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.

Aim To investigate the regulatory mecha-nism of arrhythmia of sodium channel ubiquitination af-ter MI and to study the electrophysiological remodeling mechanism of lncRNA-CCRR after MI for the preven-tion and treatment of arrhythmia after MI.Methods LncRNA-CCRR transgenic mice and C57BL/6 mice injected with lncRNA-CCRR overexpressed adeno-asso-ciated virus were used.Four weeks after infection,the left anterior descending branch of the coronary artery was ligated for 12 h to establish a mouse acute myocar-dial infarction model,and the incidence of arrhythmia was detected by programmed electrical stimulation.Ln-cRNA-CCRR overexpression/knockdown adeno-associ-ated virus and negative control were transfected into neonatal mouse cardiomyocytes(NMCMs),and the model was prepared by hypoxia for 12 h.LncRNA-CCRR expression was detected by FISH,Nav1.5 and UBA6 protein and Nav.1.5 mRNA expression were de-tected by Western blot and real-time quantitative poly-merase chain reaction(qRT-PCR),Nav1.5 and UBA6 expressions were detected by immunofluores-cence,and the relationship between lncRNA-CCRR and UBA6 was detected by RIP.INa current density af-ter CCRR overexpression and knockdown was detected by Whole-cell clamp patch.Results In MI mice,the expression of lncRNA-CCRR decreased,the incidence of arrhythmia increased,the expression of CCRR and Nav1.5 mRNA was down-regulated,the protein ex-pression of Nav1.5 was down-regulated,and the pro-tein expression of UBA6 was up-regulated compared with sham group.Overexpression of CCRR could re-verse the above changes.AAV-CCRR could reverse the down-regulated CCRR and Nav1.5 mRNA levels af-ter hypoxia,and improve the expression of Nav1.5 and UBA6 protein.The direct relationship between ln-cRNA-CCRR and UBA6 was identified by RIP analy-sis.The INa density increased after transfection with AAV-CCRR.The INa density decreased after transfec-tion with AAV-si-CCRR.Conclusions The expres-sion of lncRNA-CCRR decreases after MI,and ln-cRNA-CCRR can improve arrhythmia induced by MI by inhibiting UBA6 to increase the protein expression level of Nav1.5 and the density of INa.

Objective To explore feasibility of 3D metal printing technology combined with virtual design proximal clavicle anatomical plate.Methods A 52-year-old male healthy volunteer was retrospectively selected to design proximal clavicle anatomical plate system by using Mimics15.01,NX12.0 and other software.STL data were input into 3D printer to print 1∶1 clavicle model and proximal clavicle anatomical plate.The fit of the plate was tested in vitro and the accuracy of screw position was evaluated by imaging.Printing time of model,nail path design and fabrication time of the anatomical plate at proximal clavicle were recorded.Results The 3D metal printing proximal clavicle anatomical plate fitted well to clavicle model,orienta-tion of proximal clavicle locking screw was accurate,and X-ray and CT scan showed the screw position was good.Printing time of model,the time of nail path design,and the time of making anatomical plate of proximal clavicle were 120,15 and 300 min respectively.Conclusion The proximal clavicular anatomical plate system based on 3D metal printing technology could achieve good lamination of proximal clavicular fracture plate and precise screw placement,providing a new and accurate surgical method for the treatment of the proximal clavicular fracture.

