysfunction. Besides, the effects of combination therapy on mitochondrial dynamics were also evaluated. RESULTS: OSCC with FAT1 mutations exhibited resistance to EGFR inhibitors treatment. The combination of KN93 and EGFR inhibitors significantly inhibited the proliferation, survival, and migration of FAT1 -mutated OSCC cells and suppressed tumor growth in vivo . Mechanistically, combination therapy enhanced the therapeutic sensitivity of FAT1 -mutated OSCC cells to EGFR inhibitors by modulating the EGFR pathway and downregulated tumor stemness-related proteins. Furthermore, combination therapy induced reactive oxygen species (ROS)-mediated mitochondrial dysfunction and disrupted mitochondrial dynamics, ultimately resulting in tumor suppression. CONCLUSION Combination therapy with EGFR inhibitors and KN93 could be a novel precision therapeutic strategy and a potential clinical solution for EGFR-resistant OSCC patients with FAT1 mutations.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/metabolism* ; Cell Line, Tumor ; Animals ; Drug Resistance, Neoplasm/genetics* ; Cadherins/metabolism* ; Carcinoma, Squamous Cell/metabolism* ; Mice ; Mutation/genetics* ; Mice, Nude ; Protein Kinase Inhibitors/therapeutic use* ; Cetuximab/pharmacology* ; Afatinib/therapeutic use* ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects*

ObjectiveTaking Shengxuebao as the model traditional Chinese medicine（TCM）， this study aims to explore the methodologies and transfer patterns in the composition changes of TCM compound preparations from liquid to solid dosage forms. MethodBased on the previously established ultra-high performance liquid chromatography（UPLC） qualitative and quantitative methods for Shengxuebao， the fingerprint profiles of three preparations of Shengxuebao（homemade decoction， commercially available granules and mixtures） were established， and nineteen components were quantitatively analyzed. The similarities of fingerprint profiles among these three preparations were assessed using the Similarity Evaluation System for Chromatographic Fingerprint of TCM（2012 edition） and the total quantum statistical moment similarity method， respectively. The molecular connectivity index （MCI） and the average molar MCI of the chemical components in the three preparations were calculated. Seventy-two MCI values across 0-8 orders， including 8 subclasses， were calculated. The average molar MCI of 0-3 orders was used to study the "imprint template" structural characteristics and transfer patterns of the medicinal component groups. ResultThe similarities of the fingerprint profile of the decoction with those of the control， granules， and mixtures were 0.971， 0.888 and 0.799， respectively， indicating that the chemical composition of the granules and mixtures differed from that of the decoction， and that the 19 components were able to basically respond to the overall situation of the component group of Shengxuebao decoction. The total mass fractions of the detectable major components in Shengxuebao decoction， granules and mixtures was measured to be 5.498 6， 2.282 0， 1.416 6 mg·g^-1， respectively. The average molar MCI of 0-3 orders showed a gradual decrease across the three preparations， with the trend of decoction>granules>mixtures， indicating subtle changes in the overall structural characteristics of the chemical components in the samples of the three preparations. ConclusionAmong the three formulations of Shengxuebao， the traditional decoction contained the highest variety and quantity of components and carried the most substantial amount of component information， with a decreasing order observed in the granules and mixtures. This study further validates that traditional decoction is a reliable and fundamental dosage form， providing a reference for the modernization of TCM dosage forms and the quality consistency evaluation of compound preparations.

According to the global cancer statistics in 2022 updated by the International Agency for Research on Cancer(IARC),there were nearly 20 million new cases of cancer and 9.7 million deaths.Lung cancer was the most commonly diagnosed cancer,accounting for nearly 2.5 million new cases(12.4%of all global cancers),followed by female breast cancer(11.6%),colorectal cancer(9.6%),prostate cancer(7.3%),and stomach cancer(4.9%).Lung cancer was also the leading cause of cancer deaths,with an estimated 1.8 million deaths(18.7%),followed by colorectal cancer(9.3%),liver cancer(7.8%),female breast cancer(6.9%),and stomach cancer(6.8%).Population-based projections suggest that the number of new cancer cases will reach 35 million by 2050.Increasing the investment in prevention and control measures targeting key cancer risk factors,including smoking,obesity,and infections,could save many lives globally and bring significant economic and social returns to countries in the coming decades.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling. Here, we focused on the role of Semaphorin 3A (Sema3A), expressed by sensory nerves, in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement (OTM) model. Firstly, bone formation was activated after the 3rd day of OTM, coinciding with a decrease in sensory nerves and an increase in pain threshold. Sema3A, rather than nerve growth factor (NGF), highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM. Moreover, in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells (hPDLCs) within 24 hours. Furthermore, exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload. Mechanistically, Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway, maintaining mitochondrial dynamics as mitochondrial fusion. Therefore, Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation, both as a pain-sensitive analgesic and a positive regulator for bone formation.
Humans ; Bone Remodeling ; Cell Differentiation ; Osteogenesis ; Semaphorin-3A/pharmacology* ; Trigeminal Ganglion/metabolism*

