1.Effect and mechanism of collagen combined with microneedles in treatment of skin photoaging
Fengyi TAN ; Jiamin XIE ; Zhenfeng PAN ; Xinxu ZHANG ; Zetai ZHENG ; Zhiying ZENG ; Yanfang ZHOU
Chinese Journal of Tissue Engineering Research 2026;30(2):451-458
BACKGROUND:Collagen combined with microneedling therapy has gradually become an important means of improving skin photoaging.OBJECTIVE:To summarize and explore the main mechanism and clinical application status of collagen combined with microneedle therapy.METHODS:PubMed,China National Knowledge Infrastructure,and ScienceDirect databases were searched for Chinese and English literature published before August 2024.Chinese and English search terms were"ultraviolet radiation,photoaging,collagen,microneedling,clinical applications."Finally,74 articles were included for summary.RESULTS AND CONCLUSION:Collagen treats skin photoaging through mechanisms such as inhibiting matrix metalloproteinase expression,retaining skin moisture,and reducing melanin formation.Microneedles can better promote the penetration of collagen into deep layers of the skin,breaking down the skin's barrier and increasing the absorption rate.Collagen combined with microneedles has various beneficial effects for treating skin photoaging,such as whitening,anti-wrinkle,improving skin elasticity,shrinking pores,and repairing skin barriers.It also has the advantages of easy operation,significant effects,and high safety.Currently,the research on collagen combined with microneedling therapy is still in its early stages,and achieving clinical application may become a key research direction in the future.The clinical application of collagen combined with microneedles for the treatment of photoaging still faces many challenges,such as exploring the optimal mechanical structure and materials of microneedles,selecting appropriate microneedle types,and insufficient clinical evidence that collagen combined with microneedles can further delay the treatment of skin photoaging.
2.Clinicopathological characteristics and immune microenvironment analysis of hepatoid adenocarcinoma of the stomach: a study of 65 cases
QIAN Yuping ; ZHANG Zhengwei ; LIU Yanfang
Chinese Journal of Cancer Biotherapy 2026;33(1):77-83
[摘 要] 目的:探讨胃肝样腺癌(HAS)的临床病理特征与免疫微环境异质性,筛选预后标志物,阐释其高侵袭性与疗效差的机制,为精准诊疗策略提供理论依据。方法: 回顾性收集2013年1月至2025年5月期间海军军医大学第一附属医院与第二附属医院收治的65例HAS患者的临床信息及病理资料。采用免疫组织化学技术检测HAS肝样分化标志物、神经内分泌标志物等分子表达情况,通过Kaplan-Meier生存分析法明确与预后相关的靶点。利用多色免疫荧光技术鉴定肿瘤区域内免疫细胞亚群分布情况,以阐明其免疫微环境特征。结果:65例患者中,男性54例(83.1%),女性11例(16.9%),中位年龄68岁。肿瘤好发于贲门(40%),其次为胃窦(32.3%)和胃体(27.7%)。中位随访时间23.18个月,15例患者死亡,46例生存,4例失访。HAS的胃镜及手术标本大体观呈灰白色实性质硬肿物,镜下可见中低分化胃腺癌与肝细胞癌(HCC)样分化区交错分布。表达神经内分泌相关分子的HAS患者呈现出更多的淋巴结转移数量及更短的总生存期。免疫微环境解析显示,HAS总体缺乏免疫细胞浸润,呈现“冷肿瘤”特征;免疫细胞主要聚集于胃癌腺体周围区域,而在HCC样分化区域罕见淋巴细胞浸润。结论:HAS侵袭性强,根治性手术是主要治疗手段;神经内分泌转化提示不良预后,是个体化治疗的关键标志;免疫细胞浸润缺乏,可能是其免疫治疗响应不佳的原因。
3.Effect and mechanism of collagen combined with microneedles in treatment of skin photoaging
Fengyi TAN ; Jiamin XIE ; Zhenfeng PAN ; Xinxu ZHANG ; Zetai ZHENG ; Zhiying ZENG ; Yanfang ZHOU
Chinese Journal of Tissue Engineering Research 2026;30(2):451-458
BACKGROUND:Collagen combined with microneedling therapy has gradually become an important means of improving skin photoaging.OBJECTIVE:To summarize and explore the main mechanism and clinical application status of collagen combined with microneedle therapy.METHODS:PubMed,China National Knowledge Infrastructure,and ScienceDirect databases were searched for Chinese and English literature published before August 2024.Chinese and English search terms were"ultraviolet radiation,photoaging,collagen,microneedling,clinical applications."Finally,74 articles were included for summary.RESULTS AND CONCLUSION:Collagen treats skin photoaging through mechanisms such as inhibiting matrix metalloproteinase expression,retaining skin moisture,and reducing melanin formation.Microneedles can better promote the penetration of collagen into deep