OBJECTIVE To analyze the clinicopathological features of laryngeal squamous carcinoma tumors and their correlation with prognosis in order to improve the understanding and diagnosis of laryngeal squamous cell carcinoma.METHODS The clinical and pathological data(including gender,age,stage,differentiation,immunohistochemistry,etc.)of 179 patients with laryngeal squamous cell carcinoma[171 males,8 females,aged 30-84(61.53±8.02)years]who were treated in Department of Otolaryngology Head and Neck Surgery,The First and Second Clinical Medical Schools of Ningxia Medical University from January 2015 to December 2022 were retrospectively studied,and the effects of various factors on prognosis were analyzed.RESULTS Among the 179 patients with laryngeal squamous cell carcinoma,the male-to-female ratio was 21.4:1,and the incidence was high in the age group of 60-79 years old(58.7%),and the youngest age of onset was 30 years old.The main clinical manifestations were hoarseness 138 cases(77.1%),sore throat 16 cases(8.9%)and pharyngeal foreign body sensation 13 cases(7.3%).Glottic type was more common in the primary site 135 cases(75.4%),and 31 cases were accompanied by cervical lymph node metastasis(17.3%).The degree of differentiation was more common in the moderately differentiated type 80 cases(44.7%).The positive rates of immunohistochemistry markers p16,EGFR(epidermal growth factor receptor),PD-1/PD-L1 and VEGF(vascular endothelial growth factor)were 20.3%,96.4%,36.4%and 77.3%,respectively.Univariate Kaplan-Meier survival analysis showed that the site of disease,lymph node metastasis,and tumor stage were significantly correlated with disease recurrence.Multivariate Cox regression analysis showed that the clinical stage of the tumor was an independent risk factor for the prognosis of the disease(HR=3.715,95%CI:1.519-9.088,P=0.04).CONCLUSION The stage of laryngeal squamous cell carcinoma,the site of the disease,and the metastasis of the lymph nodes are the main factors affecting the prognosis.The high positive expression rate of immunohistochemistry markers EGFR and VEGF is worth paying attention to Targeted therapy for patients with positive PD-1/PD-L1 testing is a promising research direction.

Objective To investigate the effects of berberine on UCP2 expression and the mitochondrial dy-namics in skeletal muscle and kidney in mice with ischemia reperfusion injury (IRI) of lower limb.Methods Thirty male Kunming mice were randomly divided into the negative control group,the positive control group,and low,medium and high doses berberine intervention groups.The mice in all experimental groups constructed the ischemia reperfusion injury model of lower limbs by tourniquet,different doses of berberine solution were injected intraperitoneally for intervention,while the ischemia repeating supply group used normal saline for re-placement.The HE staining was used to detect the pathological conditions of skeletal muscle and kidney,PCR and Western blot were used to detect the gene and protein expression levels of UCP2,FIS1,DRP1,Mfn1 and Mfn2,and the changes of SOD,MDA and GSH were detected by kit.Results After IRI intervention in the both lower extremities,the inflammatory cell infiltration occurred in both skeletal muscle and kidney,and the cell structure of skeletal muscle showed the damage changes.Meanwhile,the gene and protein expressions lev-els of UCP2,FIS1 and DRP1 and the levels of GSH and SOD in skeletal muscle were significantly increased (P<0.05),while the gene and protein expression levels of Mfn1 and Mfn2 and the levels of MDA were signif-icantly decreased (P<0.05).The increase of UCP2 and DRP1 gene and protein expression levels in kidney was different (P<0.05).Berberine could up-regulate the UCP2 gene expression in skeletal muscle and the pro-tein expression in skeletal muscle (P<0.05).At the same time,DRP1 gene and protein were significantly inhibited in the kidney tissue (P<0.05),but which in skeletal muscle had no significant change.Conclusion IRI of skeletal muscle of lower extremity in mice leads to severe oxidative stress injury,mitochondrial dynamic imbalance and inflammatory damage of kidney in the injured parts.The therapeutic effect of berberine on skeletal muscle and kidney IRI may be achieved by inhibiting oxidative stress injury,in which its protective effect on kidney may also be related to inhibit the expression of DRP1 after UCP2 up-regulation,thus limit the mitochondrial fission and slow down the development of injury.

