1.Mass Spectrometry-based Antibody Sequencing Technologies
Sheng-Mei LIU ; Peng XUE ; Xiao-Jian WANG
Progress in Biochemistry and Biophysics 2026;53(4):840-854
Antibodies play a critical role in adaptive immune responses and serve as key components in disease diagnosis and treatment. These molecules exhibit dynamic post-translational modifications (PTMs), such as glycosylation and phosphorylation, which regulate their effector functions. To date, nearly all of our knowledge about antibody repertoires has come from B cell receptor (BCR) sequencing (BCR-seq), which facilitates the profiling of clonal composition and the tracing of maturation trajectories within B-cell repertoires. However, circulating antibodies found in bodily fluids—such as serum, saliva, milk, mucosal secretions, and cerebrospinal fluid—exhibit diversities and specificities beyond what BCR-seq alone can predict. Therefore, identifying and quantifying antibody clonotypes at the protein level could enhance diagnosis, prognosis, and treatment strategies in personalized medicine. The critical gap between genotype and phenotype necessitates complementary methodologies that enable the direct characterization of antibody proteins in their native functional states. Mass spectrometry (MS)-based antibody repertoire sequencing (Ab-seq) is currently the only feasible approach for this task and primarily includes database-dependent methods—such as bottom-up, middle-down, and top-down approaches—as well as database-independent de novo sequencing technology. These strategies enable multi-level, high-precision characterization ranging from peptides and domains to intact antibody molecules. Unlike the shotgun strategy commonly used in routine proteomics, obtaining full sequences of all antibodies presents unique challenges. It requires specialized methodological adaptations to address issues related to dynamic range, sequence variation, and sample complexity. This review introduces the technical principles, methodological workflows, and recent applications of various mass spectrometry-based antibody repertoire sequencing (Ab-seq) strategies, with a focus on approaches designed to improve sequence coverage and identification accuracy. These include multi-enzyme digestion, hybrid fragmentation methods, and artificial intelligence-assisted de novo sequencing. By systematically comparing database-dependent techniques—such as bottom-up, middle-down, and top-down approaches—with database-independent de novo sequencing, this review outlines their respective advantages and limitations in terms of sample throughput, sequence coverage, post-translational modification characterization, and data analysis complexity. In addition, this review discusses emerging technological trends, including the integration of ion mobility separation, native mass spectrometry, and artificial intelligence-driven data interpretation, which are expected to enhance the depth and accuracy of antibody characterization. Although current methods continue to face challenges related to sample complexity, dynamic range, and unambiguous sequence variant assignment, we emphasize the importance of integrating BCR-seq and Ab-seq data to construct gene-protein association maps. These maps help validate sequence accuracy and facilitate epitope discovery. This dual-platform strategy helps bridge the gap between genotype and phenotype, thereby enhancing both the resolution and scope of antibody repertoire studies. Such an integrative approach also offers a valuable tool for therapeutic antibody development, structure-function analysis, and precise evaluation of vaccine efficacy.
2.Olfactory Receptors Expressed in The Intestine and Their Functions
Pei-Wen YANG ; Meng-Meng YUAN ; Ying ZHOU ; Peng LI ; Gui-Hong QI ; Ying YANG ; Zhong-Yi MAO ; Meng-Sha ZHOU ; Xiao-Shuang MAO ; Jian-Ping XIE ; Yi-Nan YANG ; Shi-Hao SUN
Progress in Biochemistry and Biophysics 2026;53(3):534-549
Olfactory receptors (ORs) form the largest superfamily of G protein-coupled receptors (GPCRs). Traditionally recognized for their role in the nasal olfactory epithelium, where they mediate the sense of smell, accumulating evidence has firmly established their ectopic expression in non-olfactory tissues, including the intestine, lungs, and kidneys. The intestine, as the primary site for nutrient digestion and absorption, harbors a highly complex chemical environment. To adapt to this environment, the gut employs a sophisticated network of “chemosensors” to monitor luminal contents and maintain homeostasis. Among these sensors, intestinal ORs have emerged as crucial functional components, serving