Objective:To explore the efficacy and safety of haploidentical hematopoietic stem cell transplantation combined with umbilical cord blood infusion for the treatment of aplastic anaemia in children.Methods:Nine cases of children with aplastic anaemia treated with umbilical cord blood combined with haploidentical hematopoietic stem cell transplantation at the People's Hospital of Henan University of Chinese Medicine from January 1,2021 to September 15,2023 with a median age of 11(2-13)years and a median follow up of 18(7.5-21)months were included,and the clinical data were retrospectively analyzed.Hematopoiesis reconstitution,the incidence of graft-versus-host disease(GVHD),infections and survival of the patients were analyzed.Results:All 9 children were successfully implanted.The median time to neutrophil and platelet implantation was 11.11±1.27 d and 12.44±3.36 d,respectively.One case developed acute gastrointestinal GVHD of degree I,which was improved after treatment,and the patient developed superficial gastritis and chronic gastrointestinal GVHD at a later stage,which is currently under clinical follow-up.Acute GVHD of Ⅱ-Ⅳ degree was 0％.Hemorrhagic cystitis in 3 cases,CMV infection in 5 cases and bacterial and fungal infections in 5 cases improved with symptomatic treatment.All 9 children demonstrated complete donor chimerism within 1 month after transplantation,at two years of follow-up,all nine children survived without recurrence or development of grade Ⅱ-Ⅳ GVHD,and there were no children with transplant-related deaths.Conclusion:Haploidentical hematopoietic stem cell transplantation combined with umbilical cord blood transfusion for aplastic anaemia in children has a low incidence and mild degree of GVHD,with significant efficacy,and can be used as a therapeutic option for children without an HLA full donor chimeric match.

Objective:To summary the clinical and electrophysiological characteristics of temporal lobe epilepsy (TLE) originating from the temporal pole (TP), and to conduct brain network analysis based on stereo-electroencephalogram (SEEG) and head positron emission tomography- computed tomography (PET-CT).Methods:A retrospective analysis was conducted on patients with TLE who underwent SEEG implantation from January 1, 2019 to September 1, 2023 in Guangdong Sanjiu Brain Hospital. Based on anatomical-electrical-clinical analysis and SEEG findings, patients with seizures originating from the TP were selected. The clinical data, head magnetic resonance imaging (MRI), PET-CT, scalp electroencephalogram were reviewed, and the seizure-induced network was analyzed based on SEEG and head PET-CT.Results:A total of 108 cases of TLE were analyzed, of whom 13 cases had an epileptogenic zone located at the TP, accounting for 12% (13/108) of all TLE patients. Among them, 8 were males and 5 were females, and age of onset was (11.6±7.8) years. All of them were drug-resistant epilepsy patients, of whom 6 cases had normal cognitive function, 4 had mild cognitive abnormalities, and 3 had severe cognitive decline. A total of 59 seizures were recorded, and the occurrence rate of generalized tonic-clonic seizures (GTCS) was 42% (25/59). Seizure symptoms were classified into 3 types: the first type was hypermotor, seen in 9 patients; the second type was complex motor, seen in 2 patients; and the third type was automotor, seen in 2 patients. Head MRI showed that 9 cases had a blurring of the TP on one side, with or without hippocampal sclerosis; 2 cases had a mass at the TP without hippocampal sclerosis; 2 cases were negative on head MRI. Head PET-CT showed that 13 cases had TP hypometabolism on the lesion side, of whom 11 cases had hypometabolism involving the medial temporal lobe (mTL), posterior orbital gyrus (POG), anterior cingulate gyrus (ACG) and insular lobe at the same time, the other 2 cases combined with ipsilateral hypometabolism of the medial temporal lobe. Pathology showed that 7 cases had microcortical dysplasia of the TP; 3 had focal cortical dysplasia Ⅰ or focal cortical dysplasia Ⅱ; 2 had benign tumors. Scalp electroencephalogram showed that interictal phase was divided into 3 discharge patterns: bilateral temporal regions with prominent lesion side; bilateral anterior regions with prominent lesion side; lesion-side hemisphere with prominent temporal region. Ictal period showed 4 initial patterns: lesion-side hemispheric rhythmic spikes-slow waves or polyspikes-slow waves; lesion-side anterior region rhythmic slow waves; lesion-side anterior region low voltage fast (LVF) activities, and diffuse LVF with prominent lesion-side hemisphere. SEEG showed that 13 patients received electrode implantation with (9±2) electrodes per patient, divided into 3 seizure patterns: type 1: TP?adjacent temporal neocortex?POG, ACG and insula?mTL; type 2: TP?para hippocampal gyrus and the base of temporal lobe?ACG ,POG and insula?mTL; type 3: TP?mTL?insular lobe?POG.Conclusions:TLE originating from the TP is relatively rare, with hypermotor or complex motor as the main manifestations, and automotor being relatively less common, which is more likely to be followed by GTCS. The epileptogenic network analysis displays a tendency to spread from the TP to the frontal and insular lobes, as well as to the mTL, with the former pattern being more common. Common etiologies are cortical dysplasia and benign tumors of the TP without hippocampal sclerosis.

