1.Use of the ETV6/RUNX1 probe to verify the performance of the fluorescence in situ hybridization probe before clinical detection
Jing XIAO ; Yingchun ZHENG ; Jiawei ZHAO ; Chenghua CUI ; Huijun WANG ; Qi SUN ; Jiao MA ; Yueshen MA ; Zhen SONG ; Zhijian XIAO ; Chengwen LI
Chinese Journal of Hematology 2024;45(1):48-53
Objective:To explore the standardized performance of a FISH probe before clinical detection.Methods:The probe sensitivity and specificity of ETV6/RUNX1 were analyzed via interphase and metaphase FISH in 20 discarded healthy bone marrow samples. The threshold system of the probe was established using an inverse beta distribution, and an interpretation standard was established. Finally, a parallel-controlled polymerase chain reaction detection study was conducted on 286 bone marrow samples from patients at our hospital. The clinical sensitivity, specificity, and diagnostic coincidence rate of ETV6/RUNX1 FISH detection were analyzed, and the diagnostic consistency of the two methods was analyzed by the kappa test.Results:The probe sensitivity and specificity of the ETV6/RUNX1 probe were 98.47% and 100%, respectively. When 50, 100, and 200 cells were counted, the typical positive signal pattern cutoffs were 5.81%, 2.95%, and 1.49%, respectively, and the atypical positive signal pattern cutoffs were 13.98%, 9.75%, and 6.26%, respectively. The clinical sensitivity of FISH was 96.1%, clinical specificity was 99.6%, diagnostic coincidence rate was 99.00%, diagnostic consistency test kappa value was 0.964, and P value was <0.001.Conclusion:For FISH probes without a national medical device registration certificate, standardized performance verification and methodology performance verification can be performed using laboratory developed test verification standards to ensure a reliable and accurate reference basis for clinical diagnosis and treatment.
2.Analytic performance validation of laboratory-developed real-time quantitative PCR (RQ-PCR) test for BCR::ABL1 p210 transcript in chronic myeloid leukemia
Cong HAN ; Ge SONG ; Jiao MA ; Shicai XU ; Qi SUN ; Huijun WANG ; Zhijian XIAO ; Yao YAO
Chinese Journal of Laboratory Medicine 2023;46(10):1040-1047
Objective:To establish and validate analytic performance of laboratory-developed real-time quantitative polymerase chain reaction (RQ-PCR) test (LDT) for BCR::ABL1 p210 transcript.Methods:Using the primes and probes released by the Europe Against Cancer Program(EAC), we have established BCR::ABL1 p210 RQ-PCR test. The laboratory-specific conversion factor (CF) was determined by the WHO 1 st International Genetic Reference Panel, and a two-level internal control is developed using a mixture of K562 and HL60 cell lines was created to ensure traceability. Analytical performance, including analytical accuracy, analytical precision, linearity range, analytic sensitivity and specificity of RQ-PCR LDT test for BCR::ABL1 p210 transcript were validated according to CLIS guidelines. Furthermore, a comparison was made with an FDA-cleared RQ-PCR in vitro diagnostics (IVD) kit by Bland-Altman analysis. Results:The laboratory specific conversion factor (CF) for LDT RQ-PCR was determined to be 0.535 based on WHO 1 st International Genetic Reference Panel, which can be used to convert to the BCR::ABL international scale (BCR::ABL1 IS) reliably. The repeatability of BCR::ABL1 IS results at 4 different molecular response (MR0.5,MR1.5,MR2.5,MR3.5) levels are 7.44%, 5.33%, 9.12% and 18.06%, respectively, with total precision of 7.99%, 5.49%, 10.95% and 17.99%. The previous CAP proficiency test (PT) results from our laboratory were within the acceptable range of variation. MR results of our laboratory and MR mean value of all CAP-PT laboratory is highly correlated ( r=0.999, P<0.01), and consistent according to Bland-Altman analysis. Furthermore, the LDT method in our laboratory has a high correlation with the test results of FDA-cleared Qiagen IVD kit ( r=0.997, P<0.01). BCR::ABL1 IS results of BCR::ABL1 e13a2 transcript showed linearity within the range of 0.001%-7.454%, with a maximum coefficient of variation (% CV) 64.09%. The linearity range of e14a2 transcript BCR::ABL1 IS was 0.002%-12.398%, with a maximum % CV of 43.37%. The test has a limit of detection (LoD) of MR5.0 (0.001% IS) for e13a2 and MR4.8 (0.002% IS) for e14a2 transcript, respectively. The limit of quantitation (LoQ) for both e13a2 and e14a2 transcripts was MR4.7 (0.002% IS). The test exhibited 100% specificity, with no cross-reactivity observed between the p190 transcript and p210. Conclusions:The analytic performance of BCR::ABL1 p210 LDT RQ-PCR test from our laboratory is excellent, which can meet the clinical needs of BCR::ABL1 detection in patients with chronic myeloid leukemia.