as a molecular bridge that connects environmental chemical signals—such as food-derived odorants—to specific physiological responses. This discovery has significantly deepened our understanding of how dietary flavors and compounds influence intestinal physiology at the molecular level. This review systematically summarizes the expression profiles, ligand classification, and biological functions of ORs within the gastrointestinal tract. Studies indicate that intestinal ORs exhibit distinct spatial distribution patterns across different gut segments and display cell-type specificity, particularly within enterocytes and enteroendocrine cells. These receptors function as versatile sensors capable of recognizing a wide variety of ligands, including exogenous dietary components, gut microbiota metabolites such as short-chain fatty acids, and endogenous small molecules like azelaic acid. Upon activation by specific ligands, intestinal ORs trigger intracellular signaling cascades, primarily involving the AC-cAMP-PKA pathway or calcium influx channels. A major focus of this review is to elucidate the molecular mechanisms by which these receptors regulate the secretion of gut hormones. Activation of specific ORs in enteroendocrine cells has been shown to stimulate the release of hormones such as glucagon-like peptide-1 (GLP-1), peptide YY (PYY), and serotonin (5-HT), thereby modulating systemic energy metabolism, glucose homeostasis, and gastrointestinal motility. Furthermore, the review addresses the critical roles of ORs in immune regulation and pathology. Evidence suggests that specific ORs contribute to the maintenance of intestinal immune homeostasis and may offer protection against inflammation. Beyond their involvement in inflammatory responses, ORs such as Olfr78 have been shown to regulate the differentiation and function of intestinal endocrine cells. Similarly, Olfr544 has been demonstrated to alleviate intestinal inflammation by remodeling the gut microbiome and metabolome. These findings collectively suggest that specific ORs hold promise as therapeutic targets for mitigating intestinal inflammation and maintaining gut homeostasis. Additionally, the review explores the emerging role of ORs in cancer. Although OR expression is often downregulated in tumor tissues compared to normal mucosa, activation of specific ORs by certain ligands can inhibit tumor cell proliferation and migration and induce apoptosis via pathways such as MEK/ERK and p38 MAPK. Conversely, other receptors, such as OR7C1, may serve as biomarkers for cancer-initiating cells. In conclusion, intestinal ORs represent a vital component of the gut’s sensory network. The review also discusses the translational potential of these findings. By elucidating the precise pairing relationships between dietary components and specific ORs, novel therapeutic strategies could be developed. Intestinal ORs may thus emerge as promising targets for nutritional and pharmacological interventions in metabolic diseases, inflammatory bowel diseases, and malignancies.
3.Guideline for Adult Weight Management in China
Weiqing WANG ; Qin WAN ; Jianhua MA ; Guang WANG ; Yufan WANG ; Guixia WANG ; Yongquan SHI ; Tingjun YE ; Xiaoguang SHI ; Jian KUANG ; Bo FENG ; Xiuyan FENG ; Guang NING ; Yiming MU ; Hongyu KUANG ; Xiaoping XING ; Chunli PIAO ; Xingbo CHENG ; Zhifeng CHENG ; Yufang BI ; Yan BI ; Wenshan LYU ; Dalong ZHU ; Cuiyan ZHU ; Wei ZHU ; Fei HUA ; Fei XIANG ; Shuang YAN ; Zilin SUN ; Yadong SUN ; Liqin SUN ; Luying SUN ; Li YAN ; Yanbing LI ; Hong LI ; Shu LI ; Ling LI ; Yiming LI ; Chenzhong LI ; Hua YANG ; Jinkui YANG ; Ling YANG ; Ying YANG ; Tao YANG ; Xiao YANG ; Xinhua XIAO ; Dan WU ; Jinsong KUANG ; Lanjie HE ; Wei GU ; Jie SHEN ; Yongfeng SONG ; Qiao ZHANG ; Hong ZHANG ; Yuwei ZHANG ; Junqing ZHANG ; Xianfeng ZHANG ; Miao ZHANG ; Yifei ZHANG ; Yingli LU ; Hong CHEN ; Li CHEN ; Bing CHEN ; Shihong CHEN ; Guiyan CHEN ; Haibing CHEN ; Lei CHEN ; Yanyan CHEN ; Genben CHEN ; Yikun ZHOU ; Xianghai ZHOU ; Qiang ZHOU ; Jiaqiang ZHOU ; Hongting ZHENG ; Zhongyan SHAN ; Jiajun ZHAO ; Dong ZHAO ; Ji HU ; Jiang HU ; Xinguo HOU ; Bimin SHI ; Tianpei HONG ; Mingxia YUAN ; Weibo XIA ; Xuejiang GU ; Yong XU ; Shuguang PANG ; Tianshu GAO ; Zuhua GAO ; Xiaohui GUO ; Hongyi CAO ; Mingfeng CAO ; Xiaopei CAO ; Jing MA ; Bin LU ; Zhen LIANG ; Jun LIANG ; Min LONG ; Yongde PENG ; Jin LU ; Hongyun LU ; Yan LU ; Chunping ZENG ; Binhong WEN ; Xueyong LOU ; Qingbo GUAN ; Lin LIAO ; Xin LIAO ; Ping XIONG ; Yaoming XUE
Chinese Journal of Endocrinology and Metabolism 2025;41(11):891-907
Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.
