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 protective effect of herba artemisiae scopariae extract on multidrug resistance protein 3(MDR3)gene mutation-induced neonatal parenteral nutrition-associated cholestasis(PNAC)and its possible mechanism.Methods ①Human primary hepatocytes were treated with cell culture in vitro,CRISPR/Cas9 lentivirus infection and MDR3 mutant gene lead-in.The levels of hepatic and biliary biochemical indexes[alanine transaminase(ALT),aspartate transaminase(AST),total bilirubin(TBil),direct bilirubin(DBil),indirect bilirubin(IBil),total bile acid(TBA)]in the supernatant of hepatocytes before and after 16,32,48 hours were compared to determine the time required for fatty acid induction of PNAC hepatocyte model with MDR3 gene mutation.② Human primary hepatocytes were divided into blank control group,MDR3 gene wild type group,MDR3 gene mutation group,and herba artemisiae scopariae extract intervention group according to random number table method.The blank control group was treated with culture medium only,the MDR3 gene wild type group was infected with lentivirus and mixed with wild type MDR3 gene and culture medium,the MDR3 gene mutation group was infected with lentivirus and cultured in culture medium with the mutant genes lead-in of LV-MDR3KI(c.485T>A,c.2793insA,c.1031G>A,c.3347G>A)mutation,while the MDR3 mutant gene was lead-in by lentivirus infection and cultured in culture medium,and then pretreated with 100 g/L herba artemisiae scopariae extract in the herba artemisiae scopariae extract intervention group,then the four groups of hepatocytes were induced with 1%fat emulsion,and the treatment time was the time needed to construct the PNAC hepatocytes model with MDR3 gene mutation.The levels of ALT,AST,TBil,DBil,IBil and TBA in the supernatant of hepatocytes were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression abundance of adenosine triphosphate binding cassette proteins(ABCB4,ABCB11,ABCC2,ABCC3,ABCC4)encoding MDR3,bile salt export pump(BSEP),multidrug resistance associated protein(MRP)2-4,and tumor necrosis factor-α(TNF-α)genes were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared to the blank control group and MDR3 gene wild type group,there was no significant difference in the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 gene mutant group before and 16 hours after induction with 1%fat emulsion,however after treated with 1%fat emulsion for 32 hours and 48 hours,the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 mutant hepatocytes were significantly increased(P<0.05),consequently the time required for fatty acid induction of PNAC hepatocyte model was 32 hours.At 32 hours after treatment with fat emulsion,the levels of ALT,AST,TBil,DBil,TBA in the supernatant of hepatocytes in the herba artemisiae scopariae extract intervention group were significantly decreased[ALT(ng/L):148.3±2.3 vs.164.9±7.0,AST(ng/L):2767.4±78.8 vs.3239.4±107.1,TBil(μmol/L):7.6±0.2 vs.13.6±0.3,DBil(μmol/L):1.8±0.1 vs.5.7±0.2,TBA(μmol/L):3.4±0.2 vs.6.7±0.1,all P<0.05].The ABCB4,ABCC2,ABCC3,ABCC4 mRNA expression of MDR3,MRP2,MRP3,MRP4 in the blank control group,MDR3 wild type group,MDR3 gene mutation group and the herba artemisiae scopariae extract intervention group had no significant difference.The expression of TNF gene mRNA was highly expressed in MDR3 gene mutation group(2-??Ct:1.258±0.200 vs.1.001±0.052),and was low expressed in the herba artemisiae scopariae extract intervention group(2-??Ct:0.387±0.247 vs.1.258±0.200),and there was a significant difference between the two groups(both P<0.05).Compared to the MDR3 gene mutation group,the ABCB11 gene encoding BSEP mRNA expression in the herba artemisiae scopariae extract intervention group was significantly increased(2-??Ct:2.955±0.479 vs.1.333±0.529,P<0.05).Conclusion The herba artemisiae scopariae extract has a protective effect on PNAC induced by MDR3 gene mutation,which may be related to antagonizing inflammatory reaction,decreasing the expression of TNF mRNA and improving the expression of ABCB11 gene encoding BSEP.

Background: The incidence density of metabolic dysfunction-associated fatty liver disease (MAFLD) and the effect of a healthy lifestyle on the risk of MAFLD remain unknown. We evaluated the prevalence and incidence density of MAFLD and investigated the association between healthy lifestyle and the risk of MAFLD. Methods: A cross-sectional analysis was conducted on 37,422 participants to explore the prevalence of MAFLD. A cohort analysis of 18,964 individuals was conducted to identify the incidence of MAFLD, as well as the association between healthy lifestyle and MAFLD. Cox proportional hazards regression was used to calculate the hazard ratio (HR) and 95% confidence interval (CI) with adjustments for confounding factors. Results: The prevalence of MAFLD, non-alcoholic fatty liver disease, and their comorbidities were 30.38%, 28.09%, and 26.13%, respectively. After approximately 70 thousand person-years of follow-up, the incidence densities of the three conditions were 61.03, 55.49, and 51.64 per 1,000 person-years, respectively. Adherence to an overall healthy lifestyle was associated with a 19% decreased risk of MAFLD (HR, 0.81; 95% CI, 0.72 to 0.92), and the effects were modified by baseline age, sex, and body mass index (BMI). Subgroup analyses revealed that younger participants, men, and those with a lower BMI experienced more significant beneficial effects from healthy lifestyle. Conclusion Our results highlight the beneficial effect of adherence to a healthy lifestyle on the prevention of MAFLD. Health management for improving dietary intake, physical activity, and smoking and drinking habits are critical to improving MAFLD.

Pancreatitis is one of the most common inflammatory diseases of the pancreas caused by autodigestion induced by excessive premature protease activation. However, recognition of novel pathophysiological mechanisms remains a still challenge. Both genetic and environmental factors contribute to the pathogenesis of pancreatitis, and the gut microbiota is a potential source of an environmental effect. In recent years, several new frontiers in gut microbiota and genetic risk assessment research have emerged and improved the understanding of the disease. These investigations showed that the disease progression of pancreatitis could be regulated by the gut microbiome, either through a translocation influence or in a host immune response manner. Meanwhile, the onset of the disease is also associated with the heritage of a pathogenic mutation, and the disease progression could be modified by genetic risk factors. In this review, we focused on the recent advances in the role of gut microbiota in the pathogenesis of pancreatitis, and the genetic susceptibility in pancreatitis.

Objective:To access the independent associations between serum ferritin quintile levels and metabolic syndrome (MS) in adults of different genders.Methods:19 563 participants over the age of 18 years were recruited from "TCLSIH Cohort Study" from 2007 to 2015. Serum ferritin concentration was measured by Enzyme-linked immunoassay, while metabolic syndrome was diagnosed according to MS diagnostic criteria formulated by Chinese Diabetes Society in 2013. Multiple logistic regression analysis was used to assess the association between serum ferritin quintile levels and the prevalence of MS in males and females. Results:After adjusting the confounding factors, the overall prevalence of MS gradually increased with the increasing of serum ferritin levels, similar results were observed in males and females. Subjects were divided into 5 subgroups according to serum ferritin levels. Compared with level 1 group, logistic regression showed that the serum ferritin quintiles of males and females ranged from low to high, the OR (95% confidence interval) for metabolic syndrome were 1.142 (0.998, 1.307), 1.382 (1.210, 1.579), 1.680 (1.472, 1.917), 2.085 (1.827, 2.380), respectively ( Ptrend<0.01), and 1.147 (0.911, 1.444), 1.346 (1.075, 1.687), 1.567 (1.268, 1.941), 2.444 (1.981, 3.023), respectively ( Ptrend<0.01). Conclusion:The elevated ferritin levels were positively related to the prevalence of metabolic syndrome in adults of different genders.

Objective To investigate the effect of enhanced recovery after surgery (ERAS) on immune function and postoperative recovery in gastric cancer patients undergoing total laparoscopic radical gastrectomy.Methods Patients were randomly divided into ERAS group and control group.Blood CD4 +,CD8+,CD4 + CD25 +,C-reactive protein,postoperative recovery and complications were compared between the two groups.Results On day1,CD4 +,CD8 +,CD4 + CD25 + in the two groups were significantly lower than those before surgery (t =9.070,7.297,5.830,12.870,3.529,10.547,all P＜0.05).The ERAS group had higher CD8 +,CD4 + CD25 + levels than the control group (t =2.163,2.203,P ＜ 0.05).On day3,CD4 + CD25 + in ERAS group was not different from that before surgery (t =1.062,P ＞ 0.05) while the other indexes in the two groups raised but still were lower than preoperative level (t =3.322,5.015,3.418,9.912,all P ＜0.05);CD4 +,CDs +,CD4 + CD25 + in ERAS group were higher than control group (t =2.804,2.040,2.210,all P＜0.05).On day5,CD4+,CD4 + CD25+ in the two groups and CDs+ in ERAS group returned to the preoperative level,while CDs + of the control group was still lower than the preoperative level (t =6.862,P ＜0.05).On day1,3 and 5,the C-reactive protein levels of the two groups were higher than those before surgery(t=-13.338,-13.715,-11.319,-12.286,-13.182,-15.076,all P ＜ 0.05),and ERAS group were lower than the control group (t =-3.246,-2.100,-2.211,all P＜0.05).There was no mortality in neither groups.The time of passage gas by anus,defecation,getting out of bed,oral feeding,and postoperative hospital stay in the ERAS group were less than thoseinthecontrolgroup[(2.8±1.0)dvs.(3.9±0.9)d,t=-5.974;(3.8± 0.9)d vs.(4.3±1.0)d,t=-2.700;(19.1 ±4.0)hvs.(35.9±6.6)h,t=-16.045;(9.9 ±1.6)d vs.(11.5±2.0) d,t =-4.479,all P ＜ 0.05].Conclusions ERAS in the perioperative period of patients with total laparoscopic radical gastrectomy mitigates the stress on the cellular immune system,reduces inflammatory response,and help fast recover the postoperative gastrointestinal function.

Minimizing the burden on study subjects and assessing the general dietary nutritional status as accurately as possible are the basis of a nutritional epidemiological cohort study in the general population. While introducing the main dietary nutrition assessment methods, this paper manly describes the basic contents and principles for the development of food frequency questionnaire, and briefly illustrates the problems and solutions for the development of area specific food frequency questionnaires by taking the example of Tianjin Chronic Low-grade Systemic Inflammation and Health (TCLSIH) cohort study. Finally, discusses preliminarily the necessity and possibility of developing a national food frequency questionnaire.

Objective To analyze the influential factors of the prognosis of neonatal cholestasis. Methods The data on 106 newborns with neonatal cholestasis collected from January 1, 2009 to June 30, 2016 was collected and analyzed retrospectively. All cases were divided into cure group (99 patients) and improvement group (7 patients) according to the effect of oral ursodeoxycholic acid therapy and comprehensive medical treatment. There was no cure for invalid case. The clinical features, course of disease and curative effect were observed. The levels of total bilirubin (TB), direct bilirubin (DB), alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyl transpeptidase (γ-GT), total bile acid (TBA) were compared between two groups. The ratio of neonatal bacterial infection, cytomegalovirus (CMV) infection, sex, total intravenous nutrition (more than 7 d), perinatal hypoxia, premature birth, maternal complicated intrahepatic cholestasis of pregnancy (ICP) were compared between two groups. Multiple Logistic regression analysis was used to analyze the prognosis factors of neonatal cholestasis. Results The levels of serum ALT, AST, γ-GT, TB, DB and TBA in improvement group were significantly higher than those in cure group, the course of disease in improvement group was significantly longer than that in cure group, there were significant differences (P<0.05). A strong linear correlation was showed between course of disease and the levels of serum ALT, AST, TB, DB and TBA (tALT=13.050, tAST=14.696, tTB=12.771, tDB=13.776, tTBA=12.019, P < 0.05). Multiple Logistic regression analysis showed that the prognosis of neonatal cholestasis was associated with bacterial infection, CMV infection, total intravenous nutrition (more than 7 d) and premature ( OR=9.375, 7.909, 11.333, 11.333, P < 0.05). Conclusions The higher the level of serum TB, DB, TBA, ALT and AST, the longer the course of the disease. Bacterial infection, CMV infection, total venous nutrition( ≥ 7 d) and premature are the risk factors of the prognosis of neonatal cholestasis. Therefore, prevention of neonatal bacterial and CMV infection, reduction of the occurrence of preterm delivery and reduction of the total parenteral nutrition time are very important to improve the prognosis of neonatal cholestasis.

Objective To explore the association between complement C3 and the prevalence of simple fatty liver disease and nonalcoholic steatohepatitis. Methods A total of 2 799 health check-up participants were recruited from Tianjin Medical University General Hospital-Health Management Centre. Serum C3 levels were measured using immunoturbidimetry method and simple fatty liver disease and nonalcoholic steatohepatitis were diagnosed by liver ultrasonography and serum alanine aminotransferase concentration. Multiple logistic regression analysis was used to examine the association between quartiles of serum C3 levels and the prevalence of simple fatty liver disease and nonalcoholic steatohepatitis. Results After adjusting for covariates, the odds ratios (95% confidence interval) of simple fatty liver disease and nonalcoholic steatohepatitis for increasing quartiles of C3 were: 1.00 (reference), 2.15 (1.55, 3.01), 3.28 (2.35, 4.6) and 4.31 (3.11, 6.02); and 1.00 (reference), 4.54 (1.68, 15.91), 7.81 (3.02, 26.67) and 14.91 (5.94, 50.14) (both P for trend<0.000 1), respectively. Conclusions The study found that the elevated C3 levels were significantly associated with the prevalence of simple fatty liver disease and nonalcoholic steatohepatitis. Further study is needed to clarify whether C3 has a predictive value for the occurrence of nonalcoholic fatty liver disease.

Objective To understand the relationship between dietary patterns and breast cancer and to provide a possible scientific basis for breast cancer prevention,we performed a meta-analysis in order to evaluate the current relevant literature.Method Meta-analysis was used to pool the effect of vegetable,fruit,fish and soybean dietary patterns on the risk of breast cancer.The breast cancer risk of the highest categories compared to the lowest categories of such dietary patterns was combined using fixed effect model.Searching the database (Chinese databases included CQVIP,WANFANG medicine online,CNKI and SinoMED,and English databases included PubMed,Embase,Cochrane database,Medline and Google Scholar) for articles published between January 1997 and June 2014.All statistical analyses were performed using the Stata 11.0.Result Twelve cohort studies involving 623 536 participants and 19 707 patients with brcast cancer were found to be eligible.Our results indicate that a reasonably healthy dietary pattern (which is characterized by a high consumption of vegetables,fruit,fish and soybean) is beneficial for the prevention of breast cancer.Compared to the lowest categories of such dietary patterns,relative risk (RR) of the highest categories is 0.86 (95％ CI 0.80-0.92).Conclusion A reasonably healthy dietary pattern (which is characterized by a high consumption of vegetables,fruit,fish and soybean) is beneficial for the prevention of breast cancer.