1.Preliminary study on an improved method for constructing internal quality control framework of ELISA
Youbin DUAN ; Rui WANG ; Le CHANG ; Changwen QIU ; Zhiqiang LI ; Gengrui CHEN ; Jingjuan YANG ; Qing HE ; Lunan WANG
Chinese Journal of Blood Transfusion 2026;39(1):103-108
Objective: To propose an improved method for constructing the internal quality control (IQC) framework for ELISA assays and validate its efficacy by statistically analyzing IQC data from nine blood center laboratories. Methods: 1) IQC data was collected from nine blood centers and analyzed using a domestic HBsAg ELISA detection kit as an example. 2) Differences between IQC values across batches within Blood Center 1 were assessed. 3) Statistical analyses were performed on batch usage, number of batches used, days of use, number of QC points, batch-specific means, and coefficients of variation (CV) across all nine centers. 4) Using the improved construction method for IQC framework, provisional and permanent frames were established for batches within Blood Center 1 and Blood Center 9, followed by outlier determination. Results: 1) Statistically significant differences were observed in IQC data between batches within Blood Center 1 (P<0.01). It is recommended that both the control material/reagents and the control chart framework be replaced simultaneously. 2) There were substantial differences among 9 blood centers regarding the control material/reagent lot numbers used, the number of QC runs per batch, and the QC values for identical lots. Therefore, individual laboratories should establish their own IQC chart frameworks. 3) The improved IQC framework construction method for ELISA assays is as follows: provisional frames are established via frame-shifting, using the pre-experimental mean and cumulative coefficient of variation (CV) from the preceding batch. For batches used >20 days with >20 QC points, permanent frames are constructed by aggregating in-control data accumulated over ≥20 days with ≥20 points to calculate cumulative mean and standard deviation. The provisional and permanent frames constructed by this method identified all 26 extreme outliers across Blood Centers 1 and 9 as out-of-control. Among the 218 general outliers, 10 were classified as normal by the provisional frames, while the remainder were designated as warnings or out-of-control. This method effectively monitors assay stability. Conclusion: Based on the statistical analysis of IQC practices across blood centers of varying scales, combined with the inherent characteristics of ELISA assays and the batch-to-batch instability of reagents/QC materials, it is recommended to reconstruct QC charts upon lot changes. The proposed method—utilizing frame-shifting for provisional frames and establishing permanent frames based on cumulative data—is applicable to blood center laboratories of differing sizes and effectively monitors the stability of the ELISA assay process.
2.Evolution, Applications, and Perspectives of Technologies for Genetically Engineered Mouse Generation
Jianfeng WU ; Zhiqiang SHAO ; Ying HE ; Suqin WU
Laboratory Animal and Comparative Medicine 2026;46(5):759-770
Genetically engineered mice are important tools for elucidating gene function and disease mechanisms and for advancing drug development. With the growing demand in biomedical research for complex genetic modifications, rapid model generation, and high-throughput functional analysis, technologies for generating genetically engineered mice continue to evolve. This review systematically summarizes the evolution and applications of these technologies, with a focus on conventional approaches, including early transgenic technologies and embryonic stem cell (ESC)-based homologous recombination for gene targeting, as well as the use of random mutagenesis approaches, such as N-ethyl-N-nitrosourea (ENU) mutagenesis and transposon insertion, in forward genetic screening. It further discusses the principles and characteristics of programmable nucleases, including zinc finger nucleases, transcription activator-like effector nucleases, and CRISPR/Cas systems, as well as their applications in generating genetically engineered mice. With advances in genome editing, Cas9-derived systems, base editing, prime editing, and RNA editing further improve editing precision, expand the repertoire of genetic modifications and precise regulatory approaches, and provide additional options for generating genetically engineered mice. This review also compares the advantages, limitations, and suitability of different technologies for specific applications and summarizes quality-control strategies addressing practical issues that may affect model reliability, including off-target effects, mosaicism, genetic background, and genetic stability. Finally, future directions in technologies for generating genetically engineered mice are discussed to inform the generation and application of related models.