Objective To observe the efficacy of 577 nm subthreshold micropulse laser photocoagulation(SMLP)on central serous chorioretinopathy(CSC)and explore the baseline characteristics and prognostic features of multimodal ima-ging-based CSC.Methods A retrospective analysis was conducted on 44 patients(46 eyes)with CSC diagnosed at the Ophthalmology Department of the General Hospital of Central Theater Command,PLA from June 2017 to May 2022.The pa-tients were classified into the simple CSC group(23 eyes)and the complex CSC group(23 eyes)based on the multimodal imaging classification.All patients underwent 577 nm SMLP treatment.The best corrected visual acuity(BCVA)and cen-tral macular thickness(CMT)of patients between the two groups were compared before treatment and at 1,3 and 6 months after treatment.The intra-group and inter-group changes and the complete absorption rate of subretinal fluid(SRF)in CSC eyes of patients between the two groups were assessed before treatment and at 6 months after follow-up.Results The BCVA of patients in the simple CSC group significantly improved at 6 months after treatment compared with the base-line(P=0.010);the BCVA of patients in the complex CSC group significantly improved at 3 and 6 months after treatment compared with the baseline(both P＜0.01).The CMT of patients in the simple CSC group was significantly lower than the baseline at 1,3,and 6 months after treatment(all P＜0.05);the CMT of patients in the complex CSC group was signifi-cantly lower than the baseline at 1 and 6 months after treatment(both P＜0.05);the change in CMT from baseline after 6 months of treatment for simple CSC was(-163.74±88.10)μm,the change in CMT from baseline after 6 months of treat-ment for complex CSC was(-71.96±164.30)μm,and the difference between the two groups was statistically significant(P=0.023).During the follow-up period,the number of eyes with persistent SRF in the simple CSC group was significant-ly lower than that in the complex CSC group(P＜0.05).The complete absorption rate of SRF at 6 months after treatment in the simple CSC group was greater than that in the complex CSC group(P＜0.05).Conclusion The prognosis of sim-ple CSC treated with 577 nm SMLP is better than that of complex CSC,and the new multimodal imaging classification has a certain value in predicting the prognosis of CSC after SMLP.