3.Application of artificial intelligence assists bone marrow cytomorphology analysis in the diagnosis and treatment of acute myeloid leukemia
Jigang XIAO ; Huijun WANG ; Wenyu CAI ; Shuying CHEN ; Ge SONG ; Xulin LU ; Chenxi LIU ; Zhigang WANG ; Chao FANG ; Yanan CHEN ; Zhijian XIAO
Chinese Journal of Laboratory Medicine 2023;46(3):274-279
Objective:To investigate the value of artificial intelligence (AI) cytomorphologic analysis system in the cytomorphological diagnosis and therapeutic evaluation of acute myeloid leukemia (AML).Methods:Bone marrow smear samples were collected from 150 patients with newly diagnosed and treated acute myeloid leukemia who were inpatients and outpatients at the Department of Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from June 1, 2021 to July 31, 2022 for retrospective analysis. Among them, there were 50 patients in the newly diagnosed group, including 28 males and 22 females, with the onset age of 43.5(32.3,58.8)years. There were 100 patients in the post-treatment group, including 36 males and 64 females, with the onset age of 34.5(23.0,47.0)years. The results from cytomorphology expert were used as the gold standard and the Python 3.6.7 was used for analysis to evaluate the accuracy, sensitivity, and specificity of the AI cytomorphologic analysis system for blast cell recognition in AML diagnosis and treatment.Results:The proportion of blasts in AI analysis of 50 samples in the newly diagnosed group was≥20%, which met the diagnostic criteria of AML. AI analysis of blasts had an accuracy of 90.3%, sensitivity of 85.5%, and specificity of 98.0%. The correlation coefficient between AI and the proportion of blasts analyzed by experts was positively correlated( r=0.882, P<0.001). Meanwhile, in the post-treatment group, the sensitivity and specificity of AI analysis of blasts were 89.7% and 99.2%, respectively. The correlation coefficient between AI and the proportion of blasts analyzed by experts was positively correlated( r=0.957, P<0.001). According to AI analysis data, there are 8 samples in this group whose AI efficacy evaluation results on AML are inconsistent with expert analysis. Conclusion:AI cytomorphologic analysis system has high accuracy, sensitivity and specificity for blast cell recognition in AML morphological diagnosis and therapeutic evaluation.
4.Impact of different diagnostic criteria for assessing mild micro-hepatic encephalopathy in liver cirrhosis: an analysis based on a prospective, multicenter, real-world study
Xiaoyan LI ; Shanghao LIU ; Chuan LIU ; Hongmei ZU ; Xiaoqing GUO ; Huiling XIANG ; Yan HUANG ; Zhaolan YAN ; Yajing LI ; Jia SUN ; Ruixin SONG ; Junqing YAN ; Qing YE ; Fei LIU ; Lei HUANG ; Fanping MENG ; Xiaoning ZHANG ; Shaoqi YANG ; Shengjuan HU ; Jigang RUAN ; Yiling LI ; Ningning WANG ; Huipeng CUI ; Yanmeng WANG ; Chuang LEI ; Qinghai WANG ; Hongling TIAN ; Zhangshu QU ; Min YUAN ; Ruichun SHI ; Xiaoting YANG ; Dan JIN ; Dan SU ; Yijun LIU ; Ying CHEN ; Yuxiang XIA ; Yongzhong LI ; Qiaohua YANG ; Huai LI ; Xuelan ZHAO ; Zemin TIAN ; Hongji YU ; Xiaojuan ZHANG ; Chenxi WU ; Zhijian WU ; Shengqiang LI ; Qian SHEN ; Xuemei LIU ; Jianping HU ; Manqun WU ; Tong DANG ; Jing WANG ; Xianmei MENG ; Haiying WANG ; Zhenyu JIANG ; Yayuan LIU ; Ying LIU ; Suxuan QU ; Hong TAO ; Dongmei YAN ; Jun LIU ; Wei FU ; Jie YU ; Fusheng WANG ; Xiaolong QI ; Junliang FU
Chinese Journal of Hepatology 2023;31(9):961-968
Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.
