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.

CaMKII is essential for long-term potentiation (LTP), a process in which synaptic strength is increased following the acquisition of information. Among the four CaMKII isoforms, γCaMKII is the one that mediates the LTP of excitatory synapses onto inhibitory interneurons (LTP_E→I). However, the molecular mechanism underlying how γCaMKII mediates LTP_E→I remains unclear. Here, we show that γCaMKII is highly enriched in cultured hippocampal inhibitory interneurons and opts to be activated by higher stimulating frequencies in the 10-30 Hz range. Following stimulation, γCaMKII is translocated to the synapse and becomes co-localized with the postsynaptic protein PSD-95. Knocking down γCaMKII prevents the chemical LTP-induced phosphorylation and trafficking of AMPA receptors (AMPARs) in putative inhibitory interneurons, which are restored by overexpression of γCaMKII but not its kinase-dead form. Taken together, these data suggest that γCaMKII decodes NMDAR-mediated signaling and in turn regulates AMPARs for expressing LTP in inhibitory interneurons.
Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism* ; Hippocampus/metabolism* ; Interneurons/physiology* ; Long-Term Potentiation/physiology* ; N-Methylaspartate/metabolism* ; Receptors, AMPA/physiology* ; Receptors, N-Methyl-D-Aspartate/metabolism* ; Synapses/physiology*

OBJECTIVE: Whether the developed new type of radiotherapy auxiliary fixation device compatible with the head and neck joint coil can improve the quality of magnetic resonance images in radiotherapy and verify whether it can be applied to clinical treatment. METHODS: The clinical trial selected patients with brain metastases and nasopharyngeal cancer patients, using thermoplastic film and head and shoulder molds for posture fixation, and treatment on the ELekta Versa accelerator. SPSS 20.0 statistical software was used to analyze the data. The measurement data were expressed by RESULTS: Considering the influence of the outer contour of the device, the target dose meets the clinical requirements. The setting error is less than 2 mm in the three translation directions, and the rotation error is less than 2 CONCLUSIONS There is no statistical difference between the treatment results of patients using the new type of fixation device and the conventional method. The target area threatens the organ dose, and the positioning error meets the treatment requirements.
Humans ; Magnetic Resonance Imaging ; Magnetic Resonance Spectroscopy ; Nasopharyngeal Neoplasms ; Neck ; Posture

MRI simulation images quality of head and neck coil scanning is better than that of radiotherapy surface coil, but currently the head and neck coil is not compatible with radiotherapy positioning devices. In this paper, a new fixation device is developed based on computer reverse engineering technology, which can be used in combination with head and neck coil. This article focuses on discussing the feasibility of the new device in radiotherapy. The obtained ACR phantom and Cat phantom 504 images were used to analyze MR and CT images quality assurance indicators. The dose attenuation of 6 MV photons was measured using the ionization chamber. The results showed each index met the clinical application requirements of intracranial tumor radiotherapy, thereby it can be used in intracranial tumor radiotherapy.
Feasibility Studies ; Head and Neck Neoplasms ; diagnostic imaging ; Humans ; Magnetic Resonance Imaging ; instrumentation ; Phantoms, Imaging ; Radiotherapy Planning, Computer-Assisted

Objective To observe the influence of once pre-injection dexmedetomidine(DEX) on clinical effect in general anesthesia induction.Methods Sixty patients,ASA Ⅰ-Ⅲ,no sex limitation,undergoing elective laparoscopic cholecystectomy under general anesthesia were selected and divided into the 1ug/kg DEX group(D) and normal saline control group(C),30 cases in each group.The BIS values were recorded after medication infusion.Then the target controlled infusion(Marsh mode) of propofol was performed according to the plasma concentration.The plasma concentration was gradually increased until BIS value reaching 40.Then the effect-site concentration(Ce) of propofol was recorded at this time and sufentanil 0.5ug/kg was injected within 10 s.The bucking incidence rate within 1 min after injection of sufentanil was recorded and cisatracurium was injected again.The tracheal intubation was performed when the TOF value was 0.The time of train-of-four stimulation(TOF) to 0(effect onset time) and intubation time were recorded.The hemodynamic indexes were recorded at pre-administration(T0),post-administration(T1),after induction(T2),and after intubation(T3).The incidence rate of intraoperative awareness was recorded.Results The bucking incidence rate at 1 min after sufentanil injection in the group D was significantly lower than that in the group C(3.33％ vs.13.3％,P＜0.01).Ce of propofol in the group D was lower than that in the group C(P＜0.01).HR af T1 in the group D was decreased;HR and MAP at T2 in the group C were decreased,which at T3 were increased.No intraoperative awareness occurred in all cases.Conclusion Pre-injection of DEX 1ug/kg before induction can decrease the propofol dose and maintains hemodynamic stability.

