Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering, which limit sequence and functional diversity, thereby constraining their therapeutic and application potential. Protein design tools have significantly advanced the creation of novel protein sequences, structures, and functions. However, further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability. Here, we extended an evolution-based design protocol to create a novel minibinder, BindHer, against the human epidermal growth factor receptor 2 (HER2). It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity. Radiolabeling experiments with ^99mTc, ⁶⁸Ga, and ¹⁸F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models, with minimal nonspecific liver absorption, outperforming scaffolds designed through traditional engineering. These findings highlight a new rational approach to automated protein design, offering significant potential for large-scale applications in therapeutic mini-protein development.

Objective To explore the value of intravoxel incoherent motion(IVIM)parameters in the diagnosis of ipsilateral thalamic diaschisis(ITD)after unilateral supratentorial cerebral infarction via combination of arterial spin labeling(ASL)and IVIM.Methods A total of 111 patients with cerebral infarction were selected,including 61 patients in the acute stage(＞6 h-3 d),50 patients in the subacute stage(＞3 d-11 d).All patients were subjected to MRI examination,and the diffusion weighted imaging(DWI),ASL,and IVIM image data were uploaded to the GE workstation.The regions of interest(ROI)in the bilateral thalamus were selected,and the cerebral blood flow(CBF),D*,D,and f values were measured,and the asymmetry index(AI)was calculated.ITD was diagnosed according to AI＞10％,and all patients were divided into ITD-positive and ITD-negative groups.The differences of each parameter between the two groups and their relationship with AI were analyzed.Results In acute stage and subacute stage cerebral infarction,the CBF,D*and D values of ipsilateral thalamus were lower than those of contralateral thalamus,while the f value was higher than those of contralateral thalamus.There was no statistical significance in D*,D and f values between ITD-positive and ITD-negative groups after acute stage cerebral infarction(P＞0.05).There were significant differences in CBF,apparent diffusion coefficient(ADC),D and f values in ITD-positive group after subacute stage cerebral infarction(P＜0.05),but no significant differences in D*value of ITD-positive group and CBF,D*,D,f values of ITD-negative group(P＞0.05).AI-D*,AI-D and AI-ADC in subacute stage were positively correlated with AI-CBF,while AI-f was negatively correlated with AI-CBF.The area under the curve(AUC)of AI-D and AI-f in subacute stage were 0.749 and 0.699,respectively.Conclusion The combination of ASL and IVIM has confirmed the phenomenon of ITD after unilateral supratentorial cerebral infarction,with high diagnostic efficacy of quantitative parameters D value and f value.It can non-invasively evaluate the thalamic microstructural and perfusion changes in the early stage after cerebral infarction.

Objective:To explore the application process, efficacy and safety of MR-guided radiotherapy based on MR-linac in esophageal cancer.Methods:The clinical data of patients with esophageal cancer treated with MR-linac at Shandong Cancer Hospital and Institute from September 2021 to July 2022 were retrospectively analyzed, to investigate the treatment process of esophageal cancer with MR-linac, and to analyze the efficacy and safety of patients. All patients received MR-guided radiotherapy, underwent CT and MR localization, target area delineation, and design of the Monaco treatment planning system plan. Adaptation-to-position adjustment was conducted during the pre-treatment evaluation. The median number of fractions was 25, the median single dose of planning target volume was 1.8 Gy, and the median total dose was 50.2 Gy. Median follow-up was 16 months.Results:Among the 12 patients in the whole group, there were 1 case of cervical esophageal cancer, 3 cases of upper thoracic esophageal cancer, 4 cases of middle thoracic esophageal cancer and 4 cases of lower thoracic esophageal cancer, including 3 cases of neoadjuvant radiotherapy and 9 cases of radical radiotherapy. All patients had a smooth treatment process. The median treatment time was 33 min, and the patients had good compliance. For patients with radical radiotherapy, one month after radiotherapy, the number of objective remission cases was 3, and the number of disease-control cases was 9; six months after radiotherapy, the number of objective remission cases was 3, and the number of disease-control cases was 6. All patients treated with neoadjuvant radiotherapy underwent surgery within 2 months, and one patient achieved pathological complete remission. The most common acute adverse reactions were radiation esophagitis (7 cases) and leukopenia in bone marrow suppression (8 cases), with late-stage adverse reactions being radiation pneumonia (1 case). The adverse reactions to radiotherapy were slight, and no grade 4 or above adverse reactions were observed.Conclusion:The clinical treatment process for esophageal cancer under MR-guided radiotherapy based on MR-linac is feasible, with good curative effects and mild adverse reactions.

OBJECTIVE To evaluate the efficacy and safety of programmed death-1/programmed death-ligand 1 （PD-1/PD-L1） inhibitors combined with bevacizumab in the treatment of advanced non-small cell lung cancer （NSCLC） based on platinum- containing dual therapy. METHODS Retrieved from CNKI， Wanfang， VIP， Web of Science， PubMed and other Chinese and English databases， cohort studies or randomized controlled trial studies on the treatment of advanced NSCLC with platinum- containing double agents in combination with PD-1/PD-L1 inhibitors and bevacizumab （trial group） versus platinum-containing double agents with or without PD-1/PD-L1 inhibitor or bevacizumab （control group） were collected from the inception to April 25， 2024. After screening literature， extracting data and evaluating quality， meta-analysis and sensitivity analysis were performed by using RevMan 5.4.1 software. RESULTS A total of 15 pieces of literature were included， involving 13 clinical studies with a total of 3 282 patients. Compared with the control group， partial response rate [RR＝0.75，95%CI（0.68，0.82），P＜0.000 01]， complete response rate [RR＝0.47，95%CI（0.29，0.76），P＝0.002]， progressive disease rate [RR＝1.23，95%CI（1.11，1.37），P＜0.000 1]， objective response rate （ORR） [RR＝0.72，95%CI（0.67，0.79），P＜0.000 01] and disease control rate （DCR） [RR＝0.85， 95%CI （0.77，0.95），P＝0.003] were higher in the trial group. There was no statistically significant difference in the stable disease rate [RR＝1.25， 95%CI （0.86， 1.83）， P＝0.25] or overall adverse drug reaction incidence rate [RR＝0.95， 95%CI （0.90， 1.00）， P＝ 0.07] between the two groups of patients. Sensitivity analyses showed robust and reliable results for all outcome indicators. CONCLUSIONS PD-1/PD-L1 inhibitors combined with bevacizumab based on platinum-containing dual therapy in the treatment of advanced NSCLC can improve patients’ clinical benefits， such as ORR and DCR， without increasing the risk of adverse drug reaction.

Using high-energy proton to image the region of interest can directly obtain the accurate estimation of the proton stopping power of the lesions,which is of great significance to reduce the range uncertainty in proton therapy.As a fundamental function of proton computed tomography(CT),radiographic imaging plays a crucial role in assisting clinical positioning.The study develops a compact proton CT detector based on an active array pixel CMOS chip in Monte-Carlo simulation toolkit Geant4,and evaluates the radiographic imaging capability of the system using 180 MeV protons.The angles of tracks are successfully reconstructed.CTP404,CTP528,and the CTP515 of specific materials are used for simulation,obtaining the spatial and density resolutions,and measuring the proton relative stopping power(RSP).The image signal-to-noise ratio is improved when using 2° proton scattering angle cut-off value.The spatial resolution is 3-4 lp/cm measured using CTP528 module.The density resolution is better than 0.05 g/cm3,and the RSP resolution is within 5%when CTP404 module is used.Through the imaging of CTP515 phantom of specific material,it is demonstrated that the system has potential for imaging common human tissues.

Objective To evaluate the clinical efficacy and safety of modified Weijing Decoction in the treatment of stable stage of bronchiectasis(phlegm heat accumulating in the lung pattern).Methods A series of N-of-1 trials using bronchiectasis patients in stable stage with phlegm heat accumulation in the lung pattern were performed.Participants meeting inclusion criteria underwent two distinct treatment protocols during a therapy period.Modified Weijing Decoction was used in the experimental period and ambroxol tablets were used in the control period,washout periods were set.The medication-use order in the control period and experimental period is random.The visual analog scale(VAS)was used to assess symptom changes before and after treatment.24-hour sputum volume and COPD self-assessment test(CAT)scores were recorded,while adverse reactions were monitored during the study period.Statistical analysis and drawing were performed on SPSS 25.0 and GraphPad Prism 8.Comparison of inter-group differences before grouping was assessed by independent samples t-test.Analysis of symptom score,24-hour sputum volume and CAT scores in the experimental and control periods,as well as comparison of differences between the above two groups before and after treatment were performed using paired t-test,which enabled comprehensive exploration of clinical and statistical distinctions.Results A total of 9 patients participated in 16 rounds of analysis.Statistically significant differences(P＜0.05)before and after the treatment in the experimental period were noted in total Traditional Chinese Medicine(TCM)symptom scores,cough,sputum,sputum color,chest tightness and chest pain,bitter and dry mouth,fatigue and tiredness,24-hour sputum volume,and CAT score.Paired t-tests revealed significant variations(P＜0.05)in the total TCM symptom scores,sputum color scores,scores of chest tightness and pain,and 24-hour sputum volume between experimental and control periods.No adverse events related to the test methods or drugs occurred during the observation period.Conclusion Modified Weijing Decoction could comprehensively improve clinical symptoms of bronchiectasis patients in stable stage.It exhibits superior efficacy compared to ambroxol,and maintains a favorable safety profile.

Objective To evaluate the effect of mefformin on the human keratinocyte line HaCaT,and to explore its molecular mechanism.Methods HaCaT cells were divided into several groups to be treated with mefformin at different concentrations of 1,2,5,10,20,50 mmol/L for 24,48 and 72 hours.Cell counting kit-8 (CCK-8) assay was performed to evaluate the effect of metformin on the survival rate of HaCaT cells.After 48-hour treatment with metformin at concentrations of 0 (control group),0.5,1,2,5,10 mmol/L,flow cytometry was conducted to evaluate the effect of metformin on cell cycle and apoptosis.Western blot analysis was performed to determine the expression of cell proliferation-and differentiation-related proteins (keratin-16 [K16],K17,K1,involucrin),apoptosis-related proteins (Bax,Bcl-2) and AKT/mTOR/STAT3 pathway proteins.Enzyme-linked immunosorbent assay (ELISA) was conducted to detect the levels of interleukin (IL)-8,tumor necrosis factor (TNF)-α and IL-23 (inflammatory factors) in the culture supernatant of HaCaT cells.Statistical analysis was carried out with SPSS19.0 software using one-way analysis of variance for comparison of the above indices among the 0.5-,1-,2-,5-,10-mmol/L metformin groups and control group,repeated measures analysis of variance for comparisons among different time points or different metformin groups,least significant difference (LSD)-t test for multiple comparisons.Results CCK-8 assay showed that mefformin had inhibitory effects on the proliferation of HaCaT cells (F =116.87,P ＜ 0.05),and the cell survival rates gradually decreased along with the increase in the concentrations of mefformin.After 48-hour treatment with mefformin at concentrations of 0,0.5,1,2,5,10mmol/L,the proportion of HaCaT cells in G2/M phase gradually increased (5.55％ ± 1.03％,6.37％ ±0.93％,8.57％ ± 1.18％,10.05％ ± 0.60％,10.76％ ± 0.87％,13.63％ ± 1.41％,respectively,F =24.98,P ＜0.05),and the early apoptosis rate also gradually increased (0.78％ ± 0.71％,19.18％ ± 1.41％,25.67％ ±1.34％,28.45％ ± 0.92％,34.97％ ± 2.12％,40.41％ ± 1.49％,respectively,F =296.08,P ＜ 0.05).Along with the increase in the concentrations of metformin,there were increasing trends in the expression of K1 and the pro-apoptotic protein Bax (F =8.86,5.38 respectively,both P ＜ 0.05),while there were decreasing trends in the expression of K16,K17 and the apoptotic protein Bcl-2 (F =8.02,4.82,12.10 respectively,all P ＜ 0.05).There was no significant difference in the expression of involucrin among different metformin groups (F =0.57,P ＞ 0.05).After 48-hour treatment with mefformin at different concentrations,the levels of IL-8 and TNF-α in HaCaT cells significantly decreased (F =33.89,14.99 respectively,both P ＜0.05),while there was no significant change in the IL-23 level (F =2.12,P ＞ 0.05).Along with the increase in the concentrations of metformin,the expression of p-AKT,p-mTOR and p-STAT3 significantly decreased (F =11.38,0.35,4.38 respectively,all P ＜ 0.05),but there were no significant changes in the protein expression of AKT,mTOR and STAT3 (F =0.66,0.35,4.24 respectively,all P ＞ 0.05).Conclusion Metformin can inhibit the proliferation,promote the differentiation and apoptosis of HaCaT cells,and inhibit the secretion of inflammatory factors by regulating the AKT/mTOR/STAT3 signaling pathway.

Objective To evaluate the cause and the treatment of the vagus nerve reflex in patients with hemoptysis during bron-chial artery embolization (BAE).Methods 1 12 patients with much hemoptysis were enrolled,9 of whom represented vagus nerve reflex in the process of interventional embolization.Results In 9 patients with mixed vagal reflex,5 occurred in the process of bron-chial artery embolization,1 in removing of sheath,1 in hemostasis by compression and 2 in returning to the ward.The intraoperative vagus reflex during BAE was related to over tension and unnormolized operation,and it improved by block of vagus nerve,raising blood pressure and fluid expansion without serious complications.Conclusion Vagus nerve reflex during BAE should be noticed, and early detection and timely intervention may improve its prognosis.

Anemia, Aplastic ; therapy ; Fetal Blood ; transplantation ; Humans ; Mesenchymal Stromal Cells

Podophyllotoxin (PTOX) is an extremely important plant-derived natural product, of which derivatives, like etoposide and teniposide, have been widely applied in therapies for cancers and venereal wart. A durable, intense plant extraction of podophyllotoxin posed a severe pressure on wild resources; researchers consequently sought to explore new sources, like cultivation, plant cell or organ culture, and chemical synthesis. Understanding biosynthesis of PTOX is one of the basic necessary steps for standard cultivation of medicinal plants and metabolite engineering. An important progress has been made in this field during the last two decades, particularly in the last ten years. Although a number of reviews concerning the related topic have existed, we specifically deal with biosynthesis of podophyllotoxin with an emphasis on the literatures of the past decade, highlighting characterization of genes encoding synthetic enzymes and down-stream metabolism of PTOX. The present review focuses on several key biosynthesis processes, important metabolites, function of related enzymes, and characterization of cDNA encoding the enzymes. Finally, the author proposed a hypothetical biosynthetic scheme of podophyllotoxin and perspectives.
Plants, Medicinal ; metabolism ; Podophyllotoxin ; biosynthesis ; chemistry ; metabolism