layers of the skin,breaking down the skin's barrier and increasing the absorption rate.Collagen combined with microneedles has various beneficial effects for treating skin photoaging,such as whitening,anti-wrinkle,improving skin elasticity,shrinking pores,and repairing skin barriers.It also has the advantages of easy operation,significant effects,and high safety.Currently,the research on collagen combined with microneedling therapy is still in its early stages,and achieving clinical application may become a key research direction in the future.The clinical application of collagen combined with microneedles for the treatment of photoaging still faces many challenges,such as exploring the optimal mechanical structure and materials of microneedles,selecting appropriate microneedle types,and insufficient clinical evidence that collagen combined with microneedles can further delay the treatment of skin photoaging.
4.Allergen positivity rate and influencing factors of children with allergic rhinitis in Shanxi Region
Shuoying ZHANG ; Qinxue WANG ; Yanfang LI
Journal of Public Health and Preventive Medicine 2026;37(4):129-133
Objective To explore the distribution characteristics of allergen positivity rates and related influencing factors among children with allergic rhinitis in Shanxi Province, and to provide a scientific basis for the prevention, diagnosis, and treatment of allergic rhinitis in children in this area. Methods A total of 1 376 pediatric patients from Shanxi Province, who visited the Department of Otolaryngology of Shanxi Children's Hospital and underwent serum-specific immunoglobulin E (IgE) testing between March 2024 and March 2025, were selected. The positive rates of common allergens were analyzed, and the sensitization differences of allergens among different age cohorts were compared. The risk factors influencing the occurrence of allergic rhinitis in children were explored. Results The test results showed that 1,258 cases were positive for inhalant allergens, with a positive rate of 91.42%, while 855 cases tested positive for ingested allergens, with a positive rate of 62.14%. The top four inhalant allergens with the highest positive rates were Artemisia (38.01%), short ragweed (33.94%), willow/beech/oak/walnut (23.26%), and cat dander (18.82%). The top four ingested allergens with the highest positive rates were milk (24.56%), egg white (22.89%), peanut/soybean (22.60%), and cashew/pistachio/hazelnut/almond/walnut (18.60%). Among the inhalant allergens in children aged ≤ 16 years old, household dust mites had the highest sensitivity, followed by dust mites. Chicken protein and pistachios/cashews/almonds/hazelnuts/walnuts were more common among food allergens in children aged ≤ 5 years old, while milk and egg white were more common among food allergens in children aged 6-16 years old. The results of the multivariate logistic regression analysis showed that urban residence (OR=1.260, 95%CI: 1.009-1.574), a meat-based diet (OR=1.525, 95%CI: 1.218-1.908), presence of smoking/passive smoking (OR=2.033, 95%CI: 1.625-2.543), history of asthma (OR=4.122, 95%CI: 2.655-6.400), history of allergies (OR=2.016, 95%CI: 1.568-2.592), comorbid adenoid hypertrophy (OR=2.248, 95%CI: 1.529-3.304), and parental allergic rhinitis (OR=1.723, 95%CI: 1.298-2.289) were risk factors for allergic rhinitis in children. Conclusion The allergen spectrum of allergic rhinitis in children in Shanxi region is predominantly inhalant. Mugwort, short ragweed, willow/beech/oak/walnut, and cat dander are the main inhalant allergens, while milk, egg white, peanut/soybean, and cashew/pistachio/hazelnut/almond/walnut are the primary ingested allergens. The allergen positivity rate exhibits characteristic changes with age. Factors such as residing in urban areas, a meat-based diet, and a history of asthma are influential factors in the prevalence of allergic rhinitis among children in the Shanxi region.