4.A Prospective Randomized Controlled Study of Fascial Platform and Commercial Platform in Transumbilical Single-port Laparoscopic Transabdominal Preperitoneal Repair
Gang XIAO ; Dandan LI ; Jian SHEN ; Peng YANG ; Shuoyang HUANG ; Jing WEN ; Bo HUANG ; Hailing LIU ; Shan HE
Chinese Journal of Minimally Invasive Surgery 2025;25(9):524-529
Objective To evaluate the application value of fascial platform versus commercial platform in transumbilical single-port laparoscopic transabdominal preperitoneal repair(TAPP)for inguinal hernia.Methods One hundred patients with inguinal hernia admitted between January and December 2024 were prospectively enrolled and randomly assigned to either the fascial platform group(n=50)or the commercial platform group(n=50)by using a random number table.The following parameters were compared between the two groups:platform establishment time,operative time,intraoperative blood loss,incision suture time,operative difficulty score,camera manipulation difficulty score,postoperative 24-hour Visual Analogue Scale(VAS)score of pain,postoperative hospital stay,total hospitalization cost,comprehensive satisfaction score,and postoperative complications.Results The fascial platform group demonstrated significantly superior outcomes compared to the commercial platform group in platform establishment time[(3.4±1.0)min vs.(8.1±1.5)min,t=-18.389,P<0.001],operative time[(67.5±7.9)min vs.(72.2±11.5)min,t=-2.380,P=0.019],intraoperative blood loss[(8.3±4.3)ml vs.(11.1±5.7)ml,t=-2.798,P=0.006],incision suture time[(7.0±1.4)min vs.(11.8±2.7)min,t=-11.212,P<0.001],operative difficulty score(14.4±3.6 vs.23.1±4.1,t=-11.215,P<0.001),camera manipulation difficulty score(11.4±2.6 vs.18.3±2.5,t=-13.370,P<0.001),VAS score of postoperative 24-hour pain(2.4±1.0 vs.3.7±1.1,t=-6.139,P<0.001),total hospitalization cost[(15.8±1.6)thousand yuan vs.(18.3±1.6)thousand yuan,t=-7.841,P<0.001],and comprehensive satisfaction score(7.3±1.1 vs.6.3±1.0,t=5.271,P<0.001).However,there were no statistically significant differences in postoperative hospital stay or postoperative complication between the two groups(P>0.05).All the patients were followed up for 1-12 months postoperatively[(5.3±1.8)months vs.(5.7±2.0)months,t=-0.945,P=0.347],with no recurrence observed in either group.Conclusions Both fascial platform and commercial platform can be used for transumbilical single-port TAPP.The fascial platform offers advantages including shorter platform establishment time,reduced incision suture time,lower operative and camera manipulation difficulty,decreased cost,less postoperative pain,and higher patient satisfaction.
5.Comparative study on the short-term efficacy of transanal natural orifice specimen extraction surgery and conventional laparoscopic surgery in left-sided colon cancer
Tingting FU ; Jingwen CHANG ; Yibo CAO ; Tiantian BAO ; Tianbao XIAO ; Jiang CHEN ; Jian PENG
China Journal of Endoscopy 2025;31(7):1-10
Objective To compare the short-term efficacy and safety of transanal natural orifice specimen extraction surgery(Ta-NOSES)and conventional laparoscopic surgery in left-sided colon cancer.Methods A retrospective analysis was conducted on the clinical data of 35 patients with left-sided colon cancer admitted to the anorectal department of the hospital from January 2018 to December 2019.According to the different surgical methods,the patients were divided into experimental group(15 cases)and control group(20 cases).The observation group underwent Ta-NOSES,and the control group underwent conventional laparoscopic surgery.The perioperative related indicators,postoperative complications,postoperative pain scores,postoperative defecation control,short-term postoperative quality of life scores and 5-year postoperative follow-up of the two groups of patients were compared.Results There was no statistically significant difference in the intraoperative blood loss,stoma status and the number of lymph node dissections between the two groups of patients(P>0.05).Moreover,no permanent stoma occurred in either group of patients.The operation time of the experimental group was longer than that of the control group,the first time to get out of bed and move around,the time of the first anal exhaust,the time of the first diet intake and the hospital stay were shorter than those of the control group,the hospitalization cost was significantly lower than that of the control group,the differences were statistically significant(P<0.05).On 1 and 3 days after operation,the VAS scores of the experimental group were significantly lower than those of the control group.At 3 days after operation,the VAS scores of the two groups were significantly lower than those at 1 day after operation,and the differences were statistically significant(P<0.05).There was a statistically significant difference in postoperative Kirwan anal function grading between two groups of patients(P<0.05),with the experimental group having a better grading(higher proportion of grade Ⅰ),the control group had poor grading(with a higher proportion of grades Ⅱ,Ⅲ,and Ⅳ).There was no statistically significant difference in postoperative complications between the two groups of patients(P>0.05).The scores of each item on the Short Form-36(SF-36)in the experimental group were higher than those in the control group at 10 and 20 days after surgery(P<0.05).There was no statistically significant difference in the scores of each item on the SF-36 between the two groups at 30 days after surgery(P>0.05).The distant recurrence rate after surgery in the experimental group was 26.7%,compared with 25.0%in the control group,the difference was not statistically significant(P>0.05).There were no tumor recurrence cases with the original incision site,rectal and intestinal cavity,pelvic cavity and other specimen removal routes in both groups.The 5-year survival rate of the experimental group was 73.3%,which was not statistically significantly different from that of the control group(70.0%)(P>0.05).Conclusion Ta-NOSES in the treatment of left-sided colon cancer can alleviate postoperative pain compared with conventional laparoscopic surgery,promote the recovery of postoperative gastrointestinal function,improve the utilization rate of medical resources,reduce the economic burden of patients,improve the short-term quality of life after surgery,and does not increase the risks of postoperative complications and tumor metastasis and recurrence.It is worthy of clinical promotion and application.
6.Design and application of individually portable oral treatment device field conditions in alpine regions
Jian-xue ZHOU ; Hong XIN ; Xue-qi MENG ; Rui-hua WANG ; Xiao-ming ZHU ; Peng-fa WANG
Chinese Medical Equipment Journal 2025;46(1):108-113
Objective To design an individually portable oral treatment device to solve the problems of oral diagnosis and treatment under field conditions in alpine regions.Methods The individually portable oral treatment device had a trolley box structure and consisted of an outer box,an inner framework and an operation panel.The outer box was made of low-density polyethylene material and formed by by one-time rotational moulding process;the inner framework integrated a plateau com-pressor,an independent negative-pressure compressor,an integrated control system for programmable logic controller(PLC),an individually portable respiratory synchronized pulsed oxygen supply module for plateau application;there were several curative devices equipped in the operation panel,including a 3-way syringe,a high-speed turbine handpiece,an electric variable-speed handpiece,a water control switch,a light curing machine and an ultrasonic dental cleaning handpiece.Trials were carried out with the test-phase prototype in alpine regions so as to verify the performance of the device.Results Trials proved that the prototype gained advantages in mobility,multifunctionality and pressure supply facilitating continuous operation of power gas source for oral diagnosis and treatment in alpine regions.Conclusion The device developed solves the problems in pressure insufficiency and instability,control system integration,portability and oxygen supply for medical staffs,improves the mobility of oral diagnosis and treatment in alpine regions and enhances the oral support service and equipment effectively.[Chinese Medical Equipment Journal,2025,46(1):108-113]
