Objective: To establish a method for the genotyping of fetal M blood group antigen by extracting cell-free fetal DNA (cff-DNA) from maternal plasma, so as to guide the management of M antigen-negative pregnant women with IgG anti-M antibody during pregnancy. Methods: A realtime fluorescent quantitative PCR (realtime PCR) method was established. The specificity and sensitivity of the method were validated by dilution of genomic DNA. Subsequently, a total of 12 M antigen-negative pregnant women were enrolled. The cff-DNA was extracted from maternal plasma, and fetal M antigen genotyping was performed by realtime PCR. Fetuses were classified as M-positive or M-negative according to the presence or absence of amplification curve. The accuracy of the method was validated by comparing fetal M antigen genotyping results with the serological results using the cord or peripheral blood of the neonate at birth. Results: Among the 12 M antigen-negative pregnant women, anti-M was detected in five cases, of which four cases had IgG anti-M, and one case had fetal anemia. The results of fetal M antigen genotyping showed that 9 cases were M-positive (9/12, 75%) and 3 cases were M-negative (3/12, 25%). Serological results of blood samples collected after birth from four M-positive fetuses and one M-negative fetus were consistent with the genotyping results. Conclusion: We have, for the first time, established a non-invasive prenatal genotyping method for fetal M antigen using maternal plasma cff-DNA, and preliminarily demonstrated the feasibility of this method.

To explore the advantages of traditional Chinese medicine （TCM） and integrative TCM-Western medicine approaches in the treatment of diabetic microvascular complications （DMC）， refine key pathophysiological insights and treatment principles， and promote academic innovation and strategic research planning in the prevention and treatment of DMC. The 38th session of the Expert Salon on Diseases Responding Specifically to Traditional Chinese Medicine， hosted by the China Association of Chinese Medicine， was held in Beijing， 2024. Experts in TCM， Western medicine， and interdisciplinary fields convened to conduct a systematic discussion on the pathogenesis， diagnostic and treatment challenges， and mechanism research related to DMC， ultimately forming a consensus on key directions. Four major research recommendations were proposed. The first is addressing clinical bottlenecks in the prevention and control of DMC by optimizing TCM-based evidence evaluation systems. The second is refining TCM core pathogenesis across DMC stages and establishing corresponding "disease-pattern-time" framework. The third is innovating mechanism research strategies to facilitate a shift from holistic regulation to targeted intervention in TCM. The fourth is advancing interdisciplinary collaboration to enhance the role of TCM in new drug development， research prioritization， and guideline formulation. TCM and integrative approaches offer distinct advantages in managing DMC. With a focus on the diseases responding specifically to TCM， strengthening evidence-based support and mechanism interpretation and promoting the integration of clinical care and research innovation will provide strong momentum for the modernization of TCM and the advancement of national health strategies.

Objective: To conduct screening for rare blood types within important blood group systems for the Chinese population, such as Rh, Duffy, Kidd, P1Pk, Diego, and MNS, in the Guangzhou region, and to establish a corresponding rare blood type database and physical repository. Methods: The saline medium microplate method was used to screen blood donors with the ccDEE phenotype combined with either Jk(a-) or Jk(b-). The polybrene microplate method was employed to screen for donors with Fy(a-), s(-), Lu(b-), Di(b-), k(-), and p phenotypes. The urea lysis microplate method was applied to screen for the Jk(a-b-) phenotype. A high-resolution melting (HRM) curve method was established for screening some donors with the Di(b-) phenotype. Subsequently, expanded phenotyping of antigens in the Rh, Kidd, MNS, Duffy, P1Pk, Lewis, Kell, and Lutheran blood group systems was performed on identified rare blood type donors using monoclonal antibodies. The test results are entered into the Rare Blood Type Bank Management System of the Guangzhou Blood Center, enabling functions such as confirmation reminders and cryopreservation storage when the donor donates again. Red blood cells of rare blood types are processed into frozen red blood cells for long-term storage. Results: Among voluntary blood donors, 16 cases of the ccDEE combined with Jk(a-) phenotype were identified (0.221 7%, 16/7 216); 10 cases of the ccDEE combined with Jk(b-) phenotype (0.138 6%, 10/7 216); 78 cases of the Fy(a-) phenotype (0.169 5%, 78/46 012); 39 cases of the Lu(b-) phenotype (0.138 2%, 39/28 214); 31 cases of the s(-) phenotype (0.081 8%, 31/37 913); 22 cases of the Di(b-) phenotype (0.029 9%, 22/73 691); 30 cases of the Jk(a-b-) phenotype (0.010 1%, 30/298 250); and 1 case of the k(-) phenotype (0.001 3%, 1/77 382), which was further identified as KELnull phenotype (K0). No p phenotype donors were identified (0/88 528). A total of 228 units of frozen red blood cells were prepared. The screening results were compared and analyzed with rare blood type data from other regions. Conclusion: This study, through a combination of different screening methods, significantly improved the efficiency of rare blood type screening while remaining cost-effective. By conducting large-scale screening and performing data informatization processing, a database and physical repository of rare blood types in the Guangzhou region were successfully established. This provides a strong guarantee for the timely supply of blood to patients with difficult-to-match and rare blood types in the region, effectively enhances the level of transfusion safety in the region, and offers a practical paradigm for constructing a comprehensive blood transfusion support system.

Objective To study the effects of cognitive behavioral therapy(CBT)on nutritional status and postoperative recovery in patients after gastric cancer surgery.Methods Sixty patients diagnosed with gastric cancer between January 1,2023,and December 30,2023,at the hospital in the department of gastroin-testinal and gland surgery were included in this study.Patients were randomly divided by drawing lots into ei-ther the observation group(received routine care plus CBT—based nursing program)or the control group(received routine care),with 30 patients in each group.Nutritional indicators and postoperative recovery out-comes were compared between the two groups.Results After the intervention,patients in the observation group showed significantly higher levels of albumin and hemoglobin compared to the control group(P＜0.05).However,there was no statistically significant difference in prealbumin levels between the two groups(P＞0.05).Furthermore,patients in the observation group scored higher on the Quality of Recovery-40 scale in terms of emotional state,physical comfort,psychological support,and overall score compared to the control group(P＜0.05).After the intervention,the observation group showed significantly lower HADS scores com-pared to the control group,with a statistically significant difference(P＜0.05).Conclusion CBT demon-strates significant positive effects in improving nutritional status and postoperative recovery quality in gastric cancer patients.CBT improves patients'emotional state,thereby affecting appetite and nutritional status,and promotes postoperative physical function recovery.

Objective To evaluate the diagnostic value of preoperative positron emission computed-tomography(PET-CT)examination for lymph node metastasis in early non-small cell lung cancer(NSCLC).Methods A total of 232 early-stage NSCLC patients who underwent preoperative PET-CT examination and radical surgery for lung cancer in the hospital from January 2019 to April 2023 were selected.Clinical data,PET-CT examination results,and lymph node dissection pathology were collected to calculate the sensitivity,specificity,accuracy,positive predictive value and negative predictive value of PET-CT diagnosis of NSCLC lymph node metastasis.The maximum standard uptake value(SUVmax)of the primary lesion and the predic-tive value of lymph node SUVmax for early NSCLC lymph node metastasis were evaluated by the receiver op-erating characteristic(ROC)curve.Results The accuracy of PET-CT examination in the diagnosis of NSCLC was 95.69％,the accuracy of lymph node metastasis diagnosis was 85.78％,the sensitivity was 35.56％,the specificity was 97.86％,the positive predictive value was 80.00％,and the negative predictive value was 86.32％.There was a statistically significant difference in the detection rate of PET-CT examination and path-ological results among pulmonary nodules of different natures and sizes(P＜0.01).For predicting lymph node metastasis in NSCLC,the cutoff point of SUVmax in the primary lesion was 3.83(P＜0.001),and the cutoff point of SUVmax in lymph nodes was 3.07(P＜0.001).Conclusion In early NSCLC,preoperative PET-CT examination has a high diagnostic accuracy for ground glass nodule lymph node metastasis,but its di-agnostic value for solid nodule lymph node metastasis is limited and insufficient to guide lymph node dissec-tion.Primary lesion SUVmax≥3.83 or lymph node SUVmax≥3.07 are high-risk factors for early lymph node metastasis in lung cancer.

In order to further promote the standardization of the integrated traditional Chinese and western medicine treatment of atopic dermatitis(AD),the team of Professor Chen Dacan,who is honored as the Qihuang Scholar,developed the Clinical Practice Guidelines for Integrated Traditional Chinese and Western Medicine in Atopic Dermatitis(hereinafter referred to as Guidelines for AD).This paper detailed the process of development of Guidelines for AD,and analyzed the characteristics of Guidelines for AD,as well as the difficulties and countermeasures encountered during such a time.The development of Guidelines for AD follows the methodology of international guidelines for clinical trial.Experts specializing in various disciplines,such as traditional Chinese medicine,western medicine,integrated traditional Chinese and western medicine,and methodology,composed an expert group and a working group.On the basis of systematic literature research,the clinic experiences of frontline experts were well-summarized,and the draft of Guidelines for AD was formed after several meetings and discussions.In the process of developing Guidelines for AD,quite a number of problems were encountered,and the project team found the corresponding countermeasures after analyzing these problems.The countermeasures became the characteristics of Guidelines for AD:the integration of traditional Chinese medicine therapies and western medicine therapies,and the evaluation of the evidence of integrated traditional Chinese and western medicine solved the problem of traditional Chinese and western medicine diagnosis and treatment of AD lacking standardization;the adequately combination of the evidence with the experience of clinical practice solved the problems of the low overall level and the insufficiency of traditional Chinese medicine evidence for AD;the establishment of management goals and strategies for AD after taking the advantages of both traditional Chinese medicine and western medicine into account was in line with the international treatment and management concepts;the formulation of individualized traditional Chinese and western medicine therapy based on the integration of traditional Chinese medicine therapy and western medicine therapy met the clinical needs of patients with different characteristics of AD.The development process of Guidelines for AD and the analysis of problems and countermeasures during such a time will provide reference and reflection for the subsequent establishment of clinical guidelines for the integrated traditional Chinese and western medicine in other diseases.

Objective To analyze the prescription patterns of Professor Chen Bin in treating chronic superficial gastritis(CSG)with data mining methods.Methods Prescription data from effective medical records of outpatients with CSG treated by Professor Chen Bin at the First Affiliated Hospital of Guangzhou University of Chinese Medicine from October of 2022 to April of 2023 were collected.R language,SPSS,and IBM SPSS Modeler were used to perform frequency statistics,association rule analysis,and systematic cluster analysis on the prescription data,and then the medication and prescription patterns of Professor Chen Bin in treating CSG were explored.Results A total of 64 outpatient prescriptions formulated by Professor Chen Bin were included,involving 95 Chinese herbal medicines.The top 10 frequently-used herbs were Glycyrrhizae Radix et Rhizoma Praeparata cum Melle(Zhigancao),Bupleuri Radix(Chaihu),Pinelliae Rhizoma Praeparatum(Fabanxia),Jujubae Fructus(Dazao),Zingiberis Rhizoma(Ganjiang),Codonopsis Radix(Dangshen),Scutellariae Radix(Huangqin),Paederiae Herba(Jishiteng),Citri Reticulatae Pericarpium(Chenpi),and Taraxaci Herba(Pugongying).Most of the herbs were warm,cold,and mild in nature,and were sweet in flavor.And the herbs mainly had the meridian tropism of spleen,stomach,and liver meridians,and had the primary therapeutic effects of strengthening the spleen and replenishing qi,regulating qi,and resolving dampness.Based on the results of association rule analysis,a total of 21 core herb combinations were screened out,and cluster analysis yielded 4 clustered prescriptions.Combining the results of high-frequency herb statistics and cluster analysis and based on the traditional Chinese medicine theory,the core herbs for treating CSG was identified as Zhigancao,Chaihu,Fabanxia,Dazao,Ganjiang,Dangshen,Huangqin,Jishiteng,Chenpi,and Pugongying.Conclusion In treating CSG,Professor Chen Bin adheres to the treatment principle of replenishing qi and regulating the pivot,and the therapeutic methods of regulating qi movement,strengthening the spleen and harmonizing the stomach are applied with Xiaochaihu Decoction as the fundamental prescriptions.His view of treatment by focusing on the spleen and stomach,emphasizing the combination of herbs with cold and warm properties and based on syndrome differentiation will provide a reference for the clinical application of traditional Chinese medicine in treating CSG.

