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 effect of transjugular intrahepatic portosystemic shunt(TIPS)on gut microbiota and blood ammonia level in patients with decompensated cirrhosis,and to analyze the correlation between the gut microbiota and blood ammonia level.Methods From July 2021 to December 2023,the patients with decompensated cirrhosis and portal hypertension complicated by esophagogastric variceal bleeding and/or refractory ascites,who received TIPS at Zhuhai Hospital of Guangdong Provincial Hospital of Traditional Chinese Medicine,were collected.The blood and stool samples were collected before and after TIPS.The changes in blood routine,prothrombin time,liver function,and blood ammonia level were determined.The change of gut microbiota was detected using 16S rRNA high-throughput sequencing,and Spearman correlation analysis was used to assess the correlation between the gut microbiota and blood ammonia level.Results In 20 patients,the post-TIPS one-month levels of AST,TBil,DBiL,PT,and blood ammonia were higher than their pre-TIPS values(P＜0.05),and the albumin(Alb)level was lower than the pre-TIPS value(P＜0.05).The post-TIPS 3-month levels of TBil,DBil,PT,and blood ammonia were higher than their pre-TIPS values(P＜0.05).One month after TIPS,the beta diversity of the gut microbiota became significantly different from the pre-TIPS pattern(P＜0.05).After TIPS,the harmful bacteria such as Veillonella,Streptococcus and Haemophilus were significantly reduced,the difference was statistically significant(P＜0.05).The correlation analysis of gut microbiota and blood ammonia level showed that in T0 group Colidextribacter was positively correlated with blood ammonia level,while Roxobella was negatively correlated with blood ammonia level;and in T1 group,Colidextribacter and Streptococcus were positively correlated with blood ammonia level,while Coprococcus,Bifidobacterium and Parasutterella were negatively correlated with blood ammonia level,and the differences were statistically significant(all P＜0.05).Conclusion In patients with decompensated cirrhosis after receiving TIPS,significant changes in the pattern of gut microbiota occur.Certain correlations exist between the changes of some microbiota and blood ammonia levels.Regulating the intestinal microecology may contribute to reducing blood ammonia level after TIPS.

Objective To analyze the changes in angiopoietin-2(Ang-2)levels before and after microwave ablation(MWA)in patients with high-risk pulmonary nodule and its impact on postoperative recurrence.Method A total of 94 patients with high-risk pulmonary nodules admitted to Guizhou Aerospace Hospital from December 2019 to December 2021 were included and categorized into a recurrence group(n=30)and a non-recurrence group(n=64).Clinical data was compared between the two groups.Pearson correlation analysis was used to analyze the correlation between pre-treatment angiopoietin-2(Ang-2)levels and CT parameters.Logistic regression model,restricted cubic splines,threshold effect analysis,and receiver operating characteristic(ROC)curve were used to analyze the relationship between pre-treatment Ang-2 levels and postoperative recurrence.Kaplan-Meier survival curves were used to analyze the differences in overall survival among patients with different Ang-2 levels.Cox regression model was used to analyze the factors affecting patients'survival.Result Compared to pre-treatment levels within the same group,Ang-2 levels in both groups decreased significantly,with statistically significant differences(P<0.05).Pearson correlation analysis showed that pre-treatment Ang-2 levels were positively correlated with BF,BV,PS,and MTT.Ang-2 level was independently associated with postoperative recurrence.Results of ROC analysis indicated that pre-treatment Ang-2 had certain predictive value for postoperative recurrence(area under the curve=0.789).Restricted cubic spline analysis revealed a nonlinear dose-response relationship between pre-treatment Ang-2 and postoperative recurrence(P<0.05).Threshold effect analysis identified that the inflection point of Ang-2 affecting recurrence as 1905.41 pg/mL.Survival analysis demonstrated that the median overall survival of patients in the Ang-2<1905.41 pg/mL group was longer than that in the Ang-2≥1905.41 pg/mL group(P=0.039).Furthermore,Ang-2≥1905.41 pg/mL was an independent factor affecting patients'survival time.Conclusion The levels of Ang-2 decreased significantly in patients with high-risk pulmonary nodules after MWA,and pre-treatment Ang-2 level has certain predictive value for postoperative recurrence.

