Objective To explore the effects of hyperglycemia and oxidized low density lipoprotein (ox-LDL) on the differentiation of macrophage derived THP-1 monocytes. Methods THP-1 human monocytic leukemia cell line was cultured in vitro, and the differentiation of THP-1 cells into macrophages was induced by phorbol esters. The macrophages were then incubated with the absence of D-glucose and ox-LDL (control group), 30 mmol/L D-glucose (hyperglycemia group), 100 μg/mL ox-LDL (ox-LDL group) or 30 mmol/L D-glucose and 100 μg/mL ox-LDL(G-ox-LDL group) for 24 h. High performance liquid chromatography was used for qualitative and quantitative analysis of intracellular cholesterol and cholesteryl esters. Both light microscope with red oil O staining technique and transmission electron microscope were employed to observe the morphology of treated and control THP-1 cells. Results A large number of intracellular red oil O stained granules and lipid vacuoles were observed in ox-LDL group and G-ox-LDL group, the contents of total cholesterol and cholesteryl esters were significantly higher than those of control group (P<0.05), and the contents of cholesteryl esters were higher than 50% of total cholesterol in both groups. However, only a few intracellular red oil O stained granules and lipid vacuoles were observed in control group and hyperglycemia group, there was no significant difference in the contents of total cholesterol and choleateryl esters between control group and hyperglycemia group (P>0.05), and the contents of cholesteryl esters were less than 50% of total cholesterol in both groups. Conclusion Foam cells form when THP-1 cells are incubated with ox-LDL, while hyperglycemia alone can not convert THP-1 cells to foam cells, indicating that ox-LDL is necessary for the macrophages derived THP-1 monocytes to turn into foam cells.

Meisoindigo is an indigo natural derivative commonly used in anti-cancer therapy. In the clinical application, it was also found to have good therapeutic effect on psoriasis. In order to further understand its mechanism of action, human normal keratinocyte cell line HaCaT and RAW 264.7 were used to identify if meisoindigo could affect the inflammatory factors such as NO and other important cytokines which were highly involved in psoriasis. Our results indicated that meisoindigo decreased the production of NO in LPS-stimulated RAW 264.7 cells and reduced the expression of cytokines in LPS-stimulated HaCaT cells. And TLR4-TAK-NF-kappaB was a possible pathway mainly involved in the attenuation of iNOS and pro-inflammatory cytokine production by meisoindigo, which may take part in the therapeutic effect of meisoindigo on psoriasis.

Objective To explore the prognostic determinants and clinical treatment strategy in 142 patients with severe trauma brain injury(STBI).Methods Retrospective analysis of clinical data of 142 patients with STBI in our department from April 2006 to April 2012.All the patients were divided into good prognosis group(Ⅲ~V grade)and poor prognosis group(I~Ⅱgrade)ac-cording to the GOS classification standard.Age,gender,GCS,encephalocele,morphotogy of the basal cisterns on CT scanning,asso-ciated inj ury,shock,hyoxemia,underlying disease and hyperglycemia were chosen as the observation index.Statistical analysis was performed with Pearson Chi-square Test.Results 52.11% of patients with good prognosis,47.89% of patients with poor progno-sis and 3 1 .6 9% of patients were dead.Age,GCS,encephalocele,morphotogy of the basal cisterns on CT scanning,associated inj ury, shock,underlying disease were the prognostic determinants of STBI(all results P<0.05).Conclusion Age,GCS,encephalocele, morphotogy of the basal cisterns on CT scanning,associated inj ury,shock,underlying disease can determine the prognosis of STBI. Multidisciplinary cooperation treatment depending on the patient′s conditions is the key of improving the outcomes of STBI.

