Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective To explore the ability of different regions of interest(ROI)location settings for the detection of bronchial artery and pulmonary vascular lesions in children with hemoptysis via computed tomography angiography(CTA)examination.Methods The hemoptysis group(79 cases)who underwent chest CTA examination and the control group(79 cases)with negative CTA results were retrospectively selected.The ROI of the hemoptysis group were placed in the pulmonary trunk,left ventricle,ascending aorta and descending aorta,which were defined as groups 1,2,3 and 4 respectively.Clinical data(age,sex)and CT parameters,including bronchial artery diameter,ascending aorta CT value,pulmonary artery CT value,△CTA-PA,bronchial artery image score,pulmonary artery image score,were recorded and analyzed,respectively.Results There was a statistically significant difference in the diameter of the left and right bronchial arteries between the hemoptysis group and the control group(P＜0.05).In the hemoptysis group,there were statistically significant differences in the image scores of the left and right bronchial arteries,pulmonary artery image scores,and△CTA-PA among the four subgroups(P＜0.05).According to post-hoc comparison results,there was a statistically significant difference in the overall mean scores of the right bronchial artery image score(x2=11.333,P＜0.05)and left bronchial artery image score(x2=8.111,P＜0.05)between groups 2 and 3.Conclusion When ROI is placed in the ascending aorta,bronchial artery lesions and pulmonary vascular lesions can be detected simultaneously in CTA examination,which is helpful for the diagnosis of hemoptysis vascular etiology in children.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

Objective:To explore the relapse-related genes and their clinicopathological connections of diffuse large B cell lymphoma (DLBCL).Methods:Targeted panel sequencing was conducted on 32 eligible DLBCL samples; the patients were diagnosed, treated, and went into complete remission at the First Affiliated Hospital of Nanjing Medical University from January 2015 to December 2019, including 14 cases with recurrence (relapsed group) and 18 cases with long-term complete remission of over five years (remission group). Clinical and pathological data were further reviewed. Fisher′s exact test was employed to compare the differences in clinicopathological characteristics and mutation patterns between the two groups.Results:Among the 32 patients, there were 18 males and 14 females, with a male to female ratio of 1.3∶1.0 and a median age of 53 (45.5, 67.0) years. In the relapsed group, PIM1 (11/14), KMT2D (7/14), PRDM1 (6/14), MYD88 (6/14), DTX1 (6/14) emerged as the most frequently mutated genes. In the remission group, while recurrent PIM1, KMT2D and MYD88 mutations were also observed, the TP53 gene exhibited the highest mutation frequency (6/18). Compared to the remission group, relapsed group showed elevated mutation frequencies of PIM1 ( P=0.013) and FAT4 ( P=0.010), alongside a reduced incidence of TP53 mutations. In all 32 patients, DLBCL with CD79B, CCND3, DTX1, KMT2D and PRDM1 mutations demonstrated a propensity towards advanced clinicopathologic stage. Conclusions:Relapsed DLBCL has distinctive clinicopathological and genetic features. PIM1 and FAT4 may be served as potential biomarkers for screening relapsed DLBCL-NOS and as targets for novel therapeutic strategies.

Objective To explore the ability of different regions of interest(ROI)location settings for the detection of bronchial artery and pulmonary vascular lesions in children with hemoptysis via computed tomography angiography(CTA)examination.Methods The hemoptysis group(79 cases)who underwent chest CTA examination and the control group(79 cases)with negative CTA results were retrospectively selected.The ROI of the hemoptysis group were placed in the pulmonary trunk,left ventricle,ascending aorta and descending aorta,which were defined as groups 1,2,3 and 4 respectively.Clinical data(age,sex)and CT parameters,including bronchial artery diameter,ascending aorta CT value,pulmonary artery CT value,△CTA-PA,bronchial artery image score,pulmonary artery image score,were recorded and analyzed,respectively.Results There was a statistically significant difference in the diameter of the left and right bronchial arteries between the hemoptysis group and the control group(P＜0.05).In the hemoptysis group,there were statistically significant differences in the image scores of the left and right bronchial arteries,pulmonary artery image scores,and△CTA-PA among the four subgroups(P＜0.05).According to post-hoc comparison results,there was a statistically significant difference in the overall mean scores of the right bronchial artery image score(x2=11.333,P＜0.05)and left bronchial artery image score(x2=8.111,P＜0.05)between groups 2 and 3.Conclusion When ROI is placed in the ascending aorta,bronchial artery lesions and pulmonary vascular lesions can be detected simultaneously in CTA examination,which is helpful for the diagnosis of hemoptysis vascular etiology in children.