5.Mechanisms of Babaodan in Attenuating Acetaminophen-induced Acute Liver Injury via Metabolic Reprogramming and Inflammatory Pathways
Ying ZHANG ; Yuchang AN ; Xiang ZHU ; Mei ZHONG ; Yanfang ZHENG ; Mingqing HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):122-130
ObjectiveTo investigate the effects and potential mechanisms of Babaodan (BBD) against acetaminophen (APAP)-induced acute liver injury (ALI) based on transcriptomics. MethodsA total of 36 male C57BL/6 mice were randomly divided into 6 groups (n=6 per group): normal group, model group, N-acetylcysteine group (NAC, 120 mg·kg-1), and BBD low-, medium-, and high-dose groups (BBD-L, BBD-M, BBD-H groups, 75, 150, 300 mg·kg-1, respectively). Except for the normal group, all other groups were subjected to APAP-induced ALI. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) were measured in each group. Hepatic levels or activities of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were detected using commercial kits. Hematoxylin-eosin (HE) staining was performed to evaluate the degree of liver histopathological damage. Transcriptomic analysis was employed to screen differentially expressed genes (DEGs), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed to identify differential pathways involved in BBD intervention against ALI. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were applied to validate the expression of differential genes and related pathway proteins. Additionally, glucose (GLU) consumption, as well as lactate (LD) and adenosine triphosphate (ATP) content were assessed across all mouse groups. ResultsPharmacodynamic evaluation showed that, compared with the normal group, the model group exhibited significantly elevated serum levels of ALT, AST, TC, TG, and LDL-C (P<0.01), significantly increased MDA level (P<0.01), and significantly decreased GSH-Px level (P<0.05). Compared with the model group, BBD intervention at different doses significantly reduced the serum levels of ALT, AST, TC, TG, and LDL-C (P<0.05, P<0.01), increased GSH-Px level (P<0.05, P<0.01), significantly decreased MDA level (P<0.01), and ameliorated hepatic histopathological injury. Liver transcriptomic analysis revealed that, following high-dose BBD intervention, the core genes were mainly enriched in pathways related to inflammatory responses and energy metabolic reprogramming, including the interleukin-17 (IL-17) signaling pathway, the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway, fructose and mannose metabolism, and glycolysis. Real-time PCR validation demonstrated that, compared with the normal group, the mRNA expression levels of glycolysis-related genes [hexokinase 1 (HK1), hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4), pyruvate kinase M (PKM), and lactate dehydrogenase A (LDHA)], as well as inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β)] were significantly upregulated in the model group (P<0.05, P<0.01). Compared with the model group, the BBD-H group showed significantly decreased mRNA expression of HK1, HK2, PFKFB3, PFKFB4, PKM, LDHA, TNF-α, IL-6, and IL-1β (P<0.05, P<0.01). Western blot results indicated that, compared with the normal group, the model group had significantly increased expression of glycolysis-related proteins [glucose transporter 1 (GLUT1), HK1, PFKFB3, and PKM], inflammatory proteins [interleukin-18 (IL-18), TNF-α, and IL-1β], and phosphorylated (p)-PI3K and p-Akt proteins (P<0.05, P<0.01). Compared with the model group, the BBD-H group exhibited significantly decreased expression of GLUT1, HK1, PFKFB3, PKM, IL-18, TNF-α, IL-1β, p-PI3K, and p-Akt (P<0.05, P<0.01). Metabolic indicator measurements showed that, compared with the model group, the BBD-H group showed significantly reduced GLU consumption, LD and ATP content (P<0.05, P<0.01). ConclusionBBD may alleviate APAP-induced ALI through dual regulation of metabolic reprogramming and inflammatory responses, potentially via inhibition of the PI3K/Akt signaling pathway.
