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.

Chronic pain is a complex condition shaped by long-standing alterations in both physiological and psychological processes. Rather than representing a simple continuation of acute nociceptive signaling, chronic pain is increasingly understood as the outcome of progressive dysregulation within distributed neural systems that govern sensation, affect, motivation, and cognitive control. Neuroimaging and electrophysiological studies indicate that this state is accompanied by extensive plastic changes in deep brain structures and large-scale networks. Beyond well-described central sensitization processes, chronic pain is characterized by disrupted oscillatory rhythms and altered connectivity within large-scale brain networks, including thalamo-cortical circuits and prefrontal-limbic-reward networks. These findings support a conceptual shift from viewing chronic pain as a focal, lesion-driven phenomenon toward recognizing it as a disorder of distributed network pathology. Pharmacological treatments remain central to clinical practice, yet their long-term efficacy is often limited and frequently accompanied by substantial side effects. The ongoing concerns about opioid-related risks and the inadequate therapeutic response in a subset of patients highlight the need for safe, non-pharmacological approaches that can address not only pain but also comorbid disturbances in mood, sleep, and social functioning. Neuromodulation provides a promising path toward mechanism-based and non-pharmacological management of chronic pain by employing physical or chemical stimulation to alter the excitability and synchrony of specific neural populations within central, peripheral, and autonomic systems. While invasive deep brain stimulation demonstrates that targeting deep brain structures can be effective, its clinical application is restricted by surgical risks and cost, highlighting the importance of non-invasive techniques capable of reaching deep targets. Current non-invasive approaches, such as transcranial electric stimulation, are constrained by limited penetration depth and insufficient spatial precision. These limitations hinder reliable engagement of deep regions implicated in pain, including the thalamus and nucleus accumbens, and tend to produce broad, non-specific modulation of cross-network oscillatory activity. Temporal interference (TI) stimulation has emerged as a means of overcoming these obstacles. By delivering interacting high-frequency currents that generate a low-frequency envelope within the head, TI enables focal stimulation of deep targets while minimizing superficial current delivery. Recent multiscale modeling and animal studies indicate that TI exploits the nonlinear rectification properties of neuronal membranes in response to high-frequency carriers, as well as their phase-locked responses to low-frequency envelopes, to generate “peak-focused” electric fields in deep regions under relatively low superficial current loads. Moreover, TI appears to exhibit potential advantages in terms of cell-type selectivity and rhythm-specific engagement, including differential responses across neuronal subtypes and distinct coupling to θ-, β-, and γ-band oscillations. These features suggest a promising avenue for correcting abnormal rhythms and network dynamics that contribute to chronic pain. This review summarizes current knowledge of the neural mechanisms underlying chronic pain and recent advances in TI research. It examines functional disturbances across key pain-related regions and networks, outlines the principles and technical characteristics of TI, and discusses potential deep-brain targets and stimulation strategies relevant to chronic pain. Evidence to date indicates that TI, with its non-invasiveness, tolerability, and capacity for precise deep brain modulation, holds great promise for the management of treatment-resistant chronic pain and may evolve into a new generation of precise and efficient non-pharmacological analgesic strategies.