Objective:To investigate the expression level of methyltransferase-like 3 (METTL3) in nasopharyngeal carcinoma cells CNE2 and CNE-2R, and to evaluate the effect of METTL3 on cell invasion, metastasis and radiosensitivity.Methods:Real-time reverse transcription PCR and Western blot were used to detect the expression levels of METTL3 in normal nasopharyngeal epithelial cells NP69 and nasopharyngeal carcinoma cells CNE2 and nasopharyngeal carcinoma radioresistant cells CNE-2R cells. METTL3 in CNE2 and CNE-2R cells was silenced by lentivirus-mediated RNA interference technology. The metastasis and invasion abilities of the cells were detected by the scratch assay and Transwell assay. Clonogenic assay and CCK-8 assay were employed to detect the proliferation capacity and viability of cells irradiated with different X-ray doses (0, 2, 4, 6 and 8 Gy). Apoptosis was detected by flow cytometry. Methylated RNA immunoprecipitation (Me-RIP) assay was used to detect the difference in m6A modification level of c-Jun in CNE2 and CNE-2R cells after METTL3 silencing. The transcriptional stability of c-Jun in cells after silencing METTL3 was detected by actinomycin D assay. A nude mouse xenograft model was constructed to detect the effect of METTL3 on the radiosensitivity of nasopharyngeal carcinoma in vivo. Results:Compared with NP69 cells, the expression levels of METTL3 mRNA and protein were significantly increased in CNE2 cells, and the expression level was even higher in CNE-2R cells (all P<0.01). Lentivirus-mediated RNA interference technology was used to construct a stable METTL3-silencing CNE2 and CNE-2R cell lines (both P<0.01). Scratch assay and Transwell assay showed that the metastasis and invasion abilities of CNE2 and CNE-2R cells were decreased significantly after METTL3 silencing (all P<0.05). Clonogenic assay showed that silencing METTL3 significantly reduced the number of colonies and survival fraction of CNE2 and CNE-2R cells after irradiation with different doses of X-rays (all P<0.05). CCK-8 assay showed that the proliferation ability of CNE2 and CNE-2R cells was significantly reduced by silencing METTL3 (all P<0.05). After different doses of irradiation, silencing METTL3 significantly reduced the survival fraction of CNE2 and CNE-2R cells (all P<0.05). The apoptotic rate after METTL3 silencing was higher than that in the control group at the irradiation dose of 0 and 8 Gy (all P<0.05). The Me-RIP assay showed that the m6A modification level of c-Jun in CNE2 and CNE-2R cells was significantly reduced after METTL3 silencing (both P<0.01), and the actinomycin D assay showed that transcriptional stability of c-Jun was reduced. Nude mouse xenograft experiment showed that silencing METTL3 inhibited xenograft proliferation and improved its radiosensitivity. Conclusion:METTL3 is highly expressed in nasopharyngeal carcinoma cells, and METTL3 mediates m6A modification of c-Jun to improve the transcriptional stability of c-Jun and promote the expression of c-Jun, thereby promoting the invasion and metastasis of nasopharyngeal carcinoma cells and reducing their radiosensitivity.

Objective To observe the changes in intradiscal internal internal pressure before and after percutaneous nucleoplasty (PN) in the patients with discogenic low back pain (DLBP),and to explore the mechanism of PN for alleviating DLBP.Methods Thirty patients definitely diagnosed as DLBP undergoing PN in the orthopedic department of this hospital from June 2023 to October 2023 were selected.The internal pressure of the lumbar disc was measured before and after the surgery.The visual analog scale (VAS),Japa-nese orthopedic association score (JOA) and Oswestry disability index (ODI) were used to assess the pain level of the patients before operation and at postoperative 1 d,1 month,3 months.The surgical efficacy was e-valuated by using the modified MacNab criteria.The linear correlation analysis was used to observe the relation-ship between the internal pressure of lumbar disc and changes in each pain score,and the factors possibly affecting postoperative effect was analyzed by using the multivariate logistic regression analysis.Results Compared with be-fore the surgery,the immediate postoperative internal pressure of lumbar disc in the patients was significantly decreased.The JOA scores at postoperative 1 d,1 month and 3 months were significantly increased,while the VAS and ODI scores were significantly decreased,and the differences were statistically significant (P<0.05). The excellent rates of surgical efficacy at postoperative 1 d,1 month and 3 months were 66.7％,73.3％ and 63.3％,respectively.The internal pressure of lumbar disc difference value between before operation and on postoperative 1 d and internal pressure of lumbar disc decrease rate had no significant correlation with the change value of each pain score (P>0.05).The multivariate logistic regression analysis showed that the dis-ease duration and the decrease rate of internal pressure of lumbar disc were the influencing factors of clinical efficacy in PN (P<0.05).Conclusion The occurrence of DLBP may be related with the increase of internal pressure of lumbar disc.PN could effectively reduce internal pressure of lumbar disc and achieve the goal of pain relief.

