The identification and optimization of mutations in nanobodies are crucial for enhancing their therapeutic potential in disease prevention and control. However, this process is often complex and time-consuming, which limit its widespread application in practice. In this study, we developed a workflow, named Evolutionary-Nanobody (EvoNB), to predict key mutation sites of nanobodies by combining protein language models (PLMs) and molecular dynamic (MD) simulations. By fine-tuning the ESM2 model on a large-scale nanobody dataset, the ability of EvoNB to capture specific sequence features of nanobodies was significantly enhanced. The fine-tuned EvoNB model demonstrated higher predictive accuracy in the conserved framework and highly variable complementarity-determining regions of nanobodies. Additionally, we selected four widely representative nanobody-antigen complexes to verify the predicted effects of mutations. MD simulations analyzed the energy changes caused by these mutations to predict their impact on binding affinity to the targets. The results showed that multiple mutations screened by EvoNB significantly enhanced the binding affinity between nanobody and its target, further validating the potential of this workflow for designing and optimizing nanobody mutations. Additionally, sequence-based predictions are generally less dependent on structural absence, allowing them to be more easily integrated with tools for structural predictions, such as AlphaFold 3. Through mutation prediction and systematic analysis of key sites, we can quickly predict the most promising variants for experimental validation without relying on traditional evolutionary or selection processes. The EvoNB workflow provides an effective tool for the rapid optimization of nanobodies and facilitates the application of PLMs in the biomedical field.

Objective:To investigate the feasibility and value of participatory bilingual teaching in nuclear medicine teaching.Methods:With the same chapter of Nuclear Medicine as the teaching contents, the traditional method of cramming bilingual teaching was used for the clinical medical students in the class of 2014 of the seven-year program and the five-year program, and the method of participatory bilingual teaching was used for the students in the class of 2015. The methods of periodical achievement test, questionnaire survey, and final examination were used to evaluate the teaching effect of the above two teaching modes.Results:In the periodical achievement test, the participatory bilingual teaching group had significantly higher scores than the cramming teaching group in terms of mean score, case analysis, and English questions [seven-year program: (82.13±10.72), (35.74±4.13), and (23.03±3.40) vs. (79.21±11.31), (33.86±5.23), and (22.12±2.75), P<0.05; five-year program: (78.66±12.75), (34.30±5.59), and (22.45±2.91) vs. (75.29±10.81), (32.70±6.04), and (21.36±3.09), P<0.05]. The questionnaire survey showed that participatory bilingual teaching had a better degree of satisfaction than cramming teaching. The participatory bilingual teaching group had a better score than the cramming teaching group in the final examination, but with no significant difference between the two groups. Conclusions:In the teaching of nuclear medicine, the participatory bilingual teaching mode can significantly improve teaching effect and achieve the teaching goal efficiently.

Objective:To investigate the effect of "simulated Chinese medicine nursing clinic" teaching on syndrome differentiation ability among nursing students and the teaching effect of this method.Methods:A total of 325 students were randomly divided into conventional teaching group with 163 students (receiving conventional nursing teaching) and real situational teaching group with 162 students (receiving "simulated Chinese medicine nursing nursing clinic" teaching at the same time). A self-made questionnaire for syndrome differentiation-based nursing ability, assessment scores of traditional Chinese medicine nursing theories and skills, and a teaching satisfaction questionnaire were used for evaluation.Results:After intervention, the real situational teaching group (the treatment group and the experience group) had significantly higher socres of syndrome differentiation-based nursing ability (mastery of four diagnostic methods, diagnosis of syndromes, analysis of syndromes, determination of nursing principles, and development of nursing regimens) than the conventional teaching group (the treatment group and the conventional teaching group: 8.97±1.00/8.47±1.20/8.33±1.06/8.30±1.26/7.89±1.13 and 7.96±1.14/7.29±1.36/7.14±1.18/7.39±1.30/7.26±1.18, P<0.05; the experience group and the conventional teaching group: 8.39±1.10/8.17±1.15/8.07±1.06/7.97±1.26/7.73±1.38 and 7.96±1.14/7.29±1.36/7.14±1.18/7.39±1.30/7.26±1.18, P<0.05). The real situational teaching group had a significantly higher overall degree of satisfaction with teaching than the conventional teaching group [160 (97.6%) and 150 (92.0%), P<0.05]. Conclusions:The "simulated Chinese medicine nursing clinic" teaching program effectively enhances the thinking of syndrome differientiation and related abilities for nursing, improves the learning effect of traditional Chinese medicine nursing theories and skills, and increases the learning interest in traditional Chinese medicine nursing among nursing students.

Objective:To investigate the features and influencing factors of language in children with various types of speech disorders.Methods:A case-control study was carried out, 262 children with speech disorder had been diagnosed at the language-speech clinic of the Center of Children′s Healthcare, Children′s Hospital, Capital Institute of Pediatrics from January 2021 to November 2023, the children with speech sound disorder as the speech sound disorder group, the children with developmental stuttering as the stuttering group. There were 100 typically-developed children who underwent physical checkups at the Center of Healthcare during the same period as the healthy group. All children experienced a standardized evaluation of language with diagnostic receptive and expressive assessment of mandarin‐comprehensive(DREAM-C) and questionnaire, One-way ANOVA and LSD test were conducted to compare the differences in overall language, receptive language, expressive language, semantics, and syntax scores among 3 groups of children. According to the results of DREAM-C, the children with speech disorder were divided into language normal group and language delay group. Chi‐square test and multivariate Logistic regression were implemented to analyze the association between the linguistic development of children with speech disorder and potential influential factors.Results:There were 145 children in the speech sound disorder group, including 110 males and 35 females respectively, with an age of (5.9±1.0) years; 117 children in the stuttering group, including 91 males and 26 females, with an age of (5.8±1.0) years; 100 children in the healthy group, including 75 males and 25 females, with an age of (5.7±1.2) years. The variations in overall language, expressive language, and syntax scores among 3 groups of children were statistically significant (92±18 vs.96±11 vs. 98±11, 81±18 vs. 84±14 vs. 88±13, 87±16 vs. 89±11 vs. 91±10, F=5.46, 4.69, 3.68, all P<0.05). Pairwise comparison revealed that the speech sound disorder group had lower scores in overall language, expressive language, and syntactic compared to the healthy group, and the differences were statistically significant (all P<0.01) and the overall language score was lower than that of children with stuttering ( P<0.05). In terms of overall language and expressive language, there was a statistically significant difference in the incidence of language delay among the three groups of children (15.9% (23/145) vs. 20.5% (24/117) vs. 7.0% (7/100), 46.2% (67/145) vs. 39.3% (46/117) vs. 26.0% (26/100); χ2=7.93, 10.28; both P<0.05). In terms of overall language, the stuttering group took up the highest proportion. In terms of expressive language, the speech sound disorder group accounted for the highest amount. The incidence of language delay in children with speech disorder was 44.3% (116/262). Non-parent-child reading, daily screen time ≥1 hour and screen exposure before 1.5 years of age are risk factors for the development of language in children with speech disorder ( OR=1.87, 2.18, 2.01; 95% CI 1.07-3.27, 1.23-3.86, 1.17-3.45; all P<0.01). Negative family history are protective factors for the progress of language ability ( OR=0.37, 95% CI 0.17-0.81, P<0.05). Conclusions:Children with speech disorder tend to have easy access to language delay, especially in expressive language and syntax. The occurrence of language delay in children with speech disorder is tightly connected with factors such as the family medical history, parent-child reading, screen time, etc. Attention should be paid to the development of language in children who suffer from speech disorder.

Objective To explore the role of long non-coding RNA smad7(Linc-smad7)in invasion and migration of pancreatic cancer(PC)cells and its mechanisms.Methods The expression level of Linc-smad7 in PC tissues and cell lines was detected by qRT-PCR assays.Transwell assays were performed to observe the effect of Linc-smad7 overexpression on the migration and invasion abilities of PaCa-2 cells.Luciferase reporter analysis was made to detect the direct binding effect of Linc-smad7 on miR-125b and miR-125b on Sirtuin1(SIRT1).qRT-PCR assays were used to detect the regulatory effect of Linc-smad7 on miR-125b expression in PaCa-2 cells.qRT-PCR and Western blotting assays were used to detect the regulatory effect of miR-125b on SIRT1 expression.Results Linc-smad7 was significantly increased in PC tissues and cell lines.Paca-2 cells with overexpressed Linc-smad7 showed higher migration and invasion abilities,while miR-125b expression was decreased.After the expression of miR-125b was increased,the expression of SIRT1 was decreased significantly.Luciferase reporter assays results suggested that Linc-smad7 directly targeted with miR-125b,and miR-125b directly bound with SIRT1.Increased Linc-Smad7 or decreased miR-125b could reverse the effect of SIRT1 inhibition on invasion and migration of PaCa-2 cells.Conclusion Linc-smad7 promotes invasion and migration of PaCa-2 cells through miR-125b/SIRT1 axis.