Objective To compare the consistency between the estimated values of nine common resting energy expenditure estimation equations and the measured values among the elderly population in pension institutions. Methods A total of 181 elderly individuals from Henan Guangda Ouan Leling Nursing Center were recruited as study objects through cluster sampling. The differences between the estimated values of nine common equations and the measured values obtained by indirect calorimetry were compared, and subgroup analyses were conducted to compare their consistency across different subgroups. Results Among the nine equations, the Cunningham equation and the Owen equation showed relatively small deviations from the measured values, with no significant differences (deviation rates of -1.9% and -2.0%, root mean square errors of 222.7 kcal/d and 239.4 kcal/d, and coincidence rates of 48.4% and 47.5%, respectively). In subgroups stratified by gender and body mass index (BMI), the Cunningham equation showed no significant difference compared with the measured values. The Owen equation showed no significant difference compared with the measured values only in some subgroups [males with BMI of 20~ < 27 kg/m² and ≥27 kg/m², and females with BMI < 20 kg/m²]. Conclusion The Cunningham equation, which utilizes body composition data, can assess the resting energy expenditure of the elderly in pension institutions with relatively high accuracy and speed, demonstrating strong applicability in this population.

New drug discovery is under growing pressure to satisfy the demand from a wide range of domains, especially from the pharmaceutical industry and healthcare services. Assessment of drug efficacy and safety prior to human clinical trials is a crucial part of drug development, which deserves greater emphasis to reduce the cost and time in drug discovery. Recent advances in microfabrication and tissue engineering have given rise to organ-on-a-chip, an in vitro model capable of recapitulating human organ functions in vivo and providing insight into disease pathophysiology, which offers a potential alternative to animal models for more efficient pre-clinical screening of drug candidates. In this review, we first give a snapshot of general considerations for organ-on-a-chip device design. Then, we comprehensively review the recent advances in organ-on-a-chip for drug screening. Finally, we summarize some key challenges of the progress in this field and discuss future prospects of organ-on-a-chip development. Overall, this review highlights the new avenue that organ-on-a-chip opens for drug development, therapeutic innovation, and precision medicine.

Objective:To investigate the efficacy of postmastectomy radiotherapy (PMRT) for human epidermal growth factor receptor 2 (HER2)-positive T 1-2N 1M 0 breast cancer in the context of HER2-targeted therapy. Methods:This study collected the clinical data of 105 female patients with HER2-positive T 1-2N 1M 0 breast cancer who underwent modified radical mastectomy in the First Affiliated Hospital of Bengbu Medical College from January 2013 to December 2019. Then, the clinical outcomes of these patients were observed, and the prognostic factors and the efficacy of PMRT were analyzed. Results:The median follow-up time was 50 months (ranging from 14 to 107 months), and the 5-year overall survival (OS), local-regional recurrence-free survival(LRFS), and disease-free survival (DFS) were 81.6%, 91.9%, and 76.2%, respectively. The multivariate analysis indicated that independent prognostic factors for OS and DFS include the age, pathologic grade, and tumor size; the independent risk factors for LRFS include positive lymph node ratio (LNR) and hormone receptor (HR) status; and the independent prognostic factor for DFS was PMRT (HR: 2.85, 95% CI: 1.10-8.80, P < 0.05). The subgroup analysis suggested that PMRT significantly improved the OS of various high-risk subgroups ( χ2=4.01-9.18, P < 0.05). However, the further stratified analysis indicated that PMRT only increased the OS of the patients who did not receive HER2-targeted therapy in various high-risk subgroups ( χ2=4.50-6.70, P < 0.05), while there was no statistical difference before and after PMRT for the individuals who received targeted treatment ( P > 0.05). Conclusions:PMRT is an independent prognostic factor for the DFS of patients with HER2-positive T 1-2N 1M 0 breast cancer who underwent modified radical mastectomy. PMRT can improve the OS of high-risk patients with ages < 45 years old, pathologic grade Ⅲ, tumor diameter ≥ 3 cm, LNR > 10%, and HR (-) who received no HER2-targeted therapy. However, the efficacy may be compromised to some extent in the context of the application of HER2-targeted therapy.

Objective:To explore the effects of the deep inspiration breath-hold (DIBH) technique on cardiac dosimetry in internal mammary node irradiation with intensity-modulated radiation therapy (IMN-IMRT) for postoperative left breast cancer.Methods:Totally 23 left breast cancer patients in the First Affiliated Hospital of Bengbu Medical College from Octorber 2021 to July 2022 receiving postoperative IMN-IMRT were enrolled in this study. The changes in dosimetric parameters for their heart and left anterior descending coronary artery (LAD) in the DIBH mode were observed, and the potential factors affecting DIBH effects were analyzed.Results:Compared with the free breath (FB) mode, the DIBH mode manifested a heart volume decrease by 18% ( t = 10.47, P < 0.001), a left lung volume increase by 42% ( t = -14.55, P < 0.001), and significantly reduced dosimetric parameters ( Dmean, Dmax, V5- V30) for the heart and LAD, exhibiting statistically significant differences ( t=-13.38 to -3.30, P<0.05). As indicated by the Pearson correlation analysis, the relative ratio of cardiac dose reduction was positively correlated with that of left lung expansion ( r = 0.82, P < 0.001) and negatively correlated with the patient′age ( r = -0.56, P = 0.005). Conclusions:DIBH can effectively reduce the heart and LAD radiation doses in IMN-IMRT for postoperative left breast cancer and that the patient's age, and the DIBH effects might be affected by the vital capacity.

Objective:To observe the occurrence of epithelial-mesenchymal transition (EMT) in cervical cancer cell line Siha irradiated by X-rays with clinical conventional fraction radiotherapy model and investigate the role of exosomes in this process.Methods:Siha cells were irradiated by 6 MV-X rays with 50 Gy in 25 fractions. EMT was evaluated by cell morphology, EMT biomarkers and cell migration and invasion ability. Exosomes released from cells were detected by transmission electron microscopy and nanoparticle tracking analysis (NTA), and its function in EMT was explored by using an exosome inhibitor GW4869 (10 μmol/L).Results:After irradiation, EMT phenomenon was induced in the survived Siha cells, including the incidence of mesenchymal phenotype, upregulation of epithelial marker E-cadherin ( t=9.66, P<0.05), downregulation of mesenchymal marker N-cadherin ( t=41.61, P<0.05), and increase of cell migration and invasion abilities ( t=6.11, 13.22; P<0.05). Meanwhile, the secretion of exosomes was also increased after irradiation ( t=7.51, P<0.05). When the cells were pre-treated with GW4869, radiation-induced exosome secretion was reduced ( t=7.28, P<0.05), so that radiation-induced EMT was reversed. Conclusions:Ionizing radiation with clinical conventional fraction radiotherapy model promotes EMT of cervical cancer cells through increasing the secretion of exosomes.

Objective:To provide a new morning check method for the output dose stability of the multileaf collimator (MLC) of the CyberKnife M6 (CK-M6) system.Methods:The CT images of a verification phantom with a size of 20 cm × 20 cm × 10 cm were transmitted into the Precision Treatment Plan ning System (ver. 1.1.1.1). The high-precision alignment between the accelerator output front and the fixed position of the phantom surface was achieved using the fiducial tracking method. A 10 cm × 10 cm radiation field was formed by the MLC and a DailyCheck plan with an output of 200 MU was designed. The repeatability, sensitivity, and accuracy of the DailyCheck plan were measured, and the CK-M6 system was continuously tested for one month using the artificial fixed method and the DailyCheck plan designed in this study. Results:The average and the standard deviation of 10 repeated measurements by the DailyCheck plan were 492.28 pC and 0.09, respectively, indicating good stability. There was a linear correlation between the measured values and the output dose, with a correlation coefficient of R2 > 0.999. Moreover, there was a position deviation of 2 mm between the phantom and the accelerator output front, and the result ant effect on the measured values was equivalent to a dose deviation caused by an output of 1.24 MU. The result from the continuous measurement of both the artificial fixed method and the DailyCheck plan fell within permissible limits, showing high consistency. Conclusions:The DailyCheck plan established through the fiducial tracking of a verification phantom can achieve the convenient, quick, and accurate daily detection of the output dose stability of the MLC of CK-M6. Therefore, this method can be widely applied in the clinical quality control of the CK-M6 system.

