Recent data suggest that vascular endothelial growth factor receptor inhibitor (VEGFRi) can enhance the anti-tumor activity of the anti-programmed cell death-1 (anti-PD-1) antibody in colorectal cancer (CRC) with microsatellite stability (MSS). However, the comparison between this combination and standard third-line VEGFRi treatment is not performed, and reliable biomarkers are still lacking. We retrospectively enrolled MSS CRC patients receiving anti-PD-1 antibody plus VEGFRi (combination group, n=54) or VEGFRi alone (VEGFRi group, n=32), and their efficacy and safety were evaluated. We additionally examined the immune characteristics of the MSS CRC tumor microenvironment (TME) through single-cell and spatial transcriptomic data, and an MSS CRC immune cell-related signature (MCICRS) that can be used to predict the clinical outcomes of MSS CRC patients receiving immunotherapy was developed and validated in our in-house cohort. Compared with VEGFRi alone, the combination of anti-PD-1 antibody and VEGFRi exhibited a prolonged survival benefit (median progression-free survival: 4.4 vs. 2.0 months, P=0.0024; median overall survival: 10.2 vs. 5.2 months, P=0.0038) and a similar adverse event incidence. Through single-cell and spatial transcriptomic analysis, we determined ten MSS CRC-enriched immune cell types and their spatial distribution, including naive CD4⁺ T, regulatory CD4⁺ T, CD4⁺ Th17, exhausted CD8⁺ T, cytotoxic CD8⁺ T, proliferated CD8⁺ T, natural killer (NK) cells, plasma, and classical and intermediate monocytes. Based on a systemic meta-analysis and ten machine learning algorithms, we obtained MCICRS, an independent risk factor for the prognosis of MSS CRC patients. Further analyses demonstrated that the low-MCICRS group presented a higher immune cell infiltration and immune-related pathway activation, and hence a significant relation with the superior efficacy of pan-cancer immunotherapy. More importantly, the predictive value of MCICRS in MSS CRC patients receiving immunotherapy was also validated with an in-house cohort. Anti-PD-1 antibody combined with VEGFRi presented an improved clinical benefit in MSS CRC with manageable toxicity. MCICRS could serve as a robust and promising tool to predict clinical outcomes for individual MSS CRC patients receiving immunotherapy.
Humans ; Colorectal Neoplasms/drug therapy* ; Male ; Female ; Immunotherapy ; Middle Aged ; Aged ; Tumor Microenvironment/immunology* ; Retrospective Studies ; Microsatellite Instability ; Transcriptome ; Single-Cell Analysis ; Programmed Cell Death 1 Receptor/immunology* ; Gene Expression Profiling ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; Receptors, Vascular Endothelial Growth Factor/antagonists & inhibitors*

Gastrointestinal (GI) cancers are a leading cause of cancer morbidity and mortality worldwide. Despite advances in treatment, cancer relapse remains a significant challenge, necessitating novel therapeutic strategies. In this study, we engineered nanobody-based chimeric antigen receptor (CAR) natural killer (NK) cells targeting cadherin 17 (CDH17) for the treatment of GI tumors. In addition, to enhance the efficacy of CAR-NK cells, we also incorporated CV1, a CD47-SIRPα axis inhibitor, to evaluate the anti-tumor effect of this combination. We found that CDH17-CAR-NK cells effectively eliminated GI cancers cells in a CDH17-dependent manner. CDH17-CAR-NK cells also exhibit potent in vivo anti-tumor effects in cancer cell-derived xenograft and patient-derived xenograft mouse models. Additionally, the anti-tumor activity of CDH17-CAR-NK cells is synergistically enhanced by CD47-signal regulatory protein α (SIRPα) axis inhibitor CV1, likely through augmented macrophages activation and an increase in M1-phenotype macrophages in the tumor microenvironment. Collectively, our findings suggest that CDH17-targeting CAR-NK cells are a promising strategy for GI cancers. The combination of CDH17-CAR-NK cells with CV1 emerges as a potential combinatorial approach to overcome the limitations of CAR-NK therapy. Further investigations are warranted to speed up the clinical translation of these findings.

Autoimmune hepatitis （AIH） is an inflammatory disease caused by immune dysfunction， and its pathogenic mechanism remains unclear. In recent years， a large number of studies have shown that iron homeostasis imbalance and ferroptosis are closely associated with the pathogenesis and progression of AIH. This article reviews the pathological mechanism and impact of iron overload and ferroptosis in AIH， in order to provide new insights and theoretical bases for research on the mechanism and clinical treatment of AIH.

By sorting out the literature related to constitution and referring to the concept and method of "element"， many ancient and modern theories of constitution are deconstructed into multi-dimensional "constitution elements". These constitution elements are divided into two levels： basic and derivative. The basic level is the most basic element of constitution， including zang-fu organs/body and material. The zang-fu organs/body dimension focuses on the fixed functional units in the human body， and the material dimension focuses on the rheology functional units of human body. The derivative level is the secondary physiological or psychological state in life activities， including emotion and qi transformation. The emotional dimension focuses on the state of human psychological activities， and the qi transformation dimension focuses on the state of human physiological activities in cold， heat， dampness， and dryness. Based on this， we can further study the influence of factors such as age， gender， region， and social class， on the combination rule of "constitution element"， so as to form a more comprehensive classification program of constitution.

In order to achieve the purpose of rapid detection of duck astrovirus type 1(DAstV-1),specific primers were designed based on the conservative region of ORF1a which belonged to DAstV-1(WF1202 strain).A real-time fluorescent quantitative PCR(qPCR)detective method for DAstV-1 was established.Clinical samples were detected by the qPCR method and the positive samples were used for virus isolation and identification.Results showed that the detection limit of the established method was 4.64×103 copies/μL,which was 10 times higher than the normal RT-PCR method.In addition,no cross-reactions were found with other common infectious disease pathogens in poultry,indicating that the qPCR method had good specificity.What's more,the coef-ficient of variations(Cv)in intra-and inter-assays were 0.85％-2.85％and 0.21％-2.94％,re-spectively,both less than 3％,indicating that the qPCR method had a good repeatability.Using this method,35 tissue samples from different duck farms in 10 provinces from 2020 to 2022 were detected for DAstV-1.Results showed that the positive rate was 25.71％(9/35),and the coinci-dence rate was 94.29％when compared with the normal RT-PCR method.A positive sample ran-domly taken for the virus isolation through duck embryo passage,and the allantoic fluid was col-lected and then was verified by the qPCR method and inoculated with 1-day-old healthy ducklings for the animal regression experiment.The infected ducklings suffered from transient disease but did not die.The liver tissues were all positive with DAstV-1 when detected by qPCR.Meanwhile,autopsy showed that there were slight changes in the livers,and the histopathological observation showed that the liver cells were steatosis.These findings indicated that the isolated DAstV-1 strain had weak pathogenicity and might be a low virulent strain.To sum up,the qPCR detection method of DAstV-1 was successfully established in this work,and could provide technical support for clini-cal diagnosis,isolation and identification,and molecular epidemiology monitoring of DAstV-1.

In order to establish a rapid,specific and sensitive detection method for Streptococcus equi subspecies zooepidemicus(SEZ)and to understand the infection status of SEZ in horses ente-ring China,specific primers were designed and synthesized based on the conserved gene comB of standard strain SEZ(ATCC 43079)in this work.Then,the pMD19-T-comB recombinant plasmid was constructed and used as a standard positive template.After that,the fluorescence-based quantitative PCR(qPCR)detection method based on SYBR Green Ⅰ dye was established.Totally,477 equine entry serum samples from 6 countries,including Netherlands,Belgium,Japan,Germa-ny,Argentina and New Zealand,during 2018 to 2023,were randomly selected and detected for SEZ by the qPCR method.Results showed that the established qPCR method had specific amplification for only SEZ,which illustrated a good specificity.Sensitivity test of the method showed that the limited detection amount was 4.58 X101 copies/μL.And the repeatability test showed that the coef-ficient of variation of intra-batch repeatability was less than 0.5％,while the inter-batch repeat-ability was less than 3.0％,which indicated good repeatability and high stability.Retrospective a-nalysis showed that totally 11 of 477 positive samples were detected,with a relatively low positive rate of 2.31％(11/477).Among them,all the 40 samples from Netherlands in 2018 were negative(0/40).In the samples of 2019,one positive was detected from Belgium(1/20),while all other 36 samples which form Japan and Germany were negative.In the samples of 2021,three samples(3/34)from Japan and one sample(1/20)from Argentina were positive,and all the other 40 samples from the Netherlands were negative.In the samples of 2022,76 samples from Netherlands were all negative.While in the 2023,5(5/126)of 126 samples from Netherlands and one(1/88)of 88 from New Zealand were found positive with SEZ.To summarize,The SYBR Green Ⅰ qPCR method for the diagnosis of SEZ was successfully established,and it could provide necessary technical support for the rapid quarantine of China's entry-exit and port departments,as well as the epidemiological investigation of the disease.