Objective: To investigate the characteristics and trend of stroke incidence in Jining City, Shangdong Province from 2015 to 2022, so as to provide the reference for formulating prevention and control strategies of stroke. Methods: Data of stroke incidence in Jining City from 2015 to 2022 were collected from Shandong Provincial Chronic Disease Surveillance Information Management System. The crude incidence was estimated, standardized using the data of the sixth national population census in 2010, and analyzed by age, gender and subtype. The trend in incidence of stroke was analyzed using annual percent change (APC). Results: A total of 316 267 cases of stroke were reported in Jining City from 2015 to 2022, with a crude incidence of 474.17/105 and a standardized incidence of 371.43/105. The incidence of stroke peaked from March to May (90 409 cases, 28.59%). There were 278 901 cases of ischemic stroke (88.19%) and 37 366 cases of hemorrhagic stroke (11.81%). The crude incidence of stroke was higher in males than in females (525.45/105 vs. 420.16/105, P<0.05). The crude incidence of stroke increased with age (P<0.05), reaching a peak in the age group of 80 years and above (2 764.92/105). From 2015 to 2022, the crude incidence of stroke in the overall population, males, females, the age groups of 0-<30 years and 40-<50 years showed increasing trends (APC=6.142%, 6.992%, 5.054%, 3.693% and 6.587%, all P<0.05); the crude incidence of ischemic stroke in the overall population, males and females showed increasing trends (APC=7.489%, 6.593% and 5.456%, all P<0.05), while the crude incidence rates of hemorrhagic stroke did not show significant trends (APC=3.455%, 2.804% and 1.919%, all P>0.05). Conclusions The crude incidence of stroke increased in Jining City from 2015 to 2022, with ischemic stroke as the predominant subtype. March to May was the peak period for disease onset, and young and middle-aged men should be focused on.

Objective:A recombinant adenoviral vector vaccine based on non-replicating human adenovirus type 5 (Ad5), encoding the conserved domain of respiratory syncytial virus G protein (RSV-G) was constructed. The immunogenicity and protective efficacy of this vaccine were subsequently evaluated in mice.Methods:The recombinant Ad5 vector plasmid (Ad5-Gbcc-Gacc) was constructed by inserted conserved domains of RSV A and RSV B. The recombinant adenovirus Ad5-Gbcc-Gacc was rescued in HEK293A cells. The genome of virus Ad5-Gbcc-Gacc was identified by multi-enzyme digestion, and the expression of Ad5-Gbcc-Gacc was verified by Western blot. Recombinant adenovirus was used to immunize BALB/c mice via intramuscular injection with signal dose, and then challenged with RSV Long strain at week 6. The levels of G specific IgG and antibody subtypes in serum were detected by enzyme-linked immunosorbent assay, the level of neutralizing antibodies was determined by micro-neutralization assay. After challenge, the mice′s weight was recorded daily, the copies of RSV virus in the lung and nasal tissues were detected. Pathological changes in lung tissue were also examined.Results:Western blot and multi-enzyme digestion identification confirmed the successful rescue of the recombinant adenovirus. Ad5-Gbcc-Gacc elicit high titers of specific IgG, robust neutralizing antibodies, and a balanced Th1/Th2 immune response in mice. In comparison to unimmunized controls, mice immunized with Ad5-Gbcc-Gacc reduced the viral copies in both lung and nasal tissue, and exhibited only minimal pathological damage of lung tissue following RSV challenge. In conclusion, Ad5-Gbcc-Gacc induced robust immunogenicity and offers protective effects against RSV infection in murine models.Conclusions:Ad5-Gbcc-Gacc induce robust immunogenicity and can protect mice from RSV challenge, which lays a foundation for further development of RSV vaccine based on G protein.

Objective:To study the application of 3D printing technology in multi-center fenestrated/branched endovascular repair (F/B-EVAR) for endovascular repair of abdominal aortic diseases.Methods:From Feb 2018 to Mar 2023, The clinical and followup data of 316 cases of abdominal aortic lesions undergoing repair with F/B-EVAR at 69 medical centers nationwide using 3D printing technology to guide physician-modified stent graft were retrospectively analyzed.Results:The mean follow-up time of the patients was 23 months (2-60 months), and 24 cases were lost to follow up, the follow-up rate was 92.4% (292/316), the mean postoperative hospitalization time was (8.2±4.9) days. A total of 944 main abdominal branch arteries were reconstructed. Intraoperative reconstruction of 11 branches failed, with a success rate of 98.8% (933/944). Within 30 days after surgery, 8 patients died (2.5%), and 6 patients died during follow-up, a total of 14 patients died (4.4%). There were 11 cases (3.5%) of spinal cord ischemia and no patient suffered from permanent paraplegia. There were 19 patients (6.0%) with postoperative renal function injury. Internal leakage was found in 26 patients, and the rate of internal leakage was 8.2%.Conclusion:3D printing technology can accurately locate the location of branch arteries, simplifing the surgical process, shortening the learning curve , and improving clinical efficacy.

