Membrane proteins are integral components of cellular membranes, accounting for approximately 30% of the mammalian proteome and serving as targets for 60% of FDA-approved drugs. They are critical to both physiological functions and disease mechanisms. Their functional protein-protein interactions form the basis for many physiological processes, such as signal transduction, material transport, and cell communication. Membrane protein interactions are characterized by membrane environment dependence, spatial asymmetry, weak interaction strength, high dynamics, and a variety of interaction sites. Therefore, in situ analysis is essential for revealing the structural basis and kinetics of these proteins. This paper introduces currently available in situ analytical techniques for studying membrane protein interactions and evaluates the characteristics of each. These techniques are divided into two categories: label-based techniques (e.g., co-immunoprecipitation, proximity ligation assay, bimolecular fluorescence complementation, resonance energy transfer, and proximity labeling) and label-free techniques (e.g., cryo-electron tomography, in situ cross-linking mass spectrometry, Raman spectroscopy, electron paramagnetic resonance, nuclear magnetic resonance, and structure prediction tools). Each technique is critically assessed in terms of its historical development, strengths, and limitations. Based on the authors’ relevant research, the paper further discusses the key issues and trends in the application of these techniques, providing valuable references for the field of membrane protein research. Label-based techniques rely on molecular tags or antibodies to detect proximity or interactions, offering high specificity and adaptability for dynamic studies. For instance, proximity ligation assay combines the specificity of antibodies with the sensitivity of PCR amplification, while proximity labeling enables spatial mapping of interactomes. Conversely, label-free techniques, such as cryo-electron tomography, provide near-native structural insights, and Raman spectroscopy directly probes molecular interactions without perturbing the membrane environment. Despite advancements, these methods face several universal challenges: (1) indirect detection, relying on proximity or tagged proxies rather than direct interaction measurement; (2) limited capacity for continuous dynamic monitoring in live cells; and (3) potential artificial influences introduced by labeling or sample preparation, which may alter native conformations. Emerging trends emphasize the multimodal integration of complementary techniques to overcome individual limitations. For example, combining in situ cross-linking mass spectrometry with proximity labeling enhances both spatial resolution and interaction coverage, enabling high-throughput subcellular interactome mapping. Similarly, coupling fluorescence resonance energy transfer with nuclear magnetic resonance and artificial intelligence (AI) simulations integrates dynamic structural data, atomic-level details, and predictive modeling for holistic insights. Advances in AI, exemplified by AlphaFold’s ability to predict interaction interfaces, further augment experimental data, accelerating structure-function analyses. Future developments in cryo-electron microscopy, super-resolution imaging, and machine learning are poised to refine spatiotemporal resolution and scalability. In conclusion, in situ analysis of membrane protein interactions remains indispensable for deciphering their roles in health and disease. While current technologies have significantly advanced our understanding, persistent gaps highlight the need for innovative, integrative approaches. By synergizing experimental and computational tools, researchers can achieve multiscale, real-time, and perturbation-free analyses, ultimately unraveling the dynamic complexity of membrane protein networks and driving therapeutic discovery.

OBJECTIVE: To explore the therapeutic effects and underlying mechanisms of Dendrobium officinale (Tiepi Shihu) extract (DOE) on insomnia. METHODS: Forty-two male Sprague-Dawley rats were randomly divided into 6 groups (n=7 per group): normal control, model control, melatonin (MT, 40 mg/kg), and 3-dose DOE (0.25, 0.50, and 1.00 g/kg) groups. Rats were raised in a strong-light (10,000 LUX) and -noise (>80 db) environment (12 h/d) for 16 weeks to induce insomnia, and from week 10 to week 16, MT and DOE were correspondingly administered to rats. The behavior tests including sodium pentobarbital-induced sleep experiment, sucrose preference test, and autonomous activity test were used to evaluate changes in sleep and emotions of rats. The metabolic-related indicators such as blood pressure, blood viscosity, blood glucose, and uric acid in rats were measured. The pathological changes in the cornu ammonis 1 (CA1) region of rat brain were evaluated using hematoxylin and eosin staining and Nissl staining. Additionally, the sleep-related factors gamma-aminobutyric acid (GABA), glutamate (GA), 5-hydroxytryptamine (5-HT), and interleukin-6 (IL-6) were measured using enzyme linked immunosorbent assay. Finally, we screened potential sleep-improving receptors of DOE using polymerase chain reaction (PCR) array and validated the results with quantitative PCR and immunohistochemistry. RESULTS: DOE significantly improved rats' sleep and mood, increased the sodium pentobarbital-induced sleep time and sucrose preference index, and reduced autonomic activity times (P<0.05 or P<0.01). DOE also had a good effect on metabolic abnormalities, significantly reducing triglyceride, blood glucose, blood pressure, and blood viscosity indicators (P<0.05 or P<0.01). DOE significantly increased the GABA content in hippocampus and reduced the GA/GABA ratio and IL-6 level (P<0.05 or P<0.01). In addition, DOE improved the pathological changes such as the disorder of cell arrangement in the hippocampus and the decrease of Nissel bodies. Seven differential genes were screened by PCR array, and the GABA_A receptors (Gabra5, Gabra6, Gabrq) were selected for verification. The results showed that DOE could up-regulate their expressions (P<0.05 or P<0.01). CONCLUSION DOE demonstrated remarkable potential for improving insomnia, which may be through regulating GABA_A receptors expressions and GA/GABA ratio.
Animals ; Dendrobium/chemistry* ; Rats, Sprague-Dawley ; Male ; Sleep Initiation and Maintenance Disorders/blood* ; Plant Extracts/therapeutic use* ; Receptors, GABA-A/metabolism* ; Noise/adverse effects* ; Light/adverse effects* ; gamma-Aminobutyric Acid/metabolism* ; Sleep/drug effects* ; Rats ; Receptors, GABA/metabolism*

OBJECTIVES: To explore the correlation of ELFN1 expression level with prognosis of colorectal cancer and its regulatory role in colorectal cancer cell proliferation and metastasis. METHODS: We analyzed the expression levels of ELFN1 across 33 cancer types using publicly available databases and identified differential genes related to ELFN1 in colorectal cancer. Gene function annotation and enrichment analysis were used to identify the involved signaling pathways. Logistic analysis, Kaplan-Meier analysis and Cox regression analysis were performed to evaluate the correlation between ELFN1 expression and clinicopathological parameters and survival of colorectal cancer patients. qPCR and Western blotting were used to validate the expression levels of ELFN1 in different colorectal cancer cell lines and tissues, and Transwell and EDU experiments were carried out to assess the effect of ELFN1 knockdown on biological behaviors of SW480 cells. RESULTS: ELFN1 was highly expressed in 14 cancers, and its expression was significantly higher in colon cancer tissues than in adjacent tissues. A high expression of ELFN1 mRNA was associated with a poorer overall survival of colorectal cancer patients. Cox regression analysis indicated that ELFN1 expression was an independent prognostic factor for overall survival of the patients. ELFN1 was significantly enriched in tumor metastasis and proliferation and participated in several tumor signaling pathways. The colon cancer cell lines showed significantly higher expression levels of ELFN1 than normal cells, ELFN1 knockdown obviously inhibited proliferation and migration of SW480 cells in vitro. CONCLUSIONS ELFN1 is overexpressed in colorectal cancer and is associated with poor clinical prognosis of the patients. A high ELFN1 expression is associated with malignant phenotypes of colorectal cancer and promotes cancer cell proliferation and metastasis, suggesting its potential as a prognostic biomarker for colorectal cancer.