3.Differences in cytokines expression between mild and severe infant cases infected with respiratory syncytial virus
Guangyu XUE ; Yuting HU ; Kexin ZONG ; Qin LUO ; Shengnan YANG ; Miao FENG ; Xiaoyu YI ; Zhiqiang XIA ; Chen GAO ; Haijun DU ; Ying LI ; Ying CHEN ; Feng HE ; Yajuan WANG ; Yingli QU ; Jin CAO ; Wenyan TIAN ; Qinqin SONG ; Hailan YAO ; Jun HAN
Chinese Journal of Experimental and Clinical Virology 2025;39(3):370-377
Objective:To analyze the clinical characteristics and cytokines expression characteristics in infants with mild and severe respiratory syncytial virus (RSV) infection.Methods:From May 2023 to December 2023, plasma samples and clinical information were collected from 16 infants with RSV infection and 14 control infants. Cytek Aurora flow cytometry (Cytek, America) and Enzyme linked immunosorbent assay (ELISA) were used to detect the expression levels of 25 cytokines after mild and severe RSV infection.Results:Cough and nasal obstruction were the main clinical manifestations in infants with mild RSV infection, accompanied by polypnea, wheezing and other symptoms. The main symptoms of severe RSV infection were cough and rales, accompanied by fever and polypnea. In comparison with the control group, the expression levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-13, IL-22, TNF-α, IFN-α, IFN-β, MIP-1β, I-TAC, ENA-78, GROα, Eotaxin, and MCP-1 in the RSV infection group all exhibited an upregulation trend. Both IP-10 and MIP-3α demonstrated a downward trend in the RSV infection group; however, there was no statistically significant difference ( P>0.05). The levels of IL-10, IFN-γ, MIP-1α, and IL-8 in the RSV infection group were significantly higher than those in the control group, whereas the levels of MIG, TARC, and RANTES in the RSV infection group were significantly lower than those in the control group ( P<0.05). The levels of IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-13, IL-22, IFN-β, IFN-γ, TNF-α, IL-8, I-TAC, MIP-1β, Eotaxin, and MCP-1 in the mild RSV infection group were significantly higher than those in the severe RSV infection group ( P>0.05). Among these, the levels of MIG, RANTES, TARC, MIP-3α, and ENA-78 in the mild infection group were all lower than those in the severe infection group. The expressions of ENA-78 and MIP-1α in the severe infection group were significantly higher than those in the mild infection group and also higher than those in the control group. There was no significant difference in IP-10 and GROα between the mild and severe RSV infection groups ( P>0.05). Conclusions:The differences in clinical features and cytokines between infants with mild and severe RSV infection provide important data support for the prevention and treatment of RSV infection in infants.
4.Psoriasis-atopic dermatitis overlap phenotype: immune dysregulation mechanisms and emerging clinical insights
Zhiqiang SONG ; Yuanxiong HE ; Anqi CHEN
Chinese Journal of Dermatology 2025;58(11):1009-1014
Atopic dermatitis and psoriasis are two chronic inflammatory skin diseases with distinct immune mechanisms, primarily driven by Th2-type inflammation and Th1/Th17 pathways, respectively. However, some patients may simultaneously or sequentially exhibit overlapping phenotypes of atopic dermatitis and psoriasis (psoriasis-atopic dermatitis overlap, PAO), characterized by the coexistence of scaly erythema and eczematous lesions. Currently, the nomenclature, diagnostic criteria, and pathogenesis of PAO remain unclear, and its incidence is increasing due to the widespread use of targeted therapies. The diagnosis of PAO requires an integrated assessment of clinical features, histopathological findings, and biomarker profiles, while treatment necessitates a balance between immune modulation and individualized strategies. This article summarizes the characteristic manifestations of and genetic-immune interactions involved in PAO based on clinical practice, and explores diagnostic challenges and therapeutic options to address this complex and increasingly prominent clinical issue.
