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.

Objective Objective To analyze the potential spatial factors affecting the genetic differentiation of Oncomelania hupensis robertsoni in Yunnan Province. Methods A total of 13 administrative villages were selected from schistosomiasis-endemic areas of Yunnan Province as O. hupensis snail sampling sites. At least 200 snails were collected in each site, and the spatial variable data of each site were recorded, including longitude, latitude and altitude. Thirty active and Schistosoma japonicum uninfected O. hupensis snails were selected from each sampling site by means of the crawling method and the cercarial shedding method. Genomic DNA was extracted from O. hupensis snails. Following PCR amplification, purification of PCR amplification products and sequencing, the gene sequences of O. hupensis snail samples were spliced and edited using the DNAstar software and the NCBI database to yield the complete mitochondrial sequences of O. hupensis snails at each sampling site, and the mitochondrial genetic distance matrix of O. hupensis robertsoni was calculated at each sampling site. The geographical coordinates of each sampling site were marked using the software ArcGIS 10.2, and the straight-line geographical distance between each sampling site was calculated. The altitude difference, longitude difference and latitude difference between each sampling site were calculated using the Excel software, and the correlation between the mitochondrial genetic distance matrix of O. hupensis robertsoni and each spatial variable matrix was examined by using the Mantel test at 13 sampling sites in Yunnan Province. Results Among the 13 O. hupensis snail sampling sites in Yunnan Province, the largest mitochondrial genetic distance of O. hupensis robertsoni snail populations was seen between Anding Village, Nanjian Yi Autonomous County and Caizhuang Village, Midu County (26.244 2), and the largest geographical distance was seen between Dongyuan Village, Gucheng District and Cangling Village, Chuxiong County (272.64 km). The highest altitude difference was seen between Anding Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1 086.10 m), and the largest longitude difference was found between Qiandian Village, Eryuan County and Cangling Village, Chuxiong County (1.86°), while the largest latitude difference was measured between Leqiu Village, Nanjian Yi Autonomous County and Dongyuan Village, Gucheng District (1.81°). In addition, the mitochondrial genetic distance of O. hupensis robertsoni snail populations was positively correlated with altitude at 13 snail sampling sites in Yunnan Province (r = 0.542 8, P < 0.001), and showed no significant correlations with geographical distance (r = 0.093 4, P > 0.05), longitude (r = −0.199 5, P > 0.05) or latitude (r = 0.205 7, P > 0.05). Conclusion Altitude may be a potential spatial factor affecting the genetic differentiation of O. hupensis robertsoni in Yunnan Province.

Hepatic alveolar echinococcosis is a highly invasive zoonotic parasitic disease with poor prognosis. Surgical intervention serves as the pivotal approach to achieve radical cure and improve the prognosis of hepatic alveolar echinococcosis patients. In recent years, with the popularization of the concept of precision surgery and the development of the multidisciplinary diagnosis and treatment model, the surgical treatment strategies for hepatic alveolar echinococcosis have been continuously enriched, and the selection of surgical procedures has become increasingly diversified. Although key surgical techniques such as radical hepatectomy, autologous liver transplantation and allogeneic liver transplantation have achieved remarkable progress in clinical application, many insurmountable challenges still remain. Therefore, by sorting out the latest evidence-based advances in the field of surgical treatment for hepatic alveolar echinococcosis, this article focuses on discussing the application status and bottlenecks of radical hepatectomy, autologous liver transplantation and allogeneic liver transplantation in hepatic alveolar echinococcosis, aiming to provide a reference for the clinical treatment of hepatic alveolar echinococcosis.

