Objective To investigate the effects of protein kinase C (PKC) on proliferation, extracelluar matrix synthesis and transcriptional factor SOX9 (SRY-related high mobility group-box gene 9) expression of rat growth plate chondrocytes in vitro. Methods Rat costochondral growth plate chondrocytes (RGC) were isolated and cultured. The 1st serum free cultured passage RGCs were treated with 1, 10 and 100 nmol/L phorbol 12,13-dibutyrate (PDBu), PKC agonist, cell morphology were observed with inverted microscopy, cell proliferation, COLLAGEN and GAG synthesis were detected by isotope incorporation COLLAGEN TYPEⅡ and AGGRECAN mRNA transcription and SOX9 expression were revealed by RT-PCR and Western blot.Results 100 nmol/L PDBu treatment made the cell morphology of serum free cultured RGC closed to serum group and inhibited proliferation but promoted COLLAGEN and AGGRECAN synthesis, 3H-TdR、3H-proline and 35S-sulfate incorporation of 100nmol/L PDBu group were as 83%, 52.6% and 146.5% as those of serum free control(P

Objective To investigate the different distribution and expression of mammalian target of rapamycin complex (mTORC) in the kidney of diabetic nephropathy (DN) mice.Methods Fourteen eight-week-old male C57BL/6 mice were assigned to 2 groups: the control group ( n=7 ) and the streptozotocin ( STZ )-induced DN group ( n=7 ) . Blood and urinary variables including glucose , albumin, creatinine and albumin/creatinine ratio were assessed 2 weeks after STZ injection.Hematoxylin-eosin staining was performed for renal pathological analyses .The distributions of mTOR , phosph-ser2448-mTOR(p-mTOR), mTORC1(Raptor), mTORC2(Rictor) and phosph-ser240/244-S6K1 (p-S6K1) were determined by immunofluorescence.The expression levels of mTOR, p-mTOR, mTORC1(Raptor), mTORC2(Rictor), S6K1 and p-S6K1 were detected by Western blotting .Results Two weeks after STZ injection , the diabetic mice developed albuminuria (P<0.01) and renal hypertrophy (P<0.05).The immunofluorescence positive staining for mTOR , Raptor, and Rictor was distributed in the epithelial cells of proximal tubules , glomerular mesangium and capillary loops as well as the medullary collecting ducts of the control mouse kidney .These positive signals increased in the DN mouse kidney ( P<0.05).However, pS6K1 was not detected in the inner medulla of control mouse and p-mTOR was not found in the glomeruli of both control and DN mice .Conclusion mTORC is widely expessed in the mouse kidney and participates in the development of DN , whereas the 2448 serine phosphorylation of mTOR may be not implicated in the hyperglycemia mediated glomerular injury .

Objective To observe the effect of single intensive hyperbaric oxygenation (HBO) on cytochrome C and caspase-3 in rats af-ter permanent middle cerebral artery occlusion (MCAO) very early. Methods Forty-eight male Sprague-Dawley rats were subjected to per-manent MCAO model using the intraluminal suture method, and were divided into control group (n=24) and HBO group (n=24). The HBO group stayed in the hyperbaric cabin with a pressure of 0.2 MPa for 9 hours 3 hours after MCAO. They were measured with Garcia scores 3 hours, 13 hours and 24 hours after MCAO. Apoptosis cells of ischemic penumbra tissue were investigated with TUNEL 13 hours and 24 hours after MCAO, while the level of cytochrome C and caspase-3 were measured with ELISA. Results The Garcia scores increased 13 hours and 24 hours after MCAO in both groups, but there was no significant difference between groups (t<2.07, P>0.05). The apoptosis cells were found in both groups 13 hours and 24 hours after MCAO, and less in the HBO group than in the control group (t>6.57, P<0.01). The levels of cytochrome C and caspase-3 were less in the HBO group than in the control group 24 hours after MCAO (t>2.41, P<0.05). Conclusion A single intensive HBO in very early stage may improve neurological function after cerebral ischemia in rats, which may associ-ate with the inhibition of cytochrome C and caspase-3 to reduce cell apoptosis.

Nitrogen source is one of the important factors that affect the microalgae growth and lipid accumulation. We studied the effects of various nitrogen sources (i.e. NaNO3, CO(NH2)2, NH4Cl and CH3COONH4) and amount on the growth density, lipid yield, and eicosapentaenoic acid (EPA) content of Nannochloropsis oculata by single factor experimental method. The results show that N. oculata preferred NH4+ as nitrogen source rather than NO3- and CO(NH2)2. NH4+ could promote the growth and lipid accumulation of N. oculata. With the increase of nitrogen concentration, the biomass and the content of polyunsaturated fatty acid (PUFA) increased, but the content of lipid decreased. CH3COONH4 was the most suitable for growth, accumulation of lipid and EPA of N. oculata among the four investigated nitrogen sources. The optimal concentration was 5.29 mmol/L.
Eicosapentaenoic Acid ; metabolism ; Lipids ; biosynthesis ; isolation & purification ; Microalgae ; growth & development ; metabolism ; Nitrogen ; analysis ; metabolism ; Stramenopiles ; growth & development ; metabolism

