1.Construction, breeding, and gene identification of TREM2 knockout mice
Rong Huang ; Xinxin Zhao ; Hui Xue ; Mengjuan Zhu ; Jiajie Tu ; Xinming Wang
Acta Universitatis Medicinalis Anhui 2025;60(6):977-983
Objective :
To construct triggering receptor expressed on myeloid cells 2(TREM2) gene knockout(TREM2-/-) mice using CRISPR/Cas9 technology, to breed TREM2-/- mice and to analyze the genotype of TREM2-/- mice.
Methods :
CRISPR/Cas9 technology was used to selectively knock out exon 2-3 regions of TREM2 gene to construct a TREM2-/- mouse model, and the genetic background of all mice was C57BL/6J. Polymerase chain reaction(PCR) was used to identify the genotype of mice. Quantitative real-time PCR(qPCR) and Western blot were used to detect the expression level of TREM2 in major tissues of mice, and the authenticity and scientific nature of PCR identification results were verified from mRNA level and protein level. According to the sgRNA sequence, the possible off-target sites were predicted on the CCTop website, and the tail DNA of mice was extracted and amplified by PCR and then Sanger sequencing was performed to detect whether there was off-target effect in TREM2-/- mice.
Results :
TREM2-/- mice were successfully constructed by CRISPR/Cas9 technology, and the mice were genotyped. The results of agarose gel electrophoresis showed that the mouse genotype with only 415 bp band amplified was wild type(WT), the mouse genotype of the 449 bp band amplified only was TREM2-/-, and the mouse genotypes amplified with 415 bp and 449 bp double bands were heterozygous. qPCR results showed that compared with WT mice, the mRNA expression of TREM2 in heart and brain tissues of TREM2-/- mice was down-regulated(P-/- mice was reduced(P-/- mice.
Conclusion
TREM2-/- mice are successfully constructed and bred, a reliable genotype identification method is established, the genetic stability of the mouse model is verified, which will provide an important genetic animal model for the study of TREM2 gene function.
2.Expert consensus on clinical application of 177Lu-prostate specific membrane antigen radio-ligand therapy in prostate cancer
Guobing LIU ; Weihai ZHUO ; Yushen GU ; Zhi YANG ; Yue CHEN ; Wei FAN ; Jianming GUO ; Jian TAN ; Xiaohua ZHU ; Li HUO ; Xiaoli LAN ; Biao LI ; Weibing MIAO ; Shaoli SONG ; Hao XU ; Rong TIAN ; Quanyong LUO ; Feng WANG ; Xuemei WANG ; Aimin YANG ; Dong DAI ; Zhiyong DENG ; Jinhua ZHAO ; Xiaoliang CHEN ; Yan FAN ; Zairong GAO ; Xingmin HAN ; Ningyi JIANG ; Anren KUANG ; Yansong LIN ; Fugeng LIU ; Cen LOU ; Xinhui SU ; Lijun TANG ; Hui WANG ; Xinlu WANG ; Fuzhou YANG ; Hui YANG ; Xinming ZHAO ; Bo YANG ; Xiaodong HUANG ; Jiliang CHEN ; Sijin LI ; Jing WANG ; Yaming LI ; Hongcheng SHI
Chinese Journal of Clinical Medicine 2024;31(5):844-850,封3
177Lu-prostate specific membrane antigen(PSMA)radio-ligand therapy has been approved abroad for advanced prostate cancer and has been in several clinical trials in China.Based on domestic clinical practice and experimental data and referred to international experience and viewpoints,the expert group forms a consensus on the clinical application of 177Lu-PSMA radio-ligand therapy in prostate cancer to guide clinical practice.