Objective To investigate the distribution characteristics of Oncomelania hupensis snails in Yunnan Province fol-lowing interruption of schistosomiasis transmission, so as to provide the evidence for assessing the risk of schistosomiasis transmission and scientifically formulating the schistosomiasis surveillance program. Methods According to the requirements of the National Schistosomiasis Surveillance Scheme (2020 Edition), O. hupensis snail surveillance data were collected from 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province from 2020 to 2024, including area of snail survey, area of snail habitats, area of re-emerging snail habitats, number of frames surveyed, number of frames with O. hupensis snails, number of O. hupensis snails captured, and number of living snails, and the occurrence of frames with snails and mean density of living snails were calculated. Changes in snail status over the 5-year period from 2020 to 2024 and the differences in snail distributions specified by epidemic intensity, environmental type, and vegetation type were analyzed. Results The areas of snail survey increased from 1 727.96 hm² in 2020 to 3 894.45 hm² in 2024 (peak) across 18 schistosomiasis-endemic counties (cities, districts) in Yunnan Province during the period from 2020 through 2024. The areas of snail habitats increased from 70.36 hm² in 2020 to a peak in 2023 (172.04 hm²), followed by a reduction to 132.36 hm² in 2024, and the areas of re-emerging snail habitats increased from 42.71 hm² in 2020 to a peak in 2022 (78.43 hm²), followed by a reduction to 40.21 hm² in 2024. The occurrence of frames with snails and mean density of living snails increased from 1.24% (3 025/244 404) and (0.033 2 ± 0.038 7) snails/0.1 m² in 2020 to peaks at 2.03% (6 231/307 563) and (0.066 9 ± 0.068 4) snails/0.1 m² in 2023, followed by reductions to 1.04% (5 829/559 941) and (0.032 6 ± 0.057 7) snails/0.1 m² in 2024, respectively. There was a significant difference in the occurrence of frames with snails over the 5-year study period (χ² = 1 962.95, P < 0.05), and the occurrence of frames with snails reduced by 48.71% in 2024 relative to in 2023 (χ² = 1 411.05, P < 0.005); however, there was no significant difference in the mean density of living snails over the 5 years (H = 5.310, P > 0.05). There were significant differences in the occurrence of frames with snails (χ² = 481.27, P < 0.05) and mean density of living snails (H = 6.872, P < 0.05) in schistosomiasis-endemic areas with different epidemic intensities. The occurrence of frames with snails (χ² = 25.32 and 38.70, both P values < 0.017) and mean density of living snails (Z = 28.55 and 49.96, both P values < 0.017) were higher in schistosomiasis transmission-interrupted and eliminated areas with snails than in schistosomiasis-eliminated areas without snails, and the occurrence of frames with snails (χ² = 453.54, P < 0.017) and mean density of living snails (Z = −56.97, P < 0.017) were higher in schistosomiasis-eliminated areas with snails than in schistosomiasis transmission-interrupted areas with snails. O. hupensis snails were mainly distributed in paddy fields, dry farmlands and ditches; however, the occurrence of frames with snails (13.40%, 424/3 164) and mean density of living snails [(0.252 8 ± 0.158 7) snails/0.1 m²] were higher in ponds/weirs than in other types of environments (both P values < 0.05). Rice, dry farmland crops and weeds were main vegetations in which O. hupensis snails were distributed, and the occurrence of frames with snails (2.29%, 7 111/310 140) and mean density of living snails [(0.072 3 ± 0.018 9) snails/0.1 m²] were higher in weeds than in other types of environments (both P values < 0.05). Conclusions O. hupensis snails have been effectively controlled in Yunnan Province following implementation of integrated schistosomiasis control measures; however, there are still risk factors for schistosomiasis transmission, including reduced attention to schistosomiasis control and snail re-emergence. Improved control efforts and surveillance system construction and timely identification of risk factors of snail status and timely management are recommended to ensure the achievement of the target of schistosomiasis elimination as scheduled.

With the rapid development of large language models and related technologies, various forms of patient modeling have emerged, including virtual patients, synthetic patients, artificial intelligence patients, and digital twin patients. Although these forms all share the core objective of patient simulation, they exhibit significant differences in theoretical ontology, data sources, modeling logic, and application orientation. Based on the abstraction levels of simulation objects and system mapping logic, this study systematically compares five paradigms—standardized patients, virtual patients, synthetic patients, AI patients, and digital twin patients—to clarify their conceptual boundaries and technical positioning. The findings indicate that these paradigms represent distinct modeling pathways: behavioral reproduction, scenario simulation, data generation, cognitive interaction, and individual system mapping, respectively. By constructing a multidimensional comparative framework, this study provides a theoretical foundation for the conceptual standardization of patient modeling and the strategic deployment of intelligent healthcare systems.

