Cryo-electron tomography (cryo-ET) enables the determination of high-resolution three-dimensional structures of macromolecular complexes within cells in a near-physiological state, providing crucial structural insights into fundamental life processes. Cryo-ET has achieved landmark successes in single-cell models. However, many critical biological processes do not occur in isolated cells but emerge from intercellular coordination within tissues. Furthermore, many research subjects, including neural tissues, tumor biopsies, plant tissues, and clinical pathological samples, cannot be obtained through single-cell culture and must be directly dissected from organisms or tissue blocks. Advancing cryo-ET from single-cell to tissue-level applications is therefore crucial for capturing the full complexity of biological activities in their native context. A major technical bottleneck for tissue cryo-ET lies in the preparation of sufficiently thin (<300 nm) lamellae from vitrified tissue specimens. Although high-pressure freezing can vitrify tissues up to 200 µm thick, these samples are far too thick for direct transmission electron microscopy imaging. Among the available thinning methods, cryo-focused ion beam (cryo-FIB) milling has emerged as the most promising approach, as it avoids the mechanical artifacts inherent to cryo-ultramicrotomy. However, conventional on-the-grid cryo-FIB milling is inefficient for thick tissues, requiring excessive milling time and discarding most of the sample. To overcome these limitations, cryo-lift-out has been developed—a technique in which a micromanipulator physically extracts a chunk of interest from deep within the tissue and transfers it to a dedicated grid for final thinning. This approach bypasses the thickness barrier and enables site-specific analysis of internal structures. This review systematically traces the evolution of cryo-lift-out from its origins in materials science to its adaptation for biological tissues. In room-temperature lift-out, reliable attachment is achieved by gas-injection system (GIS)-assisted metal deposition. Transferring this approach to cryogenic conditions proved challenging because precursor gases condense on all cold surfaces, leading to contamination and poor adhesion. The development of copper-assisted redeposition marked a critical turning point: instead of relying on gas deposition, this method uses ion-beam sputtering to deposit copper atoms at the needle-chunk interface, creating a strong, low-contamination bond. This innovation has enabled robust cryo-lift-out workflows and paved the way for serial lift-out, in which multiple consecutive lamellae are prepared from a single tissue chunk, substantially increasing throughput and enabling volumetric imaging. Despite these advances, several technical challenges remain. Curtaining effects caused by uneven chunk surfaces can introduce artifacts into tomograms, requiring careful optimization of milling parameters and protective coating. The cryo-adhesion step still demands precise control of beam angle, needle positioning, and milling depth, making the process highly operator-dependent. Additionally, the choice of grid geometry is critical. Custom-designed grids with double-sided attachment improves stability and offer better compatibility with cryo-ET tilt series. Automation, which has greatly improved room-temperature lift-out, has not yet been achieved for cryo-lift-out due to the complexity of handling heterogeneous biological tissues and the need for real-time adaptation. Future progress will likely focus on integrating cryo-lift-out with volume electron microscopy to correlate ultrastructure across scales, developing intelligent control systems to reduce user intervention, and extending the technology to challenging samples such as plant tissues and some material science samples for interface study. A systematic analysis of the cryo-lift-out technique clarifies the key limiting factors for its large-scale application and lays a foundation for methodological refinement and technological innovation. By consolidating recent advances and identifying remaining bottlenecks, this review aims to support the broader adoption of cryo-lift-out and accelerate the development of tissue-scale in situ structural biology.

Objective:To investigate the survival outcomes and prognostic factors in patients undergoing radical resection for pancreatic body and tail cancer.Methods:A retrospective case series study was conducted on 992 patients who underwent radical resection for pancreatic body and tail cancer at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from January 2016 to June 2024. In this study, 577 (58.2%) were male and 415 (41.8%) were female,with an age of (65±9) years (range: 26 to 86 years). Follow-up continued until June 2024. Survival rates were estimated using the Kaplan-Meier method,and prognostic factors were identified using univariate and multivariate Cox proportional hazards models.Results:Among 992 patients,open surgery was the predominant approach (89.1%, 884/992), and radical antegrade modular pancreatosplenectomy (RAMPS) was performed in 317 patients (32.0%). Combined organ resection,venous resection,and arterial resection were performed in 23.5%, 9.3%,and 11.2% of patients,respectively. The rates of R0, R1-1 mm, and R1-direct resections were 49.8% (494/992),41.5% (412/992), and 8.7% (86/992),respectively. Stage ⅡB was the most common TNM stage (32.2%,319/992). A total of 801 patients (80.8%) received adjuvant chemotherapy. The median follow-up period was 32.0(8.8) months(range:3.2 to 105.3 months),during which 508 patients (51.2%) died. The overall median survival (OS) was 26.4 months,with 1-,3-, and 5-year survival rates of 79.0%,40.0%, and 29.0%, respectively. In the recent five years (from 2020 to 2024), the median OS improved significantly to 34.1 months compared to 20.0 months from 2016 to 2019 ( P<0.01). Histological subtype analysis showed that the median OS time was 26.7 months for pancreatic ductal adenocarcinoma (PDAC, n=855),58.9 months for invasive intraductal papillary mucinous carcinoma (IPMC, n=32),and 15.7 months for adenosquamous carcinoma of pancreas (ASCP, n=73) ( P=0.001). Among PDAC patients, adjuvant chemotherapy significantly improved survival (29.1 months vs. 14.4 months, P<0.01);in IPMC patients, adjuvant chemotherapy also extended survival (65.7 months vs. 58.9 months, P=0.047). Although ASCP patients receiving chemotherapy had a longer median OS time than those without (18.8 months vs. 8.9 months),the difference was not statistically significant ( P=0.151). Multivariate Cox regression analysis in PDAC patients indicated that adjuvant chemotherapy, R0 resection, T stage,N stage,and tumor differentiation were independent prognostic factors ( P<0.01). The median OS time by TNM stage was:not reached for stage ⅠA, 51.6 months for ⅠB, 25.5 months for ⅡA, 23.7 months for ⅡB, 23.0 months for Ⅲ, and 14.4 months for Ⅳ. The median OS time for R0,R1-1 mm,and R1-direct resections was 34.1,24.7,and 15.7 months,respectively ( P<0.01). Conclusion:Adjuvant chemotherapy,R0 resection,tumor stage,and differentiation are independent prognostic factors for pancreatic body and tail cancer.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective:To investigate the survival outcomes and prognostic factors in patients undergoing radical resection for pancreatic body and tail cancer.Methods:A retrospective case series study was conducted on 992 patients who underwent radical resection for pancreatic body and tail cancer at the Pancreatic Center of the First Affiliated Hospital of Nanjing Medical University from January 2016 to June 2024. In this study, 577 (58.2%) were male and 415 (41.8%) were female,with an age of (65±9) years (range: 26 to 86 years). Follow-up continued until June 2024. Survival rates were estimated using the Kaplan-Meier method,and prognostic factors were identified using univariate and multivariate Cox proportional hazards models.Results:Among 992 patients,open surgery was the predominant approach (89.1%, 884/992), and radical antegrade modular pancreatosplenectomy (RAMPS) was performed in 317 patients (32.0%). Combined organ resection,venous resection,and arterial resection were performed in 23.5%, 9.3%,and 11.2% of patients,respectively. The rates of R0, R1-1 mm, and R1-direct resections were 49.8% (494/992),41.5% (412/992), and 8.7% (86/992),respectively. Stage ⅡB was the most common TNM stage (32.2%,319/992). A total of 801 patients (80.8%) received adjuvant chemotherapy. The median follow-up period was 32.0(8.8) months(range:3.2 to 105.3 months),during which 508 patients (51.2%) died. The overall median survival (OS) was 26.4 months,with 1-,3-, and 5-year survival rates of 79.0%,40.0%, and 29.0%, respectively. In the recent five years (from 2020 to 2024), the median OS improved significantly to 34.1 months compared to 20.0 months from 2016 to 2019 ( P<0.01). Histological subtype analysis showed that the median OS time was 26.7 months for pancreatic ductal adenocarcinoma (PDAC, n=855),58.9 months for invasive intraductal papillary mucinous carcinoma (IPMC, n=32),and 15.7 months for adenosquamous carcinoma of pancreas (ASCP, n=73) ( P=0.001). Among PDAC patients, adjuvant chemotherapy significantly improved survival (29.1 months vs. 14.4 months, P<0.01);in IPMC patients, adjuvant chemotherapy also extended survival (65.7 months vs. 58.9 months, P=0.047). Although ASCP patients receiving chemotherapy had a longer median OS time than those without (18.8 months vs. 8.9 months),the difference was not statistically significant ( P=0.151). Multivariate Cox regression analysis in PDAC patients indicated that adjuvant chemotherapy, R0 resection, T stage,N stage,and tumor differentiation were independent prognostic factors ( P<0.01). The median OS time by TNM stage was:not reached for stage ⅠA, 51.6 months for ⅠB, 25.5 months for ⅡA, 23.7 months for ⅡB, 23.0 months for Ⅲ, and 14.4 months for Ⅳ. The median OS time for R0,R1-1 mm,and R1-direct resections was 34.1,24.7,and 15.7 months,respectively ( P<0.01). Conclusion:Adjuvant chemotherapy,R0 resection,tumor stage,and differentiation are independent prognostic factors for pancreatic body and tail cancer.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective:To investigate the application and management status of midline catheters in 1 954 hospitals, providing a basis for optimizing intravenous therapy nursing practices.Methods:This study used convenience sampling. From November 2023, members of the Intravenous Therapy Nursing Professional Committee of the Chinese Nursing Association selected 1 954 hospitals across various regions of China. Questionnaire on the Current Status of Intravenous Therapy in Hospitals at All Levels designed by the committee, based on literature review and expert discussions, was used to collect data on intravenous therapy practices in different hospitals. Multiple response analysis was applied to analysis the results of multiple-choice questions, where response numbers represent the total number of times each option was selected, and response rates refer to the proportion of selected times for each option out of all selected responses.Results:A total of 1 954 questionnaires were distributed across 31 provinces/municipalities/autonomous regions, and 1 954 valid questionnaires were returned, achieving a 100.0% valid response rate. Among the hospitals surveyed, 844 used midline catheters. Regarding the skin disinfection area for midline catheter insertion, the highest response rate was for a range of>20 cm. The highest response rate for catheter insertion techniques was ultrasound-guided Seldinger puncture. The highest response rate for maintenance interval was once a week. The top three responses for nursing documentation related to midline catheters were informed consent for intubation, puncture record, and maintenance record. The most frequently chosen processes were catheter placement, maintenance, removal, and complication management processes. The qualification for midline catheter intravenous therapy specialist nurses was mostly obtained through specialized nurse training, followed by hospital-based and department-based training.Conclusions:The application of midline catheters has rapidly developed but still reveals some deficiencies, including the choice of puncture tools, infection control, and catheter maintenance. It is recommended to improve nursing documentation and management processes related to midline catheters, establish industry standards suitable for China's national conditions, and strengthen and standardize the specialized training of intravenous therapy nurses to promote the healthy development of intravenous therapy in China.

