ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

The brain-gut interaction theory is a multidimensional integrative concept based on the brain-gut axis， involving neural， endocrine， and immune regulatory networks as well as the gut microbiota. Zang-fu organs （脏腑） theory in traditional Chinese medicine （TCM） shows a high degree of consistency with the brain-gut interaction theory， and the core functions such as the spleen and stomach governing the ascending of the clear and descending of the turbid， the liver governing the free flow of qi， and the heart governing mental and emotional activities are closely associated with the multi-level regulatory mechanisms of the brain-gut axis. TCM therapy can modulate brain-gut interactions through multiple pathways in the treatment of spleen and stomach diseases， including the regulation of gastrointestinal hormone secretion， neurotransmitter levels， the hypothalamic-pituitary-adrenal （HPA） axis， immune homeostasis and inflammatory responses， as well as the gut microecology. However， current basic research on the brain-gut interaction theory in TCM for spleen and stomach diseases still faces several challenges， such as difficulties in integrating TCM spleen-stomach theory with modern pathophysiology， lack of innovation in research concepts， and limitations in research methodologies. It is therefore proposed that multidisciplinary collaboration， multi-omics technologies， and targeted research approaches should be adopted to provide more comprehensive methods for basic research on TCM spleen and stomach diseases， thereby promoting the in-depth development of brain-gut interaction theory.

The Expert Consensus on Clinical Application of Qinbaohong Zhike Oral Liquid in Treatment of Acute Bronchitis and Acute Attack of Chronic Bronchitis （GS/CACM 337-2023） was released by the China Association of Chinese Medicine on December 13^th， 2023. This expert consensus was developed by experts in methodology， pharmacy， and Chinese medicine in strict accordance with the development requirements of the China Association of Chinese Medicine （CACM） and based on the latest medical evidence and the clinical medication experience of well-known experts in the fields of respiratory medicine （pulmonary diseases） and pediatrics. This expert consensus defines the application of Qinbaohong Zhike oral liquid in the treatment of cough and excessive sputum caused by phlegm-heat obstructing lung， acute bronchitis， and acute attack of chronic bronchitis from the aspects of applicable populations， efficacy evaluation， usage， dosage， drug combination， and safety. It is expected to guide the rational drug use in medical and health institutions， give full play to the unique value of Qinbaohong Zhike oral liquid， and vigorously promote the inheritance and innovation of Chinese patent medicines.

19 cinnamamide/ester-triazole compounds were designed, synthesized and evaluated for their anti-Alzheimer's disease (AD) activity. Among them, compound 4f displayed excellent anti-β-amyloid protein (Aβ)-mediated cytotoxicity (EC₅₀ = 2.03 ± 2.45 μmol·L^-1) in APPswe cells and acetylcholinesterase inhibiton (IC₅₀ = 4.88 ± 4.70 μmol·L^-1). Further study indicated that, at dosages of (1, 5 and 25 mg·kg^-1), compound 4f was effective in improving spatial learning and memory deficits in Aβ_1-42-impaired mice, which was achieved by promoting the nonamyloidogenic signaling and inhibiting the amyloidogenic pathway, along with the suppression of Aβ-induced Tau phosphorylation. All animal experiments in this study were approved by the Experimental Animal Care and Use Committee of the Institute of Medicinal Biotechnology (IMB-20220908D701). In conclusion, compound 4f holds promise as a lead candidate for AD treatment, and the present study lays the foundation for its subsequent development.

ObjectiveThis study leverages structural data from antigen-antibody complexes of the influenza A virus neuraminidase (NA) protein to investigate the spatial recognition relationship between the antigenic epitopes and antibody paratopes. MethodsStructural data on NA protein antigen-antibody complexes were comprehensively collected from the SAbDab database, and processed to obtain the amino acid sequences and spatial distribution information on antigenic epitopes and corresponding antibody paratopes. Statistical analysis was conducted on the antibody sequences, frequency of use of genes, amino acid preferences, and the lengths of complementarity determining regions (CDR). Epitope hotspots for antibody binding were analyzed, and the spatial structural similarity of antibody paratopes was calculated and subjected to clustering, which allowed for a comprehensively exploration of the spatial recognition relationship between antigenic epitopes and antibodies. The specificity of antibodies targeting different antigenic epitope clusters was further validated through bio-layer interferometry (BLI) experiments. ResultsThe collected data revealed that the antigen-antibody complex structure data of influenza A virus NA protein in SAbDab database were mainly from H3N2, H7N9 and H1N1 subtypes. The hotspot regions of antigen epitopes were primarily located around the catalytic active site. The antibodies used for structural analysis were primarily derived from human and murine sources. Among murine antibodies, the most frequently used V-J gene combination was IGHV1-12*01/IGHJ2*01, while for human antibodies, the most common combination was IGHV1-69*01/IGHJ6*01. There were significant differences in the lengths and usage preferences of heavy chain CDR amino acids between antibodies that bind within the catalytic active site and those that bind to regions outside the catalytic active site. The results revealed that structurally similar antibodies could recognize the same epitopes, indicating a specific spatial recognition between antibody and antigen epitopes. Structural overlap in the binding regions was observed for antibodies with similar paratope structures, and the competitive binding of these antibodies to the epitope was confirmed through BLI experiments. ConclusionThe antigen epitopes of NA protein mainly ditributed around the catalytic active site and its surrounding loops. Spatial complementarity and electrostatic interactions play crucial roles in the recognition and binding of antibodies to antigenic epitopes in the catalytic region. There existed a spatial recognition relationship between antigens and antibodies that was independent of the uniqueness of antibody sequences, which means that antibodies with different sequences could potentially form similar local spatial structures and recognize the same epitopes.