Diffuse large B cell lymphoma （DLBCL） is a malignant tumor derived from mature B cells. Currently， chemotherapy is still the main clinical treatment. However， some patients experience recurrence or refractory conditions after treatment. On June 15， 2023， the FDA approved the marketing of glofitamab， a CD3/CD20 bispecific monoclonal antibody， to provide the new treatment plan for patients with recurrent or refractory DLBCL after receiving 2-line or above systemic treatment. This article reviews pharmacological effects， clinical studies， safety， usage and dosage of glofitamab. Glofitamab mainly plays a therapeutic role in DLBCL by promoting the activation and proliferation of T cells，activating T cells to release tumor cell-killing proteins， and mediating the lysis of B cells. Clinical studies have shown that glofitamab has a better complete and objective response rate for recurrent or refractory DLBCL. Common adverse reactions caused by glofitamab include mild/moderate cytokine release syndrome， musculoskeletal pain， rash， fatigue， and so on，without significant drug interactions.

Objective:To explore the curative effects of individualized surgical method for treating pyriform sinus fistula (PSF) in children.Methods:A total of 43 PSF children treated in the Department of Pediatric Surgery of Henan Provincial People′s Hospital from June 2015 to November 2019 were selected, with 41 cases on the left side and 2 cases on the right side.There were 27 cases in the quiescent inflammatory infection stage and 16 cases in the acute inflammatory infection stage.According to patients′ condition, low-temperature plasma radiofrequency ablation, fistulectomy, abscess incision and drainage or combined operation were conducted.The postoperative complications and recurrence were observed.Results:Among 43 cases, there were 19 cases treated with low-temperature plasma radiofrequency ablation, 11 cases treated with low-temperature plasma radiofrequency ablation combined with abscess incision and drainage, 5 cases treated with low-temperature plasma radiofrequency ablation combined with fistulectomy, and 8 cases treated with fistulectomy.Four cases had postoperative hoarseness and recovered within 1-5 months.One case occurred pseudo healing after abscess incision and drainage, and healed after incision and debridement.Totally, 4 cases recurred, and the follow-up time was from 3 months to 57 months.Conclusions:Low-temperature plasma radiofrequency ablation has the advantages of minimally invasive, safe and effective, which provides a new treatment for PSF in children.Based on the condition of children, the choice of individualized treatment can significantly improve the curative effects.

OBJECTIVE: To study the expression of BIRC6 in renal cancer tissues and investigate the effect of BIRC6 silencing on apoptosis and autophagy of 786-O cells. METHODS: Twenty surgical specimens of renal cancer tissues and adjacent renal tissues were collected from Meizhou People's Hospital between February, 2016 and December, 2018 for detection of BIRC6 protein expression using immunohistochemistry. Renal cancer 786-O cells were transfected with a control small interfering RNA (siRNA) or BIRC6 siRNA RESULTS: The expression of BIRC6 protein was significantly higher in renal cancer tissues than in the adjacent renal tissues. Western blotting showed that siRNA-mediated silencing of BIRC6 significantly lowered the expression of BIRC6 in 786-O cells. In the cells with BIRC6 silencing, treatment with 12.5, 25, 50, 100 and 200 μg/mL 5-FU resulted in significantly higher proliferation inhibition rates than in the cells transfected with the control siRNA ( CONCLUSIONS Interference of BIRC6 mediated by siRNA can inhibit autophagy and promote 5-FU-induced apoptosis to enhance the sensitivity of 786-O cells to 5-FU.
Apoptosis ; Autophagy ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Inhibitor of Apoptosis Proteins/genetics* ; Kidney Neoplasms/genetics* ; RNA, Small Interfering/genetics*