5.Comparative analysis of clinical characteristics between psoriasis-atopic dermatitis overlap phenotype and psoriasis vulgaris: a single-center retrospective exploratory study
Yuanxiong HE ; Sisi DENG ; Zhiqiang SONG
Chinese Journal of Dermatology 2025;58(11):1027-1033
Objective:To analyze the clinical characteristics between patients with psoriasis-atopic dermatitis overlap (PAO) and those with psoriasis vulgaris (PSO), and to enhance the understanding of the diagnosis and treatment of this overlap phenotype.Methods:A retrospective exploratory study was conducted on clinical data from patients who were diagnosed with PAO or PSO at the Department of Dermatology, Southwest Hospital, Army Medical University between January 2018 and June 2025. Clinical characteristics, laboratory examination results, comorbidities, and treatment regimens were compared between the two groups. Categorical data were compared using the chi-square test or Fisher's exact test; for non-normally distributed measurement data, intergroup comparisons were conducted using the Mann-Whitney U test. Results:A total of 103 PSO patients and 13 PAO patients were included. Patients in the PAO group were older than those in the PSO group ( M [ Q1, Q3]:63.00 [54.00, 71.50] years vs. 50.00 [38.00, 61.00] years; Z = 2.75, P = 0.006]. No significant differences were found between the two groups in terms of gender distribution, body mass index, disease duration, personal or family history of atopic diseases (all P > 0.05). Skin lesions involved the whole body in both PAO and PSO groups, with the trunk and limbs being commonly affected sites, and no significant difference in the lesion distribution was observed ( P > 0.05). Compared with the PSO group, the PAO group had fewer plaque lesions (5/13 [38.5%] vs. 70/103 [68.0%]), but more eczematous changes such as erosions and exudation, as well as scratches and crusts due to pruritus (all P < 0.05). Laboratory tests revealed that the PAO group showed increased peripheral blood neutrophil counts, eosinophil counts, serum IgE levels, eosinophil-to-lymphocyte ratios (ELRs), and neutrophil-to-lymphocyte ratios (NLRs) compared with the PSO group (all P < 0.05) ; moreover, the proportions of patients with elevated eosinophil counts (5/13 [38.5%] vs. 8/103 [7.8%], P < 0.001) and those with elevated serum IgE levels (10/13 [76.9%] vs. 39/103 [37.9%], P = 0.014) were significantly higher in the PAO group than in the PSO group. Compared with the PSO group, the PAO group had a higher overall comorbidity rate (11/13 [84.6%] vs. 52/103 [50.5%], P = 0.035), including a higher prevalence of hypertension. Regarding topical treatments, no significant differences were found in the use frequency of topical glucocorticoids (96 [93.2%] vs. 11 [84.6%]) or vitamin D3 analogs between the two groups (both P > 0.05) ; for systemic treatments, immunosuppressants such as cyclosporine (4/13 [30.8%] vs. 2/103 [1.9%], P = 0.001) and Tripterygium wilfordii (4/13 [30.8%] vs. 7/103 [6.8%], P = 0.021) were more commonly used in the PAO group compared with the PSO group; for targeted therapies, the PSO group received interleukin (IL) -17A inhibitors (13 cases, 12.6%), IL-23 inhibitors (4 cases, 3.9%), or tumor necrosis factor-α inhibitors (5 cases, 4.9%), while the PAO group received Janus kinase inhibitors (2 cases, 15.4%) or an IL-23 inhibitor (1 case, 7.7%) . Conclusions:PAO exhibited characteristics of both PSO and atopic dermatitis, with distinct differences in skin manifestations, laboratory findings, and treatment approaches compared with PSO. Topical glucocorticoids were the primary topical treatment for PAO, while systemic treatment was centered on immunosuppressants, highlighting the need for personalized treatment strategies.