3.Feasibility and efficacy of pre-management of superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer
Shisheng LI ; Shiying ZENG ; Qinglai TANG ; Gangcai ZHU ; Danhui YIN ; Xia PENG ; Qian YANG ; Xinming YANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2024;59(2):127-132
Objective:To explore the feasibility and efficacy for the dissection and ligation of the superior laryngeal artery in endoscopic surgery for hypopharyngeal cancer.Methods:Eight cadaveric heads were selected, and the laryngopharynxes were harvested. The positions of the superior laryngeal arteries entering the larynxes were dissected and observed under endoscopic vision, and their anatomical characteristics were summarized. Twenty-nine patients (all were male, aged 39-74 years old) with hypopharyngeal cancer who underwent transoral endoscopic surgery at the Department of Otorhinolaryngology Head and Neck Surgery of the Second Xiangya Hospital, Central South University from January 2018 to December 2019 were selected, and the patients were randomly divided into two groups by drawing lots, namely, the superior laryngeal artery was actively dissected and occluded during surgery in observation group ( n=15) or not in control group ( n=14). The differences in surgical time, bleeding volume, postoperative complications, and postoperative disease-free survival rate were compared between the two groups. Statistical analysis was conducted using SPSS 25.0 software. Results:The entry point of the superior laryngeal artery into the larynx was approximately at the level of the superior edge of the thyroid cartilage, and entered the larynx at the posterior one-third of the lateral wall of the pyriform fossa. The superior laryngeal artery might be determined through endoscopic exploration in all patients of observation group. The endoscopic surgery time [(40.00±7.56) minutes] and intraoperative bleeding volume [(24.00±8.28) ml] in the observation group were respectively less than those [(48.57±14.06) minutes and (42.86±15.41) ml] in the control group, and the differences were statistically significant ( t=-2.064, P=0.049; t=-4.064, P=0.001). There was no case with postoperative bleeding in the observation group, but with one case of postoperative bleeding in the control group. Total disease free survival rate was 86.2% and there was no significant difference in disease free survival rates between the two groups during a follow-up period of at least 36 months ( P=0.986). Conclusion:Dissection of the superior laryngeal artery during endoscopic surgery for hypopharyngeal cancer is feasible, and pre-management and occlusion of the superior laryngeal artery can effectively reduce intraoperative bleeding.
4.Expert consensus on operational guideline for 68Ga-DOTATATE/18F-FDG two-day total-body PET/CT imaging of neuroendocrine neoplasms
Haojun YU ; Yushen GU ; Zhi YANG ; Ruimin WANG ; Hubing WU ; Xinming ZHAO ; Xiaohua ZHU ; Zhaoping CHENG ; Wei FAN ; Zhifang WU ; Sijin LI ; Hongcheng SHI
Chinese Journal of Clinical Medicine 2024;31(4):687-690,封3
68Ga-DOTATATE/18F-FDG two-day low-dose total-body PET/CT imaging is increasingly employed to facilitate the diagnosis,prognosis,and heterogeneity assessment of neuroendocrine neoplasms.We present a consensus on operational guideline for a two-day combined imaging from experts in low-dose/ultra-low-dose total-body PET/CT from several domestic medical institutions.
5.Comparative study of RECIL2017 and Lugano classification in prediction of prognosis at the end of treatment in patients with diffuse large B-cell lymphoma
Jianqiang ZHAO ; Xinming ZHAO ; Fenglian JING ; Huanxin ZHU ; Yifan AN ; Bingyao LIU
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(8):456-461
Objective:To compare the predictive values of response evaluation criteria in lymphoma (RECIL)2017 and Lugano classification for the prognosis of patients with diffuse large B-cell lymphoma(DLBCL) at the end of treatment.Methods:A total of 107 patients (50 males, 57 females, age (49.3±17.4) years) with DLBCL who underwent PET/CT at the end of chemotherapy in the Fourth Hospital of Hebei Medical University between February 2014 and December 2021 were analyzed retrospectively. The RECIL2017 and Lugano classification were used to evaluate the response. Kaplan-Meier survival analysis was used to evaluate progression-free survival (PFS) and overall survival (OS). The Kappa test was used to evaluate the consistency of the two criteria after chemotherapy, and ROC curve (Delong test)was used to compare the predictive values of the two criteria for PFS