To investigate the changes in peripheral blood immune cells before and after the onset of septic shock in patients with malignant tumors, and to analyze the relationship between these immune cells and patient prognosis.

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 observe the effects of Qigui Buxue Decoction combined with timed press-needle therapy on myelosuppression and cancer-related fatigue in chemotherapy patients with non-small cell lung cancer(NSCLC)of lung-spleen qi deficiency type.Methods A total of 124 NSCLC patients with lung-spleen qi deficiency type admitted to the Department of Radiotherapy and Chemotherapy at Cangzhou Integrated Traditional Chinese and Western Medicine Hospital in Hebei Province from January 2022 to June 2024 were selected as the study subjects.The patients were randomly divided into an observation group and a control group using a random number table,with 62 cases in each group.Both groups received platinum-based two-drug combination chemotherapy.The control group received timed press-needle therapy in addition to chemotherapy,while the observation group received Qigui Buxue Decoction in addition to the control group's treatment.The treatment course was 3 weeks,with a total of 2 courses.After 2 courses of treatment,changes in blood routine,lymphocyte subsets(CD3+,CD4+,CD8+,NK cells),CD4+/CD8+ratio,and the incidence of myelosuppression were observed.Changes in Piper Fatigue Scale and Karnofsky Performance Scale(KPS)scores before and after treatment were compared between the two groups.Results(1)After treatment,the white blood cell count in the observation group significantly improved(P＜0.05),and the observation group showed significantly better improvement in white blood cell count compared to the control group,with a statistically significant difference(P＜0.05).(2)After treatment,the levels of CD3+,CD4+,CD4+/CD8+,and NK cells in both groups significantly improved(P＜0.05),and the observation group showed significantly better improvement in these levels compared to the control group,with a statistically significant difference(P＜0.05).(3)The incidence of myelosuppression in the observation group was 58.06%(36/62),while it was 77.42%(48/62)in the control group.The incidence of myelosuppression in the observation group was significantly lower than that in the control group,with a statistically significant difference(P＜0.05).(4)After treatment,the Piper Fatigue Scale scores,including behavioral,emotional,physical,and total scores,significantly improved in both groups(P＜0.05),and the observation group showed significantly better improvement in these scores compared to the control group,with a statistically significant difference(P＜0.05).The cognitive scores of the Piper Fatigue Scale showed slight improvement in both groups,but there was no statistically significant difference compared to before treatment(P＞0.05).(5)After treatment,the KPS scores and TCM syndrome scores significantly improved in both groups(P＜0.05),and the observation group showed significantly better improvement in these scores compared to the control group,with a statistically significant difference(P＜0.05).Conclusion Qigui Buxue Decoction combined with timed press-needle therapy can effectively improve myelosuppression,enhance immune function,and alleviate cancer-related fatigue in NSCLC patients with lung-spleen qi deficiency type after chemotherapy,thereby improving their quality of life.This approach is worthy of clinical promotion.

The identification and optimization of mutations in nanobodies are crucial for enhancing their thera-peutic 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 work-flow,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 mu-tations to predict their impact on binding affinity to the targets.The results showed that multiple mu-tations 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 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 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 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.

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.