[摘 要] 目的：探讨肿瘤坏死因子受体相关蛋白1（TRAP1）在结肠癌组织和细胞中的表达及其与临床病理特征和患者预后的关系和相关分子机制。方法：通过TCGA和GEO数据全面分析TRAP1在结肠癌中的表达及其与临床病理特征和患者预后的关系，选取2020年10月至2021年03月间在山西医科大学第一医院手术切除的10例结肠癌组织及相应癌旁组织标本，用IHC染色法检测中国人结肠癌组织中TRAP1的表达进行验证，运行R包（survival和survminer）进行Kaplan-Meier生存分析；在线分析TRAP1蛋白的信号肽及穿膜结构域，通过基因富集分析软件进行GO分析和KEGG分析。培养结肠癌SW480和SW620细胞，将si-NC和si-TRAP1转染结肠癌细胞，实验分为空白对照组、si-NC组和si-TRAP1组，采用qPCR法检测转染后各组结肠癌细胞中TRAP1的表达，FCM检测转染后各组细胞的细胞周期和凋亡情况。结果：与癌旁组织比较，TRAP1在结肠癌组织中呈高表达（P<0.01），TRAP1表达水平与淋巴结转移有关联（P<0.05），TRAP1高表达组结肠癌患者5年OS率较低（P<0.05）。TRAP1蛋白属于细胞质蛋白，功能富集结果显示TRAP1及其相关分子与细胞周期、核糖体生物发生等信号通路有关（均P<0.01），TRAP1高表达组的结肠癌代谢重编程基因簇和线粒体蛋白输入基因簇水平升高（均P<0.01）。敲减TRAP1后，结肠癌细胞周期阻滞于G1期，细胞凋亡水平显著升高（均P<0.01）。结论：TRAP1在结肠癌组织中呈高表达，且与患者淋巴结转移和低OS率相关联，敲减TRAP1可阻滞结肠癌细胞周期并促进其凋亡。

Clinical metagenomic next-generation sequencing (mNGS) entails unbiased shotgun sequencing of all microbial and host nucleic acids present in a clinical sample. By analyzing the microbiota diversity, taxonomic, and phylogenetic relationships of clinical specimens, metagenomics related analysis provides an opportunity to investigate substantial biological significance of different microbes. According to the published paper, most studies on mNGS mainly focused on the clinical impact evaluation. However, the studies focused on the analytical performance validation of mNGS before clinical application were rare. Here, a scheme, included intended use, method establishment, assay validation and standard operating protocol, for the laboratory validation of clinical metagenomics sequencing assay was provided by summarizing experiences of clinical laboratory department of Peking Union Medical College Hospital protocol and relevant research. In this scheme, we discussed important topics of mNGS laboratory validation as below: specimen type and pathogen list, bioinformatics pipeline setup, dry lab standard preparation and performance validation, mNGS workflow setup, background nucleotide acid evaluation, wet lab standard preparation and performance validation.

Objective: The aim of this study is to explore the application and advantages of combined intrathecal and extrathecal hypothermic plasma tonsillectomy in reducing intraoperative and postoperative hemorrhage in OSA children. Method: We retrospectively reviewed 726 cases who were diagnosed as OSA. All patients were divided into two groups according to the surgical method: 320 cases by total tonsillectomy and 406 cases by combined extracapsular and intracapsular tonsillectomy. The intro operative bleeding volume, post operative haemorrhage data as time, location and degree in the two groups were compared. Result: There was no statistical difference in the intro operative bleeding volume in the two groups [（9.3±4.6） mL]vs [（7.6±3.5） mL], t=12.687, P=0.235. Two patients who underwent combined extracapsular and intracapsular tonsillectomy presented with post operative haemorrhage, the total post operative haemorrhage rate was significantly decreased that in the total tonsillectomy group（14 cases）（χ²=10.779, P=0.001）. The 2 patients in combined extracapsular and intracapsular tonsillectomy group were secondary haemorrhage, with location in the upper pole and medium, grade A haemorrhage; while in the 14 cases in in the total tonsillectomy group, there were 2 cases presented with primary haemorrhage and 12 cases with secondary haemorrhage; with regard to location of haemorrhage, 1 in the upper pole, 2 in the medium, 11 in the lower pole; 5 cases presented with grade A haemorrhage, 8 with grade B haemorrhage and 1 with grade C haemorrhage. The haemorrhage rate at 7 days after surgery （χ²=5.697, P=0.017）, at the lower pole（χ²=11.961, P=0.001） and grade B（χ²=8.097, P=0.004） were all significantly decreases in the combined extracapsular and intracapsular tonsillectomy group. Conclusion Plasma tonsillectomy combined with intrathecal and extrathecal hypothermic tonsillectomy is a safe and effective method, which has obvious advantages in reducing the postoperative hemorrhage, especially the secondary hemorrhage of Subtonsillar Pole.