Research integrity is the foundation for ensuring the sound and orderly development of scientific and technological innovation. As the main battlefield of clinical medical research, hospitals should effectively fulfill their main responsibilities and do a good job in research integrity management. The China Hospital Research Integrity Alliance, consisting of the first batch of 43 hospitals, was established in November 2021. With the aim of " complementary advantages, resource sharing, and collaborative development", the alliance has carried out construction practices from seven aspects: construction mode, cultural system construction, organizational management, institutional construction, publicity and education, early warning and supervision, and technological empowerment. It has achieved the overall improvement of the research integrity construction ability of member units of the alliance, organic linkage between government and medical institutions, and efficient combination of internal and external resources, which can provide reference for the research integrity construction of medical institutions in China.

Objective:To observe the effects of electroacupuncture(EA)combined with Western medication on lower-limb motor function and blood circulation in patients with cerebral infarction-induced hemiplegia in the acute stage. Methods:One hundred eligible patients with acute ischemic stroke accompanied by lower-limb motor dysfunction were allocated to an observation group and a control group using the random number table method,with 50 cases in each group.The control group received routine Western medications for treatment,and the observation group received additional EA intervention.After 2-week and 4-week treatments,the improvement of lower-limb motor function was assessed using the Fugl-Meyer assessment scale for lower extremity(FMA-LE),and changes in the peak blood flow velocities of the posterior tibial(PT)and dorsalis pedis(DP)arteries on the affected side were detected using Doppler. Results:Three cases dropped out during the study,so there were 48 cases in the observation group and 49 in the control group collected for statistical analysis.The FMA-LE score and the peak blood flow velocities of PT and DP arteries increased after 2-week and 4-week treatments in both groups compared with the pre-treatment baseline(P<0.05).After 2-week treatments,the FMA-LE score and the peak blood flow velocities of PT and DP arteries showed no significant differences between the two groups(P>0.05).After 4-week treatments,compared with the control group,the FMA-LE score was higher(P<0.05),and the peak blood flow velocities of PT and DP arteries on the affected side were larger in the observation group(P<0.05). Conclusion:EA combined with Western medication can significantly improve the motor function and blood flow velocity of the affected lower limb in patients with acute cerebral infarction accompanied by hemiplegia.

Objective This paper analyzed the characteristics and changes in the disease spectrum of internal medicine inpatients of a tertiary hospital in Zhongshan City.It aimed to explore the evolving trends in disease ranking and treatment charac-teristics to provide a scientific basis for enhancing hospital management.Methods This paper selected 26 053 patients dis-charged from the internal medicine department of the tertiary hospital in Zhongshan City between January 1,2018,and December 31,2022.A retrospective analysis was used to classify these patients by ages,hospitalization date,and relative weight.Results In the past five years,those discharged patients showed no significant increase in the number,but a significant increase in age and ase Mix Index(CMI).Conclusion Since the outbreak of COVID-19,intractable inpatients in the hospital has steadily in-creased in number each year,while the diseases spectrum remains stable.The CMI of circulatory system,respiratory system,and nervous system diseases is relatively high.As key tricky components of internal medicine,these sub-specialties should concentrate on enhancing diagnostic and treatment capabilities as well as service quality to offer improved healthcare services to the public.

