OBJECTIVE To investigate the risk factors for nosocomial pneumonia(NP) caused by imipenem-resistant Acinetobacter baumannii(IRAB) and its antimicrobial susceptibility in vitro. METHODS The data of 34 cases of IRAB-NP were analyzed and 68 cases of NP caused by imipenem-susceptible A.baumannii(ISAB) were randomized as control.Antimicrobial susceptibility(MIC) was determined with the method of agar dilution. RESULTS The two independent factors associated with the development of IRAB-NP: previous fluoroquinolone(OR=5.738) and imipenem/meropenem(OR=7.129) use.The drug sensitivity test in vitro showed that these strains were multiresistant to commonly used antibiotics,and only ampicillin/sulbactam and cefoperazone/sulbactam whose resistance rate was less than 30%. CONCLUSIONS Previous imipenem/meropenem and fluoroquinolone use is independent risk factors for IRAB-NP.These strains are high drug resistant.

Objective: Numerous studies have shown that bone marrow-derived mesenchymal stem cells (MSCs) enhance neurological recovery after cerebral ischemia. However, the mechanisms are still not clear. The present study aimed to investigate the beneficial effects of MSCs on global cerebral ischemia induced by cardiac arrest (CA) and the underlying mechanisms. Methods: Rats subjected to asphyxial CA were injected intravenously with MSCs (5×106 ) at 2 hours after resuscitation. Whole brain histopathologic damage scores (HDS) were assessed by histopathology at 3 and 7 days after resuscitation. The distribution of donor MSCs in the brain was evaluated. The expression of tumor necrosis factor-α-induced protein 6 (TSG-6) and pro-inflammatory cytokines in cerebral cortex was assayed. After intravenous infusion of TSG-6 siRNA-MSCs, HDS and pro-inflammatory cytokines were reevaluated at 7 days after resuscitation. Results: Intravenously administered MSCs significantly reduced whole brain HDS after global cerebral ischemia. Immunofluorescence microscopy revealed that donor MSCs were primarily found in cerebral cortex and expressed TSG-6. MSCs treatment significantly increased the expression of TSG-6 and reduced the expression of pro-inflammatory cytokines in cerebral cortex. In addition, intravenous infusion of TSG-6 siRNA-MSCs failed to attenuate brain inflammation. Conclusion: Systemically administered MSCs reduced inflammatory damage to brain in rats with global cerebral ischemia via secretion of TSG-6.
Heart Arrest ; Mesenchymal Stromal Cells

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

To study the effect of sodium aescinate in inducing human breast cancer MCF-7 cells apoptosis and its possible mechanism. MTT assay was used to detect the inhibitory effect of sodium aescinate on the proliferation of MCF-7 cells. The morphological changes were observed under inverted microscope. DAPI nuclear staining was used to detect the changes in cell nucleus. Annexin V-FITC/PI flow cytometry was adopted to test the apoptosis rate. Changes in apoptosis-related proteins (PARP, cleaved caspase-8 and pro-caspase-3), cell survival-associated signal molecules (AKT and ERK) and their common upstream kinase SRC was detected by Western blotting. The result showed that after different concentrations of sodium aescinate were used to treat breast cancer MCF-7 cells, they inhibited the proliferation of MCF-7 cells in a dose-dependent manner, induced cell apoptosis (typical morphological changes in nucleus, significant increase in cell apoptosis rate). The expressions of cleaved PARP and caspase-8 increased, while the expression of pro-caspase-3 decreased, which further verified sodium aescinate's effect in inducing cell apoptosis. Sodium aescinate significantly inhibited the phosphorylation of cell survival-related signal molecules (AKT, ERK) and down-regulate the activation of their common up-stream kinase SRC. The findings indicated that sodium aescinate can block signals transiting to downstream molecules AKT, ERK, inhibit the proliferation of breast cancer cell MCF-7 cell apoptosis and induced cell apoptosis by suppressing the activation of SRC.
Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Breast Neoplasms ; drug therapy ; enzymology ; genetics ; physiopathology ; Down-Regulation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Female ; Humans ; MCF-7 Cells ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Saponins ; pharmacology ; Signal Transduction ; drug effects ; Triterpenes ; pharmacology ; src-Family Kinases ; genetics ; metabolism

Objective To study the level of serum lipids and body-fat content of high-fat diet induced pbesity rats(DIO).explore the relationship between intracellular caleium and ventrieufar arrhythmia.Methods Sixty male Sprague-Dawley(SD)rats were divided into control group(15)and experiment group(45),high-fat diet was administrated for 12 weeks to established obesity model,15 rats were selected into obesity group according to their body weight gain.The standard 2-lead electrocardiograph was used to detect the incidence and scores of arrhythmia induced by barium chloride(BaCl2,0.1 mg/kg)for 1 hour on every 8 rats from different groups respectively.Body-fat content.the level of serum total cholesterol(TC),triglyceride(TG),low density lipopmtein cholesterol (LDL-C) and high density lipopmtein cholesterol (HDL-C)were measured.The epididymal(EP),retroperitoneal (RE) and mesenteric(ME)white adipose pads was measured to obtain the body fat content.Single ventricular myrocytes of rats were isolated by enzymatic dissociation.The confocal laser scanning microscope was used to record basic intraceUular calcium level([Ca2+]i).Results The body-fat content in obesity group[(7.71±0.74)%] was significantly higher than control group[(4.69±0.37)%](t＝3.650,P＜0.05).The level of serum TC,TGand LDL-C were significantly higher(t＝3.801,2.778,3.536.P＜0.05) in obesity group[(1.26±0.04),(0.58±0.10),(0.51±0.04)mmol/L]than those in control group[(0.92±0.08),(0.29+0.03),(0.31±0.04) mmol/L].The level of serum HDL-C wa8 decreased gradually from control group[(0.53±0.05)mmol/L] toobesity group[(0.52±0.02)mmol/L],but there waft no significant difference between them(t＝0.186,P＞0.05).The incidence of arrhythmia induced by BaCl2(0.1 mg/kg)in obesity group was significantly higher than controlgroup(X2＝5.333,P＜0.05),and the scores of arrhythmla was increased in obesity group(2.5±0.6)too.The fluorescence intensity standing for[Ca2+]i was increased significantly(t＝2.409,P＜0.05)from obesity group(247.96±20.03)to control group (174.25±23.13).Conclusion As the free cytosolic calcium begin to accumulate,the arrhythmia morbidity is increased in obesity rats.

Objective To investgate prevalence and clinical significance of antiproteinase 3(PR3)an- tibodies in Wegener's granulomatosis(WG)and other vasculitis patients.Methods One hundred and eleven systemic vasculitis patients with WG(9 cases,including 21 serums of tracking WG patients)and other systemic vasculitis(102 cases),403 secondnary vasculitis CTD(SLE 213 cases,RA 135 cases),nephritis 62 cases,30 healthy subjects were examined for anti-PR3 and anti-MPO antibody by enzyme-linked immunosorbent assay (ELISA)and ANCA by indirect immunofluorescence(IIF)was performed.Result Anti-PR3 positive were 23 in 588 serums of patients.The prevalence of anti-PR3 positive was WG(16/21,71.4 %),other systemic vas- culitis were not found anti-PR3,SLE(6/213,2.8%),RA(1/135,0.7%).In particular,the prevalence of anti- PR3 and cANCA with WG tended to be higher in the patients with other systemic and secondnary vasculitis (P＜0.05).The sensitivity and specificity of anti-PR3 for diagnosis of WG were 71.42% and 98.58%.The sensi- tivity and specificity of combination anti-PR3 and cANCA were 61.90% and 99.82%.Anti-PR3 and cANCA are associated with treament of WG.Conclusion Anti-PR3 antibody has high specificity for diagnosis of RA. Detection of anti-PR3 and cANCA at the same time can improve the specificity considerably.As sensitive markers of WG,anti-PR3 antibody may be useful for diagonosis and early treament.Anti-PR3 also may be useful for activity and relapse of WG.

By RT-PCR and TAIL-PCR, the full coding region of Batai virus isolated in China (YN92-4 strain)was sequenced for the first time. According to the results, the genome of the virus contained three segments S, M and L of 947, 4,371 and 6,860 nucleotides, respectively. The S segment coded a nucleoprotein of 234 amino acids and a nonstructural protein of 102 amino acids, the M and L segments coded a precursor protein of 1 ,435 amino acids and RNA polymerase of 2,239 amino acids, respectively. Compared with the full coding sequence of Batai viruses isolated out of China, the S and M segments of YN92-4 and ON-7/B/01 showed the highest homology in nucleotide and amino acid sequenes with similarity of 97.7% (100%) and 95.7% (98%), respectively. Since there was no full coding sequence information on the L segment in GenBank for the reference, the L segment of YN92-4 was compared with that of Bunyamwera virus and the homology of nucleotide and amino acid was 73.5%and 81.6%, respectively. Phylogenetic analysis showed YN92-4 strain was clustered into one group with the prototype of Batai virus (MM2222). The results suggested that the YN92-4 strain had no occurrance of genetic reassortment (like Ngari virus) and was close to the Batai virus (ON-7/B/01 strain) isolated from cattle serum in Japan.
Amino Acid Sequence ; Animals ; Base Sequence ; Bunyamwera virus ; genetics ; Cattle ; China ; Clinical Laboratory Techniques ; Cloning, Molecular ; Genome, Viral ; genetics ; Hemorrhagic Fever with Renal Syndrome ; Molecular Sequence Data ; Open Reading Frames ; Polymerase Chain Reaction ; Reassortant Viruses ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment ; Sequence Analysis, DNA

OBJECTIVETo reveal the relationship between the storage time of the bark of Magnolia officinalis and the content of phenols in it, and lay a theoretical foundation for the harvest, processing, management and storage.