6.Prevalence Characteristics and Disease Burden of Lung Cancer in Hebei Cancer Registration Areas from 2012 to 2020
Yanfang FU ; Daojuan LI ; Yanyu LIU ; Zhiqiang YAN ; Yaxian PANG ; Yutong HE
China Cancer 2025;34(6):452-463
[Purpose]To analyze the prevalence characteristics and disease burden of lung cancer in Hebei cancer registration areas from 2012 to 2020.[Methods]Lung cancer data were collected from the Hebei Provincial Cancer Registry from 2012 to 2020.The crude incidence/mortality rates,age-standardized incidence/mortality rates by Chinese standard population(ASIRC/ASMRC)and by world standard population(ASIRW/ASMRW)were calculated.The Joinpoint model was used to calculate the annual percentage change(APC)and average annual percentage change(AAPC).Years of life lost(YLL)and years lived with disability(YLD)and the disability-adjusted life years(DALY)were calculated.[Results]From 2012 to 2020,the ASIRW of lung cancer in Hebei can-cer registration areas was 33.13/105,44.56/105 for men and 22.54/105 for women,respectively;the incidence rates of urban and rural areas were 29.05/105 and 33.52/105,respectively.The incidence rates increased with ages,reaching a peak in the age group of 80~84 years old.There was a de-creasing trend in the ASIRW of lung cancer(AAPC=-3.99%,P<0.05).From 2012 to 2020,the ASMRW of lung cancer was 25.80/105,36.56/105 for men and 15.96/105 for women,respectively;the mortality rates of urban and rural areas were 25.14/105 and 26.12/105,respectively.The mor-tality rates increase with ages,reaching a peak in the age group of 85 and above years old.There was a decreasing trend in the mortality of lung cancer(AAPC=-4.65%,P<0.001)from 2012 to 2020.The DALY of lung cancer in Hebei Province from 2012 to 2020 was 484 194 person-years,with male accounting for 66.77%,female accounted for 33.23%,the DALY rate of lung cancer was 3.31‰,of which 35.57%in urban areas and 64.43%in rural areas.[Conclusion]Lung can-cer incidence and mortality rate in Hebei cancer registration areas from 2012 to 2020 showed a decreasing trend.The disease burden is gradually increasing with age in middle-aged and el-derly population.
7.Treatment outcome and influencing factors of rifampicin-resistant pulmonary tuberculosis patients in Shaanxi Province from 2017 to 2022
Kaikai LI ; Zhanfeng HAN ; Hongwei ZHANG ; Zhiqiang HE
Chinese Journal of Infectious Diseases 2025;43(10):599-605
Objective:To analyze the treatment outcome and its related factors of rifampicin-resistant pulmonary tuberculosis (RR-PTB) patients in Shaanxi Province from 2017 to 2022.Methods:Through the "Drug-resistant Cases" in the "Surveillance Report Management" subsystem of the "China Disease Control and Prevention System", the RR-PTB patients registered from January 1st, 2017 to December 31st, 2022 in Shaanxi Province were collected.A cross-sectional study was used to analyze the epidemiological characteristics, history of anti-tuberculosis treatment, medication status, and treatment outcomes and prognosis of the patients was performed. The trend analysis was conducted by trend chi-square test. Chi-square test was used for statistical comparison. Multivariable logistic regression model was used to analyze factors influencing the treatment outcome of RR-PTB patients.Results:From 2017 to 2022, a total of 2 582 cases of RR-PTB were registered in Shaanxi Province. Epidemiological characteristics showed that the male registration incidence (1.55/100 000) was higher than that of females (0.61/100 000), and the difference was statistically significant ( χ2=473.04, P<0.001). The population was mainly young and middle-aged (38.88%(1 004/2 582)) and farmers (66.50%(1 717/2 582)). The highest registered incidence was in Southern Shaanxi (1.71/100 000). A total of 1 567 cases were successfully treated, with a successful treatment rate of 60.69%. The successful treatment rate of RR-PTB patients increased from 58.17%(121/208) in 2017 to 66.41%(174/262) in 2022, which showed an overall upward trend with statistically significant (trend χ2=62.84, P<0.001). The multivariable logistic regression analysis showed that male (odds ratio ( OR)=1.838, 95% confidence interval ( CI) 1.392 to 2.427, P<0.001), 45 to 64 years old ( OR=2.119, 95% CI 1.361 to 3.300, P=0.001), ≥65 years old ( OR=5.070, 95% CI 3.016 to 8.521, P<0.001), living the Northern Shaanxi ( OR=1.639, 95% CI 1.087 to 2.471, P=0.018), retreatment ( OR=1.646, 95% CI 1.264 to 2.144, P<0.001), with cross-regional mobility ( OR=1.821, 95% CI 1.403 to 2.363, P<0.001), cumulative months of medication <6 months ( OR=55.310, 95% CI 40.267 to 75.974, P<0.001), family member or self-medication management ( OR=2.176, 95% CI 1.527 to 3.100, P<0.001) were risk factors for successful treatment of RR-PTB patients. Conclusions:The successful treatment rate of RR-PTB patients in Shaanxi Province has shown an upward trend.Male, age ≥45 years, living in the Northern Shaanxi, retreatment, cross-regional mobility, cumulative months of medication <6 months, family or self-medication management are risk factors for treatment outcome of RR-PTB patients.Treatment management, the supervision and propaganda education should be strengthened to further reduce the risk of adverse treatment outcomes.