Objective: The aim of this study is to investigate the effect of fibreoptic endoscopic of sallowing （FEES） in the assessment of pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma. Method: Fifty-three NPC patients with post-irradiated underwent FEES and video fluoroscopy（VF）.The results were analyzed using the Bolus Residue Scale and Rosenbek's penetration aspiration scale. Result: The agreement in the detection of penetration and aspiration between FEES and VF of liquid（κ=0.56, 95%CI 0.38-0.73） and porridge（κ=0.64, 95%CI 0.43-0.81） was "fair". The detection rates of penetration on FEES with liquid and porridge were 60% and 51%, the detection rates of aspiration on VF with liquid and porridge were 70% and 53%. There were no statistical differences. The agreement in the detection of pharyngeal residue between FEES and VF of liquid （κ=0.38, 95%CI0.12-0.62） and porridge （κ=0.66, 95%CI 0.44-0.86） was "fair". The detection rates of pharyngeal residue on FEES and VF with porridge were 43% and 45%, the difference was not statistically significant. The detection rates of pharyngeal residue on FEES and VF with liquid were 44% and 24%, and the difference was statistically significant. Conclusion FEES is an effective and valuable tool for evaluating pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma.

The aim of this study is to investigate the effect of fibreoptic endoscopic of sallowing （FEES） in the assessment of pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma. Fifty-three NPC patients with post-irradiated underwent FEES and video fluoroscopy（VF）.The results were analyzed using the Bolus Residue Scale and Rosenbek's penetration aspiration scale. The agreement in the detection of penetration and aspiration between FEES and VF of liquid（κ=0.56, 95% 0.38-0.73） and porridge（κ=0.64, 95% 0.43-0.81） was "fair". The detection rates of penetration on FEES with liquid and porridge were 60% and 51%, the detection rates of aspiration on VF with liquid and porridge were 70% and 53%. There were no statistical differences. The agreement in the detection of pharyngeal residue between FEES and VF of liquid （κ=0.38, 95%0.12-0.62） and porridge （κ=0.66, 95% 0.44-0.86） was "fair". The detection rates of pharyngeal residue on FEES and VF with porridge were 43% and 45%, the difference was not statistically significant. The detection rates of pharyngeal residue on FEES and VF with liquid were 44% and 24%, and the difference was statistically significant. FEES is an effective and valuable tool for evaluating pharyngeal dysphagia in post-irradiated patients with nasopharyngeal carcinoma.

The aim of this study is to explore the application and advantages of combined intrathecal and extrathecal hypothermic plasma tonsillectomy in reducing intraoperative and postoperative hemorrhage in OSA children. We retrospectively reviewed 726 cases who were diagnosed as OSA. All patients were divided into two groups according to the surgical method: 320 cases by total tonsillectomy and 406 cases by combined extracapsular and intracapsular tonsillectomy. The intro operative bleeding volume, post operative haemorrhage data as time, location and degree in the two groups were compared. There was no statistical difference in the intro operative bleeding volume in the two groups [（9.3±4.6） mL]vs [（7.6±3.5） mL], =12.687, =0.235. Two patients who underwent combined extracapsular and intracapsular tonsillectomy presented with post operative haemorrhage, the total post operative haemorrhage rate was significantly decreased that in the total tonsillectomy group（14 cases）（χ²=10.779, =0.001）. The 2 patients in combined extracapsular and intracapsular tonsillectomy group were secondary haemorrhage, with location in the upper pole and medium, grade A haemorrhage; while in the 14 cases in in the total tonsillectomy group, there were 2 cases presented with primary haemorrhage and 12 cases with secondary haemorrhage; with regard to location of haemorrhage, 1 in the upper pole, 2 in the medium, 11 in the lower pole; 5 cases presented with grade A haemorrhage, 8 with grade B haemorrhage and 1 with grade C haemorrhage. The haemorrhage rate at 7 days after surgery （χ²=5.697, =0.017）, at the lower pole（χ²=11.961, =0.001） and grade B（χ²=8.097, =0.004） were all significantly decreases in the combined extracapsular and intracapsular tonsillectomy group. Plasma tonsillectomy combined with intrathecal and extrathecal hypothermic tonsillectomy is a safe and effective method, which has obvious advantages in reducing the postoperative hemorrhage, especially the secondary hemorrhage of Subtonsillar Pole.

