1.Explore of nanopore sequencing technology in ambiguities of HLA genotyping
Nanying CHEN ; Wei ZHANG ; Lina DONG ; Fang WANG ; Yizhen HE ; Chen CHEN ; Faming ZHU
Chinese Journal of Blood Transfusion 2025;38(3):309-315
[Objective] To resolve the ambiguities of HLA genotyping generated by next generation sequencing (NGS) using nanopore sequencing technology. [Methods] A total of 38 samples with ambiguous HLA genotyping by NGS in our laboratory were collected, and HLA-A, -B, -C, -DRB1, -DRB3/4/5, -DQA1, -DQB1, -DPA1 and -DPB1 loci in these samples were amplified using primers in the same commercial NGS HLA genotyping kit, then subjected to third-generation library construction, and sequenced on the nanopore sequencer. The sequencing data were converted into Fastq files and analyzed by software, and the genotypes of 11 HLA loci were obtained. The ambiguities were counted directly. [Results] The high-resolution genotyping at the second domain of 11 HLA loci of 38 samples using the third generation sequencing (TGS) were consistent with the results of the NGS method at a rate of 100%. The genotypes for the HLA-A, -B, -C, -DRB3, -DRB4, -DQA1 and -DPA1 loci by TGS were all only one result, and the discrimination rate for ambiguities of the HLA-A, -B, -C, and -DQA1 loci (all caused by the difficulty in phasing due to the short NGS read length) was 100%. Among the HLA-DRB1, -DRB5, -DQB1 and -DPB1 loci, the discrimination rate of TGS for the ambiguities caused by non-amplification of exon 1 was 0% and by the short NGS read length was 100%. [Conclusion] Nanopore technology was used to identify the ambiguities of 11 HLA loci in this study, and the ambiguities caused by the short read length disadvantage of the NGS method could be solved effectively and the accuracy of HLA genotyping would be improved.
2.Design and application of clinical probation log based on subjective-objective-assessment-plan
Zhifeng WU ; Li MING ; Zhangya YANG ; Min WANG ; Yizhen SUN ; Wang YANG ; Zhiqiang CHEN ; Zhangxue HU ; Yuping ZHANG
Chinese Journal of Medical Education Research 2024;23(7):906-909
Objective:To investigate the application value of a new clinical probation log designed based on the subjective-objective-assessment-plan (SOAP) structured medical record.Methods:Quantitative and qualitative studies were conducted among 97 students in the autumn semester to evaluate the effect of the new clinical probation log based on the SOAP structured medical record on their academic performance and clinical thinking. SPSS 26.0 was used to perform the t-test and ANOVA. Results:The students using the new version had a significantly better score (96.29±1.38) than those using the old version (93.53±1.60) ( P<0.001), while the multivariate analysis showed that the students who first used the old version and then switched to the new version had a significantly better improvement in the score compared with those who first used the new version and then switched to the old version ( P<0.001). The qualitative interview showed that it was necessary to use handwritten internship log, and compared with the old version, the new version could better promote the ordered clinical thinking of students. The teacher comments could give feedbacks, and all the students interviewed thought that the new version held promise for clinical application. Conclusions:The clinical probation log based on the SOAP structured medical record can help to improve the effectiveness of probation and cultivate clinical thinking ability, and thus it holds promise for application in clinical probation teaching.
