ABSTRACT
Objective: To investigate the feasibility of isolation and culture of human adenoid-derived mesenchymal stem cells (aMSCs) in vitro, and to observe the differentiation of aMSCs into olfactory sensory neurons. Methods: Adenoid tissues surgically removed from children with adenoid hypertrophy in the Second Xiangya Hospital of Central South University from September to November of 2020 were collected. The adenoid tissues were digested and isolated by trypsin and then cultured with adhesion method. The expressions of cell surface antigens CD45, CD73 and CD90 on aMSCs of P5 generation were tested by flow cytometry, and the ability of osteogenic and adipogenic induction were used to identify cell differentiation ability. Then, aMSCs were induced into differentiation by retinoic acid (RA), sonic hedgehog (SHH), basic fibroblast growth factor (bFGF), RA+SHH, RA+bFGF, SHH+bFGF and RA+SHH+bFGF, respectively. The morphology of differentiated cells was observed under inverted microscope. The expression of β-tubulin 3, which was the specific marker of sensory neuron, the expressions of growth associated protein-43 (GAP43) and olfactory maker protein (OMP), which were the specific markers of olfactory sensory neuron, were detected by immunofluorescence antibody assay. The expression intensities were compared by Chi-square test of four-grid table data. Results: aMSCs were successively isolated and cultured from human adenoid tissues. P0 cells generation had good adhesion and proliferation performance. P2 cells were basically purified. P5 cells expressed CD73 and CD90 with the purity of 99.3% and 99.75% respectively, without CD45 expression. P5 cells had a good ability of osteogenic differentiation and adipogenic differentiation. Neuron-like morphology and expression of β-tubulin 3 were found in differentiated cells after induced by RA, SHH, or bFGF, respectively. An induction of expression of GAP43 was found in differentiated cells of bFGF+SHH group and RA+SHH+bFGF group, without expression of OMP of each group. The intensity of GAP43 expression of RA+SHH+bFGF group was stronger than that of bFGF+SHH group (χ2=17.48, P<0.005). Conclusions: aMSCs can be cultured from human adenoid tissues, with the stably passaged and good differentiation ability. As a new population of mesenchymal stem cells, aMSCs have the neuroregenerative properties and could differentiate into immature olfactory sensory neurons under the induction of RA+SHH+bFGF in vitro.
Subject(s)
Child , Humans , Hedgehog Proteins , Olfactory Receptor Neurons , Tubulin , Adenoids , Osteogenesis , Cell DifferentiationABSTRACT
BACKGROUND: In the clinic, 25%-35% heel pain is mostly caused by plantar fasciitis. Previous studies mainly focused on plantar fasciitis with heel pain caused by flat foot and the growth of calcaneus bone spur. There are no reports on other reasons for plantar fasciitis in large-sample studies. OBJECTIVE: Using the real-time shear wave elastography, CT scan and X-ray, the anatomic site and thickness of normal two-dimensional ultrasound standard flat plantar aponeurosis were identified to analyze the relationship between plantar elastic characteristics and plantar arch angle from non-weight-bearing to weight-bearing position so as to explore the correlation of plantar fasciitis with plantar elastics and plantar arch angle. METHODS: Fifty healthy volunteers (feet) were selected as the healthy control group. 100 cases of plantar fasciitis (one foot) were selected as the case group. Plantar arch angle from non-weight-bearing to weight-bearing position was obtained using X-ray and CT scan to identify anatomic site of plantar aponeurosis in both groups. Two-dimensional ultrasound and real-time shear wave elastography were utilized to obtain thickness and elastic modulus of plantar aponeurosis. RESULTS AND CONCLUSION: (1) From non-weight-bearing to weight-bearing, arch angle change value was (16.4±4.5)° in the healthy control group and (10.5±3.5)° in the case group. Significant differences in arch angle change were detected between the two groups (P <0.01). (2) Thickness of plantar fascia was obviously smaller in the healthy control group (2.4±0.3) mm than in the case group (3.5±0.9) mm. Elastic modulus of plantar fascia was obviously larger in the healthy control group (30.1±1.3) kPa than in the case group (9.1±1.2) kPa. Thickness of plantar fascia and elastic modulus of plantar fascia were significantly different between the two groups (P < 0.01). (3) In summary, real-time shear wave elastography combined with CT and X-ray images can investigate the morphological and elastic characteristics of the plantar aponeurosis from many aspects. Arch angle change is strongly associated with elastic modulus of plantar fascia. The decreased elastic modulus of plantar fascia is possibly one of the reasons for arch angle change from non-weight-bearing to weight-bearing conditions, and is probably one of reasons for plantar fasciitis with heel pain.
ABSTRACT
<p><b>OBJECTIVE</b>To prepare reference samples of Mycobacterium tuberculosis culture filtrate protein-10 (CFP-10) and CFP10-streptavidin fusion proteins (CFP10/SA) for time-resolved fluoroimmunoassay (TRFIA).</p><p><b>METHODS</b>The CFP10 gene was amplified by PCR from Mycobacterium tuberculosis strain H37Rv and cloned into pET24b, pET24b-streptavidin (SA) or pET21a-SA expression vectors. The recombinant proteins CFP10, CFP10-SA and SA-CFP10 were expressed in Rosetta cells, purified via nickel affinity chromatography and refolded by dialysis. The sensitivity and stability of the resultant proteins as reference samples were evaluated by double-antibody sandwich TRFIA.</p><p><b>RESULTS</b>CFP10-SA and SA-CFP10 fusion proteins were expressed as inclusion bodies, whereas CFP10 was expressed in a soluble form. The resultant purity of the 3 recombinant proteins all exceeded 95%. TRFIA results showed that CFP-SA fusion protein possessed the best sensitivity (0.02 µg/L) and stability.</p><p><b>CONCLUSION</b>The reference samples of CFP10 for TRFIA detection have been successfully prepared and can be used in the development of a diagnostic kit for Mycobacterium tuberculosis.</p>
Subject(s)
Bacterial Proteins , Genetics , Reference Standards , Fluoroimmunoassay , Methods , Gene Amplification , Mycobacterium tuberculosis , Reference StandardsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of early continuous hyperthermic peritoneal perfusion (CHPP) on patients of advanced gastric carcinoma after surgical resection.</p><p><b>METHODS</b>Seventy patients were divided into control and treatment group. Patients in treatment group received CHPP 1 or 2 days postoperatively. Treatment was administered daily consecutively for 4 days. Patients in control group underwent resection of gastric carcinoma without CHPP. Chemotherapy was administered of Calcium Folinate and 5-Fluorouracil (LF regimen) intravenously for 6 cycles in both groups two or three weeks postoperatively. Survival and recurrence in both groups were observed and compared.</p><p><b>RESULTS</b>One-year survival and recurrence rates were 83.3% and 8.3% in treatment group, 79.4% and 11.7% in control group, there were no significant differences between the two groups. Three-year survival and recurrence rates were 63.9% and 19.4 in treatment group, 39.8% and 44.1% in control group, there were significant differences between the two groups.</p><p><b>CONCLUSION</b>Early administration of CHPP to patients with advanced gastric carcinoma after surgery may be advantageous for preventing peritoneal metastasis and recurrence, thus may prolong survival time.</p>