6.Mechanisms of Babaodan in Attenuating Acetaminophen-induced Acute Liver Injury via Metabolic Reprogramming and Inflammatory Pathways
Ying ZHANG ; Yuchang AN ; Xiang ZHU ; Mei ZHONG ; Yanfang ZHENG ; Mingqing HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(20):122-130
ObjectiveTo investigate the effects and potential mechanisms of Babaodan (BBD) against acetaminophen (APAP)-induced acute liver injury (ALI) based on transcriptomics. MethodsA total of 36 male C57BL/6 mice were randomly divided into 6 groups (n=6 per group): normal group, model group, N-acetylcysteine group (NAC, 120 mg·kg-1), and BBD low-, medium-, and high-dose groups (BBD-L, BBD-M, BBD-H groups, 75, 150, 300 mg·kg-1, respectively). Except for the normal group, all other groups were subjected to APAP-induced ALI. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) were measured in each group. Hepatic levels or activities of malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) were detected using commercial kits. Hematoxylin-eosin (HE) staining was performed to evaluate the degree of liver histopathological damage. Transcriptomic analysis was employed to screen differentially expressed genes (DEGs), and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was performed to identify differential pathways involved in BBD intervention against ALI. Real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot were applied to validate the expression of differential genes and related pathway proteins. Additionally, glucose (GLU) consumption, as well as lactate (LD) and adenosine triphosphate (ATP) content were assessed across all mouse groups. ResultsPharmacodynamic evaluation showed that, compared with the normal group, the model group exhibited significantly elevated serum levels of ALT, AST, TC, TG, and LDL-C (P<0.01), significantly increased MDA level (P<0.01), and significantly decreased GSH-Px level (P<0.05). Compared with the model group, BBD intervention at different doses significantly reduced the serum levels of ALT, AST, TC, TG, and LDL-C (P<0.05, P<0.01), increased GSH-Px level (P<0.05, P<0.01), significantly decreased MDA level (P<0.01), and ameliorated hepatic histopathological injury. Liver transcriptomic analysis revealed that, following high-dose BBD intervention, the core genes were mainly enriched in pathways related to inflammatory responses and energy metabolic reprogramming, including the interleukin-17 (IL-17) signaling pathway, the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway, fructose and mannose metabolism, and glycolysis. Real-time PCR validation demonstrated that, compared with the normal group, the mRNA expression levels of glycolysis-related genes [hexokinase 1 (HK1), hexokinase 2 (HK2), 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3), 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4), pyruvate kinase M (PKM), and lactate dehydrogenase A (LDHA)], as well as inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β)] were significantly upregulated in the model group (P<0.05, P<0.01). Compared with the model group, the BBD-H group showed significantly decreased mRNA expression of HK1, HK2, PFKFB3, PFKFB4, PKM, LDHA, TNF-α, IL-6, and IL-1β (P<0.05, P<0.01). Western blot results indicated that, compared with the normal group, the model group had significantly increased expression of glycolysis-related proteins [glucose transporter 1 (GLUT1), HK1, PFKFB3, and PKM], inflammatory proteins [interleukin-18 (IL-18), TNF-α, and IL-1β], and phosphorylated (p)-PI3K and p-Akt proteins (P<0.05, P<0.01). Compared with the model group, the BBD-H group exhibited significantly decreased expression of GLUT1, HK1, PFKFB3, PKM, IL-18, TNF-α, IL-1β, p-PI3K, and p-Akt (P<0.05, P<0.01). Metabolic indicator measurements showed that, compared with the model group, the BBD-H group showed significantly reduced GLU consumption, LD and ATP content (P<0.05, P<0.01). ConclusionBBD may alleviate APAP-induced ALI through dual regulation of metabolic reprogramming and inflammatory responses, potentially via inhibition of the PI3K/Akt signaling pathway.