Objective To investigate the expression patterns of fibroblast growth factor 23 (FGF23) in osteoblast and its responses to calcium, phosphate, exogenous PTH and 1,25(OH)_2D~3. Methods The primary rat calvarial osteoblasts were cultured in MEM medium which containing 10% FBS, then were harvested when cells were in half-confluence, confluence, osteoid deposition and osteoid mineralization stages respectively. The procedure was monitored under microscopy. Total RNA was extracted from cells according to the Trizol procedure. FGF23 mRNA levels were determined by Real-time PCR. Further, the confluent osteoblasts were treated with 3.2 mmol/L CaCl_2, 4.4 mmol/L β-glycerophosphate, 10~(-9) mol/L rhPTH(1-34) and 10~(-8) mol/L 1,25(OH)_2D_3 respectively for 3 days, and same volume of the medium was added as the control. The gene expressions were determined by Real-time PCR. Results FGF23 expression was transiently up-regulated at cell confluent stage and down-regulated after that. The FGF23 mRNA levels were 7.5-fold higher in confluent cells compared with that in half-confluent cells (P<0.001). The markedlly stimulating effect (about 16 times) on FGF23 expression was stimulated by exogenous 1,25(OH)_2D_3 treatment while no significant effect was found on FGF23 mRNA levels by CaCl_2,β-glycerophosphate, and rhPTH(1-34) treatments when compared with the control. Conclusions The FGF23 expression in osteoblast is developmental stage-related and its powerful stimulator is 1,25(OH)_2D.

This paper summarize the practice in the teaching reform of microbiology experiment in recent years. We identify the main contents of experimental teaching systems and pay much more attention to peo-ple-oriented. Through the reform of teaching and assessment methods,students are trained to cultivate their practical ability and spirit of innovation.

Objective To investigate the effects of different doses of propofor on ATP-sensitive K~+currents(I_KATP)in human atrial myocytes and the underlying mechsnism.Methods A small piece of myocardiumwas obtained from right atrium in patients undergoing atrial septal defect or ventricular septal defect surgery.Themyocardium specimen was placed in cold Ca~(2+)-free cardioplegic solution aerated with 100% oxygen.Themyocardium specimen was cut into small chunks(1 mm~3).Atrial myocytes were isolated by enzymatic dissociationtechnique.The effects of propofol on I_KATP in atrial myocytes were studied using the whole-cell configuration ofpatch-clamp technique.Results The outward currents were recorded with a pipitte solution containing 0.3mmol?L~(-1) ATP.The currents were inhibited by glibendamide 10 ?mol?L~(-1),a specific K_ATP channel inhibitor,suggesting that the outward currents were I_KATP.I_KATP aws activited by propofol in a dose-dependent manner.Conclusion Propefol can activate the I_KATP in human myocytes in a concentration-dependent manner and themechanism of its myocardial depressant action may be partly explained.

BackgroundWhether the peeling of the inner limiting membrane (ILMP) increase the closure rate of idiopathic macular hole is still in controversy.Some ophthalmologist recommend vitrectomy combined with inner limiting membrane peeling for the treatment of idiopathic macular hole.However,the removal of ILMP is difficult because of its similar appearance to adjacent tissues.Objective This study was to investigate the efficacy of triamcinolone acetonide(TA) and indocyanine green(ICG) double staining-assisted vitrectomy combined with ILM peeling during the surgery.Methods A consecutive case- observational study was designed.The standardized vitrectomy was performed in 25 eye of 23 cases with IMH.During the vitrectomy,TA and ICG were injected into posterior pole vitreous to visualize and assist the ILM peeling.The dying effectiveness was observed,and the closure rate of macular hole,visual acuity,intraocular pressure and complications were evaluated after surgery.Written informed consent was obtained from each patient prior to operation.Results Posterior vitreous cortex and ILM were visible and the residual vitreous and cortex were removed clearly after dying of TA and ICG in all the 25 eyes.During the following-up duration of 3-8 months,the completely anatomical reattachment of the macular area was in 22 eyes ( 88.0％ ) and partially reattachment in 3 eyes( 12.0％ ).The best corrected vision was 0.07-0.60 in all of the operated eyes 2 months after surgery.Conclusions TA and ICG- assisted vitrectomy combined with ILM peeling appears to be a safe and effective method for IMH repair.

