Establishment of a cell model of insulin-resistant 3T3-L1 adipocytes / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes.</p><p><b>METHOS</b>Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10(-8), 10(-7), and 10(-6) mol·L(-1)) were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulin- resistant 3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1µmol·L(-1) DEX, and the changes of glucose concen- tration in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay.</p><p><b>RESULTS</b>Treatment of 3T3-L1 cells with DEX, IBMX and 10(-6) mol·L(-1)) insulin for 9 days resulted in the differentiation of >90% of the cells into mature adipocytes. IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 µmol·L(-1) DEX showed significantly increased glucose consumption (P=0.0003) as compared with the control group at 36 h (P<0.001).</p><p><b>CONCLUSION</b>3T3-L1 cells can be induced into mature adipocytes by exposure to 1 µmol·L(-1) DEX, 0.5 mmol·L(-1) IBMX and 10(-6) mol·L(-1)) insulin. A 96 h exposure to 1 µmol·L(-1) DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.</p>

Effects of rutaecarpine on inflammatory cytokines in insulin resistant primary skeletal muscle cells / 中国中药杂志

It is now well established that inflammation plays an important role in the development of numerous chronic metabolic diseases including insulin resistance (IR) and type 2 diabetes (T2DM). Skeletal muscle is responsible for 75% of total insulin-dependent glucose uptake; consequently, skeletal muscle IR is considered to be the primary defect of systemic IR development. Our pre- vious study has shown that rutaecarpine (Rut) can benefit blood lipid profile, mitigate inflammation, and improve kidney, liver, pan- creas pathology status of T2DM rats. However, the effects of Rut on inflammatory cytokines in the development of IR-skeletal muscle cells have not been studied. Thus, our objective was to investigate effects of Rut on inflammatory cytokines interleukiri (IL)-1, IL-6 and tumor necrosis factor (TNF)-α in insulin resistant primary skeletal muscle cells (IR-PSMC). Primary cultures of skeletal muscle cells were prepared from 5 neonate SD rats, and the primary rat skeletal muscle cells were identified by cell morphology, effect of ru- taecarpine on cell proliferation by MTT assay. IR-PSMC cells were induced by palmitic acid (PA), the glucose concentration was measured by glucose oxidase and peroxidase (GOD-POD) method. The effects of Rut on inflammatory cytokines IL-1, IL-6 and TNF-α in IR-PSMC cells were tested by enzyme-linked immunosorbent assay (ELISA) kit. The results show that the primary skeletal muscle cells from neonatal rat cultured for 2-4 days, parallel alignment regularly, and cultured for 7 days, cells fused and myotube formed. It was shown that Rut in concentration 0-180. 0 μmol x L(-1) possessed no cytotoxic effect towards cultured primary skeletal muscle cells. However, after 24 h exposure to 0.6 mmol x L(-1) PA, primary skeletal muscle cells were able to induce a state of insulin resistance. The results obtained indicated significant decrease (P < 0.05 to P < 0.001) IL-1, IL-6 and TNF-α production by cultured IR-PSMC cells when incubating 24 hours with Rut, beginning from 20 to 180.0 μmol x L(-1). IL-1, IL-6 and TNF-α in the Rut treated groups were dose-dependently decreased compared with that in the IR-PSMC control group. Our results demonstrated that the Rut promoted glucose consumption and improved insulin resistance possibly through suppression of inflammatory cytokines in the IR-PSMC cells.

Effect of activation of cellular immunity on p58+ cells expressing killer-cell-inhibitory receptor cells / 中国实验血液学杂志

<p><b>UNLABELLED</b>The purpose of this study was to evaluate the effects of cellular immunity activation on P58(+) cells expressing killer cell inhibitory receptor (KIR) and their regulatory function on cellular immunity, and provid theoretical data for preventing graft-vers-host disease (GVHD) in stem cell transplantation therapy. The mononuclear cells from human peripheral blood were incubated with IL-2, Con A and Lipostin (LP) for 72 hours. The KIR expressing cells, P58.1(+) and P58.2(+) cells, were analyzed by flow cytometry. The results showed that the percentages of CD3(+), CD4(+), CD8(+), CD16(+)CD56(+), P58.1(+) and P58.2(+) cells were greatly increased after treated with IL-2, Con A and LP, separately or in combination, and the percentages of above cells in combined treatment groups were higher than those of single stimulated groups, especially the percentage of cells in the IL-2 + LP group was significantly higher than those in IL-2 and LP singly treated groups.</p><p><b>IN CONCLUSION</b>IL-2, Con A and LP possess the ability to induce the expression of KIR and stimulate proliferation of P58.1(+) and P58.2(+) cells while to activate the celluar immunity response, the expression of P58 gene may be regulated by the activation of cellular immunity.</p>

Effect of 36 item neuro-motor examination on early diagnosis of children with cerebral palsy within 1 year-old / 中国康复理论与实践

@#Objective To evaluate the effect of 36-neuro-motorial examination on early diagnosis of children with cerebral palsy within 1 year-old.Methods36-neuro-motorial examination was analysed in 210 children with cerebral palsy from 2 to 12 months. Results4-29 items reflecting the abnormity of reflex, muscle tone, posture and motion were observeed in patients with cerebral palsy.If these abnormities were laid according as frequency among 2-6 months,reflex abnormities are knee tendon reflex, palmar grasp reflex,Babinski sign,asymmetrical tonic neck reflex,ankle clonus,sitting equilibrium reflex,stepping reflex,crossed extension reflex,tonic labyrinthine reflex,trunk incurvation reflex,Moro reflex; posture abnormities are head to back≥15°,from supine to the side position, axillar suspension reflex,posterior neck fovea≥1cm in the supine position,landau reflex,Vojta reflex,traction reflex,tonic torticollis,abnormal spontaneous posture;muscle tone abnormities are foot dorsiflexion angle,scarf sign,tonic palmar grasp,to up the foot heel≥30° in the stand position,thumb decussation to the palm,poples angle,adductor angle,heel-ear angle; others are head cycle≤-2s, squint, active movement attenuation or abnormity, auditory abnormity, visual abnormity, ophthalmodonesis, epilepsy.Conclusions 36-neuro-motorial examination is effective for early diagnosis of cerebral palsy within 1 year.