The exploration of pathogenic single nucleotide polymorphisms in the genome plays a pivotal role in the study of human disease-associated genetic mutations. However, there remains a lack of suitable high-throughput screening platforms to investigate the impact of point mutations on genomic structure and function. CRISPR/Cas9-mediated base editors has enabled large-scale annotation of the human genome and phenotypic characterization of monogenic genetic disorders. Base editors, a precise gene-editing technology capable of achieving targeted base substitutions, can be employed to induce mutations at specific functional sites, thereby observing their effects on gene expression, protein function, and cellular phenotypes. Furthermore, integrating base editors with high-throughput screening technologies allows for the large-scale evaluation of multiple candidate sites, accelerating the identification of functional loci and providing a powerful tool for disease research and therapeutic target discovery. This article aims to introduce the working principles of various base editors, including cytosine base editors, adenine base editors, and prime editors, and summarize recent advances in high-throughput screening of functional genomic sites using base-editing techniques.

The exploration of pathogenic single nucleotide polymorphisms in the genome plays a pivotal role in the study of human disease-associated genetic mutations. However, there remains a lack of suitable high-throughput screening platforms to investigate the impact of point mutations on genomic structure and function. CRISPR/Cas9-mediated base editors has enabled large-scale annotation of the human genome and phenotypic characterization of monogenic disorders. Base editors, a precise gene-editing technique capable of achieving targeted base substitutions, can be employed to induce mutations at specific functional sites, thereby observing their effects on gene expression, protein function, and cellular phenotypes. Furthermore, integrating base editors with high-throughput screening technologies allows for large-scale evaluation of multiple candidate sites, accelerating the identification of functional loci and providing a powerful tool for disease research and therapeutic target discovery. This article aims to introduce the working principles of various base editors, including cytosine base editors, adenine base editors, and prime editors, and summarize recent advances in high-throughput screening of functional genomic sites using base-editing techniques.
Humans ; Gene Editing/methods* ; CRISPR-Cas Systems/genetics* ; Genome, Human ; Polymorphism, Single Nucleotide

The exploration of pathogenic single nucleotide polymorphisms in the genome plays a pivotal role in the study of human disease-associated genetic mutations. However, there remains a lack of suitable high-throughput screening platforms to investigate the impact of point mutations on genomic structure and function. CRISPR/Cas9-mediated base editors has enabled large-scale annotation of the human genome and phenotypic characterization of monogenic genetic disorders. Base editors, a precise gene-editing technology capable of achieving targeted base substitutions, can be employed to induce mutations at specific functional sites, thereby observing their effects on gene expression, protein function, and cellular phenotypes. Furthermore, integrating base editors with high-throughput screening technologies allows for the large-scale evaluation of multiple candidate sites, accelerating the identification of functional loci and providing a powerful tool for disease research and therapeutic target discovery. This article aims to introduce the working principles of various base editors, including cytosine base editors, adenine base editors, and prime editors, and summarize recent advances in high-throughput screening of functional genomic sites using base-editing techniques.

Objective To understand the carrying situation and common variation of pathogenic genes of single gene hereditary disease in childbearing age population in Anhui province,to explore the establishment of clinical application network and referral model of carrier screening in Anhui province,and to explore the application value of expansible carrier screening(expanded carrier screening,ECS)in clinic.Methods Samples were collected from 604 individuals of childbearing age,all exhibiting a normal phenotype and a family history of inherited dis-ease.These samples were obtained during the first trimester or early stages of pregnancy(≤13+6 weeks).Based on high-throughput sequencing and special PCR analysis techniques,pathogenic variants associated with 220 disea-ses were detected,and related genes were detected in the spouses of positive carriers.Results As of May 16,2023,604 tested samples had been collected,and 340 carriers of the target disease had been detected;The posi-tive rate of pathogenic variation detection was 56.29％ ;A total of 499 pathogenic variants were detected,with each tested individual carrying 0-5 variants;216 cases,accounting for 35.76％ ,carried a single gene recessive dis-ease pathogenic variation,which was the most common.There were 95 cases carrying two types of single gene re-cessive genetic disease pathogenic variation,accounting for 15.73％ .As of now,302 couples have been reported,and a total of 7 high-risk couples have been found through screening,with a high-risk rate of 2.32％ .There are a total of 5 pairs with autosomal recessive genetic pattern(both spouses carry the same pathogenic gene),and 2 pairs with X-linked genetic pattern(the female carries the X-linked pathogenic gene).Conclusion In this study,we obtained the overall carrier and clinical application of target diseases as well as the carrier rates of causative genes of common single-gene genetic diseases in 604 subjects who underwent ECS testing,which could provide scientific guidance for the establishment of a clinical application network and referral model for carrier screening in Anhui Province.

Objective:To analyze the value of 68Ga-fibroblast activation protein inhibitor (FAPI)-04 PET/CT in the detection of myocardial injury in patients treated with anti-tumor therapy. Methods:A retrospective study was conducted on 164 patients who underwent 68Ga-FAPI-04 PET/CT to evaluate the efficacy of anti-tumor therapy in Renji Hospital, School of Medicine, Shanghai Jiao Tong University between August 2021 and March 2024. The patients were divided into 68Ga-FAPI-04-positive group ( n=63, 36 males, 27 females, age (66.7±9.6) years) and 68Ga-FAPI-04-negative group ( n=101, 42 males, 59 females, age (55.2±14.1) years) based on the uptake of left ventricular myocardium (LVM). Moreover, FAPI-04 uptake was analyzed based on different types and locations, and the corresponding SUV max differences were analyzed by Kruskal-Wallis rank sum test. The differences of SUV max between 68Ga-FAPI-04-positive group and 68Ga-FAPI-04-negative group were analyzed by Mann-Whitney U test. The clinical factors such as gender, age, body mass index (BMI), previous history of coronary heart disease, left ventricular ejection fraction (LVEF), smoking history, hypertension, diabetes, cancer types and immune checkpoint inhibitors (ICIs) treatment were collected, and their predictive values for LVM 68Ga-FAPI-04 uptake were investigated by the binary logistic regression analysis. Results:Fifty patients of the 68Ga-FAPI-04-positive group (79.4%, 50/63) showed focal uptake of LVM, 7 patients (11.1%, 7/63) showed multifocal myocardial uptake, and 6 patients (9.5%, 6/63) showed diffuse myocardial uptake. A total of 127 uptake lesions were found, and most of them were located in the septum (37.8%, 48/127). The SUV max of LVM in 68Ga-FAPI-04-positive group and 68Ga-FAPI-04-negative group were 4.00(3.10, 5.40) and 1.31(1.20, 1.40) respectively ( z=-10.82, P<0.001). Differences of the SUV max among focal uptake group, multifocal myocardial uptake group, and diffuse myocardial uptake group were not significantly different (4.00(3.00, 5.10) vs 7.60(3.60, 9.30) vs 3.95(3.05, 5.05); H=3.81, P=0.149). There is no statistically significant difference either in FAPI uptake among different sites of LVM ( H=1.51, P=0.825). Age, previous history of coronary heart disease, BMI, LVEF and ICIs treatment were independent predictive factors for positive 68Ga-FAPI-04 uptake in the LVM (odds ratio ( OR) values: 0.87-10.43, all P<0.05). Conclusion:68Ga-FAPI-04 PET/CT is a potential new imaging method for the visualization of myocardial injury in patients with anti-tumor therapy.

Objective To analyze the effects of massage therapy on the inflammatory state and lung function of pediatric refractory Mycoplasma pneumonia (RMPP). Methods A total of 320 children with RMPP treated in Hebei Seventh People′s Hospital from September 2022 to August 2023 were selected in the study. According to different treatment methods, they were divided into two groups, with 160 cases in each group. The western medicine group was given conventional anti-inflammatory and other symptomatic treatment, while the massage group was given dialectical massage treatment based on the western medicine group. The clinical efficacy, duration of disappearance of clinical symptoms, pulmonary function indexes, serum inflammatory factors, immune function indexes and total incidence of adverse reactions were compared between the two groups. Results The total effective rate of massage group was higher than that of control group (P < 0.05). The disappearance time of fever, cough, sputum and lung moist rales in observation group was shorter than that in control group (P < 0.05). After treatment, the peak expiratory flow (PEF), maximum mid-expiratory flow (MMEF) and forced vital capacity (FVC) in the massage group were higher than those in the control group (P < 0.05). The levels of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the massage group were lower than those in the control group after treatment (P < 0.05). After treatment, CD3⁺, CD4⁺, CD4⁺/CD8⁺ in the massage group were higher than those in the control group (P < 0.05), and CD8⁺ was lower than that in the control group (P < 0.05). There was nosignificant difference in the total incidence of adverse reactions between the massage group and the western medicine group (P>0.05). Conclusion Massage based on syndrome differentiation combined with western medicine can effectively relieve fever, cough and other symptoms of RMPP children, reduce inflammatory response, improve lung function and immune function, and has less adverse reactions.

Severe insulin resistance syndrome associated with mutations in the insulin receptor(INSR) gene is rare in clinical practice. We report a 13-year-old female patient with insulin resistance, acanthosis nigricans, and Class Ⅱ malocclusion, whose family history included hyperinsulinemia in both her mother and grandmother. Whole-exome sequencing and PCR-Sanger validation identified a novel INSR mutation, c. 637delA(p.S213Vfs*69), resulting in a pathogenic variant that substitutes serine at position 213 with valine. This case highlights a clinical phenotype that is challenging to differentiate between Rabson-Mendenhall syndrome and A-type insulin resistance syndrome. Long-term follow-up is crucial to assess disease progression and prognosis.