Objective To observe the effects of AnMeiDan(AMD),a representative formula of Peiyuan Guben Tranquilisation method,on the learning memory of zebrafish model of sleep deprivation(SD),and to explore the possible mechanism and the optimal apparent dose based on the Orexin signalling pathway.Methods 120 4-month-old wild-type AB line zebrafish were randomly divided into blank group,model group,AMD low-dose group(0.009 mg·mL-1·d-1),AMD medium-dose group(0.027 mg·mL-1·d-1),AMD high-dose group(0.081 mg·mL-1·d-1),and melatonin group(0.4 mg·mL-1·d-1).Zebrafish SD model was established by LED light induction method,24 h behavioural changes of zebrafish in each group were monitored by zebrafish behavioural tracking system,differences in learning and memory ability of zebrafish in each group were detected by T-maze,the state and number of neurons in the brain of zebrafish in each group were observed by Hematoxylin-Eosin(HE)staining method,the structural changes of neurons in the brain of zebrafish in each group were observed by transmission electron microscopy,and changes in neuron structure in the brain of zebrafish in each group were detected by ELISA method.The levels of Orexin A(OXA)and Orexin B(OXB)in zebrafish were detected by enzyme-linked immunosorbent assay(ELISA),and the protein and mRNA expression levels of OXA/p38 mitogen-activated protein kinase(P38 MAPK)/extracellular regulated protein kinase(ERK1/2)were detected by protein immunoblotting and real-time fluorescence quantitative assay(RT-PCR)in zebrafish.Results Compared with the blank group,the total number of rests,duration and distance were reduced in the model group(P<0.05),and the latency to enter the target region(OR)was prolonged(P<0.01);the number of neuronal cells was reduced,the nuclei were degenerated(P<0.01),cytoplasmic hyalinosis was increased(P<0.01),and light colouring was observed,the nuclei were collapsed,the nuclear membranes were ruptured and lysed,the chromatin was solidified,and the mitochondria were swollen and deformed with internal cavities,and lamellipod-like myelin-like structure was formed;the brain tissue OXA,OXA,and ERK1/2 expression levels of the mRNAs and the mRNAs of these signaling pathways were increased(P<0.01).brain tissue OXA and OXB proteins were significantly elevated,OXA mRNA and protein were significantly up-regulated,and P38 MAPK and ERK1/2 mRNA and protein were significantly down-regulated(P<0.01).Compared with the model group,the drug intervention could prolong the total resting exercise time of SD zebrafish(P<0.05);shorten the latency of zebrafish to reach OR(P<0.05);protect the structure and morphology of neuronal cells,and alleviate the damage of brain tissues;reduce the content of OXA and OXB in the brain tissues(P<0.01);and down-regulate the expression of OXA mRNA and protein(P<0.01),and up-regulate the P38 MAPK and ERK1/2 mRNA and protein expression of P38 MAPK and ERK1/2(P<0.01);and the quantity-effect relationship was obvious,and the effect of high dose of AMD was optimal.Conclusion AMD can improve the learning and memory ability in zebrafish SD model,and its mechanism may be related to the regulation of OXA/P38 MAPK/ERK1/2 signalling pathway and alleviation of neuronal damage in the brain,and the high dose group of AMD showed the best effect.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling.Here,we focused on the role of Semaphorin 3A(Sema3A),expressed by sensory nerves,in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement(OTM)model.Firstly,bone formation was activated after the 3rd day of OTM,coinciding with a decrease in sensory nerves and an increase in pain threshold.Sema3A,rather than nerve growth factor(NGF),highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM.Moreover,in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells(hPDLCs)within 24 hours.Furthermore,exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload.Mechanistically,Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway,maintaining mitochondrial dynamics as mitochondrial fusion.Therefore,Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation,both as a pain-sensitive analgesic and a positive regulator for bone formation.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling.Here,we focused on the role of Semaphorin 3A(Sema3A),expressed by sensory nerves,in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement(OTM)model.Firstly,bone formation was activated after the 3rd day of OTM,coinciding with a decrease in sensory nerves and an increase in pain threshold.Sema3A,rather than nerve growth factor(NGF),highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM.Moreover,in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells(hPDLCs)within 24 hours.Furthermore,exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload.Mechanistically,Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway,maintaining mitochondrial dynamics as mitochondrial fusion.Therefore,Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation,both as a pain-sensitive analgesic and a positive regulator for bone formation.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling.Here,we focused on the role of Semaphorin 3A(Sema3A),expressed by sensory nerves,in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement(OTM)model.Firstly,bone formation was activated after the 3rd day of OTM,coinciding with a decrease in sensory nerves and an increase in pain threshold.Sema3A,rather than nerve growth factor(NGF),highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM.Moreover,in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells(hPDLCs)within 24 hours.Furthermore,exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload.Mechanistically,Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway,maintaining mitochondrial dynamics as mitochondrial fusion.Therefore,Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation,both as a pain-sensitive analgesic and a positive regulator for bone formation.

Bone formation and deposition are initiated by sensory nerve infiltration in adaptive bone remodeling.Here,we focused on the role of Semaphorin 3A(Sema3A),expressed by sensory nerves,in mechanical loads-induced bone formation and nerve withdrawal using orthodontic tooth movement(OTM)model.Firstly,bone formation was activated after the 3rd day of OTM,coinciding with a decrease in sensory nerves and an increase in pain threshold.Sema3A,rather than nerve growth factor(NGF),highly expressed in both trigeminal ganglion and the axons of periodontal ligament following the 3rd day of OTM.Moreover,in vitro mechanical loads upregulated Sema3A in neurons instead of in human periodontal ligament cells(hPDLCs)within 24 hours.Furthermore,exogenous Sema3A restored the suppressed alveolar bone formation and the osteogenic differentiation of hPDLCs induced by mechanical overload.Mechanistically,Sema3A prevented overstretching of F-actin induced by mechanical overload through ROCK2 pathway,maintaining mitochondrial dynamics as mitochondrial fusion.Therefore,Sema3A exhibits dual therapeutic effects in mechanical loads-induced bone formation,both as a pain-sensitive analgesic and a positive regulator for bone formation.