Objective: : Baculovirus inhibitory of apoptosis repeat-containing 5 (BIRC5) is critically implicated in various types of tumors. However, the specific mechanisms by which it operates in glioma are yet to be fully understood. Methods: : The data sourced from The Cancer Genome Atlas and Gene Expression Omnibus were merged and analyzed using the R software to investigate the relationship between BIRC5 expression and prognosis and diagnosis outcomes. This exploration was conducted utilizing various biological information repositories. The correlation between BIRC5 and immunity was obtained based on TIMER and TISIDB databases. Results: : Gliomas displayed a markedly elevated level of BIRC5 expression compared to adjacent tissues. Patients with glioma who exhibit elevated levels of BIRC5 experience poorer prognoses and shorter survival times. Subgroup classification further revealed that heightened expression of BIRC5 led to diminished overall survival. Analysis of logistic regression and COX indicated that expression of BIRC5 serves as a risk factor in glioma development. Functional enrichment pathways showed that the 72 hub genes related to BIRC5 were mainly closely related to nuclear division, spindle, tubulin binding, and cell cycle in glioma patients. BBIRC5 methylation suggested that BIRC5 might influence the immune response regulation and the tumor microenvironment within gliomas. BIRC5 is associated with many chemicals. Additionally, studies conducted using cell experiments and pathological sections have consistently shown that BIRC5 expression is higher in tumor cells compared to normal cells and tissues. Conclusion : BIRC5 holds promise as a valuable tool in the diagnosis, prognosis, and management of gliomas.

Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group,model group,TLYZF low-dose (12.5 mg/kg/d) group,TLYZF medium dose (25 mg/kg/d) group and TLYZF high-dose (50 mg/kg/d) group,with 10 rats in each group. Except for the Sham group,other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention,Morris water maze experiment was used to evaluate the learning and memory abilities of rats.Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus.Nissl staining was used to detect hippocampal neuronal damage.In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis.Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue.Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins.Results Compared with the Sham group,the average escape latency of the model group was prolonged(t=14.059),the number of crossing the platform was reduced(t=8.534),the hippocampal neurons were disorderly arranged,the morphology was fuzzy,the cytoplasmic vacuolization,the nuclear pyknosis,the inflammatory cell infiltration increased,the number of Nissl bodies decreased(t=17.131),and the neuronal apoptosis rate increased(t=17.701). The contents of tumor necrosis factor-α (TNF-α),interleukin-1 β (IL-1β) and IL-6 increased(t=6.541,6.957,10.014),the expression of Bcl-2 associated X protein (Bax),cleaved caspase-3,glucose-regulated protein 78 (GRP78),phosphorylated endoplasmic reticulum kinase (p-PERK)/PERK,phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α,activating transcription factor 4 (ATF4) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) increased(t=13.548～76.468),while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased (t=39.691),and the difference were statistically significant (all P<0.05),respectively. Compared with the model group,the average escape latency of TLYZF-treated rats was shortened(t=2.476,7.266,11.306),the number of platform crossing was increased(t=2.187,4.471,6.932),the hippocampal neurons were relatively regular,the nucleus was clear,the inflammatory cell infiltration was significantly reduced,the content of Nissl bodies was increased(t=4.359,9.477,11.449),the apoptosis rate of neurons was decreased(t=3.631,6.145,7.580),the contents of TNF-α,IL-1β and IL-6 were decreased (t=2.382～8.293),the expression of Bax(t=4.696,16.250,20.250),cleaved caspase-3(t=15.205,27.000,26.833),GRP78(t=5.918,13.139,13.741),p-PERK/PERK(t=13.416,14.230,17.889),p-eIF2α/eIF2α(t=20.152,39.346,50.750),ATF4(t=12.093,24.395,22.946)and CHOP protein((t=21.592,19.207,22.136)decreased,while Bcl-2 protein increased(t=7.474,20.761,35.350),the differences were statistically significant (all P<0.05),respectively,and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats,and its mechanism may be related to inhibiting ERS mediated by PERK.