Objective:To observe the intervention effect of aerobic Baduanjin exercise combined with treadmill train-ing on chronic obstructive pulmonary disease(COPD)complicated chronic heart failure(CHF).Methods:A total of 180 COPD+CHF patients who were treated in Mianyang Central Hospital from January 2019 to June 2021 were selected.They were equally divided into control group(basic treatment and intervention)and experimental group(received aerobic Baduanjin exercise combined with treadmill training based on control group)by random number table method,both groups were intervened for 12 weeks.COPD assessment test(CAT)score,left ventricular ejec-tion fraction(LVEF),left ventricular end systolic volume(LVESV),left ventricular end diastolic volume(LV-EDV),scores of modified medical research council(mMRC)and Minnesota living with heart failure questionnaire(MLHFQ)were compared between two groups before,4,8 and 12 weeks after intervention.Results:During 12-week intervention,CAT score,LVESV,LVEDV,scores of mMRC and MLHFQ showed a gradual decreasing trend,while LVEF showed a gradual increasing trend in two groups,P＜0.05 or＜0.01.Compared with control group after 12-week intervention,there were significant reductions in CAT score[(17.47±3.96)points vs.(13.36±3.42)points],LVEDV[(139.44±7.12)ml vs.(131.74±6.47)ml],LVESV[(81.84±8.54)ml vs.(74.29±9.18)ml],scores of mMRC[(1.67±0.47)points vs.(1.42±0.50)points],each dimension and total score of MLHFQ[(40.07±6.86)points vs.(32.54±6.61)points],and significant rise in LVEF[(41.29±4.76)％vs.(44.56±4.42)％]in experimental group(P＜0.001 all).Conclusion:Aerobic Baduanjin exercise combined with treadmill training based on basic treatment can significantly improve cardiopulmonary function,re-lieve dyspnea and improve quality of life in COPD+CHF patients,which is worth promoting.

Objective To investigate the clinical characteristics and prognosis of pneumococcal meningitis(PM),and drug sensitivity of Streptococcus pneumoniae(SP)isolates in Chinese children.Methods A retrospective analysis was conducted on clinical information,laboratory data,and microbiological data of 160 hospitalized children under 15 years old with PM from January 2019 to December 2020 in 33 tertiary hospitals across the country.Results Among the 160 children with PM,there were 103 males and 57 females.The age ranged from 15 days to 15 years,with 109 cases(68.1% )aged 3 months to under 3 years.SP strains were isolated from 95 cases(59.4% )in cerebrospinal fluid cultures and from 57 cases(35.6% )in blood cultures.The positive rates of SP detection by cerebrospinal fluid metagenomic next-generation sequencing and cerebrospinal fluid SP antigen testing were 40% (35/87)and 27% (21/78),respectively.Fifty-five cases(34.4% )had one or more risk factors for purulent meningitis,113 cases(70.6% )had one or more extra-cranial infectious foci,and 18 cases(11.3% )had underlying diseases.The most common clinical symptoms were fever(147 cases,91.9% ),followed by lethargy(98 cases,61.3% )and vomiting(61 cases,38.1% ).Sixty-nine cases(43.1% )experienced intracranial complications during hospitalization,with subdural effusion and/or empyema being the most common complication[43 cases(26.9% )],followed by hydrocephalus in 24 cases(15.0% ),brain abscess in 23 cases(14.4% ),and cerebral hemorrhage in 8 cases(5.0% ).Subdural effusion and/or empyema and hydrocephalus mainly occurred in children under 1 year old,with rates of 91% (39/43)and 83% (20/24),respectively.SP strains exhibited complete sensitivity to vancomycin(100% ,75/75),linezolid(100% ,56/56),and meropenem(100% ,6/6).High sensitivity rates were also observed for levofloxacin(81% ,22/27),moxifloxacin(82% ,14/17),rifampicin(96% ,25/26),and chloramphenicol(91% ,21/23).However,low sensitivity rates were found for penicillin(16% ,11/68)and clindamycin(6% ,1/17),and SP strains were completely resistant to erythromycin(100% ,31/31).The rates of discharge with cure and improvement were 22.5% (36/160)and 66.2% (106/160),respectively,while 18 cases(11.3% )had adverse outcomes.Conclusions Pediatric PM is more common in children aged 3 months to under 3 years.Intracranial complications are more frequently observed in children under 1 year old.Fever is the most common clinical manifestation of PM,and subdural effusion/emphysema and hydrocephalus are the most frequent complications.Non-culture detection methods for cerebrospinal fluid can improve pathogen detection rates.Adverse outcomes can be noted in more than 10% of PM cases.SP strains are high sensitivity to vancomycin,linezolid,meropenem,levofloxacin,moxifloxacin,rifampicin,and chloramphenicol.[Chinese Journal of Contemporary Pediatrics,2024,26(2):131-138]