Adenoid cystic carcinoma is a malignant tumor of the head and neck, this article reports a case of a large adenoid cystic carcinoma of the skull base, with the lesion involving the sphenoid sinus, sphenoid bone wings, pterygopalatine fossa, nfratemporal fossa, hard palate, and other structures. The treatment plan consisted of surgical excision, primary reconstrction of the surgical defect,and postoperative radiotherapy, resulting in a favorable prognosis for the patient.
Humans ; Carcinoma, Adenoid Cystic/surgery* ; Maxillary Sinus/surgery* ; Maxillary Sinus Neoplasms/surgery* ; Palate, Hard/surgery* ; Skull Base Neoplasms/surgery* ; Surgical Flaps

Objective:To investigate the dynamic changes of cellular miRNA expression profiles in EBV latently infected Raji cells upon reactivation with Phorbol ester (TPA).Methods:Total RNA was extracted using TRIzol reagent from Raji cells treated with TPA at different time points (0 h, 24 h, 48 h). Small RNA libraries were constructed and sequenced on an Illumina SE50 platform. Differentially expressed miRNAs were identified, and their target genes were predicted and functionally annotated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out through online tools. Additionally, miRanda and RNAhybrid software were used to predict cellular miRNAs targeting the EBV genome. Real-time RT-qPCR was employed to validate the expression levels of differentially expressed novel miRNAs.Results:High-throughput sequencing identified 1 301 celluar miRNAs, comprising 1 189 known and 112 novel miRNAs. A total of 264 known differentially expressed cellular miRNAs and 13 novel miRNAs were identified through high-throughput miRNA sequencing. Secondary structure prediction revealed that the novel miRNAs exhibited typical pre-miRNA hairpin structures. Stem-loop quantitative real-time PCR (RT-qPCR) validation of Novel_miR_183 and Novel_miR_242 did not exhibit a statistically significant difference ( F=1.407, P=0.370 7 for Novel_miR_183; F=1.277, P=0.397 0 for Novel_miR_242) between the TPA-stimulated and untreated groups. Target gene prediction analysis revealed that the differentially expressed cellular miRNAs were involved in various important biological processes and signaling pathways. Furthermore, 1 189 known cellular miRNAs and 108 novel miRNAs were predicted to target the EBV genome. Conclusions:Treatment of Raji cells with TPA stimulation successfully reactivated Raji cells and significantly altered their miRNA expression patterns. Differentially expressed miRNAs were identified, suggesting that these miRNAs probably play crucial roles in regulating EBV infection and replication by directly targeting the EBV genome.

Objective:To investigate the effect of nuclear factor erythroid-2 related factor (Nrf2) overexpression on high-risk human papillomavirus type 16 (HPV16) E6 protein.Methods:The pRK5-FLAG-NFE2L2 plasmid was constructed, and pRK5-FLAG-NFE2L2 and pRK5-HA-HPV16E6 plasmids were co-transfected in HEK293T cells, and the expression of the two proteins was detected by Western blotting (WB). Next, pRK5-HA-HPV16E6 and pRK5-FLAG-Nrf2 plasmids were expressed using an in vitro transcription system to observe the expression of both. Finally, pEGFP-HPV16E6 and pLVX-mCherry-Nrf2 plasmids were co-transfected in HEK293T cells, and the cellular localization of the E6 protein and the Nrf2 protein was observed using fluorescence microscopy.Results:Nrf2 protein was successfully overexpressed in HEK293T cells, and the WB result showed that Nrf2 inhibited HPV16 E6 protein expression in a significant dose-dependent manner. The expression of HPV16 E6 protein in the TNT Quick Coupled Transcription/Translation Systems was affected by Nrf2, while the expression of HPV16 E6 in TnT SP6 High-Yield Wheat Germ Protein Expression System was no longer inhibited by Nrf2. Fluorescence imaging further showed no intracellular co-localization of Nrf2 and HPV16 E6.Conclusions:Overexpression of the nuclear transcription factor Nrf2 reduces HPV16 E6 protein expression, but there is no intracellular co-localization of them.

Objective:To establish a protocol for virus isolation using the mixed method, and evaluate the efficacy of the suspended method and the mixed method in isolating herpes simplex virus (HSV).Methods:Simulated HSV-infected clinical samples were prepared using HSV-1 F strain and CDC-P1 strain. Both the suspended method and the mixed method were used to isolate HSV-1 from these samples. The virus isolation efficiency of the mixed method under various conditions was assessed. These conditions included different multiplicity of infection (MOI), cell seeding densities, and virus adsorption times.The 50% tissue culture infective dose (TCID 50) assay was used for the assessment. The positive rates of virus detection under low viral load conditions were compared between the two methods. Results:Under the conditions of a MOI of 0.005, a virus adsorption time of 15 min, and a cell seeding density of 1×10 6 cell/ml, the mixed method achieved effective isolation of HSV-1. When the virus titer of the sample was 100 TCID 50/ml, the positivity rate of the mixed method reached 100.0%, while the positivity rates of the suspended method were 50.7% (38/75) and 52.0% (39/75) after cultured for 72 h and 96 h, respectively. When the virus titer of the sample was 10 TCID 50/ml, the positivity rate of the mixed method was 100.0%, while the positivity rate of the suspension method was 0. Conclusions:The mixed method exhibits significantly higher efficiency in HSV isolation compared with the suspended method. Under the conditions of high viral load, both the suspended method and the mixed method can be effective in isolating HSV-1. For clinical samples with low viral loads, the mixed method has greater applicability.