Objective To observe the effect of self-formulated Tongluo Yizhi formula on memory deficits and endoplasmic reticulum stress (ERS) in rats with vascular dementia (VD) and its mechanism. Methods Fifty SPF-grade Wistar rats were randomly divided into sham surgery group,model group,TLYZF low-dose (12.5 mg/kg/d) group,TLYZF medium dose (25 mg/kg/d) group and TLYZF high-dose (50 mg/kg/d) group,with 10 rats in each group. Except for the Sham group,other rats were subjected to modified bilateral carotid artery ligation to construct VD models. After 4 weeks of TLYZF intervention,Morris water maze experiment was used to evaluate the learning and memory abilities of rats.Hematoxylin eosin staining was used to detect the pathological structure of the hippocampus.Nissl staining was used to detect hippocampal neuronal damage.In situ end transfer enzyme labeling staining was used to detect hippocampal neuronal apoptosis.Enzyme-linked immunosorbent assay was used to detect the content of inflammatory factors in hippocampal tissue.Protein immunoblotting was used to detect apoptosis of hippocampal tissue cells and the expression of ERS related proteins.Results Compared with the Sham group,the average escape latency of the model group was prolonged(t=14.059),the number of crossing the platform was reduced(t=8.534),the hippocampal neurons were disorderly arranged,the morphology was fuzzy,the cytoplasmic vacuolization,the nuclear pyknosis,the inflammatory cell infiltration increased,the number of Nissl bodies decreased(t=17.131),and the neuronal apoptosis rate increased(t=17.701). The contents of tumor necrosis factor-α (TNF-α),interleukin-1 β (IL-1β) and IL-6 increased(t=6.541,6.957,10.014),the expression of Bcl-2 associated X protein (Bax),cleaved caspase-3,glucose-regulated protein 78 (GRP78),phosphorylated endoplasmic reticulum kinase (p-PERK)/PERK,phosphorylated eukaryotic initiation factor 2α (p-eIF2α)/eIF2α,activating transcription factor 4 (ATF4) and CCAAT/enhancer binding protein (C/EBP) homologous protein (CHOP) increased(t=13.548～76.468),while the expression of B-cell lymphoma-2 (Bcl-2) proteinwas decreased (t=39.691),and the difference were statistically significant (all P<0.05),respectively. Compared with the model group,the average escape latency of TLYZF-treated rats was shortened(t=2.476,7.266,11.306),the number of platform crossing was increased(t=2.187,4.471,6.932),the hippocampal neurons were relatively regular,the nucleus was clear,the inflammatory cell infiltration was significantly reduced,the content of Nissl bodies was increased(t=4.359,9.477,11.449),the apoptosis rate of neurons was decreased(t=3.631,6.145,7.580),the contents of TNF-α,IL-1β and IL-6 were decreased (t=2.382～8.293),the expression of Bax(t=4.696,16.250,20.250),cleaved caspase-3(t=15.205,27.000,26.833),GRP78(t=5.918,13.139,13.741),p-PERK/PERK(t=13.416,14.230,17.889),p-eIF2α/eIF2α(t=20.152,39.346,50.750),ATF4(t=12.093,24.395,22.946)and CHOP protein((t=21.592,19.207,22.136)decreased,while Bcl-2 protein increased(t=7.474,20.761,35.350),the differences were statistically significant (all P<0.05),respectively,and the high-dose group had the best therapeutic effect.Conclusion Self-formulated Tongluo Yizhi formula could reduce the apoptosis and inflammatory injury of hippocampal neurons in VD rats,and its mechanism may be related to inhibiting ERS mediated by PERK.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.

AIM: To delineate the specific mutation responsible for retinitis pigmentosa(RP)in a Yi pedigree, and to analyze the correlation of RHO gene mutation with clinical phenotype.METHODS:A comprehensive clinical evaluation was conducted on the proband diagnosed with RP and other familial members, complemented by a thorough ophthalmic examination. Peripheral blood samples were obtained from the proband and familial members, from which genomic DNA was extracte. Subsequent whole exome sequencing(WES)was employed to identify the variant genes in the proband. The identified variant gene was validated through Sanger sequencing, then an in-depth analysis of the mutation genes was carried out using genetic databases to ascertain the pathogenic mutation sites. Furthermore, an exhaustive analysis was performed to delineate the genotype and phenotype characteristics.RESULTS:The RP pedigree encompasses 5 generations with 42 members, including 19 males and 23 females. A total of 13 cases of RP were identified, consisting of 4 males and 9 females, which conforms to the autosomal dominant inheritance pattern. The clinical features of this family include an early onset age, rapid progression, and a more severe condition. The patients were found to have night blindness around 6 years old, representing the earliest reported case of night blindness in RP families. The retina was manifested by progressive osteocytoid pigmentation of the fundus, a reduced visual field, and significantly decreased or even vanished a and b amplitudes of ERG. The combined results of WES and Sanger sequencing indicated that the proband had a heterozygous missense mutation of the RHO gene c.1040C>T:p.P347L, where the 1 040 base C of cDNA was replaced by T, causing codon 347 to encode leucine instead of proline. Interestingly, this mutation has not been reported in the Chinese population.CONCLUSION:This study confirmed that the mutant gene of RP in a Yi nationality pedigree was RHO(c.1040C>T). This variant leads to the change of codon 347 from encoding proline to encoding leucine, resulting in a severe clinical phenotype among family members. This study provides a certain molecular, clinical, and genetic basis for genetic counseling and gene diagnosis of RHO.