Humans ; Colorectal Neoplasms/diagnosis* ; Cell Proliferation ; Prognosis ; Cell Line, Tumor ; Biomarkers, Tumor/metabolism* ; Neoplasm Metastasis ; Gene Expression Regulation, Neoplastic ; Nerve Tissue Proteins/metabolism* ; Female ; Male

INTRODUCTION: Lecanemab has shown promise in treating early Alzheimer's disease (AD), but its safety and efficacy in Chinese populations remain unexplored. This study aimed to evaluate the safety and 6-month clinical outcomes of lecanemab in Chinese patients with mild cognitive impairment (MCI) or mild AD. METHODS: In this single-arm, real-world study, participants with MCI due to AD or mild AD received biweekly intravenous lecanemab (10 mg/kg). The study was conducted at Hainan Branch, Ruijin Hospital Shanghai Jiao Tong University School of Medicine. Patient enrollment and baseline assessments commenced in November 2023. Safety assessments included monitoring for amyloid-related imaging abnormalities (ARIA) and other adverse events. Clinical and biomarker changes from baseline to 6 months were evaluated using cognitive scales (mini-mental state examination [MMSE], montreal cognitive assessment [MoCA], clinical dementia rating-sum of boxes [CDR-SB]), plasma biomarker analysis, and advanced neuroimaging. RESULTS: A total of 64 patients were enrolled in this ongoing real-world study. Safety analysis revealed predominantly mild adverse events, with infusion-related reactions (20.3%, 13/64) being the most common. Of these, 69.2% (9/13) occurred during the initial infusion and 84.6% (11/13) did not recur. ARIA-H (microhemorrhages/superficial siderosis) and ARIA-E (edema/effusion) were observed in 9.4% (6/64) and 3.1% (2/64) of participants, respectively, with only two symptomatic cases (one ARIA-E presenting with headache and one ARIA-H with visual disturbances). After 6 months of treatment, cognitive scores remained stable compared to baseline (MMSE: 22.33 ± 5.58 vs . 21.27 ± 4.30, P = 0.733; MoCA: 16.38 ± 6.67 vs . 15.90 ± 4.78, P = 0.785; CDR-SB: 2.30 ± 1.65 vs . 3.16 ± 1.72, P = 0.357), while significantly increasing plasma amyloid-β 42 (Aβ42) (+21.42%) and Aβ40 (+23.53%) levels compared to baseline. CONCLUSIONS: Lecanemab demonstrated a favorable safety profile in Chinese patients with early AD. Cognitive stability and biomarker changes over 6 months suggest potential efficacy, though high dropout rates and absence of a control group warrant cautious interpretation. These findings provide preliminary real-world evidence for lecanemab's use in China, supporting further investigation in larger controlled studies. REGISTRATION ClinicalTrials.gov , NCT07034222.
Humans ; Alzheimer Disease/drug therapy* ; Male ; Female ; Aged ; Middle Aged ; Cognitive Dysfunction/drug therapy* ; Aged, 80 and over ; Amyloid beta-Peptides/metabolism* ; Biomarkers ; East Asian People

Objective:To explore the expression of growth differentiation factor-15 (GDF-15) in different pathological types of prostate cancer, and to analyze the impact of GDF-15 on biochemical recurrence of prostate cancer after radical prostatectomy.Methods:A retrospective analysis was conducted on the case data of 138 patients with prostate cancer who underwent radical prostatectomy at the 363rd Hospital from January 2019 to January 2022. The patients were divided into the recurrence group ( n=46) and the non-recurrence group ( n=92) based on postoperative biochemical recurrence. Measurement data were expressed as mean±standard deviation ( ± s), and analysis of variance was used for comparisons among multiple groups, while t-test was used for comparisons between two groups. Count data were expressed as the number of cases and percentages, and the Chi-test was used for comparisons between groups. Stratified regression analysis was used to explore the relationship between GDF-15 and different clinical pathological characteristics. By combining spline functions and Logistic regression analysis, the restricted cubic spline model was used to analyze the effect of GDF-15 on biochemical recurrence of prostate cancer in different pathological types. Kaplan-Meier survival curves were drawn to visually display the recurrence differences of prostate cancer patients at different GDF-15 levels. Results:The expression level of GDF-15 was the lowest in Gleason grade group (GGG)1 type at (548.71±78.25) pg/mL, and the highest in GGG5 type at (916.75±94.33) pg/mL, and the difference was statistically significant ( F=87.39, P<0.001). The level of GDF-15 was positively correlated with prostate-specific antigen (PSA), GGG5 type, and vascular endothelial growth factor (VEGF) ( P=0.029, 0.022, 0.015). The levels of PSA, GDF-15, Gleason score, alkaline phosphatase (ALP), C-reactive protein (CRP)/albumin (ALB), VEGF, and the proportion of TNM stage Ⅲ-Ⅳ, lymph node invasion, seminal vesicle invasion, nerve invasion, capsule rupture, and GGG5 type in the recurrence group were significantly higher than those in the non-recurrence group, the differences were all statistically significant ( P<0.05); the lymphocyte-to-monocyte ratio (LMR) was significantly lower than that in the non-recurrence group, and the difference was statistically significant ( P<0.001). GDF-15 had a non-linear dose-response relationship with the biochemical recurrence risk value. The risk value is significantly positively correlated with the level of GDF-15, and this relationship was not affected by the type of prostate cancer pathology. In patients with GGG1 to GGG5 types of prostate cancer, when the GDF-15 levels were >720, 700, 690, 650, and 610 pg/mL, respectively, the risk values increased as the GDF-15 levels rose. The Kaplan-Meier survival curve analysis showed that as the level of GDF-15 increased, the recurrence rate of patients significantly rose. Conclusions:The expression level of GDF-15 expression is the lowest in GGG1 type and the highest in GGG5 type. Both GDF-15 and GGG5 type are the independent risk factors for biochemical recurrence after radical prostatectomy, and GDF-15 exhibits a significant positive nonlinear dose-response relationship with the risk of biochemical recurrence.

With the establishment of bio-psycho-social medical model,both social and psychological factors play an important role in the occurrence,development and treatment of diseases.Hypertension is a common chronic multiple disease in China,and patients are often complicated with depression and other e-motional disorders.The interaction between hypertension and depression significantly increases the risk of poor prognosis.Current studies have shown a bidirectional promoting relationship between hypertension and depression,and they have some com-mon pathogenesis.However,the specific mechanism of their co-morbidity has not been fully elucidated.Renin-angiotensin sys-tem(RAS)plays an important role in the regulation of hyperten-sion and depression and other emotions.It is composed of two antagonistic pathways.The balance is maintained by angioten-sin-converting enzyme 2(ACE2).Therefore,this article reviews the relationship and mechanism of RAS in hypertension,depres-sion and comorbid states,in order to provide new treatment ide-as for hypertension and depression.

Objective To analyze the risk factors for increased drainage volume after open transforaminal lumbar interbody fusion(TLIF),and to establish a predictive model and then validate it.Methods The clinical data of 680 patients who underwent open TLIF at the General Hospital of Northern Theater Command from January 2016 to December 2019 were collected and the patients were randomly divided into the training group(n=476)and the validation group(n=204).Taking the predictive factors screened out by LASSO regression analysis as independent variables,a multivariate Logistic regression predictive model was constructed.The model was internally validated through the receiver operating characteristic(ROC)curve,Hosmer-Lemeshow goodness-of-fit test,and calibration curve,and its clinical utility was assessed via decision curve analysis(DCA).Results LASSO regression analysis screened out four predictive variables:age,number of surgical segments,operative duration,and intraoperative blood loss.The multivariate Logistic regression predictive model demonstrated that age≥60 years,number of surgical segments≥4,operative duration≥2 hours,and intraoperative blood loss≥200 mL were independent influencing factors for the increased postoperative drainage volume in patients undergoing TLIF(P<0.05).ROC curve analysis revealed an area under the curve(AUC)of 0.816(95%CI:0.798 to 0.867)in the training group and 0.783(95%CI:0.685 to 0.823)in the validation group,indicating that the predictive model had good discriminatory ability.Additionally,the Hosmer-Lemeshow goodness-of-fit test and calibration curve indicated that the predictive model had a good degree of fit,and the predicted probability was basically consistent with the actual probability,demonstrating a good calibration.The DCA results confirmed that this predictive model could be applied in clinical practice.Conclusion The risk factors for increased drainage volume after open TLIF include age,number of surgical segments,operative duration,and intraoperative blood loss.The predictive model established based on these factors demonstrates good performance,and it can be applied in clinical guidance for the selection of drainage tube removal time after TLIF.