Objective To explore the CT and MRI characteristics of hepatic epithelioid angiomyolipomas (H EA)in order to improve recognition and diagnostic accuracy of the disease.Methods The CT and MR imaging data of 8 patients with HEA proved by pathology were analyzed retrospectively.Among these patients,4 cases underwent plain and enhanced CT scans,2 plain and enhanced MRI scans and 2 both CT and MRI scans.Results In the 8 cases,7 were female.All lesions were solitary and well-defined,with maximum diameters ranging from 42 mm to 68 mm.6 leions on plain CT displayed slightly low density,of which 1 showed fat density and other 2 showed patchy cystic areas.MRI showed 4 masses with slight hypointensity on T1 WI,slight hyperintensity or hyperintensity on T2 WI,of which one demonstrated decreased signal on out-phase of T1 WI.On enhanced MR images,7 masses were significantly inhomogeneous in arterial phase,5 showed "fast wash-in and slow wash-out",2 "fast wash-in and wash-out"and 1 "slow wash-in and wash-out".6 showed"central vascular sign"in arterial phase and 2 displayed incomplete pseudocapsule in the portal venous and delayed phases.Conclusion Inhomogeneous enhancement in arterial phase,central vascular sign,fast wash-in and slow wash-out enhancement and no pseudocapsule are imaging features of HEA,which can help to improve the diagnostic accuracy.Its diagnosis still depends on histopathology and immunohistochemistry.

Objective:To investigate the impact of Notch signaling pathway on the migration of human hepatic carcinoma cells and the expression of E-cadherin and COX-2 in these cells.Methods:Cultured hepatic carcinoma cell lines (SMMC-7721,MHCC97H),and normal non tumor liver cell line (HL-7702) in vitro.Transwell cell was used to measure the cell's capacity of invasion and migration.Western blot was used to measure the expression level ofNotch1,E-cadherin,COX-2 protein.DAPT was used to block the Notch signaling pathway,and compared the ability of invasion and migration between hepatic carcinoma cell lines and normal non tumor liver cell line,and the change of expression level of E-cadherin and COX-2 protein in hepatic carcinoma cells.Results:The migration ability of SMMC-7721 cells and MHCC97H cells were higher than HL-7702 cells,the difference was statistically significant (P＜0.05);Compared to HL-7702 cells,the expression level of Notch1 and COX-2 in MHCC97H cells and SMMC-7721 cells significantly increased,the expression level of E-cadherin decreased significantly (P＜0.05);After DAPT treatment,the migration ability of SMMC-7721 cells,MHCC97H cells were weaker than the control group,the difference was statistically significant (P＜0.05);After DAPT treatment,the expression of COX-2 and Notch1 in SMMC-7721 and MHCC97H cells decreased significantly,while the expression of E-cadherin significantly increased (P＜0.05).Conclusion:Notch signaling pathway plays an important role in the process of liver cancer cell migration and invasion,and its mechanism is related to the expression of E-cadherin and COX-2.

BACKGROUND: Adipose-derived mesenchymal stem cells (ADMSCs) can improve the liver function of rats with liver failure, which illustrates the important research value in the field of tissue engineering and cell transplantation.OBJECTIVE: To evaluate the therapeutic potential of human ADMSCs in heart failure rats and to discuss the possible biological mechanisms involved.METHODS: Heart failure rats were randomized into model and ADMSCs groups, which were given normal saline or DAPI-labeled human ADMSCs (3.0×106) via the tail vein. At 1, 3, 7 days after transplantation, we detected the biochemical indexes for liver function in rats. At 3 days after transplantation, the serum levels of cytokines, such as tumor necrosis factor α and interleukin-10, were detected, the histomorphological changes in the liver were observed by hematoxylin-eosin staining, and the protein expression of proliferating cell nuclear antigen was detected by western blot. RESULTS AND CONCLUSION: We found that human ADMSCs could migrate to the liver and lung tissues in rats after the transplantation via the tail vein. At 1 and 3 days after transplantation, the levels of serum alanine aminotransferase and aspartate aminotransferase were significantly reduced in the ADMSCs group as compared with the model group (P< 0.05); furthermore, the secretion of tumor necrosis factor α and interleukin-10 was significantly suppressed at 3 days after cell transplantation (P < 0.05). The results of hematoxylin-eosin staining indicated a significant improvement in liver degeneration and necrosis. The expression of proliferating cell nuclear antigen protein in the ADMSCs group was significantly up-regulated compared with the model group. To conclude, human ADMSCs can inhibit the inflammatory reaction and up-regulate the expression of proliferating cell nuclear antigen, to promote the regeneration of liver cells and he recovery of liver function.