7.Efficacy and safety of a facilitated percutaneous coronary intervention with half-dose recombinant staphylokinase in ST-segment elevation myocardial infarction
Tian-yu WU ; Wen-hao ZHANG ; Peng-sheng CHEN ; Chen LI ; Tian WU ; Zhan LÜ ; Tong WANG ; Kun LIU ; Zhi-wen TAO ; Xiao-xuan GONG ; Liang YUAN ; Yong LI ; Bo CHEN ; Xin CHEN ; Zeng-guang CHEN ; Nai-quan YANG ; Yuan-yuan SANG ; Xiao-yan WANG ; Bai-hong LI ; Li ZHU ; Guo-yu WANG ; Xin ZHAO ; Chuan LU ; Jun JIANG ; Rui-na HAO ; Chun-jian LI
Chinese Journal of Interventional Cardiology 2025;33(8):431-438
Objective To investigate the clinical efficacy and safety of facilitated percutaneous coronary intervention(PCI)with half-dose recombinant staphylokinase(r-SAK)in patients with ST-segment elevation myocardial infarction(STEMI)who are expected to undergo PCI within 120 minutes.Methods From October 2021 to August 2022,a total of 200 STEMI patients in eight centers were included and randomly assigned in a 1﹕1 ratio to either r-SAK group or control group.Patients received loading doses of aspirin and ticagrelor and intravenous heparin and were randomized to receive an intravenous bolus of either 5 mg r-SAK or normal saline prior to PCI.The outcomes were set as ST-segment resolution(STR)at 60-90 minutes after PCI,the proportion and transition of pathological Q waves on the 5th day after PCI,and the proportion of high-sensitivity cardiac troponin T(hs-cTnT)peaking within 12 hours of onset.The safety outcome was major bleeding events defined as Bleeding Academic Research Consortium(BARC)≥type 3 bleeding during hospitalization.Results Compared with the control group,the r-SAK group had a higher proportion of STR≥70%within 60-90 minutes after PCI(58.3%vs.40.3%,P=0.009);a lower proportion of pathological Q waves(59.1%vs.74.1%,P=0.040);a lower rate of Q wave progression(14.8%vs.43.2%,P<0.001);a higher rate of Q wave disappearance(12.5%vs.3.7%,P=0.027);and a higher proportion of hs-cTnT peaking within 12 hours of symptom onset[31/40(77.5%)vs.17/33(51.5%),P=0.027].Regarding the safety outcome,no significant difference in BARC≥type 3 bleeding was found between the two groups during hospitalization(P>0.05).Conclusions For STEMI patients who were expected to undergo primary PCI within 120 minutes of symptom onset,the facilitated PCI with half-dose r-SAK significantly increased the proportion of STR≥70%at 60-90 minutes after PCI,reduced the formation of pathological Q waves,and shortened the time to peak hs-cTnT,without increasing the risk of bleeding,which should be an alternative reperfusion strategy worthy of further study.
8.Clinical research of Zhuangdan Yanshi Decoction combined with dapoxetine hydrochloride in the treatment of premature ejaculation with cholestasis and phlegm disturbance syndrome
Xing-yun PENG ; Jian-guo LIU ; Jiao-jiao LI ; Xiao-guang MIAO ; Xu ZHANG
National Journal of Andrology 2025;31(8):732-736
Objective:To investigate the clinical efficacy of Zhuangdan Yanshi Decoction combined with dapoxetine hydrochlo-ride in the treatment of premature ejaculation with cholestasis and phlegm disturbance.Methods:A total of 120 patients diagnosed with premature ejaculation and treated in the Andrology Outpatient Department of Shaanxi Provincial Hospital of Traditional Chinese Medicine from March to December in 2022 were selected and randomly divided into treatment group and control group,with 60 cases in each group.The incubation period of intravaginal ejaculation(IELT),the Diagnostic Scale of Premature Ejaculation(PEDT),the Premature Ejaculation Assessment Scale(PEP),the 5-item Sexual Function Evaluation of Chinese Premature Ejaculation Patients(CIPE-5)and the improvement of traditional Chinese medicine symptom scores were compared before and after the treatment.And the adverse reactions were recorded as well.Results:A total of 105 cases were ultimately included,with 55 cases in the treatment group and 50 cases in the control group.Measurable improvements in IELT,PEDT scores,PEP scores,CIPE scores and TCM symptom scores had been found after treatment in both of two groups(P<0.05).Moreover,the improvement in the treatment group was superi-or to that in the control group(P<0.05).The total effective rate in the treatment group was 89.1%,which was higher than that(84%)in the control group,with no statistically significant difference between the two groups(P>0.05).The incidence of adverse reactions in the treatment group was 9.1%,which was 24%in the control group.There was significantly difference between two groups(P<0.05).Conclusion:The combination therapy with Zhuangdan Yanshi Decoction and dapoxetine hydrochloride for premature ejaculation associated with cholestasis and phlegm disturbance syndrome is definite,and it can reduce the side effects of drugs,which is better than oral dapoxetine hydrochloride alone.