【Objective】 To analyze the predictive value of serum β-defensin-3 (HBD-3) and decoy receptor 3 (DCR3) for urinary tract infection after percutaneous nephrolithotomy (PCNL) in patients with complex kidney calculi. 【Methods】 A prospective study was conducted on 112 patients treated with PCNL at our hospital during Jan.2020 and Dec.2022.The patients were divided into the non-infection group (52 cases) and infection group (60 cases).The general data, HBD-3 and DCR3 levels of the two groups were compared.Receiver operating characteristic (ROC) curve was drawn to analyze the predictive value of C reactive protein (CRP), procalcitonin (PCT), HBD-3 and DCR3 levels for postoperative urinary tract infection. 【Results】 Compared with the non-infection group, the infection group had higher levels of HBD-3 [(0.77±0.08) ng/mL vs. (1.36±0.25) ng/mL, P=0.001] and DCR3 [(4.68±0.53) ng/mL vs.(13.21±0.28) ng/mL, P=0.001].Multivariate logistic regression showed that a history of urinary tract surgery, preoperative urinary tract infection, operation time, catheterization time, stone load, type of antibiotics, concomitant renal dysfunction, intraoperative channel type, CRP, PCT, HBD-3 and DCR3 were risk factors of postoperative urinary tract infection (P<0.05).The ROC curve showed that the accuracy of CRP, PCT, and CRP plus PCT were 70.54%, 72.32%, and 78.57%, respectively; the accuracy of HBD-3, DCR3, and HBD-3 plus DCR3 were 69.64%, 75.89%, and 86.61%, respectively. 【Conclusion】 Postoperative urinary tract infection in patients with complex kidney calculi is associated with multiple factors, especially high expression levels of HBD-3 and DCR3.Combined detection has high predictive value.

Objective To construct and verify Nomogram model for individualized prediction of the risk of infectious complications in patients with complex kidney stones after percutaneous nephrolithotripsy(PCNL).Methods A total of 585 patients with complex kid-ney stones admitted to Cangzhou Hospital of Integrated Traditional and Western Medicine from March 2021 to June 2023 were collected as the study subjects,and divided into modeling group(n=410)and validation group(n=175)at 7∶3.The modeling group was further di-vided into non-complication group(n=342)and complication group(n=68)according to whether there were infectious complications after PCNL surgery.The clinical data were collected and multivariate Logistic regression analysis was applied to determine the factors that affected the occurrence of infectious complications after PCNL surgery in patients with complex kidney stones,and Nomogram model was constructed to predict the occurrence of infectious complications after PCNL surgery in patients with complex kidney stones.Results There were no statistically significant differences in gender,age,body mass index,surgical time,interleukin-6(IL-6)level,C-re-active protein(CRP)level,location and number of stone between the modeling group and the validation group(P＞0.05);compared with the non-complication group,the operation time and stone diameter,the levels of IL-6,CRP in the complication group was higher(t were 5.084,6.727,5.936,7.869,P＜0.05),and the proportions of diabetes and preoperative urinary tract infection were higher(x2 were 12.520,35.117,P＜0.05).The results of multivariate Logistic regression analysis showed that operation time(OR=1.077),stone diameter(OR=1.303),IL-6(OR=1.334),CRP(OR=1.381),diabetes(OR=3.288),preoperative urinary tract infection(OR=5.458)were all independent risk factors for infection complications after PCNL surgery in patients with complex kidney stones(P＜0.05).The area under the curve for the modeling group and validation group were 0.919(95％CI:0.889-0.949)and 0.939(95％CI:0.882-0.996),respectively.The results of the Hosmer-Limeshow goodness of fit test showed that the modeling group x2=5.484,P=0.705;validation group x2=10.101,P=0.258.Conclusion Operation time,stone diameter,IL-6,CRP,diabetes and preoperative urinary tract infection are independent risk factors for infectious complications after PCNL in patients with complex kidney stones.The Nomogram model based on these factors has high degree of discrimination and consistency.

Here we report the diagnosis and treatment of a rare IgG4-related kidney disease with nephrotic syndrome as the first manifestation. A 62-year-old male patient, presented with edema in both lower limbs and foam urine, had a history of "lung malignant tumor with brain and lymph node metastasis". The increase of IgG4 and decrease of glomerular filtration rate were detected at admission, and the pathological consideration of renal biopsy was membranous nephropathy with IgG4-related tubulointerstitial nephritis. After the combination of low-dose glucocorticoids therapy and rituximab treatment, the patient showed good prognosis in a 9 month follow-up.

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:To compare the advantages of high or low inferior mesenteric artery (IMA) ligation in laparoscopic anterior resection of rectum based on CT precise guidance.Methods:One hundred and twenty patients with rectal cancer who underwent laparoscopic anterior resection of rectum in Shanghai Sixth People′s Hospital South Campus from January 2016 to June 2020 were selected. Surgical protocol was selected according to the principle of voluntariness. Sixty-eight cases underwent high IMA ligation (high IMA ligation group), and 52 cases underwent low IMA ligation (low IMA ligation group). The perioperative conditions (operative time, intraoperative blood loss, postoperative anal exhaust time, abdominal drainage, hospitalization time and ileostomy), postoperative pathological data (proximal margin distance, distal margin distance, total number of dissected lymph nodes and number of positive lymph nodes) and postoperative recovery (anastomotic stoma fistula, urinary retention, sexual dysfunction and daily number of defecation) were recorded. The quality of life 6 months after surgery was evaluated by the quality of life instruments for colorectal cancer scale (QLQ-CR38). Patients were followed up for 6 to 36 months, and the local recurrence and distant metastasis were recorded.Results:The postoperative anal exhaust time in low IMA ligation group was significantly shorter than that in high IMA ligation group: (2.87 ± 1.04) d vs. (3.26 ± 1.00) d, and there was statistical difference ( P<0.05); there were no statistical differences in operative time, intraoperative blood loss, abdominal drainage, hospitalization time and rate of ileostomy between 2 groups ( P>0.05). There were no statistical difference in proximal margin distance, distal margin distance, total number of dissected lymph nodes and number of positive lymph nodes between 2 groups ( P>0.05). The incidences of anastomotic stoma fistula, urinary retention and sexual dysfunction in low IMA ligation group were significantly lower than those in high ligation group: 3.85% (2/52) vs. 13.24% (9/68), 3.85% (2/52) vs. 11.76% (8/68) and 5.77% (3/52) vs. 14.71% (10/68), the daily number of defecation was significantly less than that in high ligation group: (2.87 ± 0.98) times vs. (4.05 ± 1.56) times, and there were statistical differences ( P<0.05 or <0.01). The functional dimension, symptom dimension and aggregate score of QLQ-CR38 in low IMA ligation group were significantly higher than those in high ligation group: (15.46 ± 4.22) scores vs. (13.68 ± 3.56) scores, (51.82 ± 13.54) scores vs. (45.65 ± 12.42) scores and (67.28 ± 14.28) scores vs. (59.33 ± 12.85) scores, and there were statistical differences ( P<0.05 or <0.01). There were no statistical difference in the rates of local recurrence and distant metastasis between 2 groups ( P>0.05). Conclusions:The application of low IMA ligation based on CT precise guidance in laparoscopic anterior resection of rectum can obtain the same surgical effect as the high IMA ligation. It can promote the recovery of gastrointestinal function after surgery, reduce the risk of complications, have less effect on the defecation function, which can improve the life quality after the surgery.

Objective: To investigate the expression and distribution of FNDC5/Irisin in pancreatic cancer tissues and adjacent tissues, and to analyze its correlation with clinicopathological features. Methods: Collection of archived wax blocks from 64 patients diagnosed with pancreatic cancer after surgical treatment from January 2015 to December 2018 in the Department of Pathology, Affiliated Qingdao Municipal Hospital of Qingdao University, and 30 tissues collected intraoperatively from January 2016 to December 2018 Samples, all collected samples included tumor tissue and corresponding adjacent tissues (>2 cm from the tumor edge). Real-time quantitative PCR (qRT-PCR) and immunohistochemistry were used to detect the expression of FNDC5/Irisin mRNA and its positivity in pancreatic cancer tissues and adjacent tissues, and to analyze its relationship with clinicopathological features of pancreatic cancer. Results: qRT-PCR showed that the expression of FNDC5/Irisin mRNA in pancreatic cancer tissues was higher than that in the corresponding adjacent pancreatic tissues, the difference was statistically significant (P<0.05). The immunohistochemistry results showed that the positivity of FNDC5/Irisin in pancreatic cancer tissues was 59.4%, and the positivity of FNDC5/Irisin in adjacent tissues was 28.1%, and the positivity of FNDC5/Irisin in cancer tissues and adjacent pancreatic tissues was significantly different (P<0.05); the expression level of FNDC5/Irisin was not related with the gender, age, tumor size, degree of differentiation, and tumor stage.(P>0.05), but FNDC5/Irisin expression was associated with liver and lymph node metastasis (P<0.05). Conclusion The positivity of FNDC5/Irisin in pancreatic cancer tissues is significantly higher than that in adjacent pancreatic tissues, and it is correlated with liver and lymph node metastasis in pancreatic cancer.

Objective To identify the risk factors for postoperative complications after radical resection of rectal cancer, and study the influence of high ligation of the inferior mesenteric artery on postoperative complications of rectal cancer. Methods Clinical data of the patients who underwent radical resection of rectal cancer in the Affiliated Hospital of North Sichuan Medical College from January 2011 to December 2015 were analyzed. The χ 2test and t test were used for all the data. Results A total of 431 patients with rectal cancer were included, of which 80 cases were excluded, and finally 351 cases met the standard. Among them, 196 cases were in high ligation group, and 155 cases were in low ligation group. The total incidence of postoperative complications was 20.4 %(40/196) in the high ligation group and 27.1 %(42/155)in the low ligation group respectively,the difference of which was not statistically significant(χ2=1.336, P= 0.245). The incidence of anastomotic leakage was 10.2 % (20/196) in the high ligation group and 7.7 % (12/155) in the low ligation group respectively, and there was no significant difference between them (χ2=0.529, P= 0.467). Logistic regression analysis revealed that gender (OR= 2.102, 95 % CI 1.278-3.459, P=0.003), body mass index (OR= 2.492, 95 % CI 1.070-5.800, P= 0.027), with or without anemia before surgery(OR=2.203,95 % CI 1.085-4.472,P=0.029), and location of tumor(OR=2.861, 95 % CI 1.288-16.007,P=0.019) were independent risk factors for postoperative complications. Conclusions High ligation of the inferior mesenteric artery does not increase the incidence of postoperative complications after radical resection of rectal cancer. Anastomotic leakage after rectal cancer resection is related to gender, body mass index,with or without anemia before surgery,and location of tumor.