3.Efficacy and safety of prednisone combined with standard quadruple antituberculosis therapy in the treatment of tuberculous pleurisy
Hongyan XU ; Tianxiang ZHANG ; Honghong GU ; Wei MA ; Yizhen HAN ; Qiyuan WANG
Chinese Journal of Primary Medicine and Pharmacy 2024;31(10):1507-1511
Objective:To investigate the efficacy and safety of prednisone combined with standard quadruple antituberculosis therapy (HRZE) in the treatment of tuberculous pleurisy.Methods:A prospective study was conducted involving 120 patients with tuberculous pleurisy who were admitted to the Shaanxi Provincial Tuberculosis Prevention and Control Hospital from February 2021 to February 2023. The patients were randomly assigned to a study group and a control group, with 60 patients in each group, using a computer-generated randomization method. The control group received HRZE alone, while the study group received prednisone therapy and HRZE. The efficacy, clinical indicators, adverse reactions, and serum inflammatory factor levels were compared between the two groups.Results:The total response rate in the study group was significantly higher than that in the control group [93.33% (56/60) vs. 78.33% (47/60), χ2 = 5.55, P < 0.05). In the study group, the time for clinical symptom improvement was (10.34 ± 1.65) days, the time for pleural effusion absorption was (21.37 ± 4.16) days, the pleural thickness measured (2.15 ± 0.35) mm, and the duration of hospitalization was (23.19 ± 4.56) days. They were significantly shorter or smaller than those in the control group [(13.27 ± 2.30) days, (27.25 ± 4.95) days, (2.62 ± 0.40) mm, (28.42 ± 5.60) days, t = 8.02, 7.04, 6.85, 5.61, all P < 0.05]. There was no significant difference in the incidence of adverse reactions between the two groups (χ2 = 2.91, P > 0.05). After 8 weeks of treatment, all serum inflammatory factors improved in both groups compared with baseline levels. In the study group, levels of interleukin-6 [(90.37 ± 12.05) ng/L] and interleukin-18 [(270.94 ± 14.58) ng/L] were significantly lower than those in the control group [(110.59 ± 16.90) ng/L, (296.10 ± 25.29) ng/L, t = 7.55, 6.68, both P < 0.05]. Levels of interleukin-10 [(78.91 ± 8.25) ng/L] and soluble interleukin-2 receptor [(1875.82 ± 359.23) pg/L] in the study group were significantly higher than those in the control group [(70.40 ± 7.16) ng/L, (1566.87 ± 311.02) pg/L, t = -6.03, -5.04, both P < 0.05]. Conclusion:The combination of prednisone and HRZE demonstrates good efficacy and safety, and it is beneficial for improving inflammatory factors.
4.Research progress on the effects of Δ9-tetrahydrocannabinol on the blood-brain barrier
Qianyao ZHANG ; Hao CHENG ; Yizhen HUANG ; Hanxin TENG ; Yue ZHANG ; Ruilin ZAHNG
Chinese Journal of Comparative Medicine 2024;34(3):120-126
The psychoactive properties of cannabinoids are well known,and there are controversies over whether cannabinoids can be used for therapeutic purposes worldwide.Δ9-tetrahydrocannabinol(THC)is the main psychoactive substance in cannabis.The neurological mechanisms of THC were only recently discovered,and its neurological mechanism of action is still not fully understood.The blood-brain barrier(BBB)is a very important structure protecting the brain and is the first line of defense preventing foreign substances from entering the brain.THC's lipophilic nature and its interaction with the endocannabinoid system make it more likely to act on the BBB.In this paper,we review the neurotoxic effects of THC,focusing on its effect and mechanism of action on the BBB,and provide a theoretical basis for studies elucidating the neural mechanism of THC.
5.Preclinical study of a novel molecular probe 89Zr DFO-G4C2 for monitoring PD-1 expression levels
Yirong ZHU ; Weiwei KONG ; Jiaxi YOU ; Kairu NI ; Bing ZHANG ; Zengli LIU ; Yizhen SHI ; Zhihui HONG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2024;44(10):603-608
Objective:To design and synthesize 89Zr-deferoxamine(DFO)-G4C2, a novel molecular probe targeting programmed cell death receptor 1(PD-1), and evaluate its in vivo biodistribution and microPET/CT imaging characteristics in tumor-bearing mice. Methods:DFO-G4C2 was prepared by coupling DFO with G4C2, a monoclonal antibody targeting PD-1. The affinity and binding specificity of this amalgamation were subsequently assessed through the implementation of flow cytometry and surface plasmon resonance techniques. The molecular probe 89Zr-DFO-G4C2 was achieved by labeling DFO-G4C2 with the radioisotope 89Zr, and the labeling efficiency and in vitro stability of 89Zr-DFO-G4C2 were determined. Mouse models laden with CT26 colorectal cancer cells expressing PD-1 were established, followed by in vivo biodistribution and microPET/CT imaging studies, to explore the potential clinical value of 89Zr-DFO-G4C2. Additionally, the validity of this molecular probe was verified in 4T1 breast cancer models, affirming its efficacy as an imaging tool across different tumor models. Independent-sample t test was used to analyze the data. Results:DFO-G4C2 exhibited an affinity constant KD of (0.55±0.02) μmol/L, indicating a strong binding affinity. The binding rate to mouse PD-1 protein was determined to be (61.82±8.49)%. The labeling rate of 89Zr-DFO-G4C2 reached a high level of (98.76±0.51)%. Furthermore, the labeling rates in lysate and human serum after 144 h were measured to be (93.07±2.16)% and (83.42±3.21)%, respectively. MicroPET/CT imaging of CT26 tumor-bearing mice injected with 89Zr-DFO-G4C2 showcased pronounced radioactivity uptake in the tumor tissue. At 72 h post-injection, the tumor uptake value reached (10.47±0.34) percentage activity of injection dose per gram of tissue (%ID/g). The tumor uptake observed in the blocked experimental group, wherein an excess of unlabeled antibody was administered, was significantly lower at (6.26±1.03) %ID/g in comparison to the non-blocked group ( t=6.67, P=0.003). The in vivo biodistribution results were consistent with the observed microPET/CT imaging outcomes. MicroPET/CT imaging observations in the 4T1 breast cancer bearing mouse model were analogous to those obtained from the CT26 model. Conclusion:ImmunoPET based on the 89Zr-DFO-G4C2 molecular probe can non-invasively and visually assess the PD-1 expression level of tumors in vivo, and it is expected to be a new molecular imaging technique for immunotherapy monitoring of PD-1 inhibitors.
6.Comparison of clinical characteristics between first-episode and recurrent acute hypertrigly-ceridemic pancreatitis: a national multicenter clinical research
Shuai LI ; Jing ZHOU ; Guixian LUO ; Hongwei ZHANG ; Siyao LIU ; Weijie YAO ; Donghuang HONG ; Kaixiu QIN ; Lanting WANG ; Rong WEI ; Yizhen XU ; Longxiang CAO ; Zhihui TONG ; Yuxiu LIU ; Weiqin LI ; Lu KE
Chinese Journal of Digestive Surgery 2024;23(5):703-711
Objective:To investigate the clinical characteristics of first-episode and recurrent acute hypertriglyceridemic pancreatitis (HTGP).Methods:The retrospective cohort study was con-ducted. The clinical data of 313 patients with HTGP admitted to 26 medical centers in China in the Chinese Acute Pancreatitis Clinical Research Group (CAPCTG)-PERFORM database from November 2020 to December 2021 were collected. There were 219 males and 94 females, aged 38(32,44)years. Of the 313 patients, 193 patients with first-episode HTGP were allocated into the first-episode group and 120 patients with recurrent HTGP were allocated into the recurrent group. Observation indica-tors: (1) propensity score matching and comparison of general data of patients between the two groups after matching; (2) comparison of severity and prognosis in the course of disease within 14 days between the two groups; (3) the association between recurrent HTGP and the risk of persistent organ failure (POF); (4) follow-up. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M( Q1, Q3), and comparison between groups was conducted using the Wilcoxon rank sum test. Count data were expressed as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data was conducted using the Wilcoxon rank sum test. The Kaplan-Meier method was used to plot the cumulative recurrence rate curve and Log-Rank test was used for survival analysis. The Logistic regression model was used for multivariate analysis, and continuous variables were converted into categorical variables according to the mean value or common criteria. Propensity score matching was performed by 1∶1 nearest neighbor matching method, with caliper value of 0.02. Paired t test or Wilcoxon rank sum test and McNemar′s test were used for comparison between matched groups. Results:(1) Propensity score matching and comparison of general data of patients between the two groups after matching. Of the 313 patients,208 cases were successfully matched, including 104 cases in the first-episode group and 104 cases in the recurrent group. After propensity score matching, there was no significant difference in demographic characteristics, severity of illness scores and laboratory test between the two groups ( P>0.05). The elimination of gender, acute physiology and chornic health evaluation (APACHE) Ⅱ score, computed tomography severity index score, systemic inflammatory response syndrome score, sequential organ failure assessment score, apolipoprotein E, C-reactive protein, creatinine, lactic acid dehydrogenase, procal-citonin confounding bias ensured comparability between the two groups. (2) Comparison of severity and prognosis in the course of disease within 14 days between the two groups. There were signifi-cant differences in POF and local complications between the first-episode group and the recurrent group ( P<0.05). (3) The association between recurrent HTGP and the risk of POF. Results of uncor-rected univariate analysis showed that there was no association between recurrent HTGP and the risk of POF ( odds ratio=0.78, 95% confidence interval as 0.46-1.30, P>0.05). Results of multivariate analysis after adjusting for covariates such as gender, age, APACHE Ⅱ score, C-reactive protein, triglyceride and total cholesterol showed that compared with first-episode HTGP, recurrent HTGP was associated with a higher risk of POF ( odds ratio=2.22, 95% confidence interval as 1.05-4.71, P<0.05). Results of subgroup analysis showed that age<40 years was associated with an increased risk of POF ( odds ratio=3.31, 95% confidence interval as 1.09-10.08, P<0.05). (4) Follow-up. Twelve of the 313 patients died during hospitalization, including 9 cases in the first-episode group and 3 cases in the recurrent group. The rest of 301 surviving patients, including 184 cases in the first-episode group and 117 cases in the recurrent group, were followed up for 19.2(15.5, 21.9)months. Results of follow-up showed that for 184 survived patients of the first-episode group, 164 cases were followed up and 24 cases experienced recurrence, for 117 survived patients of the recurrent group,29 cases experienced recurrence, showing a significant difference between the two groups ( χ2=4.67, P<0.05). Conclusion:Compared with first-episode HTGP, patients with recurrent HTGP are more prone to POF and local complications, and are more prone to recurrence after discharge. The risk of POF in recurrent HTGP patients is 2.22 times that of those with first-episode, and the risk is higher in patients with age <40 years.