Objective: To explore the clinical manifestation and treatment strategy for descending necrotizing mediastinitis (DNM). Methods: A total of 27 cases diagnosed as DNM from January 2010 to August 2018 in the First People’s Hospital of Foshan were reviewed. There were 16 males and 11 females, age ranged from 16 to 84 years. The clinical data were collected. SPSS 16.0 software and chi square test were used for statistical analysis. Results: ALL 27 cases were diagnosed as DNM by contrast-enhanced CT scan of the neck and chest. Among the 27 cases, 13 cases resulted from peritonsillar abscess, 8 cases from esophageal foreign body perforation, 5 cases from parapharyngeal and retropharyngeal space abscess, and one case from infection of oral cavity. These 27 cases were divided into three subtypes according to the sites of mediastinitis, including 11 cases for typeⅠ, 5 cases for type ⅡA and 11 cases for type ⅡB. Of 27 cases, 20 cases underwent transcervical drainage for DNM, of which 5 cases with tracheotomy and 6 cases with thoracic drainage, and finally 19 of the 20 patients were cured, and one patient died of bacteremia; 7 cases refused to received surgery and were routinely treated with antibiotics, of which, one case was cured and 6 cases died. The curative rate in patients underwent surgery was significantly higher than that in patients treated with medication (χ²=13.638, P<0.001). Among the 20 cured cases, 4 cases were combined with diabetes mellitus and 6 cases with necrotizing fasciitis, while in the 7 died cases, 5 cases were combined with diabetes mellitus and 6 cases with necrotizing fasciitis. The comorbidity rates of diabetes mellitus (χ²=4.074, P=0.044) and necrotizing fasciitis (χ²=4.457, P=0.035) in died cases were significantly higher than those in cured cases. Conclusion DNM is a serious infection, with high mortality especially in patients with diabetes and necrotizing fasciitis. Timely cervical and chest enhanced CT scan play vital role in its diagnosis. DNM can be treated effectively with transcervical drainage.

Objective: To study the clinical outcome of transoral CO₂ laser surgery for glottic cancer involving the anterior commissure. Methods: Thirty-two cases of glottic cancer involving the anterior commissure treated by transoral CO₂ laser surgery between March 2009 and December 2013 were retrospectively reviewed. Among these cases, 27 were T1bM0M0, 5 were T2N0M0. All cases were followed-up for more than 3 years. Results: All the 32 cases were successfully treated. Perioperative complications included injuries in the soft palate mucosa(13/32, 40.63%), loose incisors(3/32, 9.38%) and subcutaneous emphysema in the neck(2/32, 6.25%). During the follow-up period, granulation was found in all cases. Three cases had local recurrence. Two patients treated by a secondary transoral CO₂ laser surgery and the other case had total laryngectomy, all three cases were followed up for 5 years without recurrence. Two cases had regional recurrence but no primary site recurrence. One patient was treated by neck dissection, and followed up for 5 years without recurrence. The other patient died of supraclavicular and mediastinal lymph node metastasis and lung metastasis 40 months after operation. The overall 5-year survival rate was 90.6%. There was no significant difference in survival rate between T1bN0M0(92.6%) and T2N0M0(80.0%) (Log Rank χ²=0.788, P=0.375). The overall 5-year local regional control rate was 84.4%. In T1bN0M0 lesions, the 5-year local regional control rate was 92.6%, which was significantly higher than that in T2N0M0 lesions(40.0%) (Log Rank χ²=9.504, P=0.002). Conclusion With appropriate surgical indication, detailed preoperative evaluation, good surgical skill, transoral CO₂ laser surgery may achieve satisfactory outcome in the treatment of glottic cancer involving the anterior commissure.

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on LPS-induced abnormal activation of platelets in peripheral blood of healthy human donors and its possible molecular mechanism.