Objective:To review the risk factors for early and medium-term complications of fenestration-branch endovascular thoracic aortic repair (F/B-TEVAR) in patients with complex aortic arch disease.Methods:The clinical and follow-up data of 202 patients undergoing F/B-TEVAR treatment from Feb 2019 to Sep 2023 in these centers were retrospectively analyzed .Results:There were 46 cases suffering from postoperative complications (22.8%). The risk factors with statistical significance included aortic atherosclerotic plaque [ OR=2.843; 95% CI (1.4-5.6); P<0.01], aortic intramural thrombosis [ OR=2.358; 95% CI (1.2-4.6), P=0.011], the aortic dilatation [ OR=4.219; 95% CI (1.6-11.3), P<0.01], the history of stroke [ OR=2.088; 95% CI (1.1-4.1), P=0.032], smoking history [ OR=2.680; 95% CI: (1.3-5.5); P<0.01], duration of surgery [ OR=1.9; 95% CI: (1.2-2.9); P=0.042].While the application of 3D printing assistive technology [ OR=0.392; 95% CI: (0.2-0.9); P=0.048] was in a negative correlation with postoperative complication. Conclusions:The independent risk factors for complications after F/B-TVAR included aortic atherosclerotic plaque, aortic intramural thrombosis, the aortic dilatation, the history of stroke, smoking history,duration of surgery.The application of 3D printing technology can effectively reduce the complication rate.

Objective:To investigate the incidence of abnormal bone mineral density (BMD) in male patients with alcoholic cirrhosis.Methods:Clinical data of 72 patients with alcoholic cirrhosis admitted in Beijing Ditan Hospital, Capital Medical University from March 2017 to July 2023 were enrolled as study group, and 40 age-and BMI-matched non-hepatopathy subjects were selected as control group.The incidence of abnormal BMD were compared between two groups. The serum levels of osteocalcin (OC), total procollagen type 1 amino-terminal propeptide (TP1NP), β-C-terminal telopeptide of typeⅠcollagen (CTX) and 25-hydroxy vitamin D [25-(OH)VD] were measured and compared among patients with different Child-Pugh grades.Results:Among 72 alcoholic cirrhosis patients, there were 54 cases (75.0%) complicated with abnormal BMD, including 21 cases (29.2%) of bone loss and 33 cases (45.8%) of osteoporosis. In control group there were 15 subjects with abnormal bone mineral density, including 9 cases (22.5%) of bone loss and 6 cases (15.0%) of osteoporosis( χ2=5.623 and 15.900,both P<0.05). The average BMDs of L1-L4, femoral neck, intertrochanteric region and trochanter of the femur in patients with alcoholic cirrhosis were significantly lower than those in control group ( t=3.574, 8.640, 7.282, 7.958, 3.755, 5.573, 5.026,all P<0.05); the average BWDs of L1-L3 and hip joint in patients with Child-Pugh C were significantly lower than those in patients with Child-Pugh A and B ( t=1.414, 1.699, 3.786, 2.590, 8.763, 2.581, 1.392, 6.232,all P<0.05). The serum levels of 25-(OH)VD in alcoholic cirrhosis patients with Child-Pugh C grade were significantly lower than those with Child-Pugh A and B ( t=3.969 and 2.911, P<0.05); the serum calcium levels in patients with Child-Pugh C grade were lower than those with Child-Pugh A( t=3.627, P<0.05); while the TP1NP levels in patients with Child-Pugh C were higher than those with Child-Pugh A and B grades( t=6.722 and 5.034, P<0.05).The Child-Pugh grade was negatively correlated with 25-(OH)VD level( β=-0.767, P<0.05)and positively correlated with TP1NP level ( β=2.186, P<0.05). Conclusion:The incidence of bone loss and osteoporosis in patients with alcoholic cirrhosis is increased significantly, and the deterioration of their liver function is closely associated with an increased TP1NP level and decreased 25-(OH)VD levels.