Objective To obtain highly purified oligodendrocyte precursor lineage cells in vitro and make identification.Methods The oligodendrocyte precursors were separated from astrocyte by orbital shaker and further purified by differential adhesion,and finally cultured in chemically defined serum-free medium,with appended neurotrophin 2(N2),platelet-derived growth factor(PDGF),basic fibroblast growth factor(bFGF).Immunofluorescence assay was applied to identify the separated cells with A2B5,O4,O1 and glial fibrillary acidic protein(GFAP) antibodies.Results Over 95% of cultured oligodendrocyte precursor cells were obtained.The oligodendrocyte progenitors were A2B5 and O4 positive,immature oligodendrocytes were O4 and O1 positive while GFAP were negative.Conclusions Separation and purification by shaking and differential adhesion and chemically defined medium are suitable and effective to obtain highly purified oligodendrocyte precursor cells.Cell output will increase notabily and rest in immature phase by appending both N2,PDGF and bFGF.

BACKGROUND: The previous experiments have conformed the traditional Chinese medicine (TCM) Jiannaaan, with the effects of tonifying kidney, promoting blood circulation and resolving phlegm, can inhibit the increased content of glucocorticoid (GC) in 2-24 hours after cerebral ischemic reperfusion (CIR), and reduce toxic effects of promoting nervous cell apoptosis induced by high GC. However, it is unclear whether this effect exists in GC receptor (GR).OBJECTIVE: To observe the intervention of TCM Jiannaoan on GR,further study protective mechanism of Jiannaoan power to hippocampal neurons after CIR, and perform positive control with compound almitrine.DESIGN: A randomized and controlled trial taken animals as subjects.SETTING: Center Laboratory of Beijing University of Chinese Medicine.MATERIALS: The experiment was conducted at the Center Laboratory of Beijing University of Traditional Chinese Medicine between July 2002 and March 2003. Eighty male SD rats were randomized into 5 groups with 16 in each: Sham group, model group, treatment group, positive control group and antagonist group. And each group was divided into 4 subgroups: 2, 6,12 and 24 hours after CIR, with 4 rats at every time point.METHODS:①Administration: Except model group, rats in other 4 groups were administrated by intragastric infusion since 7 days before model establishment, once per day, with dose of 7 μL/g per day distilled water in sham group, 7.39 mg/kg per day compound almitrine in positive control group, 6.7 g/kg per day Jiannaoan crude drug (consisted of desertliving cistanche herb, tatarinowii sweetflag rhizome and rhubarb, etc) in treatment group and 10 g/kg per day GR antagonist mifepristone in antagonist group.② After 7-day administration, the CIR models were prepared on the experimental rats with middle cerebral artery occlusion (MCAO) filament method, while the rats in sham group were sutured after common carotid artery detachment at anesthesia, without filament.MAIN OUTCOME MEASURES: All the rats were executed to take out brains at different time points of reperfusion, and the change of GR protein expression was observed with immunohistochemical method then the amount of positive cells were calculated in 3×200 sight of CA2 region.RESULTS: Totally 80 rats were entered into the result analysis. Compared with uninjured side, the protein expression of GR in model group,treatment group, positive control group and antagonist group were significantly lower than that of sham group (P ＜ 0.05), in which GR expression of injured side was equal to that of uninjured side without significant difference. No obvious change was found in the protein expression of GR among treatment group, positive group and antagonist group at different time points of reperfusion, and no significant difference was found between above groups and model group (P ＞ 0.05).CONCLUSION: Jiannaoan power is selective for adjusting GR and content of GC: Jiannaoan can not adjust expression of GR, identical as compound almitrine; But Jiannaoan can protect the neurons through decreasing the content of GC in plasm and brain tissues after CIR.

Objective To explore the effect and the possible pathway of different concentrations of QLT0267,which was the inhibitor of the integrin-linked kinase (ILK),on the process of high glucose-induced tubularepithelial-myofibroblast transdifferentiation (TEMT) in human renal tubular epithelial cells (HK-2).Methods HK-2 cells were exposed to 30 mmol/L GS,and TEMT model was established.After excluding the effect of high osmotic in TEMT,HK-2 cells were divided into 6 groups by different concentrations of GS and QLT0267 for 48 hours.The rate of the cell proliferation was calculated by MTT.The expression of ILK and α-smooth muscle actin (α-SMA) were determined by immunofluorescence and Western blot,and the expression of protein kinase B (AKT),phosphorylated protein kinase B (p-AKT),and E-cadherin were determined by Western blot.Results (1) The expression of ILK,p-AKT,and α-SMA in HK-2 cells were unregulated and the expression of E-cadherin was downregulated for 48 hours with glucose treating vs control (P ＜ 0.05);(2) The proliferation rate in high glucose group was higher than the group which concentration of QLT0267 was greater than 5 μmol/L (P ＜ 0.05);(3) With the concentrations of QLT0267 increased,the expression of p-AKT,α-SMA was gradually decreased (all P ＜ 0.05),and the expression of E-cadherin was gradually increased (all P ＜ 0.05).Conclusions 30 μmol/L of GS can lead to TEMT in HK-2 cell.The QLT0267 with concentration greater than 5 μmol/L may prevent the activation of ILK downstream proteins,then partially inhibits cell proliferation and TEMT in HK-2 cell.