METHODSVenous blood samples were collected from a healthy volunteer, and platelet-rich plasma (PRP) from the blood were isolated by differential centrifugation. The PRP was subpackaged into siliconized test tubes and then divided into control group, LPS group, inactive CORM-2 (iCORM-2) group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group according to the random number table, with 3 tubes in each group. The PRP in control group did not receive any treatment. The PRP in LPS group received LPS (20 mL, 10 µg/mL) stimulation, and the PRP in iCORM-2 group, 10 µmol/L CORM-2 group, and 50 µmol/L CORM-2 group underwent the same LPS stimulation and treatment of 50 µmol/L iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2, respectively, with the dosage of 20 mL. After being cultured for 30 min, the platelet adhesion rate was determined by glass bottle method, the number of platelet spreading on fibrinogen was determined with immunofluorescent method, and the platelet aggregation rate was measured by turbidimetric method. The platelet poor plasma (PPP) was prepared from PRP, the levels of ATP in PPP and platelets were determined by chemical fluorescein method. The expressions of platelet glycoprotein I bα (GPIbα) and GPVI were analyzed by flow cytometer. The expressions of glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β were determined by Western blotting and immunoprecipitation, respectively. Measurement of the above indices was repeated for 3 times. Data were processed with one-way analysis of variance and SNK test.

RESULTSCompared with those in control group, the platelet adhesion rates, numbers of platelets spreading on fibrinogen, platelet aggregation rates, expressions of GPIbα and GPVI in PRP, levels of ATP in PPP in LPS and iCORM-2 groups were significantly increased, while levels of ATP in platelets were significantly decreased (with P values below 0.05). Compared with those in LPS group, the former 7 indices in iCORM-2 group showed no significant differences (with P values above 0.05), while the levels of ATP in platelets in the 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly increased, and the other 6 indices in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased (with P values below 0.05). The expression levels of GSK-3β of the platelets in PRP in control, LPS, iCORM-2, 10 µmol/L CORM-2, and 50 µmol/L CORM-2 groups were 0.550 ± 0.060, 1.437 ± 0.214, 1.210 ± 0.108, 0.720 ± 0.010, and 0.670 ± 0.010, respectively, and the expression levels of the phosphorylated GSK-3β of the platelets in PRP in the above 5 groups were 0.950 ± 0.070, 1.607 ± 0.121, 1.420 ± 0.040, 1.167 ± 0.015, and 0.513 ± 0.122, respectively. Compared with those in control group, both the expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in LPS and iCORM-2 groups were significantly increased (with P values below 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP between LPS group and iCORM-2 group were similar (with P values above 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in 10 µmol/L CORM-2 and 50 µmol/L CORM-2 groups were significantly decreased compared with those in LPS group (with P values below 0.05).

CONCLUSIONSLPS stimulation can abnormally activate the platelets in peripheral blood of healthy human, but the abnormal activation can be inhibited by CORM-2 intervention, and the mechanism of the latter may involve the phosphorylation of GSK-3β mediated by GP.

Blood Platelets ; drug effects ; metabolism ; Carbon Monoxide ; metabolism ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Humans ; Lipopolysaccharides ; pharmacology ; Organometallic Compounds ; pharmacology ; Phosphorylation ; drug effects ; Platelet Activation ; drug effects ; Platelet Aggregation ; drug effects ; Platelet-Rich Plasma

OBJECTIVE: The optimal multiplicity of infection (MOI) of the recombinant adenovirus Ad-Rad50-GFP carrying a mutant Rad50 gene expression region on the cell growth of nasopharyngeal carcinoma and the viral amplification efficiency of CNE1 cell infected by this adenovirus were studied. METHOD: The biological titer of Ad-Rad50-GFP was measured by end point dilution method. The impact of recombinant adenoviral vector transfection on the growth of CNE1 cells was observed by cell growth curve. Transfection efficacy of recombinant adenoviral vector was observed and calculated through fluorescence microscope. The expression f mutant Rad50 in the Ad-Rad50-GFP transfected CNE1 cells with optimal MOI was detected by Western Blot after transfection. RESULT: The biological titer of Ad-Rad50-GFP was 1.26 x 10¹¹ pfu/ml. CNE1 cell growth was not influenced significantly as they were transfected by recombinant adenoviral vector with MOI less than 50. Transfection efficacy of recombinant adenoviral vector was most salient at 24 hours after transfection, with the high expression of mutant Rad50, and the efficiency still remained about 70% after 72 hours. CONCLUSION Recombinant adenoviral vector Ad-Rad50-GFP could transfect CNE1 cells as well as result in the expression of mutant Rad50 in CNE1 cells effectively. MOI = 50 was the optimal multiplicity of infection of CNE1 cells transfected by recombinant adenoviral vector Ad-Rad50-GFP.
Adenoviridae ; Carcinoma ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; Transfection