Objective To investigate the role of bufalin(BU)in inhibiting M2-type macrophage-mediated colorec-tal cancer metastasis.Methods Human acute leukemia mononuclear cells(THP-1)were differentiated into M0 macrophages using phorbol ester induction(PMA)for 48 hours.The M0 macrophages were then treated with IL-4 and IL-13 medium.Surface markers and morphological changes were observed through ELISA,morphology,and RT-qPCR experiments.RT-PCR and ELISA experiments were conducted to detect the surface markers TGF-β and IL-10 of M2 macrophages.The secretion level of IL-6 in the supernatant of M2 macrophages and colorectal cancer cells HCT116 was compared using ELISA.Additionally,the effect of conditioned medium on colorectal cancer cell HCT116 was assessed through Transwell,Wound healing,RT-qPCR,and Western blot experiments.Subsequent-ly,bufalin was added to the conditioned medium and the changes in AKT/PI3K protein,migration,and epithelial-mesenchymal transition ability in HCT116 were observed using Western blot,Transwell,Wound healing and RT-qPCR experiments.Results THP-1 were successfully differentiated into M2 macrophages.The activation of AKT/PI3K protein in HCT116 cells was induced by the secretion of IL-6 from M2 macrophages,which in turn promoted the migration and epithelial-mesenchymal transition ability of the HCT116 cells.The migration and epithelial-mes-enchymal transition mediated by M2 macrophages in HCT116 cells were effectively inhibited by Bufalin.Conclu-sion The release of IL-6 from M2 macrophages activates the AKT/PI3K signaling pathway in colorectal cancer cells,thereby promoting their migration and epithelial-mesenchymal transition capacity.Moreover,bufalin exhibits inhibitory effects on this effect.

Objective To explore the correlation between serum forkhead box protein O1(FOXO1)and myeloid cell leukemia protein 1(MCL1)in patients with acute ischemic stroke(AIS)and the severity and short-term prognosis of the disease.Methods A total of 359 AIS patients admitted to the hospital from Feb-ruary 2020 to February 2023 were regarded as the study subjects.According to the National Institutes of Health Stroke Scale(NIHSS)scores at admission,359 patients were grouped into mild group(131 cases),moderate group(156 cases),and severe group(72 cases),and the patients were separated into a good progno-sis group(263 cases)and a poor prognosis group(96 cases)based on the modified Rankin Scale(mRS)score after 90 days of onset.Real-time fluorescence quantitative PCR(qPCR)was applied to detect serum FOXO1 mRNA expression level.Enzyme linked immunosorbent assay(ELISA)was applied to detect the expression level of serum MCL1.Multivariate Logistic regression analysis method was applied to analyze the factors that affected the short-term prognosis of AIS patients.Receiver operating characteristic(ROC)curve was applied to analyze the predictive value of serum FOXO1 mRNA and MCL1 expression for short-term prognosis of AIS patients.Results Compared with the mild group,the serum FOXO1 mRNA level in the moderate and severe groups was obviously reduced(P<0.05),while the MCL1 level was obviously increased(P<0.05).The FOXO1 mRNA level in the severe group was lower than that in the moderate group(P<0.05),and the MCL1 level was higher than that in the moderate group(P<0.05).Compared with the good prognosis group,the serum FOXO1 mRNA level was significantly lower in the poor prognosis group,and the level of MCL1,C-reactive protein(CRP),age,NIHSS score,and proportion of diabetes were significantly higher than those in the poor prognosis group(t/χ2=11.328,7.617,5.344,2.314,16.788,4.459,all P<0.05).The re-sults of multivariate Logistic analysis showed that FOXO1 mRNA was a protective factor for the short-term prognosis of AIS patients(OR=0.726,P<0.05),while MCL1,CRP,and NIHSS score were independent risk factors affecting the short-term prognosis of AIS patients(OR=1.334,1.319,1.442,P<0.05).The results of ROC curve showed that the area under the curve(AUC)of serum FOXO1 mRNA level for predicting poor short-term prognosis of AIS patients was 0.807,with sensitivity and specificity of 69.79%and 85.93%re-spectively.The AUC of serum MCL1 level for predicting poor short-term prognosis of AIS patients was 0.824,with sensitivity and specificity of 71.87%and 84.79%,respectively.The AUC of the combination of serum FOXO1 mRNA and MCL1 for predicting poor short-term prognosis of AIS patients was 0.886,with sensitivity and specificity of 85.42%and 81.37%,respectively.Conclusion The expression of FOXO1 mRNA and MCL1 is abnormal in AIS patients,they are related to the severity of AIS and have high predictive value for the short-term prognosis of AIS.

Objective:To analyze the correlation between serum antibody titers of anti-contactin associated protein-like 2 (CASPR2) antibody and clinical features and prognosis in encephalitis.Methods:A retrospective analysis was conducted on the clinical data of 31 patients diagnosed with anti-CASPR2 antibody encephalitis at the First Affiliated Hospital of Zhengzhou University from January 2018 to April 2024. Patients were divided into low titer group (≤1∶32) and high titer group (>1∶32) based on serum anti-CASPR2 antibody titers, and their clinical characteristics, auxiliary examination results, and prognosis were compared between the two groups.Results:Among the 31 patients with anti CASPR2 antibody encephalitis (male∶female=1∶1.4), there were 16 cases in the low titer group and 15 cases in the high titer group; The age of patients in the high titer group was (33.9±17.9)years, which was lower than that of patients in the low titer group [(52.9±17.9)years], and the difference was statistically significant ( P=0.006). The proportion of patients with prodromal infection in the high titer group (6/15) was higher than that in the low titer group (1/16, P=0.037). There was no statistically significant difference in the cerebrospinal fluid related test results, imaging examination of intracranial abnormal lesions, abnormal electroencephalogram, serum abnormal tumor markers, and serum abnormal rheumatic immune indicators between the two groups of patients (all P>0.05). During hospitalization, one patient in the high titer group died; During the follow-up period, one patient died and three patients relapsed, all of whom were in the high titer group. During follow-up, the mRS scores of 6 patients ranged from 3 to 5 points (indicating functional impairment), with 4 cases in the high titer group and 2 cases in the low titer group. The proportion of patients with poor prognosis in the high titer group (9/15) was higher than that in the low titer group (2/16), and the difference was statistically significant ( P=0.021). Conclusions:Patients with high serum anti-CASPR2 antibody titers and encephalitis have a lower age of onset and are prone to pre infection triggers. High antibody titers may be associated with a higher risk of disease recurrence and poor prognosis for patients.

Objective:To explore the effect of acute kidney injury(AKI)on near-term survival after lung transplantation(LT)in patients with idiopathic pulmonary fibrosis(IPF).Methods:Through consulting electronic medical records, anesthetic modes and Chinese Lung Transplant Registration System, clinical data are retrospectively reviewed for 275 IPF patients undergoing LT at Affiliated Wuxi People's Hospital of Nanjing Medical University from January 2017 to April 2021.According to the diagnostic criteria of Kidney Disease: Improving Global Outcomes(KDIGO), they are divided into two groups of AKI(169 cases)and non-AKI(106 cases).Perioperative findings of two groups are recorded.Then univariate and multivariate Cox regression models are employed for determining whether or not inter-group differences existed in survival rates post-LT.Also AKI is staged according to the KDIGO.And the effect of stage 1/2/3 AKI on near-term postoperative prognosis is examined.Results:The differences are significantly different in recipient gender, creatinine, 6-minute walking test, forced vital capacity(FVC), lung allocation score, oxygenation index, N-terminal pro-brain natriuretic peptide(NT-Pro BNP), preoperative hormone use and volume of crystal infusion( P<0.05).After multivariate Cox regression correcting for covariates, no statistical significance exists in effect of AKI stage 1 on near-term postoperative survival rate( P<0.05).AKI stage 2/3 still has statistical significance in risk of mortality at Day 30/90/180/365 post-operation( P>0.05). Conclusions:As a common complication post-LT, AKI significantly affects near-term postoperative prognosis of transplant IPF patients.Stage 2/3 AKI impacts near-term postoperative survival while stage 1 AKI is not associated with higher mortality.

Objective:To investigate the clinical manifestation, genetic characteristics, treatment and prognosis of Crouzon-like syndrome.Methods:Clinical data of one case of Crouzon-like syndrome diagnosed in Children′s Hospital Affiliated to Zhengzhou University in May 2019 were collected, including clinical test, treatment plan, follow-up outcomes. The clinical characteristics and the mutation characteristics of IL11RA-related Crouzon-like syndrome were analyzed combined with the literature.Results:The male proband, five years and four months old, was admitted with the main clinical manifestations including headache, vomiting, exophthalmos, ocular hypertelorism, nasal root flat and scaphocephaly. CT showed that the cerebellar tonsil moved down slightly, the occipital magnum was full, the bilateral cranial plates were locally thinner, the bilateral cranial diameters were increased, and the cranial seams were closed. Magnetic resonance imaging showed ChiariⅠmalformation. The mutation c.40_63del and splice site mutation c.811-2A>G of the patient′s IL11RA gene were screened by whole exome sequencing. Sanger sequencing showed that the mutations are compound heterozygous and both are first reported. The mutation c.811-2A>G was derived from the patient′s mother, and the other one is de novo.Conclusions:The main clinical manifestations of Crouzon-like syndrome are craniosynostosis and midface hypoplasia and ocular deformity. The study identified two novel mutations in the Crouzon-like syndrome related IL11RA gene. Genetic sequencing is helpful for accurate diagnosis and timely surgical treatment.

Gut microbiota is involved in the physiological,biochemical and pathological processes of the body.It could participate in the immunoregulation and the development of enteric nervous system,prevent the invasion of pathogens,maintain normal bowel movement and inhibit the occurrence of tumor.Gut microbiota dysbiosis may also cause many diseases,such as inflammatory bowel disease,obesity and diabetes.A variety of diseases can affect gut microbiota with different ways.In addition,remote organs play a counteractive rule to gut microbiota.Maintaining the balance of the bacteria sets different requirements in different disease processes.This paper reviewed the advanced progression of gut microbiota dysbiosis after cholecystectomy and provided a new therapeutic target for gallbladder diseases.

Objective To compare endoscopic stenting with surgery for pyloric obstruction caused by unresectable gastric cancer.Methods Between June 2002 and June 2012,6 patients underwent endoscopic stenting and 70 did surgery for gastric outlet obstruction caused by gastric cancer.Results There were no significant difference in technical success rate and clinical success rate between the stenting and surgery groups (P ＞ 0.05).The stenting group had shorter time to oral intake,and shorter length of hospital stay [(2.5-± 3.1) d vs.(6.6 ± 3.5) d,t =-7.0,P ＜ 0.001].The incidence of early complications was significantly higher in the surgery group.However,the rates of late complications were significantly lower in the surgery group.Moreover,the surgery group was significantly associated with a longer patency duration [(295.8 d,95％ CI:260.7-330.8) vs.(168.2 d,95％ CI:134.7-201.7 d),P ＜0.001] and overall survival [(307.6 d,95％ CI:272.4-342.8 d) vs.(229.6 d,95％ CI:195.1-264.3 d),P =0.003].Conclusions Both endoscopic stenting and surgery are effective palliative therapy for gastric outlet obstruction caused by gastric cancer.Endoscopic stenting is associated with better shortterm outcomes.Surgery is preferable to ES in longer patency duration.

Objective To obtain alpaca single domain antibody targeting Her2.Methods An alpaca was immunized with human recombination Her2 protein mixed with Freund's adjuvant.Total RNA was extracted from the alpaca's blood and was used to synthesize first strand cDNA.Single domain antibody variable region (VHH) gene of the alpaca was amplified by PCR and cloned into pMES4 vector for library construction.After screening, E.coli BL21 (DE3) was transformed with selected clones and was induced with IPTG for the expression of recombinant proteins.The nanobody was purified by nickel ion affinity chromatography column.The affinity of the nanobodies to Her2 was tested.Results After the second round of screening, two antibody clones were selected, H3 and H5.The affinity of H5 was 8.106×10-10mol/L.Histochemistry results showed that H5 could recognize Her2 antigen in breast tumor tissue.Conclusion An Her2 specific nanobody derived from alpaca is obtained through phage display library screening, which can recognize human Her2 antibody in human breast tumor tissue.

Objective To evaluated the efficacy of biomarker?guided concurrent chemoradiotherapy in unrescetable esophageal squamous cell carcinoma patients. Methods 54 cases of unrescetable esophageal squamous cell carcinoma were enrolled in the prospective non?randomized clinical study and divided into study group and control group. All cases were treated with concurrent chemoraditherapy. Intensity?modulated radiation therapy was used with a dose of 60?66 Gy. Chemotherapy was perfromed on day 1 and d29. In the study group the selection of the chemotherapy drug was based on the excision repair cross?complementation 1 ( ERCC1) ,thymidylate synthetase ( TYMS) ,ribonucleotide reductase M1( RRM1) ,and theβ?tubulin isotypeⅢ( TUBB3) mRNA expression levels. In the control group,the regiment for chemotherapy was Cisplatin plus Fluorouracil. The objective response rate and overall survival ( OS ) were calculated using Kaplan?meier method and log?rank test was used for between?group comparison. The survival rate was calculated using Kaplan?Meier method and analyzed using log?rank method, other comparison was performed by χ2 test. Results The follow?up rate was 100% in the study group and 96% in the controll group. The objective response rate of the study group and the control group were 85% and 86 ( P=0. 483 ) , respectively. The median survival time ( MST) in the study group was 35. 5 months and that in the control group was 25. 8 months. The 1?,2?,and 3?year OS rates of the study group and the control group were 84%,68%,46% and 71%,59%,28% respectively (P=0. 047).No significant differences were observed in the incidence of side?effects in the two groups. Conclusions Selecting the chemotherapy drug according to biomarker,combined with radiation therapy,could improve survival.in unrescetable esophageal squamous cell carcinoma. The value needs further investigation.