Objective: To construct gene recombinant expression plasmids of human anti-P antibody with IgG1 and kappa chain based on the human hybridoma cell line. Methods: Starting from the specific RT-PCR products encoding the variable regions of the IgH chain and IgL chain of the anti-P monoclonal cell line, appropriate restriction enzyme digestion sites were introduced at both ends of the VH and VL fragments through nested PCR. The plasmids carrying the antibody constant region and the nested PCR products of VH and VL were ligated by the action of T4 ligase and subsequently transferred into competent E. coli DH5ɑ, and positive clones were selected in the antibiotic resistant LB medium. After sequence confirmation, recombinant plasmids DNA were transfected into HEK293T cells, and the recombinant antibody obtained in the culture supernatant. The characteristics of recombinant expression antibodies were determined by using rapid antibody isotying kit, serological agglutination tests and flow cytometry. Results: Recombinant gene expression vectors, pFUSEss-CHIg-hG1+VH, pFUSE2ss-CLIg-hκ+VL, were successfully constructed. Human IgG1 kappa light chain antibodies were detected in the supernatant of HEK293T cells transient transfected with recombinant plasmids. After the supernatant was ultra-filtered and concentrated, it could cause agglutination reactions with P antigen-positive red blood cells. The mean fluorescence intensity (MFI) of the reaction between recombinant antibodies and antigen-positive red blood cells in flow cytometry experiments was higher than that of antigen-negative red blood cells. Conclusion: The experimental study on the conversion of red blood group antibody types by genetic engineering technology represents a beneficial exploration towards establishing a feasible technical route for the development of genetic recombination and modification of antibodies reagent.

BACKGROUND:Collagen combined with microneedling therapy has gradually become an important means of improving skin photoaging.OBJECTIVE:To summarize and explore the main mechanism and clinical application status of collagen combined with microneedle therapy.METHODS:PubMed,China National Knowledge Infrastructure,and ScienceDirect databases were searched for Chinese and English literature published before August 2024.Chinese and English search terms were"ultraviolet radiation,photoaging,collagen,microneedling,clinical applications."Finally,74 articles were included for summary.RESULTS AND CONCLUSION:Collagen treats skin photoaging through mechanisms such as inhibiting matrix metalloproteinase expression,retaining skin moisture,and reducing melanin formation.Microneedles can better promote the penetration of collagen into deep layers of the skin,breaking down the skin's barrier and increasing the absorption rate.Collagen combined with microneedles has various beneficial effects for treating skin photoaging,such as whitening,anti-wrinkle,improving skin elasticity,shrinking pores,and repairing skin barriers.It also has the advantages of easy operation,significant effects,and high safety.Currently,the research on collagen combined with microneedling therapy is still in its early stages,and achieving clinical application may become a key research direction in the future.The clinical application of collagen combined with microneedles for the treatment of photoaging still faces many challenges,such as exploring the optimal mechanical structure and materials of microneedles,selecting appropriate microneedle types,and insufficient clinical evidence that collagen combined with microneedles can further delay the treatment of skin photoaging.

BACKGROUND:Collagen combined with microneedling therapy has gradually become an important means of improving skin photoaging.OBJECTIVE:To summarize and explore the main mechanism and clinical application status of collagen combined with microneedle therapy.METHODS:PubMed,China National Knowledge Infrastructure,and ScienceDirect databases were searched for Chinese and English literature published before August 2024.Chinese and English search terms were"ultraviolet radiation,photoaging,collagen,microneedling,clinical applications."Finally,74 articles were included for summary.RESULTS AND CONCLUSION:Collagen treats skin photoaging through mechanisms such as inhibiting matrix metalloproteinase expression,retaining skin moisture,and reducing melanin formation.Microneedles can better promote the penetration of collagen into deep layers of the skin,breaking down the skin's barrier and increasing the absorption rate.Collagen combined with microneedles has various beneficial effects for treating skin photoaging,such as whitening,anti-wrinkle,improving skin elasticity,shrinking pores,and repairing skin barriers.It also has the advantages of easy operation,significant effects,and high safety.Currently,the research on collagen combined with microneedling therapy is still in its early stages,and achieving clinical application may become a key research direction in the future.The clinical application of collagen combined with microneedles for the treatment of photoaging still faces many challenges,such as exploring the optimal mechanical structure and materials of microneedles,selecting appropriate microneedle types,and insufficient clinical evidence that collagen combined with microneedles can further delay the treatment of skin photoaging.

Peptide-drug conjugates (PDCs) have emerged as a promising modality in precision oncology, enabling targeted delivery of cytotoxic payloads while minimizing off-target toxicity. The integration of covalent warheads, such as those based on sulfur(VI) fluoride exchange (SuFEx) chemistry, enhances drug-target residence time and tumor accumulation. However, existing screening methods for covalent peptide (CP) libraries require post-translational warhead conjugation, limiting throughput. Here, we present an integrated mRNA display platform that incorporates covalent warheads during ribosomal synthesis, enabling efficient screening of ultra-diverse covalent macrocyclic peptide libraries (>10¹³ variants). This approach, using site-specific incorporation of N-chloroacetyl-d-phenylalanine and fluorosulfate-l-tyrosine, accelerated the discovery of irreversibly binding (K _i = 3.58 μmol/L) Nectin-4-targeting peptide CP-N1-N₃ via proximity-triggered SuFEx. The peptide was further conjugated to cytotoxic payloads, yielding the covalent PDC CP-N1-MMAE with potent cytotoxicity (IC₅₀ ≈ 43 nmol/L) against MDA-MB-468 cells. This platform establishes a new paradigm for precision covalent drug discovery.

The global incidence of head and neck cancer (HNC) is rising, with over 60% of patients presenting at a locally advanced stage. Radiotherapy remains a cornerstone of HNC treatment, and advancements in modern techniques and concurrent chemotherapy have improved local control and survival rates of HNC patients. However, these benefits also bring challenges in the management of toxicities. Due to the proximity of salivary glands and tumors, especially the highly radiosensitive parotid and submandibular glands, this condition is among the most common adverse effects of radiotherapy. Radiation damages acinar cells and ducts, causing glandular atrophy, fibrosis, and reduced saliva secretion, thereby leading to xerostomia and related complications. The risk and severity of injury are associated with the radiation dose and volume affecting the glands. Prevention and management strategies emphasize precise radiotherapy planning, target optimization, and supportive care. Emerging multimodal imaging techniques offer potential for non-invasive prediction and early diagnosis and treatment of radiation-induced salivary gland injury. Future research in regenerative medicine, tissue engineering, and molecular biology aims to elucidate molecular mechanisms, such as signaling pathways and genomics, facilitating personalized strategies to mitigate radiotherapy-induced toxicities and enhance the quality of life of patients.

Objective To evaluate the efficacy and safety of Bailemian capsule in the treatment of stroke-related sleep disorder(SSD).Methods The relevant literatures published by CNKI,VIP,Wanfang Data Knowledge Service Platform,SinoMed,PubMed,Web of Science,Cochrane Library database were retrieved from the establishment of the database to December 21,2024.Cochrane risk bias assessment tool was used to evaluate the literature quality,and RevMan 5.3 software was used to conduct Meta-analysis of the included literatures.Results A total of 11 literatures and 1054 patients were included.The results of Meta-analysis showed that cure and significant efficiency rate of patients of bservation group were significantly higher than those of control group(OR=1.93,95％CI:1.50-2.48,P＜0.000 01),clinical effective rate was significantly higher than that of control group(OR=3.70,95％CI:2.50-5.49,P＜0.000 01),incidence of adverse reactions was significantly lower than that of control group(OR=0.38,95％CI:0.22-0.67,P=0.0007).Conclusion Western medicine treatment combined with Bailemian capsule can reduce the insomnia symptoms of SSD patients,effectively improve the quality of life,and have good safety.

p21-activated kinase 5(PAK5),a critical member of the PAK family,plays a significant regulatory role in cellular signaling transduction and cytoskeleton rearrangement.Accumulating researches have revealed that PAK5 is aberrantly expressed in various human malignancies,and its expression level is significantly correlated with tumor metastasis,invasiveness,and clinical prognosis of patients.At the molecular level,PAK5 promotes tumor cells migration and invasion by activating key signaling pathways(such as Wnt/β-catenin and MAPK)to induce epithelial-mesenchymal transition(EMT),modulating tumor-associated microRNA(miRNA)networks,and remodeling actin cytoskeleton to enhance cellular motility.This review systematically dissects the molecular structural features and phosphorylation-dependent activation mechanisms of PAK5,delves into molecular mechanisms of PAK5 in tumor cells migration and invasion,and evaluates the potential of PAK5 as a therapeutic target.This work aims to provide a new perspective for developing PAK5-targeted precision therapeutic strategies in malignant tumors.

Objective:To compare the etiological characteristics between influenza-like illness (ILI) cases in outpatient and emergency departments and those with severe acute respiratory infection (SARI) in Guangdong Province, hoping to provide scientific evidence for the treatment, prevention, and control of respiratory infectious diseases.Methods:Laboratory testing for multiple respiratory pathogens was conducted on 6 090 specimens collected from ILI and SARI cases in Guangdong Province from August to December 2023. Chi-square test was used to analyze the differences in positive rates. Results:The overall positive rate of respiratory pathogens was 49.5% (3 016/6 090). The positive rate was 54.5% (2 260/4 145) in ILI cases and 38.9% (756/1 945) in SARI cases. The overall positive rate was higher in ILI cases than in SARI cases across all genders and age groups, and in most cities of the province, with statistically significant differences ( P<0.05). No significant difference was found in the overall positive rate between different genders. However, the difference between different age groups was statistically significant( P<0.001), with the highest rate identified in children aged 5-14 years (57.2%, 957/1 673). The main pathogens detected in ILI cases were influenza virus, human rhinovirus/enterovirus, and Streptococcus pneumoniae, while in SARI cases they were Mycoplasma pneumoniae, human rhinovirus/enterovirus, and influenza virus. The positive rates of adenovirus, human parainfluenza virus, SARS-CoV-2, human coronavirus, influenza virus, and Streptococcus pneumoniae were significantly higher in ILI cases than in SARI cases, while the positive rate of Mycoplasma pneumoniae was significantly higher in SARI cases ( P<0.05). Moreover, ILI cases were characterized by a higher risk of coinfection compared with SARI cases, especially in males or those aged 25-59 years. Conclusions:There are differences in the detection rates and spectrum of respiratory pathogens between ILI and SARI cases in Guangdong Province. Case features should be considered when developing strategies for preventing and treating respiratory infections.

Objective: To investigate the association between monocyte to high-density lipoprotein cholesterol ratio (MHR) and periodontitis and to provide new epidemiologic evidence on the factors affecting periodontitis. Methods: Data on MHR, periodontitis, and other covariates were selected from the NHANES(National Health and Nutrition Examination) database for 3 cycles of subjects in 2009-2010, 2011-2012, and 2013-2014, and a total of 8 456 study subjects were included. The study participants were grouped according to the prevalence of periodontitis (presence or absence), and three regression models (unadjusted covariates, partially adjusted covariates, and fully adjusted covariates) were constructed to analyze the relationship between MHR and periodontitis by using a weighted logistic regression method with stepwise adjustment for confounders. MHR was divided into four groups from Q1 to Q4 according to quartiles from small to large for weighted trend analysis, and the nonlinear relationship between MHR (continuous) and periodontitis was analyzed using a restricted cubic spline with subgroup analysis and sensitivity analysis. Results: All three logistic regression models showed a positive association between MHR and periodontitis (OR = 2.92, 95%CI: 2.14-3.99, P<0.001 (not adjusted); OR = 1.97, 95%CI: 1.39-2.78, P<0.001 (partially adjusted); OR = 1.62, 95%CI: 1.10-2.39, P = 0.017 (fully adjusted)). Trend analysis showed a significantly higher risk of developing periodontitis in the Q4 group compared with the Q1 group in both single (OR = 1.92, 95% CI: 1.58-2.33, P<0.001) and multifactorial analyses (OR = 1.30, 95% CI: 1.03-1.64, P = 0.029). Restricted cubic spline results did not support a nonlinear relationship between MHR and periodontitis (P for nonlinear>0.05), subgroup analysis showed no significant interaction between the covariates and MHR (P>0.05), and sensitivity analysis also showed a positive correlation between MHR and periodontitis (OR = 1.67, 95%CI: 1.31-2.14, P<0.001). Conclusion MHR is positively associated with the risk of developing periodontitis.

ObjectiveTo investigate the pharmacodynamic effects of Houshihei San （HSHS） recorded with the effects of treating wind and limb heaviness on muscle tissue injury after middle cerebral artery occlusion （MCAO） in rats through the ferroptosis pathway. MethodsThirty SD male rats were selected and randomly grouped as follows： sham， MCAO， deferoxamine mesylate， high-dose HSHS （HSHS-H， 0.54 g·kg^-1）， and low-dose HSHS （HSHS-L， 0.27 g·kg^-1）， with 6 rats in each group. A laser scattering system was used to evaluate the stability of the MCAO model， and rats were administrated with corresponding agents by gavage for 7 days. During the administration period， behavioral， imaging and other methods were used to systematically evaluate the skeletal muscle tissue injury after MCAO and the therapeutic effect in each administration group. Hematoxylin-eosin staining was employed to evaluate the cross-section of muscle cells. Subsequently， immunohistochemistry was used to detect tumor suppressor p53 and glutathione peroxidase 4 （GPX4） in the soleus tissue. Western blot was employed to determine the protein levels of p53， GPX4， myogenic differentiation 1 （MyoD1）， nuclear factor E₂-related factor 2 （Nrf2）， Myostatin， solute carrier family 7 member 11 （SLC7A11）， muscle ring-finger protein-1 （MuRF1）， and muscle atrophy F-box protein （MAFbx） to verify the therapeutic effect in each group. ResultsCompared with the MCAO group， HSHS enhanced the locomotor ability and promoted muscle regeneration， which suggested that the pharmacological effects of HSHS were related to the inhibition of muscle tissue ferroptosis to reduce the expression of muscle atrophy factors. Behavioral and imaging results suggested that compared with the MCAO group， HSHS ameliorated neurological impairments in rats on day 7 （P<0.01）， enhanced 5-min locomotor distance and postural control （P<0.01）， strengthened grasping power and promoted muscle growth （P<0.01）， stabilized skeletal muscle length and weight （P<0.01）， and increased the cross-section of muscle cells （P<0.01）. Compared with the MCAO group， HSHS promoted the increases in glutathione and superoxide dismutase content and inhibited the increase in malondialdehyde content （P<0.05，P<0.01）. Ferroptosis pathway-related assays suggested that HSHS reduced the p53-positive cells and increased the GPX4-positive cells （P<0.01）. HSHS ameliorated muscle function decline after stroke by promoting the expression of GPX4， Nrf2， SLC7A11， and MyoD1 and inhibiting the expression of p53， Myostatin， MurRF1， and MAFbx to reduce ferroptosis in the muscle （P<0.01）. ConclusionHSHS， prepared with reference to the method in the Synopsis of Golden Chamber， can simultaneously reduce the myolysis and increase the protein synthesis in the skeletal muscle tissue after ischemic stroke by regulating the ferroptosis pathway.