Objective To explore the clinical efficacy of single unicompartmental knee arthroplasty(UKA)and total knee arthroplasty(TKA)for the treatment of knee osteoarthritis.Methods A total of 21 patients who underwent TKA and 15 who underwent UKA were randomly recruited.Biomechanical tests were performed before surgery and at 6th and 12th month after surgery.A Vicon infrared motion capture system and Kistler three-dimensional force plate were used to simultaneously acquire the kinematic and kinetic data of the patients during stair walking.Results During stair ascent,the peak knee flexion moment in the TKA group was significantly lower than that in the UKA group;the time to peak knee flexion/adduction moment,knee flexion moment impulse,and load rate of the peak knee adduction moment in the UKA group were significantly lower than those in the UKA group.During stair descent,the peak knee extension power in the UKA group was significantly lower before surgery and at 6th month after surgery;the load rate of the peak vertical ground reaction force was significantly higher before surgery and the peak knee extension moment was significantly greater at 6th month after surgery;at 12th month after surgery,there was no significant difference in the biomechanical characteristics during stair ascent and descent.Conclusions The TKA and UKA groups showed similar knee joint function after surgery;however,compared with the UKA group,the TKA group may adopt a different lower extremity biomechanical pattern.The UKA group showed better quadriceps control after surgery and improved postural control during stair descent,whereas the TKA group adopted a conservative stair gait strategy to reduce the knee load.Compared with the peak moment,the time to peak moment and load rate of the peak moment were more sensitive indicators for determining the difference in the knee load.

Objective To investigate the predictive value of a decision tree-based machine learning model for prognosis in acute paraquat(PQ)poisoning(APP)patients.Methods A retrospective study was conducted.The clinical data of APP patients from Cangzhou Central Hospital between May 2012 and August 2021 were collected,including gender,age,time from ingestion to gastric lavage,proportion of hemoperfusion,serum PQ concentration,biochemical indicators[white blood cell count(WBC),alanine aminotransferase(ALT),serum creatinine(SCr),serum amylase,and serum potassium],and blood gas indicators[arterial blood lactic acid(Lac),base excess(BE),and arterial partial pressure of oxygen(PaO2)].Patients were divided into a survival group(n = 56)and a death group(n = 74)based on 90-day prognosis,and the clinical data between the two groups were compared.The multivariate Logistic regression analysis was conducted to analyze the risk factors of prognosis in APP patients,and two decision tree models(i.e.,with/without serum PQ concentration)were constructed based on the risk factors.The predictive value was evaluated by the receiver operator characteristic(ROC)curve,and the area under the ROC curve(AUC)of two decision tree models was compared by Hanley&McNeil method.Results The 90-day survival rate of the patients was 43.1%(56/130).Compared with death group,patients in the survival group had lower WBC[×109/L:8.9(7.0,11.6)vs.17.4(11.9,23.1)],ALT[U/L:25.3(21.2,31.8)vs.29.3(23.2,40.3)],SCr[μmol/L:64.0(53.0,74.0)vs.91.0(72.5,141.5)],Lac[mmol/L:2.5(1.4,4.0)vs.7.1(3.7,11.0)],and serum PQ concentration[ng/L:0.3(0.1,0.9)vs.2.9(1.9,8.1)],the difference were statistically significant(all P<0.05),higher BE[mmol/L:-2.5(-4.2,-1.1)vs.-7.2(-10.9,-4.7)]and serum potassium[mmol/L:3.7(3.5,4.0)vs.3.2(2.8,3.7)],the difference were statistically significant(all P<0.05),and patients were younger[years:33.5(26.0,47.8)vs.42.5(26.0,58.0),P<0.05].Univariate Logistic regression analysis showed that age,WBC,ALT,SCr,serum potassium,Lac,BE and serum PQ concentration were independent risk factors of 90-day survival[odds ratio(OR)and 95%confidence interval(95%CI)were 1.03(1.01-1.05),1.30(1.18-1.44),1.04(1.01-1.07),1.02(1.01-1.04),7.59(3.25-17.70),1.64(1.35-1.99),1.51(1.29-1.76),7.00(3.41-14.37),P values were 0.018,<0.001,0.011,<0.001,<0.001,<0.001,<0.001,<0.001].Multivariate Logistic regression analysis with serum PQ concentration showed that WBC,serum potassium,and serum PQ concentration were independent risk factors for 90-day survival[OR and 95%CI were 1.17(1.03-1.33),7.29(1.66-32.01),5.49(2.48-12.13),P values were 0.014,0.008,<0.001].Multivariate Logistic regression analysis without serum PQ concentration showed that age,WBC,serum potassium and BE were independent risk factors for 90-day survival[OR and 95%CI were 1.05(1.01-1.08),1.20(1.07-1.34),3.12(1.01-9.66),1.41(1.16-1.72),P values were 0.008,0.002,0.049,0.001].The decision tree model based on serum PQ concentration and serum potassium showed an AUC of 0.94(95%CI was 0.89-0.98),along with 91.9%sensitivity,89.3%specificity,and 90.0%accuracy.The decision tree model based on WBC,BE,and age showed an AUC of 0.89(95%CI was 0.84-0.95),with 86.5%sensitivity,91.1%specificity,and 88.5%accuracy.Pairwise comparison of the AUC using Hanley&McNeil method demonstrated that no statistical difference between the two decision tree models(Z = 1.34,P = 0.180).Conclusion The decision tree-based models can provide quantitative and intuitive prediction tools for the early detection of prognosis in APP patients in clinical practice.

Objective:To evaluate the immunogenicity of a quadrivalent influenza virus subunit vaccine in a healthy population aged 3-64 years.Methods:Healthy people aged 3-64 years old were selected as the study subjects, and a randomized, blind, positive controlled, non-inferiority test was adopted. The subjects were randomly inoculated with one dose of the corresponding experimental vaccine or control vaccine in a ratio of 1∶1. Blood samples were collected from all subjects before and at 28 d after immunization, and hemagglutination inhibition (HI) test was used to measure the levels of antibodies against H1N1, H3N2, B/Victoria (BV) and B/Yamagata (BY) in serum. The geometric mean titers (GMT), geometric mean increase (GMI), positive conversion rates and protection rates of antibodies against the four types of viruses were analyzed.Results:A total of 2 157 subjects aged 3-64 years were included, including 1 074 in the experimental group and 1 083 in the control group. There were no significant differences in the GMT or protection rates of antibodies against H1N1, H3N2, BV or BY before immunization between the two groups ( P>0.05), and the two groups were balanced at baseline. After full immunization, the GMI of antibodies to H1N1, H3N2, BV and BY in the experimental group was 11.16, 17.77, 9.61 and 15.13, respectively; the positive conversion rates were 84.08% (903/1 074), 92.46% (993/1 074), 86.03% (924/1 074) and 91.71% (985/1 074), respectively; the protection rates were 96.74% (1 039/1 074), 97.58% (1 048/1 074), 88.08% (946/1 074) and 94.97% (1 020/1 074), respectively. In the experimental group, the GMT of each antibody increased by more than 2.5 times after immunization; the lower limit of the 95%CI of the positive conversion rate was higher than 40%, and the lower limit of the 95%CI of the protection rate was higher than 70%. The lower limit of the 95%CI of the difference in the positive conversion rate of each antibody between the experimental group and the control group was >-10%, and the lower limit of the 95%CI for GMT (experimental group)/GMT (control group) was over 2/3. Conclusions:The experimental vaccine had good immunogenicity and was non-inferior to the control vaccine in the population aged 3-64 years.

Objective To explore the effect of mucle force on contact force, peak pressure and contact area of foot joint in in vitro biomechanical experiment of foot and ankle, so as to provide references for choosing appropriate loading modes. Methods In neutral position of the ankle joint, fresh calf and foot specimens were simulated with or without mucle force loading. The contact force, peak pressure and contact area of the 1st metatarsophalangeal joint, the 2nd metatarsophalangeal joint, the 1st tarsometatarsal joint, the 2nd tarsometatarsal joint, the medial cuneonavicular joint, the intermediate cuneonavicular joint, the talonavicular joint, the calcicocuboid joint, the subtalar joint ( posterior articular surface) and the tibiotalar joint of normal foot under loading were measured, the results are compared and analyzed. Results Under muscle force loading, the contact force of the 1st metatarsophalangeal joint, the 2nd metatarsophalangeal joint, the 1st tarsometatarsal joint,the 2nd tarsometatarsal joint, the medial cuneonavicular joint, the intermediate cuneonavicular joint, the talonavicular joint and the tibiotalar joint were significantly greater than those without muscle force loading (P<0. 05), and the change percentages were 719. 28% , 311. 37% , 128. 67% , 50. 82% , 54. 89% , 57. 63% ,79. 98% and 50. 34% , respectively. The peak pressures of the 1st metatarsophalangeal joint , the 1st tarsometatarsal joint and the talonavicular joint under muscle force loading were significantly higher than those without muscle force loading ( P < 0. 05), and the change percentages were 176. 14% , 62. 91% and 40. 07% ,respectively. The contact area of the 1st metatarsophalangeal joint, the 1st tarsometatarsal joint, the intermediate cuneonavicular joint and the subtalar joint ( posterior articular surface) under muscle force loading increased significantly (P<0. 05), and the change percentages were 132. 20% , 55. 41% , 30. 97% and 26. 87% , respectively. Conclusions In biomechanical experiment of foot and ankle specimens, muscle force loading has a significant effect on contact force, peak pressure and contact area of each foot joint, especially the forefoot.Therefore, it is necessary to consider the effect of muscle force loading on stress of foot and ankle in the study ofrelated in vitro specimens