Objective To observe the effects of different concentrations of SB203580, the inhibitor of P38MAPK, in process of high glucose (GS)-induced renal tubular epithelial-myofibroblast transdifferentiation (TEMT). Methods The cultured human renal tubular epithelial cells (HK-2) were divided into control group (5.5 mmol/L GS), GS (30 mmol/L GS) group and different concentrations of SB203580 (30 mmol/L GS +5, 10, 20 and 30 μmol/L SB203580) groups. The treat?ments were for 48 hours. MTT assay was used to observe cell proliferation. The median inhibiting concentration (IC50) was cal?culated. Western blot assay was used to detect the expressions of P38MAPK, P-P38MAPK andα-smooth muscle actin (α-SMA) in control group, high-glucose group and S30 group. The expression ofα-SMA was also detected by the method of im?munofluorescence. Results 1.Compared with control group, there was no significant inhitory effect on proliferation rate in DMSO group (P>0.05). There were increased HK-2 cells in high glucose group and S5group (P<0.05). Proliferation rates were significantly decreased in S20 and S30 groups (P<0.05). Compared with high glucose group, the proliferation rates of HK-2 cells were inhibited in S5, S10, S20 and S30 groups (P<0.05). 2. The expression of P-P38MAPK was significantly higher in high glucose group and S30 group than that of control group (P<0.05). Compared with high glucose group, the ex?pression of P-P38MAPK was significantly decreased in S30 group (P<0.05), whereas no significant difference in the expres?sion of P38MAPK between the two groups (P>0.05). 3. Compared with control group, the expression ofα-SMA was signifi?cantly increased in high glucose group and S30 group (P<0.05). Compared with high glucose group, the expression of α-SMA was significantly decreased in S30 group (P < 0.05). Conclusion The 30 mmol/L GS can lead to TEMT in HK-2 cells. The more suitable inhibitory concentration of SB203580 in the process of TEMT is 30μmol/L. SB203580 can slow down the process of TEMT by inhibiting P38MAPK activation and inhibiting proliferation and the expression ofα-SAM s of HK-2 cells.

Animals ; Cell Proliferation ; DNA ; genetics ; immunology ; Female ; Hepatitis B Core Antigens ; genetics ; Hepatitis B virus ; genetics ; Interferon-gamma ; blood ; genetics ; Killer Cells, Natural ; immunology ; Lymphocyte Activation ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; immunology ; Random Allocation ; Recombination, Genetic ; Spleen ; cytology ; immunology ; metabolism ; T-Lymphocytes ; immunology

OBJECTIVE To investigate the epidemic,drug resistance and gene distribution of ESBLs-producing Klebsiella pneumoniae (KPN) from Jiangxi TCM Hospital. METHODS The susceptibility of KPN was detected by MIC PCR was used to detect ESBLs gene. RESULTS There were 42 strains with ESBLs isolated,the positive rate was 35.0%. The drug resistance rate of KPN with ESBLs was higher than that without ESBLs,PCR typing result:TEM 33 (78.6%),SHV 8 (19.0%) and CTXM 29 (69.0%). CONCLUSIONS The ESBLs-producing bacteria have multiple drug resistant genes;TEM and CTXM are the main drug resistant genes in our hospital.

Objective To investigate the effects of 1,25-dihydroxyvitamin D3[1,25 (OH)2D3] on cell proliferation in hu?man glomerular mesangial cells and it′s effects on the regulation of mTOR/p70s6K signaling pathway in this cell line. Meth?ods The cultured human mesangial cells at passage 3-7 were divided into four groups:control group,VD group (addition of 10-8 mol/L of 1,25-dihydroxyvitamin D3 ),R group (addition of 5 mg/L of rapamycin) and R+VD group(addition of 5 mg/L ra?pamycin combined with 10-8 mol/L of 1,25-dihydroxyvitamin D3). Drug incubation last 48 h. The effect of mesangial cell pro?liferation was measured by CCK-8 colorimetric assay. The cell cycles were measured by flow cytometry. The expression of mTOR and p70s6K were detected by immunofluorescence. Results (1) The absorbance of A450 was higher in control group than that in VD group than that in R group than that in R+VD group. But the inhibition rate (IR) was lower in control group than that in VD group than that in R group than that in R+VD group. All comparisons were of statistic significance. ( 2) Cells in G1 phase were higher while cells in G2/M and S phases as well as proliferation rate (PI) were lower in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance except in?dexes between group R and group VD. (3) mTOR and p30s6K expressions in mesangial cells were higher in control group than those in VD group than those in R group than those in R+VD group. All comparisons were of statistic significance ex?cept indexes between group R and group VD. Conclusion 1,25-dihydroxyvitamin D3 might inhibit mesangial cell prolifera?tion significantly through mTOR/p70s6K signaling pathways.

Objective To study the influence of different body serum iron levels on follicle stimulating hormone (FSH),luteinizing hormone (LH) and uterine ovarian index,and explore the body serum iron level effects on female mice sexual maturity further.Methods 36 three weeks C57BL/6J female mice were randomly divided into the normal control group,strengthen-iron 1 group and strengthen-iron 2 group,12 cases in each group.The normal control group with normal diet(310mgFe/kg),strengthen-iron 1 group and strengthen-iron 2 group were given high -iron diets(respectively with 465mgFe/kg and 620mgFe/kg) after born 3 weeks.each group body,uterus and ovary mass after 2 weeks were checked;colorimetric method was used to detect the serum iron level;the serum follicle stimulating hormone (FSH) and luteinizing hormone(LH) were detected by chemiluminescent immunoassay.Results The body of mice serum iron levels,uterine ovarian index (uterine ovarian wet mass/body mass) and follicle stimulating hormone(FSH) level of strengthen-iron 1 group and strengthen-iron 2 group were higher than those of normal control group (F =9.11,11.11,7.09,all P ＜ 0.01).Luteinizing hormone (LH) level of the strengthen-iron group was higher than those of normal control group (F =3.89,P ＜ 0.05).Conclusion Uterine ovarian index,follicle stimulating hormone level and luteinizing hormone level were affected with increased serum iron level;It suggests that the increasing serum iron level can cause female mice sexual maturity in advance.

Objective To explore the factors influencing serum trough concentration of vancomycin in pediatric patients with severe gram-positive cocci pneumonia. Methods The general information, the biochemical test results, and plasma concentration of vancomycin were collected from 93 pediatric patients with severe gram-positive cocci pneumonia. The relative factors influencing trough concentration of vancomycin were analyzed retrospectively. Results With the dosage of 40-60 mg/(kg·d), serum trough concentration of vancomycin were between 10-20 mg/L in 26 patients, <10 mg/L in 54 cases, ≥20 mg/L in 13 cases. The ALT, AST, GFR, and γ-GT were significantly different among three groups (P<0.05); the 10-20 mg/L group had the highest levels of AST and γ-GT, the ≥20 mg/L group had the highest level of ALT and the lowest level of GFR. Multiple linear regression analysis showed that GFR was negatively linearly correlated with the serum trough concentration of vancomycin (R2=0.039, P<0.05). The median serum trough concentration of vancomycin in pediatric patients with GFR≥90, 60–90, 30–60 mL/(min·1.73m2) were 8.66, 18.21, 8.45 mg/L respectively, and the difference is statistically significant (P<0.05). Conclusions The serum trough concentration of vancomycin is negatively linearly correlated with GFR in pediatric patients with severe gram-positive cocci pneumonia. The patients with impaired renal function are easier to reach the target serum trough concentration of vancomycin. Clinical use of vancomycin should follow the low doses in the range the guideline recommended, and the serum trough concentration should be closely monitored.

BACKGROUND: The study aimed to investigate the therapeutic benefits of intravenous Xuebijing on acute kidney injury (AKI) in rats with paraquat intoxication. METHODS: Male Sprague-Dawley rats were randomly divided equally into three groups:sham group (n=8), paraquat group (n=8) and Xuebijing-treated group (n=8) using a random number table. The rats were intraperitoneally injected with 50 mg/kg of paraquat. One hour after paraquat administration, the rats were treated intravenously with Xuebijing (8 mL/kg). At 12 hours after paraquat administration, serum was collected to evaluate kidney function, then the rats were sacrificed and kidney samples were immediately harvested. AKI scores were evaluated by renal histopathology and pro-inflammatory cytokines mRNA levels in kidney were assayed using real-time RT-PCR. RESULTS: Serum urea nitrogen, creatinine and AKI scores were significantly higher in the paraquat group, compared with the sham group (P<0.05, respectively). Moreover, interleukin (IL)-1β, IL-6 and TNF-α mRNA levels were significantly higher in the paraquat group (P<0.01, respectively). However, intravenous Xuebijing significantly decreased serum urea nitrogen, creatinine, AKI scores and IL-1β, IL-6 and TNF-α mRNA levels, compared with the paraquat group (P<0.05, respectively). CONCLUSION: Intravenous Xuebijing attenuates AKI fol owing paraquat poisoning by suppressing inflammatory response.