7.Exploration on the Onset of Acute Lymphoblastic Leukemia in Children Based on the Five-Circuit and Six-Qi Theory
Yunyun SUN ; Liang LI ; Leilei CHU ; Ke YANG ; Zhexiu JIANG ; Tian LIU ; Jianmin ZHANG ; Jian PAN ; Yanfang TAO
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(1):32-38
Objective To explore the five-circuit and six-qi features of birth time and onset time of children with acute lymphoblastic leukemia(ALL).Methods A total of 877 cases of children with ALL from Children's Hospital of Soochow University from June 2021 to February 2023 were collected,and their five-circuit and six-qi features of birth time and onset time were analyzed.And then the correlation of five-circuit and six-qi features of birth time and onset time with ALL was explored preliminarily,and the pathogenic characteristics of congenital factors and acquired pathogenic factors were revealed.Results(1)The children who were born in the year with the heavenly stems being bing(the 3rd of the ten heavenly stems)and ding(the 4th of the ten heavenly stems)and with the earthly branches being shen(the 8th of the twelve earthly branches)and you(the 9th of the twelve earthly branches)are prone to suffer from ALL,and the birth year of children with ALL had the five-circuit and six-qi features of the joining of guest circuit with dominant circuit being rebellious.ALL is commonly seen in the year with the heavenly stems being geng(the 7th of the ten heavenly stems)and xin(the 8th of the ten heavenly stems)and with the earthly branches being zi(the 1st of the twelve earthly branches)and chou(the 2nd of the twelve earthly branches),and the onset year of ALL in children had the five-circuit and six-qi features of the yearly circuit being gold-circuit and water-circuit,sitian-zaiquan yearly circuit qi being shaoyin monarch-fire with yangming dryness-gold,taiyin damp-earth with taiyang cold-water,and the qi-circuit assimilation relationship being celestial correspondence,same celestial correspondence,celestial correspondence in convergent year,disharmony,mildly-rebellious,and celestial restriction.Conclusion Gold-dryness and water-cold are the congenital factors and acquired pathogenic factors of ALL.The onset of ALL in children is closely related to qi insufficiency and qi stagnation of wood and fire in five-circuit and six-qi theory.
8.Efficacy Observation of Therapy Focusing on Regulating Spleen and Stomach for the Treatment of Acoustic Hypersensitivity
Haixin ZHANG ; Peng LIU ; Jinguang LIU ; Jieheng LIU ; Yanfang CHEN ; Wenzhi LIN ; Weiping HE
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(1):131-139
Objective To observe the clinical efficacy of therapy focusing on regulating spleen and stomach for the treatment of acoustic hypersensitivity(AH),which also named as auditory hyperaesthesia,and to explore the relevant factors influencing the efficacy.Methods From January 2018 to December 2023,the patients admitted to the outpatient department of otolaryngology-head and neck surgery of the First Affiliated Hospital of Guangzhou University of Chinese Medicine and diagnosed as AH with complete medical data were included as study subjects.Follow-up through telephone or network was carried out for those patients who with uncertain prognosis.The changes in the pre-and post-treatment scores of Indicators of Acoustic Hypersensitivity Severity(IAHS)were used as the reference index for efficacy evaluation,and the cure rate and effective rate were used as the efficacy analysis indexes for statistical analysis.And then the efficacy of traditional Chinese medicine(TCM)medical care combined with health care focusing on regulating spleen and stomach for the treatment of AH was evaluated.Furthermore,multivariate logistic regression was used to explore the relevant factors influencing the efficacy of AH.Results A total of 298 cases of AH patients meeting the inclusion criteria were collected,among which 151 cases(50.67%)were cured,22 cases(7.38%)were markedly effective,75 cases(25.17%)were effective,and 50 cases(16.78%)were ineffective,with a total effective rate of 83.22%(248/298).The univariate analysis results showed that eight factors(including post-treatment lifestyle score,range of lifestyle adjustment,severity of AH,duration of the disease,presence of tinnitus,presence of hearing drop,presence of vertigo,and age)had an influence on the efficacy,and the differences were statistically significant(P<0.05 or P<0.01).However,the factors of gender,occupation,education level,and history of noise exposure had no influence on the efficacy(P>0.05).The multivariate binary logistic regression analysis showed that five factors,namely post-treatment lifestyle score,range of lifestyle adjustment,severity of AH,presence of vertigo and duration of the disease,had a significant correlation with the cure rate of AH(P<0.05 or P<0.01),and four factors,namely post-treatment lifestyle score,severity of AH,range of lifestyle adjustment and age,had a significant correlation with the effective rate of AH(P<0.05 or P<0.01).Conclusion The method of TCM medical care and health care focusing on regulating spleen and stomach exerts a better efficacy for AH patients.And the lifestyle,severity of AH,duration of the disease,age,and presence of vertigo are the relevant factors affecting the outcomes,which is worthy of further in-depth study.
