This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.

This studyobtained the proteins of virulence factors EsxA and EsxB from Mycobacterium haemophilum and explored their interactions both in vitro and in vivo.The aim was to lay a foundation for further analysis of the functional mechanisms of EsxA and EsxB,and to further the development of drugs targeting Mycobacterium haemophilum.On the basis of bioinformatics analysis of the virulence factors EsxA and EsxB from Mycobacterium haemophilum,we performed molecular cloning,using Mycobacterium haemophi-lum as a template to construct recombinant plasmids EsxA-pGl01,EsxB-pGl01,and EsxB-pET-29b.The proteins of EsxA,EsxB,and their complex were expressed with a prokaryotic system,then purified through nickel-affinity chromatography and gel filtration chromatography.Intracellular colocalization was determined through fluorescence colocalization.Prokaryotic expression systems for EsxA,EsxB,and their complex were successfully constructed,and purified proteins were obtained.Fluorescence colocalization indi-cated that EsxA and EsxB interacted in vivo.In vivo fluorescence colocalization and in vitro complex formation suggested that EsxA and EsxB interacted both intracellularly and extracellularly.Our findings lay a foundation for studying the pathogenic mechanisms of the virulence factors EsxA and EsxB in Mycobacterium haemophilum,and performing structural analysis of their complex.

To further explore the current situation of disease communication between cancer patients and their families in clinical practice, and provide decision-making reference for the construction of harmonious doctor-patient relationship. From September to November, 2021, 10 oncologists from a third-class A general hospital in X city were selected for semi-structured interviews by objective sampling method, and Colaizzi seven-step analysis was used to analyze, refine and summarize the interview data. The results showed that the current situation of disease communication between cancer patients and their families can be summarized into three themes: concealing information during the diagnosis period, embarrassing decision-making during the treatment period, and difficult in autonomy at the end stage of the disease. Based on this situation, this paper suggested that a clinical communication mode with medical staff as a bridge and patients and their families actively facing and participating together can be constructed from different dimensions such as disease notification, win-win cooperation and psychological care.

Objective With a computer-assisted program,retinal vascular calibers were measured quantitatively.In this study the relationship between retinal vascular calibers and components of the metabolic syndrome was examined.Methods A total of 450 hypertensive patients were collected.Medical history,physical examination,blood tests,and retinal photographs were taken.Retinal vascular calibers were measured quantitatively from digital retinal photographs.In the hypertensive population the associations of retinal vascular calibers with components of the metabolic syndrome were described,and the factors that influenced retinal vascular calibers were analyzed.Results In the enrolled population,mean age was (57.53 ± 10.01) years,mean systolic blood pressure (138 ± 17) mm Hg(1 mm Hg =0.133 kPa),diastolic blood pressure (84 ± 10) mm Hg.Mean central retinal arteriolar equivalent(CRAE) was(129.26 ± 12.68) μm,and mean central retinal venular equivalent (CRVE) (198.25 ± 18.37) μm.After adjusting for age,gender,etc,CRAE in group with poor blood pressure control was smaller than that in the group with good blood pressure control [(126.45 ± 15.74) μm vs (130.30 ± 11.30) μm,P =0.029].CRAE tended to be narrower with worsened blood pressure control (P =0.075).CRVE was smaller in patients with normal high density lipoprotein-cholesterol (HDL-C) than in those with abnormal level [(197.36 ±17.62) μm vs (203.07 ± 21.52) μm,P =0.040].The diastolic blood pressure was raised along with the decreasing CRAE(P=0.009).And the HDL-C level was reduced as CRVE was increasing(P=0.042).Old age (r =-0.090,P=0.013) and poor blood pressure control(r=-0.098,P=0.038) were independent risk factors for narrow CRAE,while lowered HDL-C (r =0.105,P =0.024) and smoking (r =0.141,P =0.010) were independent risk factors for wide CRVE.Conclusions Narrow CRAE was related to poor blood pressure control,while wide CRVE was related to lowed HDL-C.Aging and poor blood pressure control were independent risk factors for narrow CRAE,while lowed HDL-C and smoking were independent risk factors for wide CRVE in the hypertensive patients.