Child, Preschool ; Humans ; Male ; Neonatology ; Research Report ; Sarcoma, Myeloid ; physiopathology

Aged ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; pathology ; Urinary Bladder ; pathology

BACKGROUND:Infrapatelar fat pad is often partialy resected in the knee surgery, which can be used as an important source of adipose-derived mesenchymal stem cels. OBJECTIVE: To explore the strategies of isolation, culture, and identification of adipose-derived mesenchymal stem cels from the infrapatelar fat pad and to detect the expression of cel surface markers of human adipose-derived stem cels. METHODS: Infrapatelar fat pad was obtained from patients undergoing knee arthroscopy surgery, and attached cels were obtained from adipose tissue by using colagenase I. Cels were cultured in 10% low-sugar DMEM. Stem cels proliferation was detected by means of MTT and then, cel growth curve was made. The obtained cels were induced and differentiated into adipocytes and osteocytes. Expressions of cel surface markers CD29 and CD44 were detected. RESULTS AND CONCLUSION:A few of attached cels were observed after cultured 24 hours. Cels proliferated faster and exhibited spindle shape after 1 week. Cel adherence and proliferation were speeded up after subculture. Growth curve of cels exhibited that the passages 5 and 2 cels had higher reproductive activity than passage 8 cels. The obtained cels can be induced and differentiated into adipocytes and osteocytes. Results from flow cytometry showed that 96.8% passage 5 cels expressed CD29 and 97.6% expressed CD44. These findings indicate that high-purity adipose-derived mesenchymal stem cels with high reproductive ability are easy to be isolated from the infrapatelar fat pad, which may be a kind of ideal seed cels for cartilage tissue engineering.

BACKGROUND:Because insulin-like growth factor I has the ability to induce mesenchymal stem cells into chondrocytes, we hypothesized that the chondrogenic diferentiation of adipose-derived stem cels can be improvedvia insulin-like growth factor I transfection. OBJECTIVE:To investigate the influence of insulin-like growth factor I transfection on chondrogenic potential of adipose-derived stem cels in vitroand tumor necrosis factor-like weak inducer of apoptosis (TWEAK)/fibroblast growth factor-inducible 14 (Fn14) signaling pathway. METHODS:Recombinant lentivirus plasmid pLVX-IGF-I-IRES-ZsGreenl was constructed and transferred into passage 3 adipose-derived stem cels, and then these stem cels were induced to diferentiate into chondrocytes (experimental group 1). Meanwhile, the cels transfected with pLVX-IRES-ZsGreenl were taken as green fluorescent protein/adipose mesenchymal stem cel group (experimental group 2), and those with no transfection acted as control group. RESULTS AND CONCLUSION:The mRNA expression of TWEAK was reduced in the experimental group 1 as compared with the other two groups, but the mRNA expressions of insulin-like growth factor I, Col2a1 and Sox9 were up-regulated in the experimental group 1. At the same time, the protein expression of matrix metaloproteinase-3 and TWEAK were down-regulated, while the protein expression of Col2a1 was increased in the insulin-like growth factor I-transfected cels in contrast to the cels modified with pLVX-IRES-ZsGreenl or with no transfection. These findings indicate that pLVX-IGF-I-IRES-ZsGreenl transfection of adipose-derived stem cels results in a higher expression of insulin-like growth factor I, and down-regulates the expression of TWEAK mRNA and protein, which improves the diferentiation of adipose-derived mesenchymal stem cels into chondrocytes.

Objective To approach the effect of fluoride on the expression of thyroid peroxidase(TPO)activity and TPO mRNA in primary porcine thyrocytes.Methods Purified cultured porcine thyrocytes waft made into sodium fluoride model,and were divided according to the final concentration of NaF into 0(control group),40,80,160 mg/L.After exposed to NaF for 48 h,the morphology of the porcine thyrocytes was investigated with acridine orange staining method,TPO activity was measured with upgrade guaiacol method and RT-PCR method was used to detect the ratio of TPO/β-actin.Results The major changes included apoptotic bodies and cell fragments in the 80,160 mg/L groups under phase contrast microscope.With the increasing dose of fluoride.TPO activity,being(3.103±0.090),(1.944±0.025),(1.361±0.008),(0.668±0.026)U/L,respectively,had obviously lowered with a statistical significance compared between the groups(F=1563.864,P＜0.05).The TPO activity had a negative correlation with the dose of fluoride(r=-0.955,P＜0.05).With the dose of fluoride increasing,the expression of TPO mRNA had obviously lowered,being(0.947±0.013),(0.634±0.018),(0.448±0.028)and (0.210±0.009)with a statistical significance in group comparison(F=2713.855,P＜0.05).Conclusion Fluoride affects the thyroid via inhibiting TPO activity and expression of TPO mRNA.