Objective To improve treatment success rate and prognosis for patients with bifrontal contusions by intracranial pressure monitoring.Methods A retrospective analysis was conducted on 79 cases of bifrontal contusions admitted between October 2004 and April 2012.The patients were divided into intracranial pressure monitoring group (n =40) and group without intracranial pressure monitoring (n =39),according to the treatments.Significance of high coronary craniotomy timing,surgical strategy and intracranial pressure monitoring in the diagnosis,treatment and prognosis was analyzed.Results The intracranial pressure monitoring group showed a significantly shorter period concerning osmotic dehydration [(14.24 ± 7.93) days vs (21.61 ± 11.97)days,P＜0.01],ICU stay [(14.38 ±7.56)days vs (24.71-± 17.94)days,P＜0.01] and total hospital stay [(17.20 ±8.09)days vs (33.92 ± 21.70)days,P＜0.01] as well as a better GOS [(4.15 ± 1.22) points vs (3.69 ± 1.56) points,P ＜ 0.05],as compared with group without intracranial pressure monitoring.Conclusions Craniotomy,especially decompressive craniectomy,is one of the most important treatment means to control cranial pressure and ensure cerebral perfusion pressure in patients with bifrontal contusions (in particular the moderate and severe ones).Besides,intracranial pressure monitoring is conducive to selection of surgery timing and is instructive to combined treatment,such as osmotherapy,intracranial pressure controlling and assurance of cerebral perfusion pressure.

Animals ; Cadmium Chloride ; toxicity ; Dose-Response Relationship, Drug ; Estradiol ; toxicity ; Female ; Organ Size ; drug effects ; Rats ; Rats, Sprague-Dawley ; Uterus ; drug effects ; pathology

OBJECTIVETo study the proliferation and differentiation of neural stem cells in the subventricular zone (SVZ) in neonatal rats after bilateral common arteries occlusion.

METHODSNinety-six 3-day-old Sparuge-Dawley rats were randomly divided into two groups: ischemia and control. Rats in the ischemia group were subjected to bilateral common arteries occlusion and the rats in the control group were sham-operated. All rats were administrated with 5-bromodeoxyuridine (BrdU) (50 mg/kg) via intraperitoneal injection. Rats were sacrificed and their brains were removed 1, 4, 7, 10, 14 and 35 days after ischemia. Using brain paraffin sections and immunofluorescence assays, the number of newborn cells in the SVZ was counted. Newborn neural stem cells and oligodendrocytes in the SVZ were observed, and then double marked with BrdU and nestin or osmium tetroxide (O4).

RESULTSThe number of BrdU+ cells (neural stem cells) in the SVZ in the ischemia group was greater than in the control group 4, 7, 10 and 14 days after ischemia, and reached a peak at 4 days after ischemia (253.1+/- 49.3 vs 133.5+/- 17.7; P< 0.01). By 35 days after ischemia, the number of BrdU+/O4+ cells (oligodendrocytes) in the corpus callosum (56.0+/- 7.2 vs 17.0+/- 6.4; P< 0.01), the septal nuclei (45.0+/- 11.9 vs 20.5+/- 5.0; P< 0.01), the striatum (34.5+/- 4.2 vs 14.5+/- 5.8; P< 0.01) and the olfactory bulb (46.5+/- 6.6 vs 23.5+/- 8.4; P< 0.01) in the ischemia group increased significantly as compared to the control group (P< 0.01).

CONCLUSIONSBrain ischemia can activate the proliferation of neural stem cells in the SVZ and promote neural stem cells differentiation into oligodendrocytes. The immature brain may have the capacity for self-repair after ischemic brain injury.

Animals ; Animals, Newborn ; Brain Ischemia ; physiopathology ; therapy ; Bromodeoxyuridine ; metabolism ; Cell Differentiation ; Cell Proliferation ; Cerebral Ventricles ; physiopathology ; Female ; Male ; Neurogenesis ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation