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.

Through an analysis of the pathogenesis of cholinergic urticaria and the therapeutic principle of "treating fire constraint by dispersing it"， it is proposed that the disease is located between the sweat pore and interstitial space. The key pathogenesis lies in fire constraint in the interstitial space and stagnation of the sweat pore. The manifestation is rash due to fire-heat constrained in the interstitial space， while the root is the obstruction of qi and fluids in the sweat pore， and the transformation involves constraint fire damaging yin and entering the collaterals. Treatment should consistently follow the principle of "diffusing constraint and dissipating fire"， with staged approaches based on disease progression. In the acute stage， treatment emphasizes activating sweat pore and rapidly dispersing constraint fire through modified Shengjiang Powder （升降散）. In the remission stage， therapy should focus on harmonizing ying-wei （营卫） and clearing residual fire using modified Zhuye Shigao Decoction （竹叶石膏汤） combined with Xiaofeng Powder （消风散）. In the chronic stage， treatment should nourish yin， penetrate collaterals and clear latent fire through modified Qinghao Biejia Decoction （青蒿鳖甲汤）.

Through an analysis of the pathogenesis of cholinergic urticaria and the therapeutic principle of "treating fire constraint by dispersing it"， it is proposed that the disease is located between the sweat pore and interstitial space. The key pathogenesis lies in fire constraint in the interstitial space and stagnation of the sweat pore. The manifestation is rash due to fire-heat constrained in the interstitial space， while the root is the obstruction of qi and fluids in the sweat pore， and the transformation involves constraint fire damaging yin and entering the collaterals. Treatment should consistently follow the principle of "diffusing constraint and dissipating fire"， with staged approaches based on disease progression. In the acute stage， treatment emphasizes activating sweat pore and rapidly dispersing constraint fire through modified Shengjiang Powder （升降散）. In the remission stage， therapy should focus on harmonizing ying-wei （营卫） and clearing residual fire using modified Zhuye Shigao Decoction （竹叶石膏汤） combined with Xiaofeng Powder （消风散）. In the chronic stage， treatment should nourish yin， penetrate collaterals and clear latent fire through modified Qinghao Biejia Decoction （青蒿鳖甲汤）.

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.

Objective To investigate the prevalence of Schistosoma japonicum infections in wild murines in schistosomiasisendemic areas of Yunnan Province, so as to provide the evidence for improving the schistosomiasis surveillance and early warning capability and implementing precise interventions in the province. Methods A total of 56 natural villages were sampled from 14 schistosomiasis-endemic counties (cities, districts) in Yunnan Province as surveillance sites from 2024 to 2025. Wild murines were captured using traps at night and characterized for species. Liver samples were collected from wild murines, S. japonicum eggs and miracidia were simultaneously detected using liver tissue compression microscopy, liver tissue homogenate microscopy, and liver tissue homogenate hatching. In addition, stool samples were collected from wild murine rectum, with S. japonicum eggs detected using a modified Kato-Katz thick smear technique and S. japonicum miracidia detected using the hatching test with nylon gauzes. Results A total of 15 577 valid murine traps were recovered, and 891 wild murines belonging to 16 species were captured, with a capture rate of 5.72%(891/15 577). The captured wild murines were predominantly districted in paddy fields, dry lands, ditches, ponds and marshlands, with Apodemus chevrieri, Rattus norvegicus, Crocidura attenuata, R. rattus sladeni, R. flavipectus, and Eothenomys miletus as dominant species. No S. japonicum eggs or miradicia were in laboratory; however, other parasites were identified in 25 wild murines. The prevalence rates of Schistosoma sinensis, Capillaria hepatica and Hymenolepis nana were 2.24% (20/891), 0.90% (8/891) and 0.56% (5/891), and the prevalence of S. sinensis and C. hepatica co-infection was 0.90% (8/891), respectively. Conclusions S. sinensis, C. hepatica and H. nana are predominant parasite species in wild murines in schistosomiasis-endemic areas of Yunan Province. No S. japonicum infection was detected during the study period; however, there is still a risk of wild murines as a potential source of infection for schistosomiasis transmission. Continuous intensified surveillance of S. japonicum infections in wild murines is a critical programme in schistosomiasis surveillance and early warning in Yunnan Province.