Objective:To investigate the effects of fluoride exposure on proliferation, apoptosis and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in mice.Methods:BMSCs were isolated and cultured from femur bone marrow of C57BL/6 mice (6 - 8 weeks). The cells in passage 3 were used to detect the surface markers of stem cells by flow cytometry. The cells were cultured in media with a final fluoride concentration of 0.0, 0.1, 1.0, 5.0, 10.0, 15.0, 20.0 and 40.0 mg/L, respectively. The effects of different fluoride concentrations on BMSCs cell proliferation (CCK8 method), apoptosis (flow cytometry analysis), osteogenic differentiation ability [alizarin red and alkaline phosphatase (ALP) staining] were detected. Western blot was applied to detect the levels of apoptosis-related proteins [poly ADP-ribose polymerase (PARP)], mitogen-activated protein kinase (MAPK) pathway member proteins [extracellular regulated protein kinase 1/2 (ERK1/2), c-Jun amino-terminal kinase (JNK), p38 and phosphorylated ERK, JNK, p38 (p-ERK, p-JNK, p-p38)], osteogenic differentiation-related protein [Runt-related transcription factor 2 (Runx2), ALP] and Wnt/β-catenin pathway member proteins [glycogen synthase kinase-3β (GSK3β), phosphorylated GSK3β (p-GSK3β) and β-catenin]. Immunocytofluorescense staining was applied to evaluate the expression levels of p-GSK3β and β-catenin. The two pathways (MAPK and Wnt/β-catenin) were blocked by SP600125 and DKK-1, respectively, to testify their involvement in mechanisms of apoptosis and osteogenic differentiation.Results:The mouse BMSCs were successfully isolated and cultured. Flow cytometry analysis showed that the mesenchymal stem cell surface biomarkers (CD73, CD90 and CD105) were positively expressed. The comparison of cell proliferation at three time points (24, 48 and 72 h) in each concentration group was statistically significant ( F = 65.36, 160.04 and 365.32, P < 0.001), and the comparison of early apoptosis (24 h) in each concentration group was statistically significant ( F = 214.04, P < 0.001); compared with the 0.0 mg/L group, the cell proliferation in 15.0, 20.0 and 40.0 mg/L groups decreased, and the early apoptosis rate in 10.0, 15.0 and 20.0 mg/L groups increased ( P < 0.05). When cells were treated with 15.0 mg/L fluoride for 0 - 24 h, the p-JNK/JNK ratio was higher at 2, 4, 8, 12, 18 and 24 h compared with that at 0 min ( P < 0.05); compared with the fluoride group (15.0 mg/L), the early apoptosis rate of cells after SP600125 block decreased ( P < 0.05), and the protein expression levels of PARP and p-JNK decreased ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, in 0.1 and 1.0 mg/L groups ALP staining was enhanced and the number of calcified nodules increased, and the protein expression levels of Runx2 and ALP in the 0.1 and 1.0 mg/L groups were higher ( P < 0.05). After osteogenic induction, compared with the 0.0 mg/L group, the p-GSK3β/GSK3β ratio and β-catenin protein level were significantly higher in the 0.1 and 1.0 mg/L groups ( P < 0.05); and compared with the fluoride group (1.0 mg/L), addition of DKK-1 significantly decreased the protein expression levels of p-GSK3β and β-catenin and reduced the nuclear entry of β-catenin, and ALP staining decreased and the number of calcified nodules decreased. Conclusions:High concentration of fluoride (> 10.0 mg/L) inhibits the proliferation and promotes apoptosis of BMSCs, while low concentration of fluoride (0.1, 1.0 mg/L) promotes osteogenic differentiation. The MAPK/JNK pathway and the classical Wnt pathway are involved in the above cellular processes, respectively.

Objective:To evaluate the safety and effectiveness of a new Chinese-made surgical biopatch for atrial septum under the establishment of atrial septal defect animal model in miniature pigs.Methods:From June 2018 to April 2019, 26 pigs were divided into experimental group (15 pigs) and the control group (11 pigs). Animal models of atrial septal defect were established by traditional surgical methods. The to-be-evaluated and listed surgical biological patches (with a diameter of 10 mm) were implanted in the experimental group and the control group to repair the atrial septal defect. Cardiac ultrasound and blood examination of all animals were performed before and at 7, 30, 90, 180 days after operation, the results were analyzed with repetitive measurement and analysis of variance. At 90 days and 180 days after the operation, tissue samples were taken from animals after euthanasia. Pathological examination of heart and major organs were conducted. The independent sample t test and rank sum test were used to compare the data between the two groups, and the nonparametric was used to compare the patch calcification score between the two groups. Results:In total of 26 animals, 14 animals in the experimental group(6 at 90 days, 8 at 180 days) and 9 animals in the control group(4 at 90 days, 5 at 180 days) reached the end of the experiment. The other 3 animals (1 in the experimental group and 2 in the control group) died of arrhythmia, whole heart failure and right heart failure, the results of pathological examination showed that the causes of death were unrelated to the experimental materials. Cardiac ultrasound showed no patch leakage in all animals. There was no statistically significant difference in cardiac ultrasound and blood examination between the two groups at different time points after operation (all P>0.05). The pathological results showed that all the implants were intact and had good biocompatibility. There was no significant difference in the mean endothelialization rate between the experimental group and the control group at 90 and 180 days after operation ((80.8±29.1)% vs. (82.5±23.6)%, t=0.095, P=0.927; (78.8±36.4)% vs. (82.0±19.2)%, t=0.182, P=0.859) on 90 and 180 days, there was no significant difference in the patch calcification score between the two groups (1.00(1.25) vs. 2.00(0.75), Z=6.500, P=0.214; 0(0.75) vs. 1.00(2.00), Z=12.000, P=0.139). Conclusion:The new Chinese-made surgical biopatch for atrial septum has comparable safety and efficacy to that of the marketable patch in miniature pig atrial septal defect animal model.