Objective:To explore the changes in nutritional status and body composition of cervical cancer patients during concurrent chemoradiotherapy (CCRT) and their correlation with CCRT toxicities.Methods:In this prospective and observational clinical study, eligible treatment -na?ve patients with stage IB-IV primary cervical cancer were consecutively enrolled in the Department of Radiotherapy of Peking Union Medical College Hospital from September 2022 to August 2023. The patients were screened for nutritional risks, received dietary assessment, and were measured for body composition using multi-frequency bioelectrical impedance at baseline (prior to treatment), 4 weeks, and 8 weeks since treatment initiation. Insufficient muscle mass was diagnosed ccording to the Asian Working Group for Sarcopenia 2019 criteria. The severity of nausea, vomiting, abdominal pain, diarrhea, and hematological toxicity was assessed by the U.S. National Cancer Institute Common Terminology Criteria for Adverse Events (version 5.0).Results:A total of 109 patients were included. At baseline, there were 11 (10.1%) patients who were lean, 17 (15.6%) patients with insufficient muscle mass, and 28 patients (25.7%) at nutritional risk; at Week 8 of CCRT, patients at nutritional risk increased to 61 (56.0%). Compared to baseline, weight [(59.34±9.67) kg vs. (61.30±9.64) kg, P<0.001], skeletal muscle index [SMI, (6.15±0.74) kg/m 2vs. (6.39±0.74) kg/m 2, P<0.001], body fat percentage [(31.13±7.67) % vs. (32.07±7.70) %, P=0.004] were significantly decreased at Week 8 of CCRT. Besides, ≥10% SMI loss was only related to baseline body fat percentage ( HR=0.216, 95% CI: 0.001-0.724, P=0.038), but not related to age, nutritional status, or muscle mass (all P>0.05). At baseline and 8 weeks since CCRT, 8 (28.6%) and 40 (65.6%) patients at nutritional risk received nutritional support, respectively. During CCRT, the rates of grade ≥2 nausea and vomiting, diarrhea, and grade 3/4 hematological toxicity were 37.6%, 28.4% and 44.0%, respectively. Baseline nutritional risk was a risk factor for diarrhea ( HR=2.447, 95% CI: 1.017-6.068, P=0.047), and an advanced International Federation of Gynecology and Obstetrics (FIGO) stage was a risk factor for severe nausea and vomiting ( HR=1.735, 95% CI: 1.005-2.995, P=0.048). Patients presenting with severe nausea and vomiting had more significant reductions in body mass index [(-1.44±1.29) kg/m 2vs. (-0.59±0.84) kg/m 2, P<0.001] and SMI [(-0.37±0.41) kg/m 2vs. (-0.12±0.27) kg/m 2, P=0.013] compared to those without nausea and vomiting, while there was no significant difference in visceral fat area between these two groups [(-9.95±19.48) cm 2vs. (-5.12±15.79) cm 2, P=0.161]. Conclusions:Patients with cervical cancer have increased nutritional risk and more loss of body weight and muscle mass during CCRT. The presence of nutritional risk at baseline is a risk factor for diarrhea, while nausea and vomiting exacerbate the losses of body weight, muscle, and fat. Close monitoring, intensive symptomatic therapy, and appropriate nutritional interventions should be performed in the clinical setting to improve patients' tolerance of treatment and maintenance of body weight.