9.Dual-modal Magnetic Resonance Imaging Contrast Agents Based on Polymetallic Nanoclusters for Targeted Diagnosis of Prostate Cancer
Qing-Dong LI ; Peng WANG ; Jian-Min XIAO ; Wen-Juan GAO ; Zhen-Hong XIA ; Gui-Long ZHANG ; Zheng-Yan WU
Chinese Journal of Analytical Chemistry 2025;53(4):602-611
Fe/Mn/Gd polymetallic nanooxide(FMGN)were prepared by one-step solvent thermal reaction by using Fe(acac)3,Mn(acac)2 and Gd(acac)3 as reaction precursors.Next,hyaluronic acid(HA)was used to modify FMGN to fabricate tumor-targeting T 1-T 2 dual-mode magnetic resonance imaging(MRI)contrast agent(HA-FMGN)for accurate diagnosis of prostate cancer.The structure and morphology of FMGN were observed by transmission electron microscope(TEM).It was found that FMGN exhibited a uniform nanocluster spherical structure when the feeding ratio of iron acetylacetonate,manganese acetylacetonate,and gadolinium acetylacetonate was 3:2:1.X-ray diffraction(XRD)analysis showed that FMGN had a typical inverse spinel structure of Mn doped Fe 3O 4,with Gd existing in the form of amorphous gadolinium oxide.The longitudinal relaxivity(r 1)and transverse relaxivity(r 2)of FMGN were 13.395 and 428.535 L/(mmol·s),respectively,measured by 0.5 T MRI analyzer,which proved that FMGN had excellent T 1-T 2 dual-mode MRI contrast capability.The cytotoxicity and hemolysis test found that HA-FMGN didn't damage red cells and induce toxicity for normal cells,indicating that HA-FMGN had excellent cell biocompatibility.The internalization efficacy of HA-FMGN was observed by CLSM,and the results showed that HA-FMGN possessed excellent prostate tumor-targeting ability.In vivo MRI experiment showed that HA-FMGN significantly enhanced T 1 and T 2 weighted MRI signal to noise ratio(SNR)of prostate tumor,which promoted the accurate diagnosis of orthotopic prostate cancer.
10.Validation and Forensic Application of a Domestic Human DNA Quantitative De-tection Kit
Jing CHEN ; Ya-Ping WANG ; Yun-Peng FENG ; Xiao-Xin HU ; Zhen-Jun JIA ; Hong-Di LIU ; An-Xin YAN ; Yong-Jiu LI ; Zhu PENG ; Zhi-Fang LIU ; Jian-Gang CHEN
Journal of Forensic Medicine 2025;41(3):252-259
Objective To verify the efficacy of a domestic human DNA quantification kit based on real-time fluorescence quantitative PCR in detecting the total human DNA concentration,male DNA concen-tration in mixed male/female DNA samples,the degree of DNA degradation and inhibitor tolerance.Methods Samples with different concentrations,different male/female ratios,different concentrations of inhibitors,and different degradation degrees were tested using the domestic human DNA quantification kit based on real-time fluorescence quantitative PCR.This kit was compared with a similar product on the market and was applied to the detection of DNA from real cases.Results This human DNA quan-tification kit can effectively detect human DNA as low as 0.001 65 ng/μL,and 6.25 pg/μL of male DNA in mixed samples with a male-to-female ratio of 1∶15 000.Even when the sample contains as high as 400 ng/μL of humic acid or 1 000 μmol/L of hemin alone,the DNA concentration can still be accurately detected.The degradation index can effectively characterize the degradation degree of the sample.This kit has been successfully applied in forensic practice.Conclusion This human DNA quan-tification kit is accurate and reliable in detection.It can accurately reflect the degradation of DNA and inhibitor tolerance.It has good performance in quantitative accuracy,determination of the male/female ratio in mixed samples,and inhibitor tolerance.It has application potential in forensic case examination.

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