8.Development and application of a digital PCR-based assay for rapid diagnosis of common fetal chromosomal aneuploidies.
Xuejiao CHEN ; Yanfeng YANG ; Yuanyuan YING ; Feiyan PAN ; Zhiqiang GU ; Weimeng JIAO ; Zehang HE ; Huihui XU
Chinese Journal of Medical Genetics 2025;42(5):592-596
OBJECTIVE:
To assess the clinical value of digital PCR (dPCR) for the prenatal diagnosis of common fetal aneuploidies.
METHODS:
A dPCR-based assay was developed for detecting trisomies 21, 18, and 13. A retrospective analysis was carried out on 173 amniotic fluid samples collected by the Prenatal Diagnosis Center of Taizhou Hospital between January 2017 and December 2023. By using chromosomal karyotyping as the gold standard, the diagnostic performance of the multiplex dPCR system was evaluated in a double-blind manner. This study has been approved by the Ethics Committee of Taizhou Hospital (Ethics No. K20250339).
RESULTS:
Chromosomal karyotyping has identified 59 cases of trisomy 21, 5 cases of trisomy 18, 2 cases of trisomy 13, 6 cases with chromosomal structural abnormalities or mosaicisms, and 101 cases with a normal karyotype. The dPCR results (Z-score cutoff = 4.0, CI = 99.997%) showed full concordance with karyotyping (sensitivity = 100%, specificity = 100%, Kappa = 1). Among the 6 structurally abnormal or mosaicism samples, dPCR has accurately detected 4 cases, but mis-classified 2 cases of trisomy 21 with very low-level mosaicisms (3.3%, 6.9%, respectively) as normal.
CONCLUSION
The established multiplex dPCR system demonstrated high diagnostic accuracy for common chromosomal aneuploidies, with results available within 24 hours. It can serve as an efficient supplementary tool to conventional chromosomal karyotyping, providing reliable support for time-sensitive clinical decision-making in prenatal diagnosis.
Humans
;
Female
;
Pregnancy
;
Aneuploidy
;
Prenatal Diagnosis/methods*
;
Karyotyping
;
Retrospective Studies
;
Polymerase Chain Reaction/methods*
;
Chromosome Disorders/genetics*
;
Adult
;
Trisomy 13 Syndrome/diagnosis*
;
Trisomy 18 Syndrome/genetics*
;
Down Syndrome/genetics*
9.Psoriasis-atopic dermatitis overlap phenotype: immune dysregulation mechanisms and emerging clinical insights
Zhiqiang SONG ; Yuanxiong HE ; Anqi CHEN
Chinese Journal of Dermatology 2025;58(11):1009-1014
Atopic dermatitis and psoriasis are two chronic inflammatory skin diseases with distinct immune mechanisms, primarily driven by Th2-type inflammation and Th1/Th17 pathways, respectively. However, some patients may simultaneously or sequentially exhibit overlapping phenotypes of atopic dermatitis and psoriasis (psoriasis-atopic dermatitis overlap, PAO), characterized by the coexistence of scaly erythema and eczematous lesions. Currently, the nomenclature, diagnostic criteria, and pathogenesis of PAO remain unclear, and its incidence is increasing due to the widespread use of targeted therapies. The diagnosis of PAO requires an integrated assessment of clinical features, histopathological findings, and biomarker profiles, while treatment necessitates a balance between immune modulation and individualized strategies. This article summarizes the characteristic manifestations of and genetic-immune interactions involved in PAO based on clinical practice, and explores diagnostic challenges and therapeutic options to address this complex and increasingly prominent clinical issue.
10.Comparative analysis of clinical characteristics between psoriasis-atopic dermatitis overlap phenotype and psoriasis vulgaris: a single-center retrospective exploratory study
Yuanxiong HE ; Sisi DENG ; Zhiqiang SONG
Chinese Journal of Dermatology 2025;58(11):1027-1033
Objective:To analyze the clinical characteristics between patients with psoriasis-atopic dermatitis overlap (PAO) and those with psoriasis vulgaris (PSO), and to enhance the understanding of the diagnosis and treatment of this overlap phenotype.Methods:A retrospective exploratory study was conducted on clinical data from patients who were diagnosed with PAO or PSO at the Department of Dermatology, Southwest Hospital, Army Medical University between January 2018 and June 2025. Clinical characteristics, laboratory examination results, comorbidities, and treatment regimens were compared between the two groups. Categorical data were compared using the chi-square test or Fisher's exact test; for non-normally distributed measurement data, intergroup comparisons were conducted using the Mann-Whitney U test. Results:A total of 103 PSO patients and 13 PAO patients were included. Patients in the PAO group were older than those in the PSO group ( M [ Q1, Q3]:63.00 [54.00, 71.50] years vs. 50.00 [38.00, 61.00] years; Z = 2.75, P = 0.006]. No significant differences were found between the two groups in terms of gender distribution, body mass index, disease duration, personal or family history of atopic diseases (all P > 0.05). Skin lesions involved the whole body in both PAO and PSO groups, with the trunk and limbs being commonly affected sites, and no significant difference in the lesion distribution was observed ( P > 0.05). Compared with the PSO group, the PAO group had fewer plaque lesions (5/13 [38.5%] vs. 70/103 [68.0%]), but more eczematous changes such as erosions and exudation, as well as scratches and crusts due to pruritus (all P < 0.05). Laboratory tests revealed that the PAO group showed increased peripheral blood neutrophil counts, eosinophil counts, serum IgE levels, eosinophil-to-lymphocyte ratios (ELRs), and neutrophil-to-lymphocyte ratios (NLRs) compared with the PSO group (all P < 0.05) ; moreover, the proportions of patients with elevated eosinophil counts (5/13 [38.5%] vs. 8/103 [7.8%], P < 0.001) and those with elevated serum IgE levels (10/13 [76.9%] vs. 39/103 [37.9%], P = 0.014) were significantly higher in the PAO group than in the PSO group. Compared with the PSO group, the PAO group had a higher overall comorbidity rate (11/13 [84.6%] vs. 52/103 [50.5%], P = 0.035), including a higher prevalence of hypertension. Regarding topical treatments, no significant differences were found in the use frequency of topical glucocorticoids (96 [93.2%] vs. 11 [84.6%]) or vitamin D3 analogs between the two groups (both P > 0.05) ; for systemic treatments, immunosuppressants such as cyclosporine (4/13 [30.8%] vs. 2/103 [1.9%], P = 0.001) and Tripterygium wilfordii (4/13 [30.8%] vs. 7/103 [6.8%], P = 0.021) were more commonly used in the PAO group compared with the PSO group; for targeted therapies, the PSO group received interleukin (IL) -17A inhibitors (13 cases, 12.6%), IL-23 inhibitors (4 cases, 3.9%), or tumor necrosis factor-α inhibitors (5 cases, 4.9%), while the PAO group received Janus kinase inhibitors (2 cases, 15.4%) or an IL-23 inhibitor (1 case, 7.7%) . Conclusions:PAO exhibited characteristics of both PSO and atopic dermatitis, with distinct differences in skin manifestations, laboratory findings, and treatment approaches compared with PSO. Topical glucocorticoids were the primary topical treatment for PAO, while systemic treatment was centered on immunosuppressants, highlighting the need for personalized treatment strategies.

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