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 investigate the expression characteristics of mucin 5B (MUC5B) protein and programmed cell death factor 4 (PDCD4) protein in patients with intrahepatic cholangiocarcinoma (ICC), and to construct a nomogram model for prognosis prediction.Methods:Clinical data of 100 patients who underwent radical surgical resection and were diagnosed as ICC by postoperative pathology from September 2009 to September 2020 in the Third Affiliated Hospital of Wenzhou Medical University were retrospectively selected, including 46 males and 54 females, aged (56.9±12.2) years old. Immunohistochemistry was used to detect the expression of MUC5B and PDCD4 protein in 100 cases of ICC and corresponding adjacent tissues respectively, and the relationship between them and clinicopathological factors of ICC patients was analyzed. Univariate and multivariate Cox regression analysis were performed to analyze the influencing factors on postoperative prognosis of ICC patients. The nomogram model was constructed using rms package and performed internal verification.Results:The positive expression rate of MUC5B protein in ICC was 76.0% (76/100), which was higher than that in para-cancer tissues 27.0%(27/100), and the difference was statistically significant ( χ2=11.33, P=0.015). While the positive expression rate of PDCD4 protein in ICC was 21.0%(21/100), which was lower than that in normal tissues 73.0% (73/100), and the difference was statistically significant ( χ2=15.57, P=0.007). Multivariate Cox regression analysis showed that ICC patients with carbohydrate antigen 19-9>37 kU/L, tumor length>5 cm, tumor TNM stage Ⅱ/Ⅲ, tumor medium/low differentiation, MUC5B positive expression, and PDCD4 negative expression had a high risk of short survival after resection (all P<0.05). The nomogram model was constructed based on the above indicators, and the C-index was 0.801. The postoperative survival calibration curve showed that the high predictive survival fit of the nomogram model, and the area under the receiver operating characteristic curve was 0.862. Conclusions:Positive expression of MUC5B protein and negative expression of PDCD4 protein in ICC tissue suggest poor prognosis of ICC patients. The nomogram model constructed on the basis of MUC5B and PDCD4 protein is well distinguished and has ideal predictive efficacy.

Objective To observe the influence of different energy windows of the medium-energy general-purpose(MEGP)collimator on image quality,so as to optimize the energy window of yttrium-90(90Y)bremsstrahlung SPECT imaging.Methods 90Y bremsstrahlung spectrum was acquired,and the sensitivity,percentage of the source counts in useful field of view(S/FOV%)and signal-to-background ratio(S/B)of 90Y bremsstrahlung SPECT imaging at MEGP under different energy windows were compared.Results The energy spectrum of 90Y bremsstrahlung was a continuous curve,with the peak of 76.2 keV with MEGP collimator.The images obtained with MEGP collimator were clear,and no significant differences of S/FOV%nor S/B was found between 10%and 20%window width groups(both P＞0.05),but the sensitivities of the latter was higher than the former(P＜0.05).The sensitivity of 70-90 keV images was relatively high,while the S/FOV%and S/B had decreased.The S/FOV%and S/B of images ranging from 40-60 keV were high,but the sensitivity was low.Images acquired with 100 keV±20%showed fairly high sensitivity,S/FOV%and S/B,which was 69.73%,0.62 and 1.64,respectively.Conclusion When performing 90Y bremsstrahlung SPECT with MEGP collimator,the image quality at 20%window width was better than at 10%window width,and 100 keV±20%showed fairly high sensitivity and not significantly decreased S/FOV%and S/B.

Objective:To investigate the expression of nuclear matrix protein 4 (NMP4) in hepatocellular carcinoma (HCC), and its relationship with clinicopathological features and survival prognosis of patients.Methods:The clinical data of 100 HCC patients who were treated with radical resection of liver cancer in the Department of Hepatobiliary Surgery of the Third Affiliated Hospital of Wenzhou Medical University from July 1, 2014 to July 1, 2019 were retrospectively analyzed. There were 63 males and 37 females, aged (58.5±10.4) years old. Immunohistochemical method was used to detect the expression of NMP4 protein in HCC cancer tissue and the corresponding adjacent normal tissue. According to the expression of NMP4 in HCC tissues, 100 patients were divided into two groups: the NMP4-positive expression group ( n=62) and the NMP4-negative expression group ( n=32). Univariate analysis was performed on the relationship between NMP4 expression and clinical pathological features as well as overall survival of HCC patients. Cox multivariate analysis was performed on the factors influencing postoperative prognosis of HCC patients. Results:Immunohistochemistry results showed that NMP4 was primarily expressed in the nucleus, the positive expression rate of NMP4 in HCC tissues was higher than that in adjacent non-cancerous tissues [62.0% (62/100) vs. 8.0%(8/100)], and the difference was statistically significant ( χ2=2.12, P=0.003). Univariate analysis revealed that the overall survival of HCC patients was correlated with the degree of tumor differentiation, tumor length, BCLC stage, number of tumor foci, vascular tumor thrombus and expression of NMP4 (all P<0.05). Cox multivariate analysis revealed that low differentiation, high BCLC stage (stage C), number of tumor foci (≥3), and positive expression of NMP4 were independent risk factors affecting postoperative survival and recurrence-free survival of HCC patients. The median overall survival and median recurrence-free survival of HCC patients in the NMP4-positive expression group were 22.3 months and 11.5 months, respectively. In contrast, that in the NMP4-negative expression group were 40.6 months and 19.4 months, respectively. The cumulative survival rate and recurrence-free survival rate of HCC patients in the NMP4-positive expression group were lower than those in the NMP4-negative expression group, and the differences were statistically significant (both P<0.05). Conclusion:Positive NMP4 expression was closely correlated with malignant biological progression and poor prognosis of HCC patients.