and OS. Results:The median follow-up time was 47.5(33.4, 57.5) months. Kaplan-Meier analysis showed that the 5-year PFS rates (74.5%(35/47), 71.4%(15/21), 57.1%(12/21), 4/18; χ2=38.85, P<0.001) and OS rates (89.4%(42/47), 81.0%(17/21), 61.9%(13/21), 7/18; χ2=29.52, P<0.001) in complete metabolic response (CMR), partial metabolic response (PMR), no metabolic response (NMR) and progressive metabolic disease (PMD) groups evaluated by Lugano classification were statistically different, as well as those in complete response (CR), partial response (PR), minor response (MR), stable disease (SD) and progressive disease (PD) groups evaluated by the RECIL2017 (5-year PFS rates: 76.9%(40/52), 8/12, 6/11, 6/12, 30.0%(6/20), χ2=29.05, P<0.001; 5-year OS rates: 90.4%(47/52), 8/12, 6/11, 9/12, 45.0%(9/20), χ2=23.63, P<0.001). The RECIL2017 and Lugano classification had good consistency in the efficacy evaluation of DLBCL patients at the end of chemotherapy (70.1%(75/107); Kappa=0.57, P<0.001). The AUCs of Lugano classification for predicting PFS and OS were 0.730 (95% CI: 0.625-0.834, P<0.001) and 0.908 (95% CI: 0.845-0.970, P<0.001) respectively, and those of RECIL2017 were 0.717 (95% CI: 0.612-0.822, P<0.001) and 0.880 (95% CI: 0.812-0.949, P<0.001). The AUCs of the Lugano classification for PFS and OS were slightly higher than those of RECIL2017, without significant differences ( z values: 0.44, 1.09, both P>0.05) . Conclusion:Both RECIL2017 and Lugano classification can evaluate the prognosis of patients with DLBCL at the end of treatment, and Lugano classification is more accurate.
6.Analysis of characteristics related to the disease activity of systemic lupus erythematosus and construction of an evaluation model.
Hongyan WANG ; Xinming LI ; Kechi FANG ; Huaqun ZHU ; Rulin JIA ; Jing WANG
Journal of Peking University(Health Sciences) 2024;56(6):1017-1022
OBJECTIVE:
To stratify systemic lupus erythematosus (SLE) patients clinically, to analyze the clinical characteristics of patients with and without disease activity, and to explore the application va-lue of key clinical indicators in assessing disease activity, as well as to construct an evaluation model.
METHODS:
A retrospective analysis was conducted on clinical data of the SLE patients diagnosed at Peking University People' s Hospital from May 1995 to April 2014. Demographic information, clinical manifestations, laboratory tests, and antibody detection results were collected. The patients were divided into active and inactive groups based on systemic lupus erythematosus disease activity index 2000(SLEDAI-2000)scores. t-tests, Mann-Whitney U tests, and χ2 tests were used to compare the differences between the groups. Spearman correlation analysis was used to evaluate the relevant clinical indicators associated with SLE activity in the active disease group. Based on the results of statistical analysis, a Logistic regression model was constructed, and the performance of the model was evaluated.
RESULTS:
No significant differences were found in demographic characteristics between the two groups. In the active disease group, positive rates of antinuclear antibodies (ANA) and anti-double-stranded DNA antibodies (anti-dsDNA) were increased; white blood cell count (WBC), red blood cell count (RBC), hemoglobin (HGB), lymphocytes (LY), total protein (TP), albumin (ALB), and complement 3(C3) levels were significantly decreased; while immunoglobulin A and G levels were markedly elevated. The correlation analysis results showed that hemoglobin, albumin, C3, and complement 4(C4) had higher correlation indices compared with other clinical indicators. Among these, C3 exhibited a certain negative correlation with disease activity. The Logistic regression model based on 12 significantly different indicators (P < 0.05) achieved an accuracy of 91.4%, sensitivity of 94.4%, specificity of 81.0%, and the area under curve (AUC) of the receiver operating characteristic (ROC) was 0.944.
CONCLUSION
This study comprehensively evaluated a range of clinical indicators related to SLE disease activity, providing a thorough understanding of both laboratory and clinical markers. The Logistic regression model, which was primarily constructed using laboratory test indicators, such as inflammatory markers, immune response parameters, and organ involvement metrics, demonstrated a high degree of accuracy in assessing the disease activity in SLE patients. Consequently, this model might provide a new basis for the diagnosis and treatment of SLE patients, offering significant clinical diagnostic value.
