ObjectiveTo study the effect of titanium particles on the proliferation, differentiation, and cytomorphology of osteoblasts, and to explore the possible internal relations and mechanism. MethodsCalvarial osteoblasts were separated from 10 newborn Sprague Dawley rats by repeated enzyme digestion, and were cultured in vitro. The cells were identified by alkaline phosphatase (ALP) staining and alizarin red staining. The cells at passage 3 were cultured with titanium particles culture medium at concentrations of 0.01, 0.05, 0.1, 0.5, and 1 mg/mL (0.01, 0.05, 0.1, 0.5, and 1 mg/mL groups). The absorbance (A) values were detected by cell counting kit 8 at 7 days after cultured to compare the effect of titanium particles at different concentrations on proliferation, and median lethal concentration was screened out. The expression of collagen type I was detected by ELISA to observe the effect of titanium particles on differentiation. The osteoblasts co-cultured with titanium particles of median lethal concentration (experimental group) for 7 days, and double fluorescence staining with FITC-phalloidine and propidium iodide was performed. The cytomorphology variation of osteoblasts after swallowing titanium particles was observed under laser scanning confocal microscope. The osteoblasts at passage 3 cultured with culture medium without titanium particles served as control group. ResultsThe cultured cells were identified as osteoblasts by ALP staining and alizarin red staining. Different concentrations of titanium particles could inhibit osteoblasts proliferation and differentiation in varying degrees, showing significant difference when compared with the control group at 7 days after culture (P<0.05). The cell proliferation and differentiation were decreased with increased titanium particles concentration; significant differences were found between the other groups (P<0.05) except 0.01 and 0.05 mg/mL groups (P>0.05). The median lethal concentration of titanium particles was 0.5 mg/mL. Laser scanning confocal microscope showed cellular shrinking, microfilaments distortion, pseudopodia contraction of osteoblasts that swallowed titanium particles in the experimental group. ConclusionTitanium particles can inhibit proliferation and differentiation of osteoblasts. The effect may be related to variation of cytomorphology after swallowing titanium particles.
ObjectiveTo explore the clinical diagnosis and treatment of Talaromyces marneffei (TM) infection by bone marrow examination, and to clarify the important role and significance of bone marrow smear and pathogenic examination.MethodsRetrospective analysis was conducted on a case of disseminated TM infection that was clearly diagnosed through bone marrow related examination. Literature review of TM infection was conducted by retrieving relevant case reports at home and abroad from 1990 to 2018.ResultsThe patient was a 23-year-old man with recurrent cough and onset of fever, superficial lymph node enlargement in multiple parts of the body, no abnormal chest CT sign, and poor efficacy in anti-infection treatment. The patient developed progressive abdominal pain, accompanied by systemic papulosis, decreased consciousness, and progressive decline of blood cells. The patient underwent bone marrow puncture surgery in our hospital, and developed circulatory and respiratory failure half an hour after surgery. TM was confirmed bybone marrow smear and pathogenic culture. In the literature review, 2 855 cases of TM infection were retrieved, among which the majority of cases were confirmed through blood and bone marrow related examination. The positive rate of bone marrow culture was significantly higher than that of blood culture (72.4% and 66.8%, respectively, P=0.007). Compared with bone marrow culture, the misdiagnosis and missed diagnosis rate of bone marrow smear microscopy was 27.6%. Patients diagnosed with TM infection by bone marrow examination had the highest HIV positive rate (95.7%).ConclusionsThe examination of bone marrow cells and the culture are of great clinical significance for the diagnosis of TM infection. TM infection should be identified in patients suspected of HIV positive with fever, lymph node enlargement and abnormal blood routine.