Objective To explore the value of diffusion-weighted magnetic resonance imaging(MRI-DWI)and arterial spin labeling magnetic resonance imaging(ASL)in evaluating the efficacy of chemoradiotherapy in patients with locally advanced nasopharyngeal carcinoma.Methods A total of 103 patients with locally advanced nasopharyngeal carcinoma who received synchronous radiotherapy and chemotherapy in Qinghai Armed Police Corps Hospital from January 2021 to December 2023 were selected as research subjects.The apparent diffusion coefficient(ADC)of the primary tumor,the volume of primary tumor(GTVnx),the volume of the primary tumor+the volume of retropharyngeal lymph nodes(GTVnx+RLN),and the tumor blood flow(TBF)of ASL quantitative perfusion parameters were compared between effective 73 case and ineffective patients(30 case)before and after chemoradiotherapy.The receiver operating characteristics(ROC)curves were used to evaluate the value of ADC,GTVnx,GTVnx+RLN and TBF in predicting the efficacy of chemoradiotherapy in nasopharyngeal carcinoma patients.Univariate method was used to analyze the basic data of patients,and Logistic regression model was used to analyze the related factors of chemoradiotherapy of locally advanced nasopharyngeal carcinoma.Results After concurrent chemoradiotherapy,73 patients were included in the effective group,including 36 patients achieving complete remission(CR)and 37 patients achieving partial remission(PR);30 patients were included in the ineffective group,including 25 patients achieving stable disease(SD)and 5 patients achieving disease progression(PD).Before and after treatment,ADC,GTVnx,GTVnx+RLN in the effective group were significantly lower than those in the ineffective group,and TBF in the effective group was higher than that in the ineffective group(P<0.05).The changes of ADC,GTVnx,GTVnx+RLN,and TBF before and after concurrent chemoradiotherapy in the effective group were significantly higher than those in the ineffective group(P<0.05).The area under the curve(AUC)values of ADC,GTVnx,GTVnx+RLN,and TBF for predicting the efficacy of concurrent chemoradiotherapy in patients with locally advanced nasopharyngeal carcinoma were 0.727,0.555,0.753,and 0.791,respectively.The results of logistic regression model showed that high levels of ADC,GTVnx and GTVnx+RLN,low level of TBF,TNM stage Ⅳ,and low tumor differentiation before treatment were independent risk factors for poor efficacy of concurrent chemoradiotherapy in nasopharyngeal carcinoma patients(P<0.05).Conclusion Elevated baseline of ADC,GTVnx and GTVnx+RLN and reduced TBF are correlated with poorer outcomes following chemoradiotherapy in patients with locally advanced nasopharyngeal carcinoma,serving as potential predictive biomarkers.

Objective:To establish a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products,screen critical quality markers,and provide methodological support for their intelligent quality control.Methods:Ultra-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS)was employed to quantitatively analyze 34 bioactive components(e.g.,tanshinone Ⅱ A and alvianol-ic acid B)in 20 batches of fermentation products.The key markers were screened through hierarchical cluster anal-ysis and partial least squares discriminant analysis,and an intelligent discriminant model was constructed with sup-port vector machine machine learning algorithm to digitally analyze the characteristics of quality differences between batches.Results:Thirty-four common peaks were calibrated across all batches.Combined with partial least squares analysis,six key difference markers were further screened,including terpenoids such as isotanshinone Ⅱ A and tan-shinone Ⅱ A and phenolic acids such as salvianolic acid G.The support vector machine model can achieve 100％accuracy of origin discrimination by optimizing parameters jointly with genetic algorithm and grid search.Conclusion:This study developed a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products through a three-step analytical strategy("chemical feature exploration-marker screening-model valida-tion"),providing a transferable technical pathway for the intelligent transformation of traditional Chinese medicine quality control.

Objective:To establish a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products,screen critical quality markers,and provide methodological support for their intelligent quality control.Methods:Ultra-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry(UPLC-Q-Orbitrap-MS)was employed to quantitatively analyze 34 bioactive components(e.g.,tanshinone Ⅱ A and alvianol-ic acid B)in 20 batches of fermentation products.The key markers were screened through hierarchical cluster anal-ysis and partial least squares discriminant analysis,and an intelligent discriminant model was constructed with sup-port vector machine machine learning algorithm to digitally analyze the characteristics of quality differences between batches.Results:Thirty-four common peaks were calibrated across all batches.Combined with partial least squares analysis,six key difference markers were further screened,including terpenoids such as isotanshinone Ⅱ A and tan-shinone Ⅱ A and phenolic acids such as salvianolic acid G.The support vector machine model can achieve 100％accuracy of origin discrimination by optimizing parameters jointly with genetic algorithm and grid search.Conclusion:This study developed a digital quality evaluation system for Salvia miltiorrhiza-Monascus fermentation products through a three-step analytical strategy("chemical feature exploration-marker screening-model valida-tion"),providing a transferable technical pathway for the intelligent transformation of traditional Chinese medicine quality control.

