Gene editing technology utilizes artificial nucleases to insert, replace, or delete specific sequences in desired genomic regions. The discovery of CRISPR/Cas9 nucleases was a milestone in the development of advanced gene editing tools, which revolutionized the field due to their simplicity and versatility. However, the limited precision of Cas9 nucleases remains a notable obstacle. Recently, derivative technologies such as prime editing have earned considerable attention for their enhanced efficiency and precision. The prime editing system consists of two components: the SpCas9 nickase (H840A) fused with reverse transcriptase (MLV-RT) and an engineered prime editing guide RNA (pegRNA). This system can irreversibly introduce various types of genetic changes into the genome, including 12 possible types of point mutations, as well as insertions, deletions and their combinations, without the need for DNA double-strand breaks (DSBs) or donor DNA templates. Prime editing offers several advantages in terms of editing accuracy, versatility, PAM constraints, and off-target effects. The editing results of prime editing system is highly accurate and can be tailored to specific needs. In addition, the system can be edited near or far from PAM sites, making it less constrained by PAM site restrictions. Moreover, it demonstrates high genome-wide specificity. The system also supports a variety of edits, demonstrating immense potential, especially in large DNA fragment editing—an area that relied heavily on CRISPR/Cas9 nucleases before. The development of prime editing, especially bi-direction prime editor, shed new light on large DNA fragment manipulations, including deletions, insertions, replacements, gene integration, as well as chromosomal translocations, inversions, and tandem duplications. Despite the significant progress made with prime editing technology, its application still faces challenges, especially low editing efficiency, which limits its potential in broader research and clinical settings. Consequently, researchers are exploring strategies to enhance the efficiency of prime editing. This review highlights several approaches to improving prime editing efficiency. These include optimizing pegRNA by refining PBS and RT parameters, increasing pegRNA stability and expression levels, and developing automated pegRNA design software. Additionally, efforts are being made to optimize the prime editing system proteins, such as screening for Cas9 and reverse transcriptase variants and performing codon optimization. The final aspect is the regulation of endogenous factors, including the inhibition of mismatch repair mechanisms and the modulation of chromatin environment. These approaches significantly enhance the practicality of prime editing in research and clinical contexts. In conclusion, prime editing represents a major advancement in the field of gene editing, offering powerful tools and methods for both basic research and clinical applications. This review will introduce the discovery, improvement and applications of prime editors, with a focus on prime editing mediated large DNA fragment manipulations. Hopefully, these insights will serve as valuable references for future research and applications of prime editing technology.

Humans ; Clinical Relevance ; Leukemia, Myeloid, Acute ; Transcription Factors ; Neoplasm Proteins ; Antigens, CD ; GPI-Linked Proteins

Humans ; Lymphangioma/surgery* ; Female ; Breast Neoplasms

Platelets are reprogrammed by cancer via a process called education, which favors cancer development. The transcriptional profile of tumor-educated platelets (TEPs) is skewed and therefore practicable for cancer detection. This intercontinental, hospital-based, diagnostic study included 761 treatment-naïve inpatients with histologically confirmed adnexal masses and 167 healthy controls from nine medical centers (China, n = 3; Netherlands, n = 5; Poland, n = 1) between September 2016 and May 2019. The main outcomes were the performance of TEPs and their combination with CA125 in two Chinese (VC1 and VC2) and the European (VC3) validation cohorts collectively and independently. Exploratory outcome was the value of TEPs in public pan-cancer platelet transcriptome datasets. The AUCs for TEPs in the combined validation cohort, VC1, VC2, and VC3 were 0.918 (95% CI 0.889-0.948), 0.923 (0.855-0.990), 0.918 (0.872-0.963), and 0.887 (0.813-0.960), respectively. Combination of TEPs and CA125 demonstrated an AUC of 0.922 (0.889-0.955) in the combined validation cohort; 0.955 (0.912-0.997) in VC1; 0.939 (0.901-0.977) in VC2; 0.917 (0.824-1.000) in VC3. For subgroup analysis, TEPs exhibited an AUC of 0.858, 0.859, and 0.920 to detect early-stage, borderline, non-epithelial diseases and 0.899 to discriminate ovarian cancer from endometriosis. TEPs had robustness, compatibility, and universality for preoperative diagnosis of ovarian cancer since it withstood validations in populations of different ethnicities, heterogeneous histological subtypes, and early-stage ovarian cancer. However, these observations warrant prospective validations in a larger population before clinical utilities.
Humans ; Female ; Blood Platelets/pathology* ; Biomarkers, Tumor/genetics* ; Ovarian Neoplasms/pathology* ; China

Objective: To understand the current status of diagnosis and treatment of chronic lymphocytic leukemia (CLL) /small lymphocytic lymphoma (SLL) among hematologists, oncologists, and lymphoma physicians from hospitals of different levels in China. Methods: This multicenter questionnaire survey was conducted from March 2021 to July 2021 and included 1,000 eligible physicians. A combination of face-to-face interviews and online questionnaire surveys was used. A standardized questionnaire regarding the composition of patients treated for CLL/SLL, disease diagnosis and prognosis evaluation, concomitant diseases, organ function evaluation, treatment selection, and Bruton tyrosine kinase (BTK) inhibitor was used. Results: ①The interviewed physicians stated that the proportion of male patients treated for CLL/SLL is higher than that of females, and the age is mainly concentrated in 61-70 years old. ②Most of the interviewed physicians conducted tests, such as bone marrow biopsies and immunohistochemistry, for patient diagnosis, in addition to the blood test. ③Only 13.7% of the interviewed physicians fully grasped the initial treatment indications recommended by the existing guidelines. ④In terms of cognition of high-risk prognostic factors, physicians' knowledge of unmutated immunoglobulin heavy-chain variable and 11q- is far inferior to that of TP53 mutation and complex karyotype, which are two high-risk prognostic factors, and only 17.1% of the interviewed physicians fully mastered CLL International Prognostic Index scoring system. ⑤Among the first-line treatment strategy, BTK inhibitors are used for different types of patients, and physicians have formed a certain understanding that BTK inhibitors should be preferentially used in patients with high-risk factors and elderly patients, but the actual use of BTK inhibitors in different types of patients is not high (31.6%-46.0%). ⑥BTK inhibitors at a reduced dose in actual clinical treatment were used by 69.0% of the physicians, and 66.8% of the physicians had interrupted the BTK inhibitor for >12 days in actual clinical treatment. The use of BTK inhibitors is reduced or interrupted mainly because of adverse reactions, such as atrial fibrillation, severe bone marrow suppression, hemorrhage, and pulmonary infection, as well as patients' payment capacity and effective disease progression control. ⑦Some differences were found in the perceptions and behaviors of hematologists and oncologists regarding the prognostic assessment of CLL/SLL, the choice of treatment options, the clinical use of BTK inhibitors, etc. Conclusion: At present, a gap remains between the diagnosis and treatment of CLL/SLL among Chinese physicians compared with the recommendations in the guidelines regarding the diagnostic criteria, treatment indications, prognosis assessment, accompanying disease assessment, treatment strategy selection, and rational BTK inhibitor use, especially the proportion of dose reduction or BTK inhibitor discontinuation due to high adverse events.
Female ; Humans ; Male ; Aged ; Middle Aged ; Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy* ; Prognosis ; Lymphoma, B-Cell ; Immunohistochemistry ; Immunoglobulin Heavy Chains/therapeutic use*

Objective: To compare the predictive efficacy of the two thrombosis risk assessment scores (Padua and IMPEDE scores) in venous thromboembolism (VTE) within 6 months in patients with newly diagnosed multiple myeloma (NDMM) in China. Methods: This study reviewed the clinical data of 421 patients with NDMM hospitalized in Beijing Jishuitan Hospital from April 2014 to February 2022. The sensitivity, specificity, accuracy, and Youden index of the two scores were calculated to quantify the thrombus risk assessment of VTE by the Padua and IMPEDE scores. The receiver operating characteristics curves of the two evaluation scores were drawn. Results: The incidence of VTE was 14.73%. The sensitivity, specificity, accuracy, and Youden index of the Padua score were 100%, 0%, 14.7%, and 0% and that of the IMPEDE score was 79%, 44%, 49.2%, and 23%, respectively. The areas under the curve of Padua and IMPEDE risk assessment scores were 0.591 and 0.722, respectively. Conclusion: IMPEDE score is suitable for predicting VTE within 6 months in patients with NDMM.
Humans ; Venous Thromboembolism/etiology* ; Multiple Myeloma/diagnosis* ; Risk Assessment ; Risk Factors ; ROC Curve ; Retrospective Studies

Objective: To examine the association between smoking status and related mortality among elderly people aged 60 and above in urban and rural areas of Beijing City. Methods: Based on Beijing City Elderly Comprehensive Health Cohort Study from 2009 to 2014, a total of 4 499 eligible older adults included in the baseline survey were followed up and investigated to collect information on survival and death. The Cox proportional hazards regression model was used to estimate hazard ratios (HRs) and corresponding 95% confidence intervals (CIs), and the dose-response relationship was estimated between the smoking index, the years of quitting and mortality. Results: The median (IQR) age of 4 499 subjects was 70.00 (10.00) years old, including 1 814 (40.32%) males. The proportion of non-smokers, former smokers and current smokers was 69.50% (3 127/4 499), 13.20% (594/4 499) and 17.30% (778/4 499), respectively. After adjusting for confounding factors such as demographic and sociological characteristics, lifestyle, etc., the results of multivariate Cox regression analysis showed that, compared to non-smokers, former smokers had a 30.6% increased risk of all-cause mortality [HR (95%CI): 1.306 (1.043-1.636)] and the HR (95%CI) of all-cause, malignant tumor and lung cancer mortality among current smokers has increased by 50.0% [HR (95%CI): 1.500 (1.199-1.877)], 80.3% [HR (95%CI): 1.803 (1.226-2.652)] and 212.6% [HR (95%CI): 3.126 (1.626-6.012)], respectively. The smoking index was positively associated with the increased risk of all-cause, malignant tumor and lung cancer mortality, while the years of smoking cessation were negatively associated with that risk (P<0.05). Conclusion: Smoking is associated with tobacco-related mortality among elderly people in Beijing City.
Aged ; Male ; Humans ; Child ; Female ; Beijing ; Cohort Studies ; Lung Neoplasms ; Smoking ; Tobacco Smoking

Objective: To examine the association between smoking status and related mortality among elderly people aged 60 and above in urban and rural areas of Beijing City. Methods: Based on Beijing City Elderly Comprehensive Health Cohort Study from 2009 to 2014, a total of 4 499 eligible older adults included in the baseline survey were followed up and investigated to collect information on survival and death. The Cox proportional hazards regression model was used to estimate hazard ratios (HRs) and corresponding 95% confidence intervals (CIs), and the dose-response relationship was estimated between the smoking index, the years of quitting and mortality. Results: The median (IQR) age of 4 499 subjects was 70.00 (10.00) years old, including 1 814 (40.32%) males. The proportion of non-smokers, former smokers and current smokers was 69.50% (3 127/4 499), 13.20% (594/4 499) and 17.30% (778/4 499), respectively. After adjusting for confounding factors such as demographic and sociological characteristics, lifestyle, etc., the results of multivariate Cox regression analysis showed that, compared to non-smokers, former smokers had a 30.6% increased risk of all-cause mortality [HR (95%CI): 1.306 (1.043-1.636)] and the HR (95%CI) of all-cause, malignant tumor and lung cancer mortality among current smokers has increased by 50.0% [HR (95%CI): 1.500 (1.199-1.877)], 80.3% [HR (95%CI): 1.803 (1.226-2.652)] and 212.6% [HR (95%CI): 3.126 (1.626-6.012)], respectively. The smoking index was positively associated with the increased risk of all-cause, malignant tumor and lung cancer mortality, while the years of smoking cessation were negatively associated with that risk (P<0.05). Conclusion: Smoking is associated with tobacco-related mortality among elderly people in Beijing City.
Aged ; Male ; Humans ; Child ; Female ; Beijing ; Cohort Studies ; Lung Neoplasms ; Smoking ; Tobacco Smoking

Objective To evaluate the feasibility,safety and efficacy of three-port laparoscopic sleeve gastrectomy(LSG).Methods Clinical data of 130 patients with obesity who underwent LSG from April 2020 to January 2021 and completed follow-up in time were analyzed retrospectively.All patients were divided into two groups as surgical method,there were 52 cases in three-port group and 78 cases in four-port group.Perioperative indicators and improvement of obesity in two groups were compared.Results All procedures were performed via LSG.Compared with four-port group,the mean operative time of three-port group was shorter[(88.35±22.64)min vs.(98.29±26.25)min,P=0.027].There were no significant differences in amount of bleeding intraoperative,postoperative pain,postoperative drainage,hospital stay after operation,postoperative complication rate and weight loss 6 months and 2 years after surgery(P>0.05).Conclusions Under the premise of no increase in surgical complications and operative time,three-port LSG can reduce the number and size of surgical scars,which is conducive to skin beauty,and obtain the same weight loss effect as four-port LSG,and can be used as a routine operation for sleeve gastrectomy.

Child ; Endoscopy ; Histiocytosis, Langerhans-Cell/therapy* ; Humans ; Mandible ; Tongue