Chronic pain is a complex condition shaped by long-standing alterations in both physiological and psychological processes. Rather than representing a simple continuation of acute nociceptive signaling, chronic pain is increasingly understood as the outcome of progressive dysregulation within distributed neural systems that govern sensation, affect, motivation, and cognitive control. Neuroimaging and electrophysiological studies indicate that this state is accompanied by extensive plastic changes in deep brain structures and large-scale networks. Beyond well-described central sensitization processes, chronic pain is characterized by disrupted oscillatory rhythms and altered connectivity within large-scale brain networks, including thalamo-cortical circuits and prefrontal-limbic-reward networks. These findings support a conceptual shift from viewing chronic pain as a focal, lesion-driven phenomenon toward recognizing it as a disorder of distributed network pathology. Pharmacological treatments remain central to clinical practice, yet their long-term efficacy is often limited and frequently accompanied by substantial side effects. The ongoing concerns about opioid-related risks and the inadequate therapeutic response in a subset of patients highlight the need for safe, non-pharmacological approaches that can address not only pain but also comorbid disturbances in mood, sleep, and social functioning. Neuromodulation provides a promising path toward mechanism-based and non-pharmacological management of chronic pain by employing physical or chemical stimulation to alter the excitability and synchrony of specific neural populations within central, peripheral, and autonomic systems. While invasive deep brain stimulation demonstrates that targeting deep brain structures can be effective, its clinical application is restricted by surgical risks and cost, highlighting the importance of non-invasive techniques capable of reaching deep targets. Current non-invasive approaches, such as transcranial electric stimulation, are constrained by limited penetration depth and insufficient spatial precision. These limitations hinder reliable engagement of deep regions implicated in pain, including the thalamus and nucleus accumbens, and tend to produce broad, non-specific modulation of cross-network oscillatory activity. Temporal interference (TI) stimulation has emerged as a means of overcoming these obstacles. By delivering interacting high-frequency currents that generate a low-frequency envelope within the head, TI enables focal stimulation of deep targets while minimizing superficial current delivery. Recent multiscale modeling and animal studies indicate that TI exploits the nonlinear rectification properties of neuronal membranes in response to high-frequency carriers, as well as their phase-locked responses to low-frequency envelopes, to generate “peak-focused” electric fields in deep regions under relatively low superficial current loads. Moreover, TI appears to exhibit potential advantages in terms of cell-type selectivity and rhythm-specific engagement, including differential responses across neuronal subtypes and distinct coupling to θ-, β-, and γ-band oscillations. These features suggest a promising avenue for correcting abnormal rhythms and network dynamics that contribute to chronic pain. This review summarizes current knowledge of the neural mechanisms underlying chronic pain and recent advances in TI research. It examines functional disturbances across key pain-related regions and networks, outlines the principles and technical characteristics of TI, and discusses potential deep-brain targets and stimulation strategies relevant to chronic pain. Evidence to date indicates that TI, with its non-invasiveness, tolerability, and capacity for precise deep brain modulation, holds great promise for the management of treatment-resistant chronic pain and may evolve into a new generation of precise and efficient non-pharmacological analgesic strategies.

Objective:To understand the molecular evolutionary characteristics of the recombinant strain GII.3[P12] of norovirus acute gastroenteritis outbreaks and aggregated outbreaks in China from 2022 to 2023.Methods:Epidemiological information, case information, clinical samples, as well as detection and genotyping information of norovirus outbreaks and aggregated outbreaks from 2022 to 2023 were collected; positive samples of the GII.3[P12] recombinant strain were subjected to nucleic acid extraction, whole-genome amplification, and sequence analysis; and homology simulation method were used to construct a three-dimensional structure and predict antigenic epitopes.Results:From January 2022 to December 2023, a total of 1 136 norovirus outbreaks and aggregated outbreaks were reported in China′s norovirus outbreak surveillance network, and genotyping result were successfully obtained for 942 outbreaks, with GII dominating, accounting for 76.0% (716/942), and the proportion of GI and mixed genotypes being 15.8% (149/942) and 8.2% (77/942). Norovirus outbreaks caused by GII were dominated by GII.3[P12] (22.5%, 161/716), while other major genotypes included GII.17[P17] (18.7%, 134/716), GII.4_Sydney 2012[P16] (11.6%, 83/716) and GII.6[P7] ( 11.3%, 81/716). 2022-2023 Outbreaks caused by GII.3[P12] were concentrated in February-March (54.0%, 87/161), with the main outbreaks occurring in nursery and primary schools (87.5%), the mode of transmission was mainly human-to-human (68.9%), and the main susceptible population was children aged 3-7 years (93.3%). In this study, the genome sequences of 25 GII.3[P12] recombinant strains were obtained, and according to the phylogenetic analysis, it was shown that the GII.3[P12] recombinant strains in China in 2022-2023 belonged to the Cluster IV cluster of sublineage b (7 strains) and sublineage c (18 strains). A total of 11 linear and 8 conformational epitopes were predicted by epitope prediction analysis, and the predicted linear and conformational epitopes had overlapping positions, and each conformational epitope was part of the predicted linear epitope with conserved potential antigen-binding and receptor-binding sites.Conclusions:The recombinant strain GII.3[P12] is one of the epidemic strains that will cause outbreaks and clusters of norovirus in China in 2022-2023, and its genome did not undergo significant mutation.

Objective:To analyze the impact of serum alpha-fetoprotein (AFP) and carbohydrate antigen 19-9 (CA19-9) on the prognosis of patients with combined hepatocellular-cholangiocarcinoma (CHC) after radical surgery.Methods:Clinical data of 100 patients diagnosed with CHC by postoperative pathology in Hunan Provincial People's Hospital from January 2009 to January 2019 were retrospectively analyzed, including 73 males and 27 females, aged (52.0±10.9) years. Univariate and multivariate Cox regression analysis were conducted to try to find out the effects of AFP and CA19-9 on postoperative disease-free survival (DFS) of patients with CHC. Subgroup analysis was also performed to analyze the DFS among patients with different levels of AFP and CA19-9. Kaplan-Meier method and log-rank test were used to plot and compare the survival curves between groups.Results:Univariate analysis showed that levels of AFP, CA19-9, alanine transaminase, aspartate transaminase, American Joint Committee on Cancer (AJCC) TNM staging, portal vein invasion, tumor number are associated with postoperative DFS in CHC patients (all P<0.05). The multivariate Cox analysis showed that AFP≥20.6 μg/L ( HR=6.686, 95% CI: 1.985-9.582), CA19-9≥35.2 U/L ( HR=5.790, 95% CI: 1.197-8.855), AJCC tumor TNM staging stage Ⅱ( HR=6.384, 95% CI: 2.069-11.532), and portal vein invasion ( HR=2.384, 95% CI: 1.100-10.125) were risk factors for a shortened DFS in CHC patients after surgery (all P<0.05). Subgroup analysis showed that patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L ( n=14) had a lower DFS compared to patients with AFP<20.6 μg/L and CA19-9≥35.2 U/L ( n=32), and patients with AFP≥20.6 μg/L and CA19-9<35.2 U/L ( n=20) ( χ2=6.23, 4.98, P=0.014, 0.043). Conclusion:AFP and CA19-9 are risk factors for DFS in CHC patients. Patients with AFP≥20.6 μg/L and CA19-9≥35.2 U/L showed a worse prognosis.

Objective:To implement an overview of systematic reviews on the effects of brain-computer interface training on upper limb functional rehabilitation in stroke patients.Methods:The systematic review/Meta-analysis of the effect of brain-computer interface training on upper limb functional rehabilitation in stroke patients was electronically retrieved in PubMed, Web of Science, Cochrane Library, Embase, CINAHL, Embase, Joanna Briggs Institute Evidence-Based Health Care Center Database, China National Knowledge Infrastructure, Wanfang Data, VIP and China Biology Medicine disc. The search period was from database establishment to November 2024. Two researchers independently performed literature screening and data extraction and evaluated the quality of methodology, reporting, and evidence.Results:A total of 14 systematic reviews/Meta-analyses were included. The results showed that brain-computer interface training helped to improve upper limb motor function, muscle strength and activities of daily living in stroke patients, but the rehabilitative effect on muscle spasm needed to be further confirmed.Conclusions:Brain-computer interface training helps to improve upper limb motor function, muscle strength and activities of daily living in stroke patients. However, the methodological quality and reporting quality of the current studies are poor, and there is still a need for high-quality studies with rigorous design and standardized process to provide reference for clinical practice.

Acute respiratory distress syndrome(ARDS)is a common respiratory emergency,but current clinical treatment remains at the level of symptomatic support and there is a lack of effective targeted treatment measures.Our previous study confirmed that inhalation of hydrogen gas can reduce the acute lung injury of ARDS,but the application of hydrogen has flammable and explosive safety concerns.Drinking hydrogen-rich liquid or inhaling hydrogen gas has been shown to play an important role in scavenging reactive oxygen species and maintaining mitochondrial quality control balance,thus improving ARDS in patients and animal models.Coral calcium hydrogenation(CCH)is a new solid molecular hydrogen carrier prepared from coral calcium(CC).Whether and how CCH affects acute lung injury in ARDS re-mains unstudied.In this study,we observed the therapeutic effect of CCH on lipopolysaccharide(LPS)induced acute lung injury in ARDS mice.The survival rate of mice treated with CCH and hydrogen inhalation was found to be comparable,demonstrating a significant improvement compared to the untreated ARDS model group.CCH treatment significantly reduced pulmonary hemorrhage and edema,and improved pulmonary function and local microcirculation in ARDS mice.CCH promoted mitochon-drial peripheral division in the early course of ARDS by activating mitochondrial thioredoxin 2(Trx2),improved lung mitochondrial dysfunction induced by LPS,and reduced oxidative stress damage.The results indicate that CCH is a highly efficient hydrogen-rich agent that can attenuate acute lung injury of ARDS by improving the mitochondrial function through Trx2 activation.

Objective To explore the molecular mechanism of circFARSA targeting miR-4319 to modulate the prolifera-tive,migratory and invasive abilities of Huh-7 liver cancer cells.Methods The qRT-PCR method was used to detect the expres-sion of circFARSA and miR-4319 in liver cancer tissues and adjacent tissues.Liver cancer cells Huh-7 were cultured in vitro,qRT-PCR method was used to detect the expression of circFARSA and miR-4319 in Huh-7 cells.CCK-8 assay and Transwell chamber assay were used to detect proliferation,migration,and invasion abilities,respectively.Dual-luciferase reporter assay was performed to analyze the interaction between circFARSA and miR-4319.Western blot was used to detect Ki-67,MMP-2 and MM P-9 protein levels.Results The level of circFARSA was increased,and the level of miR-4319 was decreased in liver cancer tissues(bothP＜0.05).Cell viability,migration,and invasion were significantly inhibited after circFARSA silencing or miR-4319 overexpression(all P＜0.05).It was verified by double luciferase reporter gene experiment that circFARSA could competi-tively bind to miR-4319.Co-transfection of circFARSA siRNA with miR-4319 inhibitor enhanced cell viability,migration,and invasion when compared with transfection of circFARSA siRNA alone(P＜0.05).Conclusion circFARSA knockdown sup-pressed liver cancer cell tumor property by increasing the expression level of miR-4319.

Objective To explore the molecular mechanism of circFARSA targeting miR-4319 to modulate the prolifera-tive,migratory and invasive abilities of Huh-7 liver cancer cells.Methods The qRT-PCR method was used to detect the expres-sion of circFARSA and miR-4319 in liver cancer tissues and adjacent tissues.Liver cancer cells Huh-7 were cultured in vitro,qRT-PCR method was used to detect the expression of circFARSA and miR-4319 in Huh-7 cells.CCK-8 assay and Transwell chamber assay were used to detect proliferation,migration,and invasion abilities,respectively.Dual-luciferase reporter assay was performed to analyze the interaction between circFARSA and miR-4319.Western blot was used to detect Ki-67,MMP-2 and MM P-9 protein levels.Results The level of circFARSA was increased,and the level of miR-4319 was decreased in liver cancer tissues(bothP＜0.05).Cell viability,migration,and invasion were significantly inhibited after circFARSA silencing or miR-4319 overexpression(all P＜0.05).It was verified by double luciferase reporter gene experiment that circFARSA could competi-tively bind to miR-4319.Co-transfection of circFARSA siRNA with miR-4319 inhibitor enhanced cell viability,migration,and invasion when compared with transfection of circFARSA siRNA alone(P＜0.05).Conclusion circFARSA knockdown sup-pressed liver cancer cell tumor property by increasing the expression level of miR-4319.

Objective:To investigate the practical value of pancreatic duct bind with supporting tube combined with single-needle suture for pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy (LPD).Methods:The clinical data of 65 patients with pancreatic head or periampullary tumors who underwent LPD at the First Affiliated Hospital of Kunming Medical University between January 2021 and June 2024 were retrospectively analysed. Among them, 31 were male and 34 were female, aged (57.7±13.3) years. The patients were divided into two groups based on the method of pancreaticojejunostomy: 22 patients who underwent pancreatic duct bind with supporting tube single-needle suture pancreaticojejunos-tomy were included in the study group, 43 patients who underwent traditional pancreatic duct-to-mucosa pancreaticojejunostomy were included in the control group. The clinical data of the patients, including gender, age, operation time, pancreaticojejunostomy time, and postoperative complications, etc. were recorded.Results:Compared with the control group, the pancreaticojejunostomy time [(25.3±2.4) min vs. (47.0±3.9) min] and operation time [(425.2±52.1) min vs. (453.7±30.8) min] of the study group were both shortened, and the differences were statistically significant (both P<0.05). There was no statistically significant difference in the postoperative complication rate between the study group and the control group [18.2% (4/22) vs. 34.9% (15/43), P=0.161]. The incidence of clinically relevant pancreatic fistula in the study group was lower than that in the control group [4.5% (1/22) vs. 30.2% (13/43)], and the difference was statistically significant ( P=0.024). Conclusion:Pancreatic duct bind with supporting tube combined with single-needle suture pancreaticojejunostomy is safe and effective in LPD. Compared with traditional pancreatic duct-to-mucosa pancreaticojejunostomy, pancreatic duct bind with supporting tube combined with single-needle suture pancreaticojejunostomy has advantages in terms of clinically relevant pancreatic fistula rate, operation time, and pancreaticojejunostomy time.