Objective To investigate the effects of dexmedetomidine plus ropivacanie on lumbar plexus-sciatic nerve blocks and sedation. Methods One hundred and twenty patients( ASA Ⅰ-Ⅲ) scheduled for unilateral arthroscopy of the knee received lumbar plexus-sciatic nerve blocks. The patients were randomly divided into 4 groups(n = 30 in each). In group R,dexmedetomidine was not used for nerve. In group RLD,dexmedetomidine was only used for lumbar-plexus block. In group RSD,dexmedeto-midine was only used for sciatic nerve block. In group RD,dexmedetomidine was used for both lumbar-plexus block and sciatic nerve block. Onset time and maximum time of sensory and motor block,duration of analgesia,Ramsay scores,HR,and the incidence of anesthetic toxicity were recorded at different time points(T0-T4). Results There were no significant differences in the onset time of sensory and motor block among the groups(P > 0. 05). However,there were significant differences in the maximum time of sensory and motor block for the lumbar plexus among the RLD group[(1008. 00 ± 104. 99)min and (800. 00 ± 97. 56)min],RD group[(922. 00 ± 149. 05)min and(732. 00 ± 139. 52)min],RSD group [(768. 00 ± 108. 48)min and(602. 00 ± 84. 09)min],and R group[(742. 00 ± 129. 44)min and (612. 00 ± 109. 62)min]. There were significant differences in the maximum time of sensory and motor block for the sciatic nerve among the RLD group[(1006. 00 ± 117. 58)min and(810. 00 ± 105. 41) min],RD group[(932. 00 ± 144. 18)min and(744. 00 ± 136. 09)min],RSD group[(738. 00 ± 120. 16)min and(582. 00 ± 96. 04)min],and R group[(708. 00 ± 126. 45)min and(548. 00 ± 111. 12)min]. Compare with the R group,the Ramsay scores of at the time point of T1-T4 were higher and the HRs were lower in the RLD group,RSD group and RD group. There were significant differences in the analgesia duration among the RLD group[(1006. 00 ± 117. 58)min],RD group[(918. 00 ± 83. 60)min],RSD group[(898. 00 ± 131. 34)min],and R group[(808. 00 ± 119. 72)min]. No local anesthetic intoxication was noticed. Conclusion Perineural dexmedetomidine plus ropivacaine increase the effects of lumbar plexus-sciatic nerve blocks in a dose-dependent manner. Dexmedetomidine provides a good sedative effect,however,it may lead to bradycardia.

Objective To investigate the optimal b value of diffusion-weighted imaging (DWI)in predicting the grade of cerebral gliomas.Methods 38 patients with pathologically-proved brain gliomas (24 high-grade gliomas and 14 low-grade ones)were studied retrospectively.All patients received conventional MRI and DWI examination with three different b values (1 000 s/mm2 ,2 000 s/mm2 and 3 000 s /mm2 ).The signal features of the tumor on DWI with three different b values were evaluated and compared.The minimum ADC values of the tumors were calculated and compared between each other.Through drawing the ROC curve of different b values, the best diagnostic threshold was found.The sensitivity and specificity in predicting the grade of brain gliomas were assessed using Chi-square test.Results On DWI with b value of 3 000 s/mm2 ,91.6% (22/24)of high-grade tumors showed hyper-intensity, while 85.7%(12/14)of low-grade tumors presented hypo-intensity signals.Regarding hyper-intensity signals as diagnostic criterion for high-grade tumors,the sensitivity and specificity were 91.6% and 100% respectively.When the b value was similar,the minimum ADC value of high-grade glioma was significantly lower than that of low-grade glioma,and statistically significant differences in differentiating high-grade glioma from low-grade glioma existed among three b values (P <0.05).When the ADC 3 000 value<0.74×10 -3 mm2/s was regarded as a standard for the identification of high-grade and low-grade glioma,the sensitivity and specificity were 100% and 87.3%respectively.Conclusion DWI with high b value is more useful than standard moderate b value in preoperative grading the gliomas.When DWI with b value of 3 000 s/mm2 is used,the minimum ADC value will provide quantitative indicators in preoperatively precisely predicting grading glioma.