Severe insulin resistance syndrome associated with mutations in the insulin receptor(INSR) gene is rare in clinical practice. We report a 13-year-old female patient with insulin resistance, acanthosis nigricans, and Class Ⅱ malocclusion, whose family history included hyperinsulinemia in both her mother and grandmother. Whole-exome sequencing and PCR-Sanger validation identified a novel INSR mutation, c. 637delA(p.S213Vfs*69), resulting in a pathogenic variant that substitutes serine at position 213 with valine. This case highlights a clinical phenotype that is challenging to differentiate between Rabson-Mendenhall syndrome and A-type insulin resistance syndrome. Long-term follow-up is crucial to assess disease progression and prognosis.

Objective To study the human papillomavirus(HPV)infection and genotype distribution characteristics in male outpatients,and compare with female infection status,in order to provide scientific basis for the clinical development of prevention and treatment measures for male HPV infection related diseases.Methods Polymerase chain reaction(PCR)amplification followed by directed hybridization was used to detect 37 HPV genotypes in 258 male outpatients of Yangzhou Maternal and Child Health Hospital from July 2018 to December 2022.The detection results were further compared with the detection results of 1436 female physical examinees and 931 cervical exfoliated cell samples of gynecological patients suspected of HPV infection at the same time.Results There were 103 of the 258 male outpatients were positive,with an infection rate of 39.92%.Among the 103 positive samples,high-risk,low-risk,and mixed high-risk HPV infections accounted for 58.25%,20.39%,and 21.36%,respectively.Among them,59 were infected with single infection,accounting for 57.28%,44 were infected with more than double infection(multiple infection),accounting for 42.72%,and the most one had ten types infections.Single infection was mainly high-risk type,while multiple infection was mainly high-risk type and mixed high-risk type.There was no statistically significant difference in HPV infection rate,infection type,and infection status between male patients and female patients(P=0.456,0.192,0.102),but there was a statistically significant difference compared with female physical examinees(P<0.001,0.032,<0.001).The peak age of HPV infection is 20-39 years old,accounting for 74.75%,and there was no statistically significant difference in HPV detection rate among different age groups(P=0.297).33 HPV genotypes were detected in both male and female groups.The top five subtypes were HPV52,58,51,54 and 61 in male patients,while HPV57,67,69 and 83 were not detected.The top five subtypes were HPV52,16,58,53 and 61 in female groups,while HPV57,69,72 and 26 were not detected.Physical examination is the main reason of 103 male infected patients seeking medical treatment.Conclusion The HPV infection status in male is similar to that of female patients,with high-risk infection being the main type and single subtype infection being the main infection.The peak age of male patients with HPV infection is 20-39 years old.HPV52,58,51,54,61 are the most common types,and most of them are asymptomatic.Therefore,men are high-risk groups of HPV infection.It is necessary to carry out HPV detection for male outpatients.

Background:This study aimed to investigate the changes in the clinical indicators and influencing factors of treatment duration among human immunodeficiency virus (HIV) patients in whom antiretroviral therapy (ART) was unsuccessful.Methods:In this retrospective study,a total of 9,418 HIV patients who failed in ART during 2004-2016 were included and divided into two treatment groups-Group 1 (treatment time ≤ 3 years,n1 =5,218) and Group 2 (treatment time ＞ 3 years,n2 =4,200).Patient follow-up data,including age,cluster of differentiation 4 (CD4) count,and viral load,glucose,creatinine,and triglyceride levels,were extracted from electronic health record databases.Covariance analysis for repeated measures was used to analyze the biochemical indicators,and multiple logistic regression modeling was used to compare relevant data extracted from the Group 1 and Group 2 HIV patient cohorts with different treatment time.Results:The median initial CD4 count was 175.0 cells/μl (interquartile range,77.0-282.0),while the initial CD4 counts for Group 1 were lower than those for Group 2 (P ＜ 0.05).A significant interaction between group and time effects was observed (P ＜ 0.05) in total cholesterol (TC).Changes in hemoglobin level among HIV patients were also significantly associated with treatment time (P =0.001).The initial CD4 count (odds ratio [OR] =0.756),female sex (OR =0.713),Zerit (d4T) (OR =1.443),TC (OR =1.285),and aspartate aminotransferase level (OR =1.002) were significantly associated with the survival time of dead patients with HIV (P ＜ 0.05).Additionally,the initial CD4 count (OR =1.456),age (OR =1.022),time interval (OR =0.903),patient's living status (OR =0.597),d4T (OR =2.256),and triglyceride (OR =0.930) and hemoglobin levels (OR =0.997) were significantly associated with the treatment time of HIV patients with drug withdrawal (P ＜ 0.05).Conclusion:The initial biochemical parameters can affect the survival and treatment time of HIV patients.With a comprehensive understanding of the physiological and biochemical indicators of patients,we can reduce the probability of drug withdrawal and prolong the survival time of HIV patients.