Humans
;
Lupus Erythematosus, Systemic/diagnosis*
;
Retrospective Studies
;
Antibodies, Antinuclear/blood*
;
Complement C3/metabolism*
;
Complement C4/metabolism*
;
Logistic Models
;
Severity of Illness Index
;
Leukocyte Count
;
Female
;
Male
;
Serum Albumin/analysis*
7.Feasibility and efficacy of preserving internal branch of superior laryngeal nerve in endoscopic surgery for hypopharyngeal squamous cancer: an observational study
Qian YANG ; Shisheng LI ; Qinglai TANG ; Xinming YANG ; Zi′an XIAO ; Xia PENG ; Gangcai ZHU ; Danhui YIN ; Peiying HUANG ; Shiying ZENG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(12):1463-1469
Objective:This study was performed to investigate the feasibility of preservation of internal branch of superior laryngeal nerve(ibSLN) during transoral endoscopic surgery for hypopharyngeal squamous cancer(HSCC) and the influence on patient′s swallowing function after operation.Methods:From May 2020 to June 2021, the data of 29 HSCC patients who required for transoral endoscopic surgery in the Department of Otorhinolaryngology Head and Neck Surgery, the Second Xiangya Hospital of Central South University were prospectively included, and the included patients were divided into two groups randomly by lottery. According to whether ibSLN was actively dissected during operation, they were divided into ibSLN preservation group ( n=15) and control group ( n=14, without ibSLN preservation). Operation time, intraoperative hemorrhage, intraoperative neck dissection, postoperative radiotherapy, postoperative recurrence within 1 year, retention and swallowing function, the recovery of oral soft diet and the quality of life were compared between two groups. SPSS 25.0 software was used for statistical analysis. Results:The study included 29 eligible patients, including 25 males and 4 females.The age ranged from 42 to 67 (56.07±5.93) years. There were no significant differences( P>0.05) between 2 groups in the following data,including age( t=-0.56), gender( χ2=0.01), TNM stage(T stage χ2=0.29, N stage χ2=0.02), pathological diagnosis( χ2=0.03), preoperative swallowing function( χ2=0.00) and M. D. Anderson Dysphagia Inventory(MDADI) score(global t=0.55, emotional t=0.16, functional t=0.60, physical t=0.64), operation time( t=1.62) and intraoperative hemorrhage( t=-1.46), intraoperative neck dissection( χ2=0.01), postoperative radiotherapy( χ2=0.32), postoperative recurrence within 1 year( P>0.050). The swallowing function was evaluated by water swallowing test after operation. The swallowing function of ibSLN preservation group was better than control group, and the difference between two groups was statistically significant on the 1st ( χ2=4.44, P=0.035), 5th ( χ2=4.24, P=0.039) and 7th ( χ2=4.55, P=0.033) day after operation. On the 14th day after operation, the MDADI scores of patients in the ibSLN preservation group were higher than those in the control group in global ( t=2.45, P=0.021), functional ( t=2.54, P=0.017) and physical ( t=2.24, P=0.034) dimensions, except for emotional dimension ( t=1.89, P=0.070). The median time of oral soft diet( U=23.00, P<0.001), normal oral diet( U=21.00, P<0.001) and the nasogastric tube removal time ( U=18.50, P<0.001) in ibSLN preservation group was 2 days, 5 days and 6 days respectively, earlier than that in control group, which had statistically significant difference. Conclusion:Our results show that it is feasible to preserve the ibSLN during HSCC transoral endoscopic surgery, which can achieve rapid recovery of postoperative swallowing function.
8.Clinical analysis of severe/critical 2019 novel coronavirus Omicron variant infection combined with atrial fibrillation
Zhihong ZHAO ; Saihua WANG ; Luoning ZHU ; Qiang HUAN ; Bei TIAN ; Shuwen HAO ; Zhongping NING ; Xiang SONG ; Xinming LI
Chinese Critical Care Medicine 2022;34(9):900-904
Objective:To investigate the clinical characteristics and prognosis of coronavirus disease 2019 (COVID-19) patients with Omicron variant combined with atrial fibrillation (AF).Methods:From March 23, 2022 to May 15, 2022, 2 675 aged ≥ 50 years old COVID-19 patients with AF were admitted to Zhoupu Hospital, the designated hospital for COVID-19 in Shanghai. Patients were divided into mild symptoms group, normal group, and serious/critical group according to the symptoms. The clinical data, imaging examination and laboratory results and prognosis of the three group patients were compared.Results:The median age of 2 675 COVID-19 patients was 69.0 (60.0, 81.0) years old, the incidence of AF was 5.05% (135/2 675), the age range of AF patients were from 55 to 101 years old, with a median age of 84.0 (74.0, 89.0), and the number of mild symptoms, normal, serious/critical patients were 68, 30, 37, respectively, including 9 of serious and 28 of critical patients. In the serious/critical patients, aged 55-75 years old accounted for 43.2%, the rate of 2019 novel coronavirus vaccination was 32.4%. The identified new-onset AF was the highest among the three groups, but the rate of persistent AF was the highest in the mild symptoms group (58.8%). The severe/critical group complicated with fever (29.7%), hepatic insufficiency (13.5%), renal insufficiency (46.0%), type 2 diabetes (46.0%), and heart failure were higher in NYHA classification [compared with the mild symptoms and normal group (score): 1.8±1.1 vs. 1.1±0.8, 1.2±0.7, respectively, all P < 0.05]. In term of laboratory examinations, C-reactive protein (CRP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were significantly higher in serious/critical patients compared to the mild symptoms and normal groups [CRP (mg/L): 27.2 (6.0, 60.8) vs. 7.6 (3.1, 19.3), 12.8 (4.9, 26.3), ALT (U/L): 31.3±15.4 vs. 15.4±9.3, 19.3±11.7, AST (U/L): 78.0±21.7 vs. 34.7±15.6, 38.1±24.4, all P < 0.05]. The hemoglobin (Hb) and albumin (ALB) levels were significantly lower than those in the mild symptoms and normal groups [Hb (g/L): 105.3±22.5 vs. 125.8±25.4, 123.0±20.4, ALB (g/L): 33.7±6.0 vs. 39.0±5.5 and 39.6±13.1, all P < 0.05]. In addition, MB isoenzyme of creatine kinase (CK-MB) was significantly higher in the serious/critical group than that in the mild symptoms group [μg/L: 2.5 (1.5, 3.4) vs. 2.2 (1.2, 2.8), P < 0.05]. In terms of the treatment, the percentage of antiplatelet agents and low-molecular heparin ratio compared among the three groups were statistically significant, with the serious/critical group using the lowest percentage of antiplatelet agents (27.0%) and a higher percentage of low-molecular heparin usage than that in mild symptoms group [81.1% (30/37) vs. 51.5% (35/68), P < 0.05]. In terms of prognosis, the mortality of patients with AF was 18.5% (25/135), all of whom were critical ill, including 32.0% (8/25) with cerebral embolism, pulmonary embolism and cerebral hemorrhage. Among them, 40.0% (10/25) died of multiple organ failure (40.0% combined with gastrointestinal hemorrhage), 20.0% (5/25) died of heart failure, and 12.0% (3/25) died of respiratory failure; while there were no death cases recorded in the mild symptoms, normal group and 9 serious patients. Conclusions:The serious/critical patients infected with COVID-19 Omicron variant with AF, have a worse prognosis and high mortality. Multiple organ failure, heart failure, sudden cardiac death, respiratory failure and embolic disease are the major causes of death.
9.Value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid enhanced magnetic resonance imaging and diffusion-weighted MR imaging in predicting microvascular invasion in hepatocellular carcinoma and the prognostic significance
Yongjian ZHU ; Bing FENG ; Bingzhi WANG ; Shuang WANG ; Feng YE ; Xiaohong MA ; Xinming ZHAO
Chinese Journal of Oncology 2021;43(3):312-317
Objective:To investigate the combined value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced magnetic resonance imaging (MRI) and diffusion-weighted imaging (DWI) in predicting pathological microvascular invasion (pMVI) preoperatively, and to determine the relationship between prediction results and prognosis in hepatocellular carcinoma (HCC) patients.Methods:A total of 181 newly diagnosed HCC patients were enrolled in this study. Imaging characteristics and the apparent diffusion coefficient (ADC) value of DWI were analyzed. The differences of imaging characteristics and ADC values between different pMVI groups were analyzed.Multivariate logistic regression and receiver operating characteristic (ROC) curve were used to analyze the value for pMVI prediction by using significant parameters. The patients were grouped based on MRI predicted MVI (mrMVI), and the relationship between mrMVI and recurrence free survival time (RFS) was analyzed.Results:Fifty-one patients were pMVI positive and 130 patients were pMVI negative. The ADC value in pMVI positive group were (1.10±0.17)×10 -3 mm 2/s, significantly lower than (1.27±0.22)×10 -3 mm 2/s of pEMVI negative group ( P<0.001). The incidence rates of incomplete enhancing "capsule" , non-smooth tumor margin, arterial peritumoral enhancement, mosaic architecture and peritumoral hypointensity on hepatobiliary phase (HBP) in pMVI positive group were significantly higher than those of negative group (all P<0.05). Multivariate logistic regression analysis showed that tumor margin, arterial peritumoral enhancement, peritumoral hypointensity on HBP and ADC value were independently associated with pMVI. ROC analysis showed that the area under curve, sensitivity and specificity of pMVI predicted by combined parameters were 0.830, 76.5% and 81.5%, respectively. The median RFS of mrMVI positive group was 23.6 months, significantly lower than 38.2 months of mrEMVI negative group ( P=0.004). Conclusion:Tumor margin, arterial peritumoral enhancement, peritumoral hypointensity on HBP and ADC value are independent predictors of pMVI in HCC, and mrMVI is related with RFS.
10.Value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid enhanced magnetic resonance imaging and diffusion-weighted MR imaging in predicting microvascular invasion in hepatocellular carcinoma and the prognostic significance
Yongjian ZHU ; Bing FENG ; Bingzhi WANG ; Shuang WANG ; Feng YE ; Xiaohong MA ; Xinming ZHAO
Chinese Journal of Oncology 2021;43(3):312-317
Objective:To investigate the combined value of gadolinium ethoxybenzyl diethylenetriamine pentaacetic acid (Gd-EOB-DTPA) enhanced magnetic resonance imaging (MRI) and diffusion-weighted imaging (DWI) in predicting pathological microvascular invasion (pMVI) preoperatively, and to determine the relationship between prediction results and prognosis in hepatocellular carcinoma (HCC) patients.Methods:A total of 181 newly diagnosed HCC patients were enrolled in this study. Imaging characteristics and the apparent diffusion coefficient (ADC) value of DWI were analyzed. The differences of imaging characteristics and ADC values between different pMVI groups were analyzed.Multivariate logistic regression and receiver operating characteristic (ROC) curve were used to analyze the value for pMVI prediction by using significant parameters. The patients were grouped based on MRI predicted MVI (mrMVI), and the relationship between mrMVI and recurrence free survival time (RFS) was analyzed.Results:Fifty-one patients were pMVI positive and 130 patients were pMVI negative. The ADC value in pMVI positive group were (1.10±0.17)×10 -3 mm 2/s, significantly lower than (1.27±0.22)×10 -3 mm 2/s of pEMVI negative group ( P<0.001). The incidence rates of incomplete enhancing "capsule" , non-smooth tumor margin, arterial peritumoral enhancement, mosaic architecture and peritumoral hypointensity on hepatobiliary phase (HBP) in pMVI positive group were significantly higher than those of negative group (all P<0.05). Multivariate logistic regression analysis showed that tumor margin, arterial peritumoral enhancement, peritumoral hypointensity on HBP and ADC value were independently associated with pMVI. ROC analysis showed that the area under curve, sensitivity and specificity of pMVI predicted by combined parameters were 0.830, 76.5% and 81.5%, respectively. The median RFS of mrMVI positive group was 23.6 months, significantly lower than 38.2 months of mrEMVI negative group ( P=0.004). Conclusion:Tumor margin, arterial peritumoral enhancement, peritumoral hypointensity on HBP and ADC value are independent predictors of pMVI in HCC, and mrMVI is related with RFS.


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