Objective:Comparative study on the application value of bronchial ultrasound combined with different biopsy methods under thin-layer CT navigation in the diagnosis of malignant peripheral lung lesions.Methods:A retrospective analysis of patients with suspected malignant peripheral lung lesions identified by chest CT from January 2019 to September 2024 at the Cancer Hospital of the Chinese Academy of Medical Sciences and Peking Union Medical College, and Xianyang Central Hospital, who underwent routine bronchoscopy with negative results (209 cases). These patients were diagnosed using bronchial ultrasound under thin-layer CT navigation. The cases were divided into a cryobiopsy group (127 cases) and a conventional forceps biopsy group based on the biopsy method (82 cases). The diagnostic rates of the two groups were statistically analyzed, along with factors influencing the diagnostic rates. The tissue size obtained from both groups was compared, and the occurrence of complications was summarized.Results:This study included 209 cases with 216 peripheral lung lesions. A total of 209 cases with 210 lesions were successfully located through thin-slice CT guidance, resulting in a guiding success rate of 97.2% (210/216). Among the 130 lesions in the cryobiopsy group, 78 lesions were diagnosed as lung malignancies, with a diagnostic rate of 82.1% (64/78) for cryobiopsy in lung malignant lesions. In the forceps biopsy group, 46 of the 86 lesions were diagnosed as lung malignancies, with a diagnostic rate of 87.0% (40/46) for forceps biopsy in lung malignant lesions. There was no statistically significant difference between the two diagnostic rates ( P=0.473). The average longest diameter of tissue obtained by cryobiopsy was (6.11±0.23) mm, while the average longest diameter of tissue obtained by forceps biopsy was (1.58±0.43) mm. There was a statistically significant difference in tissue longest diameter between the two groups ( P＜0.001). When the distance from the bronchoscopic tip to the lesion was ≥3 cm and the most distal bronchus visible under bronchoscopy was ≤5th generation, the diagnostic rate of forceps biopsy was higher [83.3%(25/30) and 94.1%(32/34)] than that of cryobiopsy [79.3%(23/29) and 78.0%(46/59)], and the difference was statistically significant ( P＜0.05). Regarding complications, one case (1.3%, 1/78) of clinically significant complications occurred in the cryobiopsy group, while no complications occurred in the forceps biopsy group. Conclusions:Under thin-layer CT navigation, bronchial ultrasound combined with different biopsy methods demonstrates a high diagnostic rate for malignant peripheral lung lesions and is safe to operate. Cryobiopsy allows for the collection of larger tissue specimens.

Objective To explore the association between plasma fibrinogen(FBG)levels and the risk of in-hospital mortality among patients with septic shock.Methods The clinical data of 563 patients diagnosed with septic shock in the Intensive Care Unit(ICU)of Shenzhen Second People's Hospital from August 1,2018,to December 31,2020,were collected.Patient demographic information,basic vital signs,and blood routine and biochemical indices upon admission were gathered.Moreover,the Acute Physiology and Chronic Health Evaluation Ⅱ(APACHEⅡ)scores were calculated.Binary logistic regression analysis was conducted to explore the correlation between plasma fibrinogen levels and in-hospital mortality in patients with septic shock.Additionally,a generalized additive model(GAM)and smoothed curve fitting were employed to investigate the nonlinear relationship between plasma fibrinogen and in-hospital mortality.Receiver operating characteristic(ROC)curves were constructed for FBG and APACHEⅡ scores to predict in-hospital mortality in septic shock patients.The area under the curve(AUC)was computed to compare the predictive efficacies of the two.Furthermore,a segmented linear regression model was utilized for quantification.Results Binary logistic regression analysis demonstrated a significant negative correlation between plasma fibrinogen levels and in-hospital mortality among patients with septic shock(P<0.05).GAM modeling and smoothed curve fitting disclosed a nonlinear association between plasma fibrinogen levels and in-hospital mortality,with an inflection point at 5.54 g/L.The segmented linear regression model indicated that,to the left of the inflection point(FBG≤5.54 g/L),for every 1 g/L decrease in plasma fibrinogen,the risk of death increased by 24.5%(OR=0.755,P=0.003).Conversely,to the right of the inflection point(FBG>5.54 g/L),the relationship was not statistically significant(OR=1.049,P=0.685).The findings of the subgroup analyses indicated that the characteristics of the subgroups did not alter the relationship between blood fibrinogen levels and in-hospital mortality.Conclusion There is a nonlinear relationship between FBG levels and in-hospital mortality in patients with septic shock,which has predictive value for evaluating the risk of in-hospital mortality in this patient cohort.

