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.

Background:Following the short-term outbreak of coronavirus disease 2019(COVID-19)in December 2022 in China,clinical data on kidney transplant recipients(KTRs)with COVID-19 are lacking.Methods:We conducted a single-center retrospective study to describe the clinical features,complications,and mortality rates of hospitalized KTRs infected with COVID-19 between Dec.16,2022 and Jan.31,2023.The patients were followed up until Mar.31,2023.Results:A total of 324 KTRs with COVID-19 were included.The median age was 49 years.The median time between the onset of symptoms and admission was 13 d.Molnupiravir,azvudine,and nirmatrelvir/ritonavir were administered to 67(20.7%),11(3.4%),and 148(45.7%)patients,respectively.Twenty-nine(9.0%)patients were treated with more than one antiviral agent.Forty-eight(14.8%)patients were treated with tocilizumab and 53(16.4%)patients received baricitinib therapy.The acute kidney injury(AKI)occurred in 81(25.0%)patients and 39(12.0%)patients were admitted to intensive care units.Fungal infections were observed in 55(17.0%)patients.Fifty(15.4%)patients lost their graft.The 28-d mortality rate of patients was 9.0%and 42(13.0%)patients died by the end of follow-up.Multivariate Cox regression analysis identified that cerebrovascular disease,AKI incidence,interleukin(IL)-6 level of>6.8 pg/mL,daily dose of corticosteroids of>50 mg,and fungal infection were all associated with an increased risk of death for hospitalized patients.Conclusions:Our findings demonstrate that hospitalized KTRs with COVID-19 are at high risk of mortality.The administration of immunomodulators or the late application of antiviral drugs does not improve patient survival,while higher doses of corticosteroids may increase the death risk.

Objective:To investigate the feasibility and safety of endoscopic ultrasound (EUS)-guided intrahepatic portal vein puncture through jugular vein implantation in transjugular intrahepatic portosystemic shunt (TIPS).Methods:As research subjects, 5 beagle dogs were anesthetized, and EUS was placed through the jugular vein to observe the intrahepatic portal vein. Under real-time guidance, the portal vein was punctured and a stent was placed to complete the TIPS.Results:Among the 5 beagles, EUS was unable to be placed in 1 due to the small diameter of the external jugular vein, and it was implanted successfully in 4 others through the external jugular vein who underwent real-time guidance of portal vein puncture. Subsequent stent placement and balloon dilation were completed. All animals survived after the experiment.Conclusion:EUS-guided intrahepatic portal vein puncture through jugular vein implantation is safe and feasible in TIPS.

Objective:To explore the necessity of anticoagulation therapy and influencing factors of stent occlusion after transjugular intrahepatic portosystemic shunt.Methods:The basic information, laboratory test results, preoperative portal venous pressure, postoperative anticoagulation time, postoperative stent stenosis or occlusion, followed-up and other data of all patients who underwent TIPS surgery in Shandong Provincial Hospital from 2010 to 2019 were retrospectively analyzed. Data were analyzed using t-test, χ2 test, and multivariate analysis (logistic regression and Cox-regression-analysis). Results:A total of 280 cases were finally included in the study, of which 110 (39.3%) had stent stenosis or occlusion, and 170 (60.7%) had stent patency. New or worsening ascites were identified in 194 cases during the follow-up period, including 14 (31.1%) cases in the stent stenosis or occlusion group and 19 (12.8%) cases in the stent patency group. Univariate analysis showed that presence or absence of platelet ( P=0.037) and total bilirubin ( P=0.038) were correlated with stent stenosis or occlusion. Postoperative continuous anticoagulation was correlated with stent blockage ( P=0.029) in patients with partial portal vein thrombosis. Postoperative continuous anticoagulation and stent occlusions were not significantly correlated in patients with preoperative portal cavernoma and preoperative portal vein patency ( P=0.848; P=0.744). Multivariate analysis results showed that whether long-term anticoagulation ( P=0.017), all-cause rebleeding ( P<0.001), postoperative significant hepatic encephalopathy ( P<0.012), and postoperative new or worsening ascites ( P<0.001) was significantly associated with stent occlusion ( P<0.05), while platelets ( P=0.134), total bilirubin ( P=0.229), international normalized ratio ( P=0.436), and portal vein pressure ( P=0.230) were not significantly associated with stent occlusion after surgery. Conclusion:In patients with partial portal vein thrombosis before surgery, continuous anticoagulation for 30 days post-TIPS therapy can effectively prevent stent stenosis or occlusion; while in patients with portal vein patency, portal cavernoma and complete portal vein blockage before surgery, postoperative anticoagulation has no significant effect on stent stenosis or occlusion.

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 of FNDC5 in hepatocellular carcinoma (HCC) and its relationship with clinicopathological characteristics and prognosis of HCC patients.Methods Immunohistochemistry and qRT-PCR were used to detect the expression of FNDC5 in HCC tissues,and the relationship between its expression and clinicopathological parameters and prognosis.Results FNDC5 was mainly expressed in the cytoplasm of HCC cells,weakly expressed in the nucleus.Among the 30 liver cancer tissues,FNDC5 was weakly positive in 5,positive in 19,and strongly positive in 6 cases.QRT-PCR assay showed that FNDC5 was highly expressed in HCC tissues with vascular invasion.The incidence of vascular invasion in the high-expression of FNDC5 was 30％ (9/30),which was significantly higher than that of the FNDC5 low-expression group [6.7％ (2/30),] and the difference was significant (x2 =15.026,P ＜0.05).There was no significant correlation between FNDC5 level and age,sex,HBsAg,and alpha-fetoprotein (AFP) level in HCC (P ＞ 0.05).Conclusion FNDC5 level in the liver cancer tissues is closely related to the occurrence of vascular invasion.

BACKGROUND: Mesenchymal stem cells can protect and repair the liver of rats with liver failure, but the mechanisms are not completely clear. OBJECTIVE: To explore the protective effects and related mechanisms of intravenous injection of human adipose-derived mesenchymal stem cells on acute liver failure in rats. METHODS: Thirty-six Sprague-Dawley rats (provided by Qingdao Daren Fucheng Animal Husbandry Co., Ltd. in China) were randomly divided into control group, model group and transplantation group. Animal models of acute liver failure were established by intraperitoneal injection of D-galactosamine in the model group and the transplantation group. One day after modeling, the rats in the transplantation group were injected with human adipose-derived mesenchymal stem cell suspension, and those in the model group were injected with the same amount of saline. After 1 and 3 days of cell transplantation, the serum levels of alanine aminotransferase, aspartate aminotransferase, and total bilirubin were measured. Three days after cell transplantation, the serum levels of tumor necrosis factor-α, interleukin-6 and interleukin-10 were detected, the pathological changes of the rat liver were observed by hematoxylin-eosin staining, and the activity of glycogen synthase kinase-3β protein in the liver tissue was detected by western blot. RESULTS AND CONCLUSION: Compared with the model group, there was a significant reduction in the serum levels of alanine aminotransferase, aspartate aminotransferase, total bilirubin, tumor necrosis factor-α, interleukin-6 and interleukin-10 in the transplantation group (P < 0.05). Inflammation and necrosis of liver tissues in the transplant group were alleviated compared with the model group. The activity of glycogen synthase kinase 3β in the liver tissue of the transplanted group was lower than that of the model group (P < 0.05). Overall, these results indicate that human adipose-derived mesenchymal stem cells can alleviate hepatic inflammation and pathological injury, and improve the liver function in rats with acute hepatic failure. Moreover, the mechanism may be related to the inhibition of glycogen synthase kinase 3β activity.

Objective To evaluate the efficacy and safety of X-ray guided endoscopic gastrojejunostomy using stent in treatment of malignant gastric outlet obstruction ( GOO ) . Methods Six hospitalized patients with malignant GOO underwent X-ray guided endoscopic gastrojejunostomy using stent in the department of gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University between March 2017 and June 2017. The technical success rate, clinical success rate, procedure time, adverse events and follow-up were recorded and analyzed in this retrospective study. Results The stent was successfully placed in the 6 patients with 100％ ( 6/6) technical success rate. The mean procedure time was 91. 7±51. 8 min. After the procedure, all patients were fed liquid or semi-liquid diet, and the GOO score system was increased from 0-1 before operation to 2-3 after operation. The clinical success rate was 100％(6/6). Peritonitis was observed in 2 patients during operation, and resolved by abdominal drainage. Gastrointestinal bleeding occurred in 1 patient after operation, which was resolved with conservative treatment. During a mean follow-up period of 78. 6 days (range 32-100 days), there was no recurrence of obstruction symptoms except that 1 patient died because of tumor progress 60 days after procedure. Conclusion The X-ray guided endoscopic gastrojejunostomy using stent is feasible and safe to treat malignant GOO with a reliable short-term efficacy.

Objective To investigate the effect of Irisin on proliferation,apoptosis,migration and invasion of human cholangiocarcinoma cell line Hucct-1.Methods After treatment with Irisin,CCK-8 assay and flow cytometry assay were conducted to investigate the effect of Irisin on proliferation and apoptosis of cholangiocarcinoma cells.Scratch test and transwell invasion assay were used to studythe effect of Irisin on the migration and invasion ability of cholangiocarcinoma cells.Western blot was utilized to detect the expression of E-cadherin,N-cadherin and Vimentin in cholangiocarcinoma cells.Results CCK-8 assay showed that Irisin inhibited cholangiocarcinoma cell proliferationin a dose-dependent manner.Flow cytometry assay showed that the apoptosis rate of Irisin group [(14.8 ±0.9)％] was higher than that in the control group [(5.4±0.6)％],(P＜0.05).The scratch test showed that the rate of cell scratch healing in Irisin group [(15.0± 1.0)％] was significantly lower than that in the control group [(28.0±2.0)％] (P＜0.05).Transwell invasion test showed that the number of cells in Irisin group was (96.0±7.0),which was significantly lower than that in control group (155.0± 9.0) (P＜0.05).Western blot showed that the expression of E-cadherin increased and N-cadherin and Vimentin decreased after Irisin treatment.Conclusion Irisin inhibits proliferation,migration and invasion and promote apoptosis of cholangiocarcinoma cells.

To investigate the changes of myocardial glucose metabolism in rabbit cardiac arrest models and the effect of hydrogen intervention by 18F-fluroro-2-deoxyglucose (18F-FDG) positron emission tomography (PET) imaging.
 Methods: Fifteen male New Zealand white rabbits were randomly divided into a hydrogen group (n=6), a control group (n=6) and a sham group (n=3). Cardiac arrest (CA) was induced by intravenous injection of potassium chloride. Conventional cardiopulmonary resuscitation (CPR) was initiated after five-minutes CA. The hydrogen group and the control group were mechanically ventilated into mixed gas with 4% hydrogen+96% oxygen and pure oxygen, respectively, for 30 minutes after CPR. Rats in the sham group was performed the same surgical procedure and was injected adrenaline and potassium chloride but did not induce CA. The vital signs at basic state and 30 min after return of spontaneous circulation (ROSC) were recorded in each group. The parameters of CPR were recorded in two CA groups. Myocardial glucose metabolism was assessed by positron emission tomography (PET) at basic state, 2 h and 24 h after ROSC. The maximum standardized uptake value (SUVmax) of 18F-FDG was measured.
 Results: There were no significant differences in the basal body weight and vital signs among the three groups. There was no significant difference in the blood glucose level before PET examination. The 18F-FDG SUVmax in the sham group at three time points was not significantly changed. In the hydrogen group and the control group, the 18F-FDG SUVmax at 2 h after ROSC were significantly higher than the basic level (1.89±0.47 vs 3.47±1.24 and 1.90±0.36 vs 4.26±0.80, respectively). Compared with the control group, the 18F-FDG SUVmax in the hydrogen group was lower at the point at 2 h after ROSC. The 18F-FDG SUVmax in the 2 CA group were down to the basic level at 24 h after ROSC (hydrogen group 2.02±0.64, control group 2.07±0.61).
 Conclusion: Myocardial glucose metabolism in CA rabbits was increased significantly after ROSC, and hydrogen intervention can reduce the degree of glucose metabolism.
Animals ; Cardiopulmonary Resuscitation ; Glucose ; metabolism ; Heart Arrest ; physiopathology ; surgery ; Male ; Myocardium ; metabolism ; Positron-Emission Tomography ; Rabbits ; Random Allocation ; Rats