5.Research progress on environmental concentration levels,population exposure and health risk of triclosan
XIE Huasen ; SONG Yang ; WANG Xiaofeng ; CHEN Zhijian ; XU Peiwei ; LOU Xiaoming
Journal of Preventive Medicine 2021;33(10):1013-1016
Triclosan, widely used in various personal care products, is one of the most common environmental endocrine disruptors in our life. It can be detected from water, soil and dust, and its environmental exposure level increased with consumption. Human body are exposed to triclosan through food and water, but the evidence that triclosan exposure leads to adverse health effects is not sufficient. This paper summarizes the progress of studies related to environmental concentration, population exposure and health risks of triclosan in recent years.
6.Protocatechuic Aldehyde Represses Proliferation and Migration of Breast Cancer Cells through Targeting C-terminal Binding Protein 1
Yu DENG ; Wanjun GUO ; Guancheng LI ; Shuang LI ; Hong LI ; Xinyan LI ; Bei NIU ; Mingzhu SONG ; Yamei ZHANG ; Zhijian XU ; Fulun LI
Journal of Breast Cancer 2020;23(1):20-35
PURPOSE:
C-terminal binding protein 1 (CtBP1) is a transcriptional co-repressor that is overexpressed in many cancers. CtBP1 transcriptionally represses a broad array of tumor suppressors, which promotes cancer cell proliferation, migration, invasion, and resistance to apoptosis. Recent studies have demonstrated that CtBP1 is a potential target for cancer therapy. This study was designed to screen for compounds that potentially target CtBP1.
METHODS:
Using a structure-based virtual screening for CtBP1 inhibitors, we found protocatechuic aldehyde (PA), a natural compound found in the root of a traditional Chinese herb, Salvia miltiorrhiza, that directly binds to CtBP1. Microscale thermophoresis assay was performed to determine whether PA and CtBP1 directly bind to each other. Further, clustered regularly interspaced short palindromic repeats associated Cas9 nuclease-mediated CtBP1 knockout in breast cancer cells was used to validate the CtBP1 targeting specificity of PA.
RESULTS:
Functional studies showed that PA repressed the proliferation and migration of breast cancer cells. Furthermore, PA elevated the expression of the downstream targets of CtBP1, p21 and E-cadherin, and decreased CtBP1 binding affinity for the promoter regions of p21 and E-cadherin in breast cancer cells. However, PA did not affect the expression of p21 and E-cadherin in the CtBP1 knockout breast cancer cells. In addition, the CtBP1 knockout breast cancer cells showed resistance to PA-induced repression of proliferation and migration.
CONCLUSION
Our findings demonstrated that PA directly bound to CtBP1 and inhibited the growth and migration of breast cancer cells through CtBP1 inhibition. Structural modifications of PA are further required to enhance its binding affinity and selectivity for CtBP1.
7.Trend of collaboration and sharing in digitalized restorative dentistry
Chinese Journal of Stomatology 2020;55(12):932-937
To digitalize restorative dentistry means to digitalize all the approaches involved in the clinical diagnosis and treatment process, in the production of individualized restorations, in the information transmission and communication between dentists and patients respectively between dentists and technicians, and even in the personalized understanding of esthetics. This is a cross-disciplinary issue that interconnects a wide range of topics. The way of understanding the digital tendency and of integrating the-already-digitalized steps into a system have a strategic impact on the development of the dental industry. By following the trace of the development of digital procedures in restorative dentistry, in this paper, we intend to classify the digitalization into two categories as the special and general ones, and will point out the limitations in understanding the digitalization in prosthodontics merely from the perspective of diagnosis or the production of individualized restorations. The role of digitalization in leading the conceptual development and the new possibilities opened up by the integrated digital technologies will be emphasized. Digitalization has changed the way of interaction between dentists and technicians. It is beneficial to the reorganization of medical and technological resources. Its potential in upgrading the dental industry deserves attention. Digitalization promotes the establishment of a collaborative and sharing ecosystem in restorative dentistry, which would pave the way towards artificial intelligence.