Aim To establish the pyroptosis model of rat chondrocytes induced by tumor necrosis factor α(TNF-α)in order to study the effect of lysine acetyl-transferase 7(KAT7)on pyroptosis of chondrocytes.Methods Chondrocytes of rat knee joint were isolated by type Ⅱ collagenase digestion,and were identified by toluidine blue staining and Col Ⅱ immunofluorescence.CCK-8 was used to evaluate cell viability.Western blot was used to detect the expression of pyroptosis-related proteins NLRP3,GSDMD,caspase-8 and KAT7 in cells intervened with TNF-α,adenovirus overexpression of KAT7(KAT7-oe)and KAT7 inhibitor WM-3835.The microstructure of the cells was observed by scanning e-lectron microscopy.Pyroptosis was detected by TUNEL staining,and the expression of pyroptosis-related pro-tein and KAT7 was detected by immunofluorescence.Results Compared with the empty virus group,KAT7-oe inhibited cell viability,promoted the expression of pyroptosis-related proteins,and TNF-α enhanced this effect.At the same time,the expression of KAT7 and pyroptosis-related proteins in the TNF-α stimulation group increased,and WM-3835 reduced the related proteins expression.Electron microscopy showed that KAT7-oe caused cell swelling,deformation,membrane perforation and rupture,while WM-3835 could restore cell morphology.TUNEL staining and immunofluores-cence results also confirmed that KAT7-oe induced chondrocyte pyroptosis,and WM-3835 could down-reg-ulate the fluorescence of pyroptosis-related proteins.Conclusions The expression of KAT7 increases in rat chondrocyte pyroptosis model,and the intervention of KAT7 expression affects signal molecules related to py-roptosis pathway,suggesting that KAT7 may be related to chondrocyte pyroptosis.

Objective To summarize the potential risk factors that may arise in the national drug sampling and testing inspection process in recent years,to focuse on the operation of the quality management system,and to put forward proposals on how to do a good job under the new drug regulatory model of sampling and testing work.Methods Focusing on the investigation of data integrity and authenticity,the analysis focuses on the analysis of risk points such as reagent management,standard substance management,instrument,and facility management,electronic data management and other issues,and carries out a comprehensive verification of the effectiveness of the operation of the quality management system and so on.Results National drug sampling and testing institutes should strengthen the overall quality management,according to the operation of the laboratory,combined with their respective risk characteristics,reagent management,standard substance management,instrument and facility management,electronic data management and other aspects of the risk of systematic sorting and the establishment of the risk alert function,the development of risk warning lists,and the implementation of the corresponding risk control strategy.Conclusion National drug sampling and testing institutes must strengthen the operation of the process of influencing factors in the effective control of the emphasis on the testing of the work of the key control points and continue to standardize and improve the inspection process of the quality system to ensure that the quality of the various activities in a controlled state.

Based on the theory of"the spleen and stomach as the pivot",Professor Xiong Lei's clinical thinking of"moving the pivot and removing stale qi"to treat depression in children and adolescents is summarized.It is believed that the key mechanism of de-pression in children and adolescents is the unfavorable pivot of the spleen and stomach,and the disharmony between qi and blood of the five organs,which leads to the growth of stale qi,the clouding of the clear orifices by turbid evils,and the malfunction of the spirit.If the pivot is unfavorable,move it,and if the stale qi is endogenous,remove it.Professor Xiong,based on the concept of"treating with orchids and removing stale qi"from The Yellow Emperor's Inner Classic,utilizes aromatherapy in Chinese medicine,taking"moving the pivot and removing stale qi"as the basic treatment,and proposes Xiangzhi Shuxin Formula as the basic formula,which applies five methods of removing dampness,resolving phlegm,eliminating accumulation,removing stasis,and dispersing fire and relieving depres-sion.An experimental case is attached to explain his clinical thinking.