7.Clinical application value of a predictive model for the efficacy of third-generation cephalosporin in treatment of community-acquired spontaneous bacterial peritonitis
Longchuan ZHU ; Wei WU ; Dakai GAN ; Wei ZHANG ; Yizhen XU ; Molong XIONG
Journal of Clinical Hepatology 2024;40(2):306-311
ObjectiveTo investigate the clinical application value of a predictive model for the efficacy of third-generation cephalosporin in the treatment of community-acquired spontaneous bacterial peritonitis (CASBP). MethodsThis prospective study was conducted among 50 patients with liver cirrhosis and CASBP who were admitted to The Ninth Hospital of Nanchang from January 2021 to June 2022, and the patients were randomly divided into optimized treatment group and traditional treatment group, with 25 patients in each group. The patients in the optimized treatment group received ceftazidime or imipenem for initial treatment based on the above predictive model, and those in the traditional treatment group received ceftazidime for initial treatment, with the subsequent use of antibiotics adjusted based on the efficacy of initial treatment. The two groups were compared in terms of the response rate of initial treatment, cure rate on day 5, and 30-day mortality rate. The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. ResultsAll patients completed the study. The optimized treatment group had a significantly higher response rate of initial treatment than the traditional treatment group (88.0% vs 60.0%, χ2=5.094, P=0.024), while there was no significant difference in the cure rate on day 5 between the two groups (80.0% vs 56.6%, χ2=3.309, P=0.069). As for the patients who received ceftazidime for initial treatment, the optimized treatment group had a significantly higher response rate of initial treatment than the traditional treatment group (88.9% vs 60.0%, χ2=4.341, P=0.037), while there was no significant difference in the cure rate on day 5 between the two groups (83.3% vs 56.0%, χ2=2.425, P=0.119). There was no significant difference in 30-day mortality rate between the two groups (8.0% vs 20.0%, χ2=0.664, P=0.415). For all patients, there was a significant association between response of initial treatment and cure on day 5 (odds ratio [OR]=9.643, 95% confidence interval [CI]: 2.292 — 40.564) and between cure on day 5 and 30-day mortality (OR=0.138, 95%CI: 0.023 — 0.813). ConclusionThis predictive model for efficacy helps clinicians to identify the patients who can benefit from third-generation cephalosporin treatment and improve the efficacy of third-generation cephalosporin in the initial empirical treatment of CASBP.
8.Key Information Research and Contemporary Research Progress of Classical Formula Jinlingzi San
Yizhen WANG ; Lyuyuan LIANG ; Jialei CAO ; Yihang LIU ; Rongze MA ; Zhengshao ZHANG ; Jing TANG ; Bingqi WEI ; Chongyi HUANG ; Hejia WAN ; Bingxiang MA ; Wenli SHI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(24):215-223
Jinlingzi San is a formula frequently used in treating pain syndrome, first recorded in the Collection of Writings on the Mechanism of Disease, Suitability of Qi, and the Safeguarding of Life as Discussed in the 'Basic Questions' written by LIU Wansu in the Jin Dynasty. Jinlingzi San is composed of 2 Chinese medicinals Toosendan Fructus and Corydalis Rhizoma with a concise composition and exact clinical efficacy, having been included in the Catalogue of Ancient Classical Formulas (Second Batch: Han Chinese Medicine). The formula name, historic evolution, medicine origins, composition, dosage, decocting methods, and ancient and modern clinical application were sorted out and analyzed with the bibliometric method. A total of 209 pieces of information were collected from ancient books and literature. After screening, 49 pieces of effective data involving 45 ancient books were included. Results showed that the name of Jinlingzi San was first recorded in Secret Formulas of the Yang Family written by Yang Tan in the Southern Song Dynasty and developed into 3 other versions of the decoction. The Jinlingzi San included in the Collection of Writings on the Mechanism of Disease, Suitability of Qi, and the Safeguarding of Life as Discussed in the 'Basic Questions' written by LIU Wansu invariably plays a dominant role. As for the 3 other versions, although they have the same name of Jinlingzi San, their composition and indications are different from those of the original formula, which were therefore viewed as prescriptions based on Jinlingzi San and also included in the research. The medicine origins and processing of Jinlingzi San are suggested: Toosendan Fructus is the dry mature fruit of Melia toosendan of Meliaceae, and the crude is used after cleansing without putamen. Corydalis Rhizoma is the dry tuber of Corydalis yanhusuo of Papaveraceae, which is used after impurity removal, cleaning, and drying. Depending on the conversion from the measurement system in the Jin Dynasty to modern measurement, it