Objective To achieve arabinose-controlled expression of HtrA strain and detect the expression of HtrA protein.Methods Arabinose promoter with htrA100 was amplified from pACD-htrA vector by PCR and cloned into pGP704 vector.Then, Shigella flexneri 2a strain 301 was transferred with the recombinant plasmid pGD-htrA and an AraC-expression vector .The expressions of HtrA in whole-cell and periplasmic space were detected by Western blotting .Results The suicide plasmid-mediated homologous recombinant vector and the inducible HtrA expression strain were successfully constructed.Without arabinose,HtrA protein was hardly detected ,but in the presense of arabinose , HtrA protein could be detected in whole-cell lysate and in periplasmic space lysate by Western blot .Conclusion Homologous recombination using suicide plasmid can significantly knock down the expression of HtrA protein .After being induced with arabinose , HtrA protein can be expressed normally .

OBJECTIVE: To investigate the computed tomography findings, clinicopathological features, genetic characteristics and prognosis of in situ adenocarcinoma (AIS) and minimally invasive adenocarcinoma (MIA) of the lung. METHODS: We retrospectively analyzed the data including computed tomography (CT) images, histopathological findings, Ki-67 immunostaining, and genetic mutations in patients with lung adenocarcinoma undergoing surgery at our hospital between 2014 and 2019. RESULTS: Of the total of 480 patients with lung adenocarcinoma we reviewed, 73 (15.2%) had AIS (=28) or MIA (=45) tumors. The age of the patients with MIA was significantly younger than that of patients with AIS ( < 0.02). CT scans identified pure ground-glass nodules in 46.4% of AIS cases and in 44.4% of MIA cases. Multiple GGOs were more common in MIA than in AIS cases ( < 0.05), and bluured tumor margins was less frequent in AIS cases ( < 0.05). No significant difference was found in EGFR mutations between MIA and AIS cases. A Ki-67 labeling index (LI) value ≥2.8% did not differentiate MIA from AIS. The follow-up time in MIA group was significantly shorter than that in AIS group, but no recurrence or death occurred. CONCLUSIONS Despite similar surgical outcomes and favorable survival outcomes, the patients with AIS and MIA show differences in terms of age, CT findings, EGFR mutations and Ki-67 LI.
Adenocarcinoma of Lung ; diagnostic imaging ; pathology ; ErbB Receptors ; genetics ; Humans ; Ki-67 Antigen ; genetics ; Lung Neoplasms ; diagnostic imaging ; pathology ; Mutation ; Prognosis ; Retrospective Studies ; Tomography, X-Ray Computed