8.Application of total mesoesophageal excision combined with sanye lymph node dissection in radical resection of esophageal carcinoma under thoracoscopy and laparoscopy
Changyun MA ; Fang WU ; Yinfeng SONG ; Zhijian HUANG ; Jinwen LIAO
Clinical Medicine of China 2018;34(4):312-317
Objective To evaluate the clinical efficacy of total mesoesophageal excision and sanye lymph node dissection in the radical resection of esophageal carcinoma under thoracoscopy and laparoscopy,and to investigate its safety and feasibility, and to find a more reasonable and effective surgical treatment of esophageal carcinoma. Methods One hundred and twenty-six cases of esophageal cancer who underwent the minimally invasive surgery under thoracoscopy and laparoscopy for esophageal cancer in Central Hospital of Hengyang from October 2015 to September 2017 were retrospectively analyzed. Among them,Sixty-four patients accepted total mesoesophageal excision and sanye lymph node dissection under thoracoscopy and laparoscopy (observation group ), Sixty-two cases accepted with conventional esophagectomy under thoracoscopy and laparoscopy ( control group) . The operation time, blood loss, indwelling time of thoracic drainage tube, postoperative drainage volume,postoperative hospitalization time,number of lymph node dissection,lymph node metastasis degree,perioperative complications of the two groups were analyzed and compared. The number of lymph node dissection and lymph node metastasis degree in different regions were compared between the two groups. The number of recurrence and death were recorded in the two groups. Results Compared with the control group,the operation time was longer in the observation group((264. 9±32. 9) min vs. (233. 5±30. 4) min,t= -5. 56,P<0. 001),but blood loss was less((152. 7±26. 4) ml vs. (235. 5± 30. 6) ml,t = 16. 27,P<0. 001). There was no significant difference in the indwelling time of thoracic drainage tube, postoperative drainage volume or postoperative hospitalization time between the two groups (P>0. 05). The number of lymph nodes in the observation group was significantly higher than that in the control group ((32. 7±15. 5) pieces vs. (20. 9±11. 2) pieces,t = - 4. 93,P< 0. 001),and lymph node metastasis degree in the observation group was smaller than that of the control group ( 6. 7% vs. 9. 3%, χ2 = 7. 22, P < 0. 01) . There were no significant differences in perioperative complications such as pulmonary complications, arrhythmia, anastomotic fistula, chylothorax,hemorrhage,recurrent laryngeal nerve injury,tracheal injury and perioperative death (P>0. 05). Left and right recurrent laryngeal nerve,thoracic esophagus,celiac artery lymph node dissection of the number of observation group was higher than that of the control group ((4. 7 ± 3. 2) pieces vs. (1. 5 ± 1. 4) pieces, t= -7. 25;(6. 0±2. 7) pieces vs. (3. 1±1. 7) pieces,t = -7. 12;(5. 7± 2. 4) pieces vs. (3. 2± 1. 9) pieces,t= -6. 48;P<0. 001). Left and right recurrent laryngeal nerve,thoracic esophagus lymph node metastasis degree in the observation group was smaller than that in the control group (8. 7%(26/ 300) vs. 18. 1%(17/ 94),χ2= 6. 53;8. 9%(34/ 382) vs. 17. 9%(35/ 195),χ2 = 10. 04;P<0. 05) . There were no significant differences in the recurrence rate of tumor recurrence at 1 and 24 months after operation in the observation group and the control group(3 cases(4. 7%) vs. 4 cases(6. 5%),χ2 = 0. 92,P > 0. 05) . There were no deaths in the two groups. Conclusion Total mesoesophageal excision and three-field lymph node dissection in radical resection of esophageal carcinoma under thoracoscopy and laparoscopy is safe and feasible,the recent effect does not increase the surgical complications, but its long-term effect need a lot of long-term follow-up. A relatively thorough cleaning of the esophageal mesentery and its lymph nodes can minimize the tumor in the subendothelial micrometastasis,and is beneficial for the prognosis of patients with esophageal cancer.
9.Study on the Analgesic Effect and Mechanism of Gutongtie Paste on Model Rats with Formaldehyde-in-duced Pain
Yufei LI ; Jieying DU ; Sen ZENG ; Lu WANG ; Jianping SONG ; Qi WANG ; Qiaobo YE ; Zhijian ZHANG ; Jie YUAN
China Pharmacy 2017;28(13):1766-1769
OBJECTIVE:To study the analgesic effect and mechanism of Gutongtie paste on model rats with formaldehyde-in-duced pain. METHODS:60 SD rats were randomly divided into blank group,model group,Gutongtie paste low-dose,medi-um-dose,high-dose groups(0.594,1.188. 2.376 g/paste,containing crude drug 0.48,0.96,1.92 g)and prednisone acetate group (ig,0.0054 g/kg,external bonding matrix). Model rats with pain was induced by formaldehyde method and immediately adminis-trated after modeling. Electronic tenderness instrument was adopted to determine the pain threshold of rats'ankle joint after adminis-tration of 1,2,3,4,6 h. After 6 h,blood sample 0.3 mL was taken from abdominal aorta then rats were sacrificed. Enzyme linked immunosorbent assay (ELISA) was conducted to determine the β-endorphin (β-EP),prostaglandin E2 (PGE2) contents;spectrophotometry was used to determine nitric oxide(NO)content in rats'serum and inflammatory tissue;and radioimmunoassay was adopted to detect the substance P content in rats'serum,inflammatory tissue and brain tissue. RESULTS:Compared with be-fore modeling,pain thresholds in model group at each period were significantly decreased (P<0.05 or P<0.01). Compared with blank group,PGE2,NO of rats,substance P content in inflammatory tissue and brain tissue in model group were significantly in-creased (P<0.05 or P<0.01). Compared with model group,pain thresholds in Gutongtie paste groups at corresponding time points were increased,PGE2 and substance P contents in inflammatory tissue and brain tissue were decreased (P<0.05 or P<0.01);β-EP and NO contents in serum in Gutongtie paste medium-dose,high-dose groups(P<0.05 or P<0.01),NO contents in serum in Gutongtie paste high-dose group were decreased(P<0.05). CONCLUSIONS:Gutongtie paste has a certain analgesic and anti-inflammatory effect,and the mechanism may be related to reducing PGE2, NO, substance P contents, increasing β-EP content.
10.Optic coherence tomography-guided repeated intravitreal injections of conbercept for macular edema of retinal vein occlusion
Zhijian HUANG ; Yanping SONG ; Qing DIN ; Ming YAN
Chinese Journal of Ocular Fundus Diseases 2017;33(2):124-128
Objective To evaluate the effectiveness of repeated intravitreal conbercept injection in patients with macular edema (ME) of retinal vein occlusion (RVO),guided by optic coherence tomography (OCT).Methods It is a retrospective case study.Forty patients (40 eyes) diagnosed as ME secondary to RVO were enrolled in this study.There were 19 males (19 eyes) and 21 females (21 eyes),with the mean age of (53.58 ± 13.19) years and the mean course of 1.5 months.The best corrected visual acuity (BCVA),indirect ophthalmoscopy,fundus fluorescein angiography (FFA) and OCT were performed.The mean baseline of BCVA,central macular thickness (CMT) were 0.25 ± 0.18 and (509.48 ± 170.13) μm respectively.All the patients were treated with 10.00 mg/ml conbercept 0.05 ml (including conbercept 0.5 mg).Follow-up of these patients was 1 to 6 months after treatments,the BCVA,fundus manifestations,OCT were retrospectively observed by every month,the FFA was retrospectively observed by every 3 months.When there was retinal edema or CMT ≥ 50 μm by OCT during follow-up,those patients were retreated with intravitreal conbercept injection.The changes of the BCVA,CMT were evaluated before and after treatment.Meanwhile,complications in eyes related to medicine and treatment methods were evaluated too.Results At the 6 months,the BCVA was improved (increase≥2 lines) in 25 eyes (62.50%),stabilized (± 1 line) in 13 eyes (32.50%) and decreased 2 lines in 2 eyes (5.00%).Retinal hemorrhage and exudates were absorbed in most patients.FFA showed no fluorescein leakage in 1 1 eyes (27.50%),minor fluorescein leakage in 26 eyes (65.00%),and retinal capillary non-perfusion in 3 eyes (7.50%).OCT showed absorption of the subretinal fluid.The mean CMT were (235.20± 100.44) μm at 6 months.Intravitreal injection of conbercept was applied for 4 times in 8 eyes (20.00%),3 times for 18 eyes (45.00%),and 2 times for 14 eyes (35.00%).The mean number of intravitreal injection was 2.85 times.There were no ocular or systemic adverse events observed in all patients.Conclusion Intravitreal conbercept injection is an efficacy and safe treatment for the patients with ME of RVO guided by OCT.It can stabilize and improve the visual acuity.


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