is suggested that Toosendan Fructus and Corydalis Rhizoma (41.3 g each) are ground into fine powder, and one dose includes 12.39 g of the powder, which should be taken with an appropriate amount of wine. If wine is not suitable for the patient, the decoction can also be taken with warm water. Jinlingzi San has the effects of soothing the liver, discharging heat, and activating blood to stop pain. As recorded in ancient books, Jinlingzi San is specialized in treating heart pain caused by reversal heat, chest and abdominal pain, hypochondriac pain, jaundice, hernia, and other diseases. Modern studies have shown that modified Jinlingzi San can be used in treating diseases involving the digestive system, the integumentary system, the gynecological system, the reproductive system, and other systems and has wide clinical application in treating epigastric pain, herpes zoster, dysmenorrhea, and other diseases. This study has made clear the key information of Jinlingzi San by textual research of ancient books and literature in the hope of providing a theoretical reference for the clinical application, set prescriptions, and new drug development.
9.Establishment and Preliminary Application of Competence Framework for Clinical Teaching Management Positions Based on Nominal Group Technique
Yizhen WEI ; Duoduo ZHANG ; Qiang WANG ; Xiao CHANG ; Meini CHEN ; Linzhi LUO
Medical Journal of Peking Union Medical College Hospital 2024;15(3):718-723
To develop a framework for clinical teaching management (CTM) position competencies, and to initiate the preliminary application of this framework. Key groups comprising clinical teaching coordinators, clinical trainees, teaching administrators, and the educational reform practice and research group at Peking Union Medical College Hospital (PUMCH) were selected as research samples. The nominal group technique (NGT) was employed to generate, integrate, and establish a framework, as well as to describe CTM competencies. Additionally, rankings of competency importance were gathered. The competency framework was then utilized to conduct a self-assessment of CTM through a questionnaire survey, scored using the Likert method (ranging from 1 to 5 points), followed by an analysis of the results. The initial NGT discussion round among key groups led to a consensus on the primary CTM competency framework, which included professionalism, coordinating ability, teaching ability, and teaching innovation. Divergent views emerged regarding the connotations and importance rankings of each dimension. Following the second NGT discussion by the educational reform practice and research group, a self-assessment was carried out using the CTM competency framework. This second discussion round resulted in a competency framework for clinical teaching management positions that encompassed seven dimensions: medical knowledge and skills, learner-centeredness, communication and cooperation, professionalism and role modeling, reflection and improvement, teaching theory and methods, and teaching coordination and management. The self-assessment revealed that, aside from the dimensions of teaching coordination and management and teaching theory and methods, the scores for the other five dimensions were above 80% (4 points). A preliminary CTM competency framework has been established and applied. The results provide significant insights for guidiny the enhancement and evaluation of competencies.However, specific items within this framework require further validation and refinement through practical application.
10.Establishment and application of drug use evaluation criteria of fibrinogenase for injection
Ruiqin REN ; Jing ZHANG ; Yizhen GONG ; Jinhua ZHANG
Chinese Journal of Pharmacoepidemiology 2024;33(2):145-151
Objective To establish the drug use evaluation(DUE)standard of fibrinogenase for injection and provide a reference for the rational clinical application of fibrinogenase for injection.Methods Based on the specification of fibrinogenase for injection,the DUE standard was established from three aspects:drug indication,drug process and drug results,with reference to relevant guidelines and literature,and through discussion with clinical experts.A retrospective survey was conducted to evaluate the inpatients using fibrinogenase for injection from January 2021 to December 2021 in Ningde Hospital of Traditional Chinese Medicine,Fujian Province.Results A total of 256 patients were included,with a medication reasonable rate of 61.72%.The irrational use of drugs was mainly including the inappropriate usage and dosage(3.91%),off-label medication(1.95%),no skin test(8.98%),too long or inadequate course of medication(25.00%).Conclusion The DUE standard established of fibrinogenase for injection is scientific,practical and feasible.The use of fibrinogenase for injection in contraindications and high blood coagulation state,and off-label medication can be further optimized.

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