9.Liu Peng's Clinical Experience in Treating Tinnitus with Yishi Therapy
Yanfang CHEN ; Haixin ZHANG ; Peng LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2025;42(10):2572-2578
Tinnitus is a refractory condition in otology.It is widely accepted that patients with tinnitus can only adapt to it,for there is no universally recognized effective pharmacotherapy till now.Through the interpretation of classical Chinese medical texts and after decades of clinical practice,Professor Liu Peng proposes that the primary pathogenesis of tinnitus is due to spleen-stomach dysfunction,largely caused by unhealthy life styles.Based on the holistic perspective of traditional Chinese medicine(TCM),he developed the Yishi therapy for tinnitus management.Yishi therapy emphasizes the integration of medical treatment and self-care via prioritizing mentorship and assisting with medical techniques.The mentorship is embodied by identifying and correcting patients' unhealthy living habits,guiding the patients to maintain dietary moderation,regular living,and emotional regulation,thereby activating their self-recuperative powers and promoting tinnitus.Medical techniques refer to the utilization of oral use of herbal medicine and external therapies on the basis of adjustment of lifestyle,aiming to restore the spleen-stomach function of ascending of lucidity and descending of turbidity,thereby alleviating tinnitus.Oral use of herbal medicine is performed with Guzhong Decoction(mainly composed of Astragali Radix,Codonopsis Radix,Cinnamomi Ramulus,Zingiberis Rhizoma,Atractylodis Macrocephalae Rhizoma,Pinelliae Rhizoma Praeparatum,Amomi Fructus,Acori Tatarinowii Rhizoma,and Glycyrrhizae Radix et Rhizoma Praeparata cum Melle)as the basic formula,through modified use based on syndrome differentiation.External treatment mainly refers to auricular acupoint pressing and occipital-knocking therapy.By combining the medical treatment and self-care,Yishi therapy exerts reliable efficacy for the treatment of tinnitus through regulating spleen-stomach function and activating the self-recuperative powers,and will provide a novel therapeutic approach to tinnitus.
10.Establishment of duplex quantitative real-time PCR detection method for gyrovirus galga1 and gyrovirus homsa1
Dan YU ; Zhixun XIE ; Junke ZHAO ; Yanfang ZHANG ; Zhiqin XIE ; Liji XIE ; Wen-qiao YIN ; Huaying YU
Chinese Journal of Veterinary Science 2025;45(1):59-65,73
Gyrovirus galga1(GyVg1)and gyrovirus homsa1(GyH1)are two newly discovered cir-coviruses that can cause symptoms related to transmissible viral proventriculitis of chickens.These viruses have been reported in various regions worldwide.This research aims to establish a duplex real-time PCR assay capable of identifying and detecting GyVg1 and GyH1.Specific primers and probes were designed based on the conserved regions of GyVg1 and GyH1 respectively using all genome sequence data currently available in GenBank.After optimizing reaction conditions,the du-plex real-time PCR detection method was established and further validated by comparing it with a conventional PCR assay and sequencing results from an analysis of 256 clinical samples collected in 2023 across eight regions of Nanning,Guangxi.The results showed that GyVg1 and GyH1 could be identified in 1 h by the duplex real-time PCR assay and two pairs of primer probes can amplify effectively but there is no any cross reaction with other pathogens.Besides,the detection limit was determined to be 7.5 copies/μL.The correlation coefficient of standard curves exceeded 0.99,and CV for intra-and inter-assay was less than 0.45%.Based on clinical performance,when the quanti-ty of template was greater than or equal to 100 copies,the agreements between the duplex real-time PCR assay and the conventional PCR assay were 94.3%(GyVg1)and 100%(GyH1).In con-clusion,the newly developed duplex real-time PCR assays exhibited good specificity,sensitivity and repeatability,which could contribute to the rapid detection and differentiation of GyVg1 and GyH1.


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