Objective:To explore the changes in nutritional status and body composition of cervical cancer patients during concurrent chemoradiotherapy (CCRT) and their correlation with CCRT toxicities.Methods:In this prospective and observational clinical study, eligible treatment -na?ve patients with stage IB-IV primary cervical cancer were consecutively enrolled in the Department of Radiotherapy of Peking Union Medical College Hospital from September 2022 to August 2023. The patients were screened for nutritional risks, received dietary assessment, and were measured for body composition using multi-frequency bioelectrical impedance at baseline (prior to treatment), 4 weeks, and 8 weeks since treatment initiation. Insufficient muscle mass was diagnosed ccording to the Asian Working Group for Sarcopenia 2019 criteria. The severity of nausea, vomiting, abdominal pain, diarrhea, and hematological toxicity was assessed by the U.S. National Cancer Institute Common Terminology Criteria for Adverse Events (version 5.0).Results:A total of 109 patients were included. At baseline, there were 11 (10.1%) patients who were lean, 17 (15.6%) patients with insufficient muscle mass, and 28 patients (25.7%) at nutritional risk; at Week 8 of CCRT, patients at nutritional risk increased to 61 (56.0%). Compared to baseline, weight [(59.34±9.67) kg vs. (61.30±9.64) kg, P<0.001], skeletal muscle index [SMI, (6.15±0.74) kg/m 2vs. (6.39±0.74) kg/m 2, P<0.001], body fat percentage [(31.13±7.67) % vs. (32.07±7.70) %, P=0.004] were significantly decreased at Week 8 of CCRT. Besides, ≥10% SMI loss was only related to baseline body fat percentage ( HR=0.216, 95% CI: 0.001-0.724, P=0.038), but not related to age, nutritional status, or muscle mass (all P>0.05). At baseline and 8 weeks since CCRT, 8 (28.6%) and 40 (65.6%) patients at nutritional risk received nutritional support, respectively. During CCRT, the rates of grade ≥2 nausea and vomiting, diarrhea, and grade 3/4 hematological toxicity were 37.6%, 28.4% and 44.0%, respectively. Baseline nutritional risk was a risk factor for diarrhea ( HR=2.447, 95% CI: 1.017-6.068, P=0.047), and an advanced International Federation of Gynecology and Obstetrics (FIGO) stage was a risk factor for severe nausea and vomiting ( HR=1.735, 95% CI: 1.005-2.995, P=0.048). Patients presenting with severe nausea and vomiting had more significant reductions in body mass index [(-1.44±1.29) kg/m 2vs. (-0.59±0.84) kg/m 2, P<0.001] and SMI [(-0.37±0.41) kg/m 2vs. (-0.12±0.27) kg/m 2, P=0.013] compared to those without nausea and vomiting, while there was no significant difference in visceral fat area between these two groups [(-9.95±19.48) cm 2vs. (-5.12±15.79) cm 2, P=0.161]. Conclusions:Patients with cervical cancer have increased nutritional risk and more loss of body weight and muscle mass during CCRT. The presence of nutritional risk at baseline is a risk factor for diarrhea, while nausea and vomiting exacerbate the losses of body weight, muscle, and fat. Close monitoring, intensive symptomatic therapy, and appropriate nutritional interventions should be performed in the clinical setting to improve patients' tolerance of treatment and maintenance of body weight.

With the establishment of bio-psycho-social medical model,both social and psychological factors play an important role in the occurrence,development and treatment of diseases.Hypertension is a common chronic multiple disease in China,and patients are often complicated with depression and other e-motional disorders.The interaction between hypertension and depression significantly increases the risk of poor prognosis.Current studies have shown a bidirectional promoting relationship between hypertension and depression,and they have some com-mon pathogenesis.However,the specific mechanism of their co-morbidity has not been fully elucidated.Renin-angiotensin sys-tem(RAS)plays an important role in the regulation of hyperten-sion and depression and other emotions.It is composed of two antagonistic pathways.The balance is maintained by angioten-sin-converting enzyme 2(ACE2).Therefore,this article reviews the relationship and mechanism of RAS in hypertension,depres-sion and comorbid states,in order to provide new treatment ide-as for hypertension and depression.