Objective To investigate the risk factors for hemorrhagic transformation (HT) and poor outcomes in patients with acute ischemic stroke after mechanical thrombectomy.Methods The patients with acute ischemic stroke received mechanical thrombectomy were enrolled retrospectively.The demography,vascular risk factors and other clinical data of the patents were collected.The modified Rankin scale (mRS) was used to evaluate the clinical outcomes at day 90.Good outcome was defined as mRS score 0-2.The patients were divided into either a HT group or a non-HT group according to their HT conditions.Multivariate logistic regression analysis was used to identify the independent risk factors for HT and poor outcomes.Results A total of 48 patients with acute ischemic stroke received mechanical thrombectomy were enrolled,including 25 males (52.1％).Their mean age was 64.77± 9.14 years.The mean National Institutes of Health Stroke Scale (NIHSS) score was 17.70 ± 3.77.Twenty-two patients (45.8％) occured HT,of which 9 were symptomatic HT;24 (50.0％) had good outcomes.The proportion of males in the HT group was significantly lower than that in the non-HT goup (30.4％vs.72.0％;x2 =8.293,P =0.004),while the proportions in patients with diabetes (65.2％ vs.36.0％;x2 =4.090,P =0.043) and atrial fibrillation (78.3％ vs.44.0％;x2 =5.880,P =0.015),as well as the baseline fasting blood glucose level (8.514 ± 4.400 mmol/L vs.6.354 ± 1.472 mmol/L;t =2.319,P =0.025) were significantly higher than those in the non-HT group.Multivariate logistic regression analysis showed that the atrial fibrillation (odds ratio [OR] 6.136,95％ confidence interval [CI] 1.617-23.291;P =0.042) was a risk factor for the occurrence of HT after mechanical thrombectomy.The proportion of diabetic patients (29.2％ vs.70.8％;x2 =8.333,P=0.04) and baseline NIHSS score (16.050±4.865 vs.19.210±4.423);t=2.310,P=0.026) of the good outcome group were significantly lower than those of the poor outcome group,while the proportions of patients in atrial fibrillation (75.0％ vs.45.8％;x2 =4.269,P =0.039),anterior circulation stroke (87.5％ vs.62.5％;x2 =4.000,P =0.046) middle cerebral artery (75.0％ vs.29.2％;x2 =10.113,P =0.006),vertebral basilar artery (37.5％ vs.12.5％;x2 =10.113,P =0.006) occlusion and parenchymal hematoma (33.3％ vs.4.1％;P=0.011) were significantly higher than the poor outcome group.Multivariate logistic regression analysis showed that diabetes (OR 5.898,95％ CI 1.699-20.479;P=0.005),baseline NIHSS score (OR 1.167,95％ CI 1.011-1.347;P =0.035),and parenchymal hematoma (OR 1.295,95％ CI 1.099-1.875;P=0.028) were the independent risk factors for poor outcomes.Conclusions Atrial fibrillation is an independent predictor of HT risk in patients with acute ischemic stroke after mechanical thrombectomy.Diabetes mellitus,higher baseline NIHSS score,and concurrent brain parenchymal hematoma are the independent predictors of poor outcomes.Therefore,the risk of HT and adverse outcomes should be fully assessed before mechanical thrombectomy in patients with acute ischemic stroke.

Exosomes are vesicles released into extracellular cells by fusion of intracellular polyvesicles and cell membranes. Exosomes are secreted by a variety of cells and exist in a variety of body fluids. Exosomes contain proteins, lipids and nucleic acids, which can participate in the information exchange between cells. Exosomes secreted by CNS cells are involved in the occurrence and development of CNS diseases. Recent studies have shown that exosomes can not only serve as early diagnostic markers for CNS diseases, but also have therapeutic potential for CNS diseases. In this paper, the development, diagnosis and treatment of CSF exosomes in CNS diseases were reviewed.

Objective To observe the effect of hyperbaric oxygenation (HBO) on apoptosis-inducing factor (AIF) and Caspase-3 levels in rats with permanent middle cerebral artery occlusion (MCAO),and to elucidate the apoptosis pathways.Methods Sixty Sprague-Dawley rats were subjected to permanent MCAO and then randomly divided into a control group and an HBO group,each of 30.Three hours later the rats of the HBO group were put into a hyperbaric cabin held at a pressure of 0.2 MPa for 9 hours.They inhaled supplementary oxygen at the 1st,3rd,5th,7th and 9th hour while the rats in the control group inhaled air at normal pressure.The neurological outcome was measured at the 3rd,13th and 72nd hour after the MCAO using Garcia scores.Apoptosis in the tissue of the ischemic penumbra,nuclear and mitochondrial AIF and Caspase-3 levels were measured at the 13th and 72nd hours after the modeling.Results The scores were significantly higher at the 13th hour than after the 3rd hour in both groups,and then even higher at the 72nd hour.Apoptosis was evident in the ischemic penumbra at the 13th and 72nd hours in both groups,but the number of cells was less at the 72nd hour than at the 13th hour in the control group.There was significantly less apoptosis in the HBO group than in the control group at the 13th hour.The average AIF level had significantly decreased in the nuclei and increased in the mitochondria by the 72nd hour compared with the 13th hour in both groups.The average levels of nuclear AIF at the 13th hour and the 72nd hour were lower than those in the mitochondria.But they were significantly higher in the HBO group than in the control group at the same time points.The levels of Caspase-3,normally zero,had increased by the 13th hour in both groups.The average level of Caspase-3 was significantly lower in both groups at the 72nd hour than at the 13th hour.Conclusions HBO can improve neurological function,inhibit the transfer of AIF from the mitochondria to the nucleus and reduce Caspase-3 levels.The mechanism may involve reducing apoptosis through caspase-dependent and caspase-independent pathways in the mitochondria.

By searching the literature on the application of entrustable professional activities in college education in China and globally, this article comprehensively analyzes the concept of entrustable professional activities, the development of evaluation items, the effectiveness of clinical application, the problems to be improved, and research prospects, so as to provide a useful reference for the reform and evaluation of competency-oriented medical education in China and the application of entrustable professional activities that can be repeated and promoted in clinical teaching.