OBJECTIVE: CRISPR-Cas protects bacteria from exogenous DNA invasion and is associated with bacterial biofilm formation and pathogenicity. METHODS: We analyzed the type I-F CRISPR-Cas system of Legionella pneumophila WX48, including Cas1, Cas2-Cas3, Csy1, Csy2, Csy3, and Cas6f, along with downstream CRISPR arrays. We explored the effects of the CRISPR-Cas system on the in vitro growth, biofilm-forming ability, and pathogenicity of L. pneumophila through constructing gene deletion mutants. RESULTS: The type I-F CRISPR-Cas system did not affect the in vitro growth of wild-type or mutant strains. The biofilm formation and intracellular proliferation of the mutant strains were weaker than those of the wild type owing to the regulation of type IV pili and Dot/Icm type IV secretion systems. In particular, Cas6f deletion strongly inhibited these processes. CONCLUSION The type I-F CRISPR-Cas system may reduce biofilm formation and intracellular proliferation in L. pneumophila.
Legionella pneumophila/pathogenicity* ; CRISPR-Cas Systems ; Biofilms/growth & development* ; Phenotype ; Bacterial Proteins/metabolism* ; Gene Deletion

Glucose transporters (GLUTs) are pivotal membrane proteins that facilitate the passive transport of glucose into cells. However, their aberrant overexpression is closely linked to the Warburg effect and chemotherapy resistance of tumors. GLUTs are complex multi-pass transmembrane proteins that require detergents for extraction from the cell membrane during preparation. The persistent presence of detergents in the sample can disrupt lipid-protein interactions, potentially leading to conformational distortion and functional losses of GLUTs, severely hindering the research into their structures and transport mechanisms. To eliminate detergent interference and preserve its authentic conformation, this study employs nanodisc technology and utilizes the self-assembly of the membrane scaffold protein MSP1E3D1 and phospholipids to produce a biomimetic membrane environment, thereby overcoming the limitations of conventional methods. The C-terminal His10-tagged GLUT1 was heterologously expressed in the insect cell Sf9/Bac-to-Bac system, and the GLUT1-nanodisc complex was obtained after detergent solubilization, affinity chromatography purification via anti-His antibody resin, and self-assembly. The successfully reconstituted nanodisc complex was further purified by Ni-NTA affinity chromatography. Nanodisc reconstitution produced monodisperse GLUT1 particles that retained native secondary structure, as confirmed by far-UV circular dichroism (CD) spectroscopy and dynamic light scattering (DLS). Unlike conventional detergent micelles, which lack a true lipid bilayer, distort transmembrane-helix topology, and occlude ligand-binding sites, the nanodisc platform embeds GLUT1 in a phospholipid bilayer that preserves its authentic conformation while eliminating detergent interference. The resulting GLUT1-nanodisc complex is therefore a superior scaffold for high-resolution cryo-EM structural analysis, permitting detailed interrogation of the transporter's conformational cycle, its interactions with partner proteins, and downstream structure-guided, high-throughput drug screening.
Nanostructures/chemistry* ; Glucose Transporter Type 1/biosynthesis* ; Humans ; Animals ; Phospholipids/chemistry* ; Detergents/chemistry*

Dopamine, as a catecholamine neurotransmitter widely distributed in the central nervous system, is involved in physiological functions such as motivation, arousal, reinforcement, and movement through various dopamine signaling pathways. The hippocampus receives dopaminergic neuron projections from regions such as the ventral tegmental area, locus coeruleus, and substantia nigra. Through D1-like and D2-like receptors, dopamine exerts significant regulatory effects such as spatial navigation, episodic memory, fear, anxiety, and reward. This review mainly summarizes the research progress on the functions of dopamine in the hippocampus from aspects including the sources of dopamine, receptor distribution and function, and the association of hippocampal dopamine system dysregulation with neurodegenerative diseases. The aim is to provide insights into the involvement of the dopamine system in hippocampal functions and the diagnosis and treatment of related diseases.
Hippocampus/physiology* ; Dopamine/physiology* ; Humans ; Animals ; Receptors, Dopamine D2/physiology* ; Memory/physiology* ; Signal Transduction/physiology* ; Neurodegenerative Diseases/physiopathology*