Objective:To establish a protocol for virus isolation using the mixed method, and evaluate the efficacy of the suspended method and the mixed method in isolating herpes simplex virus (HSV).Methods:Simulated HSV-infected clinical samples were prepared using HSV-1 F strain and CDC-P1 strain. Both the suspended method and the mixed method were used to isolate HSV-1 from these samples. The virus isolation efficiency of the mixed method under various conditions was assessed. These conditions included different multiplicity of infection (MOI), cell seeding densities, and virus adsorption times.The 50% tissue culture infective dose (TCID 50) assay was used for the assessment. The positive rates of virus detection under low viral load conditions were compared between the two methods. Results:Under the conditions of a MOI of 0.005, a virus adsorption time of 15 min, and a cell seeding density of 1×10 6 cell/ml, the mixed method achieved effective isolation of HSV-1. When the virus titer of the sample was 100 TCID 50/ml, the positivity rate of the mixed method reached 100.0%, while the positivity rates of the suspended method were 50.7% (38/75) and 52.0% (39/75) after cultured for 72 h and 96 h, respectively. When the virus titer of the sample was 10 TCID 50/ml, the positivity rate of the mixed method was 100.0%, while the positivity rate of the suspension method was 0. Conclusions:The mixed method exhibits significantly higher efficiency in HSV isolation compared with the suspended method. Under the conditions of high viral load, both the suspended method and the mixed method can be effective in isolating HSV-1. For clinical samples with low viral loads, the mixed method has greater applicability.

Objective:To investigate the dynamic changes of cellular miRNA expression profiles in EBV latently infected Raji cells upon reactivation with Phorbol ester (TPA).Methods:Total RNA was extracted using TRIzol reagent from Raji cells treated with TPA at different time points (0 h, 24 h, 48 h). Small RNA libraries were constructed and sequenced on an Illumina SE50 platform. Differentially expressed miRNAs were identified, and their target genes were predicted and functionally annotated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out through online tools. Additionally, miRanda and RNAhybrid software were used to predict cellular miRNAs targeting the EBV genome. Real-time RT-qPCR was employed to validate the expression levels of differentially expressed novel miRNAs.Results:High-throughput sequencing identified 1 301 celluar miRNAs, comprising 1 189 known and 112 novel miRNAs. A total of 264 known differentially expressed cellular miRNAs and 13 novel miRNAs were identified through high-throughput miRNA sequencing. Secondary structure prediction revealed that the novel miRNAs exhibited typical pre-miRNA hairpin structures. Stem-loop quantitative real-time PCR (RT-qPCR) validation of Novel_miR_183 and Novel_miR_242 did not exhibit a statistically significant difference ( F=1.407, P=0.370 7 for Novel_miR_183; F=1.277, P=0.397 0 for Novel_miR_242) between the TPA-stimulated and untreated groups. Target gene prediction analysis revealed that the differentially expressed cellular miRNAs were involved in various important biological processes and signaling pathways. Furthermore, 1 189 known cellular miRNAs and 108 novel miRNAs were predicted to target the EBV genome. Conclusions:Treatment of Raji cells with TPA stimulation successfully reactivated Raji cells and significantly altered their miRNA expression patterns. Differentially expressed miRNAs were identified, suggesting that these miRNAs probably play crucial roles in regulating EBV infection and replication by directly targeting the EBV genome.

Objective:To investigate the effect of nuclear factor erythroid-2 related factor (Nrf2) overexpression on high-risk human papillomavirus type 16 (HPV16) E6 protein.Methods:The pRK5-FLAG-NFE2L2 plasmid was constructed, and pRK5-FLAG-NFE2L2 and pRK5-HA-HPV16E6 plasmids were co-transfected in HEK293T cells, and the expression of the two proteins was detected by Western blotting (WB). Next, pRK5-HA-HPV16E6 and pRK5-FLAG-Nrf2 plasmids were expressed using an in vitro transcription system to observe the expression of both. Finally, pEGFP-HPV16E6 and pLVX-mCherry-Nrf2 plasmids were co-transfected in HEK293T cells, and the cellular localization of the E6 protein and the Nrf2 protein was observed using fluorescence microscopy.Results:Nrf2 protein was successfully overexpressed in HEK293T cells, and the WB result showed that Nrf2 inhibited HPV16 E6 protein expression in a significant dose-dependent manner. The expression of HPV16 E6 protein in the TNT Quick Coupled Transcription/Translation Systems was affected by Nrf2, while the expression of HPV16 E6 in TnT SP6 High-Yield Wheat Germ Protein Expression System was no longer inhibited by Nrf2. Fluorescence imaging further showed no intracellular co-localization of Nrf2 and HPV16 E6.Conclusions:Overexpression of the nuclear transcription factor Nrf2 reduces HPV16 E6 protein expression, but there is no intracellular co-localization of them.