Development of new radiopharmaceuticals is crucial for the advance of nuclear medicine and molecular imaging. In recent years, significant progress has been made in the development of radiopharmaceuticals based on radioactive metal nuclides such as ⁶⁴Cu. Copper is a transition metal element necessary for human physiology. Disorders of copper metabolism not only cause hereditary Wilson disease and Menkes disease, but also play an important role in the development of malignant tumors. In addition to radiolabeling for probes, ⁶⁴Cu can be used for PET diagnostic imaging of copper metabolism disorders. ⁶⁴CuCl₂ is a useful radiotracer that can be used to track the dynamic flow of copper ions in human body copper metabolism by PET imaging. In recent years, many preclinical and clinical studies of ⁶⁴CuCl₂ radiotracer have achieved good results, and have attracted a lot of attention in the field of nuclear medicine and molecular imaging. In this article, the progress in the development of ⁶⁴CuCl₂ as a radiotracer for PET diagnostic imaging of hereditary copper metabolism disorders and copper hypermetabolic tumors is reviewed, and further research studies and potential clinical application of ⁶⁴CuCl₂ PET/CT imaging is prospected.

As an important branch of systematic biology, metabonomics is an emerging discipline following genomics, proteomics and transcriptomics. It focuses on the quality and quantity changes of the small molecular metabolites in appointed physiological or pathological state, and provides important informa?tion for studies of diseases mechanism, clinical early diagnosis and prognosis prediction. Compared with tra?ditional clinical diagnostic method ( single index) , metabonomics method can more comprehensively analyze metabolites changes in the body. This review introduces the metabonomics and summarizes its application in thyroid diseases.

The radioactive iodine-refractory differentiated thyroid carcinoma (RIR-DTC) is a complex process that involves multiple genetic changes and multiple signaling pathways.Radionuclide imaging, genomics and proteomics are effective to clarify the mechanism and helpful in clinical diagnosis and therapy.The treatment of RIR-DTC includes the removal of distant metastases, drug therapy, external radiotherapy and radiofrequency ablation.This review mainly focuses on the pathogenesis, diagnosis and treatment of RIR-DTC.

Norepinephrine transporter (NET) transfection leads to significant uptake of iodine-131-1abeled metaiodobenzylguanidine (131I-MIBG) in non-neuroendocrine tumors.However,the use of 131I-MIBG is limited by its short retention time in target cells.To prolong the retention of 131I-MIBG in target cells,we infected hepatocarcinoma (HepG2) cells with Lentivirus-encoding human NET and vesicular monoamine transporter 2 (VMAT2) genes to obtain NET-expressing,NET-VMAT2-coexpressing,and negative-control cell lines.We evaluated the uptake and efflux of 131I-MIBG both in vitro and in vivo in mice bearing transfected tumors.NET-expressing and NET-VMAT2-coexpressing cells respectively showed 2.24 and 2.22 times higher 131I-MIBG uptake than controls.Two hours after removal of 131I-MIBG-containing medium,25.4％ efflux was observed in NET-VMAT2-coexpressing cells and 38.6％ in NET-expressing cells.In vivo experiments were performed in nude mice bearing transfected tumors;results revealed that NET-VMAT2-coexpressing tumors had longer 131I-MIBG retention time than NET-expressing tumors.Meanwhile,NET-VMAT2-coexpressing and NET-expressing tumors displayed 0.54％ and 0.19％,respectively,of the injected dose per gram of tissue 24 h after 131I-MIBG administration.Cotransfection of HepG2 cells with NET and VMAT2 resulted in increased 131I-MIBG uptake and retention.However,the degree of increase was insufficient to be therapeutically effective in target cells.

Objective Some drugs can effectively improve the uptake of 131 I in dedifferentiated thyroid cancer .This study was to investigate the effect of all-trans retinoic acid (ATRA) combined with interferon-α2a (IFN-α2a) on the iodine uptake and re-tention rate of FTC-133 follicular thyroid carcinoma cells . Methods FTC-133 cells were cultured in the presence of 2 μmol/L AT-RA and/or 1500 IU/mL IFN-α2a for 72 hours.The the cells were collected for measurement of the uptake and retention rate of Na 125 I. Results The Na125 I uptake of the FTC-133 cells was significantly increased after 72-hour incubation with 2μmol/L ATRA and 1500 IU/mL IFN-α2a (2423.1 ±237.5) as compared with those of the control (1279.5 ±102.8), ATRA (1438.2 ±149.6), and IFN-α2a groups (1355.3 ±198.4) (P<0.05).Statistically significant differences were observed in the retention rate of Na 125I in the FTC-133 cells at different time points in the each of the blank control , 2μmol/L ATRA, 1500 IU/mL IFN-α2a, and combination groups (P<0.01), but not in that of Na125I among these groups (P<0.05). Conclusion ATRA in combination with IFN-α2a can promote the Na125 I uptake but cannot improve the Na 125 I retention rate in FTC-133 cells.

Cancer development is a complex process that involves multiple genetic changes and multiple signaling pathways . Recent findings show that the GRIM-19 is a novel apoptosis regulation gene , and its gene mutations and loss of protein expression have been observed in many tumor types such as urinarysystem tumor , digestive system neoplasm , which are closely related to cancer devel-opment.Thus, GRIM-19 may be a potential target for gene therapy .Pro-apoptotic mechanisms of GRIM-19 and its related proteins such as STAT3,GW112,p16INK4aare overviewed in this article.