Objective To construct PPAR?and PPAR?response element (PPRE)-controlled luciferase expression vectors,and to determine whether the traditional Chinese medicine emodin activates PPAR?and improves the glucose uptake by HepG2 hepatocytes.Methods (1) PPAR?and PPRE luciferase expression vectors were constructed and were applied to screen more than 20 ingredients of the traditional Chinese medicine. (2) HepG2 cells were incubated with emodin which can activate PPAR?and PPRE luciferase activity,and the PPAR?mRNA expression level was evaluated by RT-PCR/Southern blot.(3) PPAR?and glucose transporter 2 (Glut2) proteins were determined by Western blot analysis in HepG2 cells treated with emodin.(4) The glucose uptake rate was measured using 2-deoxy-[~3H]-D-glucose in HepG2 cells after treatment with emodin.Results (1) Emodin stimulated luciferase activity controlled by PPRE in dose-dependent manner at concentrations of 0.04 to 180?mol/L in COS-7 cells.The highest value was about 4 folds of control in the cells treated with 90?mol/L emodin (P

Objective To investigate the osteogenesis capability of different frequency extracorporeal shock wave.Methods 39 rabbits received different frequency extracorporeal shock wave at the middle potion of tibia for 3 or 7 days,these rabbits were then sac rificed and the tibia bones were collected to process for HE and toluidine blue staining,the pathological changes were observed under the light microscope.Results After different frequency extracorporeal shock wave treatment,the typical periosteal reaction were observed,external periosteum bleeded and thickened but there was no reaction at internal periosteum,marrow cavity opened and fibrosed.the osteoblast-1iking cell proliferated,however,no cartilage cells were observed;The rabbits received 7 days shock wave treatment showed more severe reaction than those for 3 days.The shock wave at lower frequency showed more severe reaction than higher frequency.Conclusion shock wave induced osteogenesis through the periosteal reaction of external periosteum;the osteogenesis capability of different frequency extracorporeal shock wave were affected by the frequency.Higher frequency of shock wave was not the ideal way to promote osteogenesis.

Background: Creatine transporter (CRTR) deficiency is the most common creatine deficiency syndrome, of which the final diagnosis relies on mutation in the X-linked CRTR gene. To date, more than 90 mutations in the SLC6A8 gene have been reported. This paper discusses a novel mutation detected via the thorough sequencing of all the X-chromosome-specific exons investigated in a four and a half year old boy with an intellectual disability, speech and language delay and motor disturbance. Methods: A brain magnetic resonance imaging (MRI) and a proton magnetic resonance spectroscopy (MRS) were carried out, the creatine and creatinine concentrations in the urine were checked and all exons were sequenced. Results: A detailed clinical investigation revealed a reduction in the cerebral creatine levels in the brain by the MRS, elevated creatine and creatinine concentrations in the urine and signal abnormalities in the left frontal cortex of the brain by the MRI. A novel change was identified in the heterozygosity of the exon 10: c.1395-c.1401 deletion. Conclusion: The use of a combination of powerful new technologies, such as thorough exome-nextgeneration sequencing and a brain MRS, should be considered, in order to determine any neurometabolic diseases, especially when the signal abnormalities in the brain MRI cannot be explained by any other factors. This mutation results most likely in a dysfunction of the creatine transport and synthesis, hence causing central nervous system symptoms.
Carrier Proteins

Objective To analyse the high-dose dexamethasone suppression test(HDDST)-related differences in the clinical and biochemical features of the patients with Cushing's disease Methods Cases were drawn from 60 consecutive patients with Cushing's disease,who were then divided into two groups according to the response to the HDDST.The clinical and biochemical features between two groups were compared.Results(1) Of the 60 patients with Cushing's disease,23.3%(14/60)of patients(group A)did not yield results of suppression with the HDDST,and the others(group B)did.No difference was found in the age[(33.8?10.4 vs 36.2?11.2)years]and duration of illness[(2.1?1.6 vs 3.9?3.1)years]between two groups.(2)In clinical features,the patients in group A were more likely to have edema of lower limbs(64.3% vs 32.6%),hypokalemia (71.4% vs 28.3%),secondary diabetes(57.1% vs 26.1%)and purple striae(85.7% vs 54.3%,all P

Proteomic analysis is an effective way to identify protein constituent in Lewy bedy-like inclusions (or aggresome) in vitro. Exposure to synthetic proteasome inhibitor (PSI, 10 μmol/L) for 48 hours was used to induce the formation of cytoplasmic proteineous inclusions (termed as PSi-induced inclusions) in PC12 cells.The proteomic approaches of biochemical fractionation, two-dimensional electrophoresis (2-D) and identification via peptide mass fingerprints (PMF) were deployed, and 20 protein components of LBs were identified,i ncluding 2 proteins involved in the production of synaptic neurotransmitter, 6 subunits of the 26 S proteasome,2 cytoskeleton proteins, 2 subunits of mitochondrial complexes, 1 anti-oxidant protein, and 7 chaperone proteins and (or) chaperone-like proteins. The results suggested that these LB protein components might had been recruited in PSI-induced inclusions formed in PC12 cells under the condition of proteasome inhibition.