Laboratory animals are increasingly utilized as in vivo models for life sciences, medical research, and drug development, and the associated ethical and welfare issues have attracted increasing attention. The "3R" principle (replacement, reduction, refinement) is a globally recognized guideline for conducting animal experiments, providing direction for balancing scientific research with animal welfare. New approach methodologies refer to technologies that mimic human physiological systems and can replace or reduce traditional animal experiments, mainly including in vitro (e.g., organoids, organ-on-a-chip,3D bioprinting models), in chemico , and in silico tools. This review systematically examines the technical characteristics, development history, and applications of new approach methodologies in disease modeling, drug development, toxicological evaluation, etc., and focuses on the implementation path of the "3R" principle. New approach methodologies can replace some animal experiments through in vitro biomimetic models, reduce the number of experimental animals used through high-throughput screening and computational simulation, optimize experimental design and alleviate animal suffering through accurate dose prediction and early biomarker monitoring. New approach methodologies still face challenges such as the inability to fully recapitulate the complex human physiological system, insufficient standardization and reproducibility, and underdeveloped validation and acceptance pathways. With continuous technological innovation, rigorous scientific validation, proactive international cooperation, and sound policy support, new approach methodologies are expected to gradually reduce the use of traditional animal experiments and advance biomedical research toward the coordinated pursuit of scientific rigor, animal welfare, and cost-effectiveness.

Real-world study (RWS), based on multi-source data from real medical environments, is gradually becoming an important supplement to traditional randomized controlled trials, and its application in the field of transfusion medicine is becoming increasingly widespread. This article systematically reviews the definition and methodological system of RWS, examines its application cases in clinical blood transfusion research, and discusses the advantages, limitations, and future research directions of RWS, aiming to provide a reference for evidence-based research in blood transfusion medicine.

Real-world study (RWS), based on multi-source data from real medical environments, is gradually becoming an important supplement to traditional randomized controlled trials, and its application in the field of transfusion medicine is becoming increasingly widespread. This article systematically reviews the definition and methodological system of RWS, examines its application cases in clinical blood transfusion research, and discusses the advantages, limitations, and future research directions of RWS, aiming to provide a reference for evidence-based research in blood transfusion medicine.

Objective To examine the changing prevalence and antimicrobial resistance profiles of Burkholderia cepacia in 52 hospitals across China from 2015 to 2021.Methods A total of 9 261 strains of B.cepacia were collected from 52 hospitals between January 1,2015 and December 31,2021.Antimicrobial susceptibility of the strains was tested using Kirby-Bauer method or automated antimicrobial susceptibility testing systems according to a unified protocol.The results were interpreted according to the breakpoints released in the Clinical & Laboratory Standards Institute(CLSI)guidelines(2023 edition).Results A total of 9 261 strains of B.cepacia were isolated from all age groups,especially elderly patients.The proportion was 11.1％(1 032 strains)in children,significantly lower than the proportion in adults.About half(46.5％,4 310/9 261)of the strains were isolated from patients at least 60 years old and 42.3％(3 919/9 261)of the strains were isolated from young adults.Most isolates(71.1％)were isolated from sputum and respiratory secretions,followed by urine(10.7％)and blood samples(8.1％).B.cepacia isolates were highly susceptible to the five antimicrobial agents recommended in the CLSI M100 document(33rd edition,2023).B.cepacia isolates showed relatively higher resistance rates to meropenem and levofloxacin.However,the resistance rates to ceftazidime,trimethoprim-sulfamethoxazole,and minocycline remained below 8.1％.The percentage of B.cepacia strains resistant to levofloxacin was the highest compared to other antibiotics in any of the three age groups(from 12.4％in the patients＜18 years old to 20.6％in the patients aged 60 years or older).Conclusions B.cepacia is one of the clinically important non-fermenting gram-negative bacteria.Accurate and timely reporting of antimicrobial susceptibility test results and ongoing antimicrobial resistance surveillance are helpful for rational prescription of antimicrobial agents and proper prevention and control of nosocomial infections.