Body weight abnormalities, including overweight, obesity, and underweight, have become a dual public health challenge in Chinese adults: overweight and obesity lead to a variety of chronic complications, while underweight increases the risks of malnutrition, sarcopenia, and organ dysfunction. To systematically address these issues, multidisciplinary experts in endocrinology, sports science, nutrition, and psychiatry from various regions have held multiple weight management seminars. Based on the latest epidemiological data and clinical evidence, they expanded the guideline to include assessment and intervention strategies for underweight, in addition to the core content of obesity management. This guideline outlines the etiological mechanisms, evaluation methods, and multidimensional management strategies for overweight and obesity, covering key areas such as diagnosis and assessment, medical nutrition therapy, exercise prescription, pharmacological intervention, and psychological support. It is intended to provide a scientific and standardized approach to weight management across the adult population, aiming to curb the rising prevalence of obesity, mitigate complications associated with abnormal body weight, and improve nutritional status and overall quality of life.

Objective To explore the optimization and effect analysis method of rational drug use pre-prescription review rule base for traditional Chinese medicine(TCM).Methods The rules for the modules including indications,dosage range,incompatibility,administration route and repeated drug use of TCM decoction pieces and Chinese patent medicine were set to improve pre-prescription review rule base.The prescription rationality rate and intervention effectiveness rate to evaluate effect by χ2 test before and after the system optimization.Results After the optimization of pre-prescription review rule base for TCM,the rate of prescription rationality of the hospital increased from approximately 78.19%to 94.50%,and the rate of intervention effectiveness increased from approximately 82.80%to 97.15%.There were statistically significant differences in prescription rationality and intervention effectiveness rate between pre-and post-optimization groups(P＜0.05).Conclusion According to the current pre-prescription status of the hospital,optimizing pre-prescription review rules of TCM can improve the applicability of the review system for TCM prescriptions,and enhance the pre-prescription accuracy and the level of rational medication.However,rules optimization on other modules still needs to be further improved.

This study investigated the disease characteristics and clinical traits of Nocardia infection,and analyzed the antibiotic resistance phenotypes,antibiotic resistance genes,and evolutionary characteristics of the strains,to provide a basis for Nocardia diag-nosis and treatment.A total of 31 Nocardia strains from a hospital in Hebei Province were collected from 2020 to 2023.The strains were identified through mass spectrometry and whole genome sequencing.Antibiotic susceptibility testing was conducted with the broth macrodilution method,and whole genome sequencing data were used to predict antibiotic resistance genes and comprehensively ana-lyze antibiotic resistance phenotypes.The 31 strains of Nocardia comprised 21 strains of Nocardia farcinica,3 strains of Nocardia ter-penica,three strains of Nocardia brasiliensis,two strains of Nocardia cyriacigeorgica,and two strains of Nocardia nova.The ceftriaxone susceptibility of 21 Nocardia farcinica strains was 85.7%,and all 31 strains were susceptible to imipenem,except for three strains of Nocardia brasiliensis.Rifampicin,aminoglycoside,and β-lactam resistance genes were found in Nocardia farcinica.Pathogenic tests should be carried out in a timely manner for suspected Nocardia infections.In clinical treatment of Nocardia infection,infected strains should be confirmed,and antibiotics should be used rationally according to the antibiotic susceptibility test results.

This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation