Objective: To isolate a lytic phage targeting K. pneumoniae type K39 from untreated sewage and systematically analyze its biological characteristics and genomic information. Methods: The bacteriophage of K. pneumoniae K39 Kp 1000 was identified based on the sequence of the capsular polysaccharide gene. It was isolated and purified using the double agar plate method. The morphology of the phage was observed through negative staining and transmission electron microscopy,and its bacteriophage spectrum was evaluated by phagocytosis. The biological characteristics of the bacteriophage were assessed by determining the optimal multiplicity of infection(MOI),constructing a one-step growth curve,and conducting temperature and pH tolerance tests. Finally,the phage DNA was extracted,and its whole genome was sequenced using the Illumina Hi Seq2000 sequencing platform. The sequencing results were then annotated and analyzed. Results: A lytic phage specifically targeting K. pneumoniae type K39was successfully isolated and named IME330. Transmission electron microscopy showed that the head diameter of the phage was(75 ± 1) nm and the tail length was(185 ± 1) nm. The lytic spectrum analysis revealed that IME330 lysed K. pneumoniae type K39,demonstrating a narrow host range and high specificity. The optimal multiplicity of infection(MOI) was 0. 1,The lysis amount reached 168 PFU/cell. Physicochemical experiments indicated that IME330 exhibited strong tolerance to high temperatures and a broad p H range(pH 4-10). Genomic analysis demonstrated that the IME330 genome had a length of 144,245 bp,a molecular weight of 46,463 MDa,G +C content of 44. 8%,and encoded 320 open reading frames(ORFs). Conclusion IME330 is a novel lytic K.pneumoniae phage belonging to the order Caudovirales. This phage exhibits strong tolerance to physicochemical factors(e. g.,temperature,acidic/alkaline conditions) and demonstrates high lytic activity,while its lytic spectrum remains narrow with strict host specificity.

Lung cancer is one of the most lethal malignancies in the world, with non-small cell lung cancer (NSCLC) accounting for approximately 80%-85% of all pathological types. Approximately 30%-55% of NSCLC patients develop brain metastases. It has been reported that 5%-6% of patients with brain metastases harbor anaplastic lymphoma kinase (ALK) fusion. ALK-positive NSCLC patients have shown significant therapeutic benefits after treatment with ALK inhibitors. Over the past decade, ALK inhibitors have rapidly evolved and now exist in three generations: first-generation drugs such as Crizotinib; second-generation drugs including Alectinib, Brigatinib, Ceritinib, and Ensartinib; and third-generation drugs like Lorlatinib. These drugs have exhibited varying efficacy in treating brain metastases in ALK-positive NSCLC patients. However, the numerous options available for ALK inhibition present a challenge for clinical decision-making. Therefore, this review aims to provide clinical guidance by summarizing the efficacy and safety of ALK inhibitors in treating NSCLC brain metastases.
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Humans ; Carcinoma, Non-Small-Cell Lung/drug therapy* ; Lung Neoplasms/drug therapy* ; Brain Neoplasms/drug therapy* ; Protein Kinase Inhibitors/adverse effects* ; Crizotinib

OBJECTIVETo compare the releasing of growth factors between platelet-rich fibrin (PRF) and platelet-rich plasma (PRP) as well as their effects on the proliferation and differentiation of adipose tissue-derived stem cells (ADSCs) in vitro.

METHODSBlood was taken from central artery of rabbits, acquiring PRF was acquired through one time centrifuge and PRP through two times centrifuge. Five milliliters of fresh alpha-MEM was added to PRF and PRP and incubated at 37 degrees C. The time points to collect exudates was in day 1, 7, 14, 21, 28 and the mass concentrations of transforming growth factor-beta1 (TGF-beta1) and platelet derived growth factor-AB(PDGF-AB) were quantified in PRF and PRP. Then the exudates of PRF and PRP were used to culture ADSCs and evaluate the effects of PRF and PRP on proliferation and differentiation of ADSCs.

RESULTS1) Growth factor release: In the PRF exudates at different time points, the mass concentration of TGF-beta1 was the highest at day 14 and the highest mass concentration of PDGF-AB at day 7, the mass concentration of both TGF-beta1 and PDGF-AB was the highest at the first day and then gradually declined. 2)The effect on proliferation and differentiation: PRF exudates of day 14 expressed the maximum proliferation and alkaline phosphatase (ALP) activity. PRF exudates of day 1 demonstrat the maximum proliferation and ALP activity.

CONCLUSIONComparing to PRP, PRF releases growth factors gradually and expressed stronger and more durable effect on proliferation and differentiation of ADSCs in vitro.

Adipose Tissue ; Blood Platelets ; Cell Differentiation ; Fibrin ; In Vitro Techniques ; Intercellular Signaling Peptides and Proteins ; Organic Chemicals ; Platelet-Derived Growth Factor ; Platelet-Rich Plasma ; Stem Cells ; Transforming Growth Factor beta1