METHODThe contents of magnolol and honokoiol in 15 bark samples, collected from the main producing areas in China, were determined in the time of freshly harvest and 3 and 10 years after respectively by HPLC method.

RESULTIt showed that within a certain period of time, bark storage was favorable to conversion and accumulation of phenols, that the content of magnolol tended to increase from year 0 to year 3, then followed by slight decrease with years on account of volatilization of phenols, but was still higher when the bark was stored for 10 years than that that when the bark was freshly harvested, and the content of honokoiol still tended to increase when the bark had been stored for 10 years.

CONCLUSIONThe phenols in bark of M. officinalis is quite stable and the bark can be stored for 10 years or longer.

Biphenyl Compounds ; analysis ; Drug Storage ; methods ; Drugs, Chinese Herbal ; chemistry ; Lignans ; analysis ; Magnolia ; chemistry ; Plant Bark ; chemistry ; Time Factors

To investigate the distribution of mitochondria DNA 4 977 bp deletion, a common deletion (CD), in normal populations of Chinese, human peripheral blood samples from sixty healthy donors were collected, and levels of the CD in genomic DNA from the samples were detected using real-time PCR. The results showed that the CD was found in 27 health donors, with its positive rate being 45% (27/60). The CD ratio was between 0 and 0.00308%, and not affected by age and gender in sixty healthy donors. Our studies indicate that the CD ratio is low, and do not show the age-dependent accumulation and any gender difference in peripheral whole blood from the normal Chinese population.
Base Sequence ; DNA Primers ; DNA, Mitochondrial ; genetics ; Female ; Humans ; Male ; Real-Time Polymerase Chain Reaction ; Sequence Deletion

OBJECTIVETo study the effect of oleic acid (OA) on expression of aquaglyceroporin genes, AQP3 and AQP9, in hepatocyte steatosis and to investigate the underlying molecular mechanisms using an in vitro system.

METHODSHepG2 cells were treated with OA at different concentration to establish in vitro models of nonalcoholic hepatocyte steatosis. The corresponding extents of hepatic steatosis modeling were assessed by oil red O staining and optical density (OD) measurements of the intracellular fat content. The model lines were then treated with inhibitors of the PI3K/Akt and p38 MAPK signaling pathway factors and effects on AQP3/9 expression was measured by real time RT-PCR and western blotting.

RESULTSThe fat concentration, indicative of hepatic steatosis, increased in conjunction with increased concentrations of OA (0 less than 250 less than 500 mumol/L). OA exposure also down-regulated AQP3 mRNA and up-regulated AQP9 mRNA levels in a concentration-dependent manner. The most robust changes in expression occurred in response to the 500 mumol/L concentration of OA for both AQP3 (0.47+/-0.18; t = 4.5450, P less than 0.05) and AQP9 (1.57+/-0.21; t = 3.0306, P less than 0.05). Treatment with OA + PI3K pathway inhibitor (LY294004) significantly decreased AQP9 mRNA expression (4.55+/-0.62) as compared to the control group (1.00+/-0.10; t = 9.7909, P less than 0.01), that 500 mumol/L OA group (2.43+/-0.53; t = 4.5018, P less than 0.05), and the LY294002 group (1.90+/-0.16; t = 7.1683, P less than 0.01). Treatment with p38 MAPK pathway inhibitor (SB230580) significantly increased the OA-suppressed level of AQP3 mRNA to the level detected in the control group (1.27+/-0.11; t = 5.7455, P less than 0.01) and decreased the OA-stimulated AQP9 mRNA (0.38+/-0.09; t = 6.5727, P less than 0.01). No significant changes in mRNA expression of AQP3/9 were observed with inhibition of the ERK1/2 and JNK signal transduction pathways. The OA-induced changes in protein expression levels of AQR3 and AQP9 followed a similar trend of the genes. Finally, OA suppressed the level of phosphorylated Akt (from 0.21+/-0.02 to 0.13+/-0.03; t = 3.8431, P less than 0.05) but elevated the level of phosphorylated p38 (from 0.58+/-0.06 to 1.02+/-0.10; t = 12.5289, P less than 0.01). Again, OA treatment produced no significant affect on ERK1/2 and JNK phosphorylation.

CONCLUSIONOA down-regulates AQP3 expression by stimulating the p38 MAPK signaling pathway, and up-regulates the AQP9 by blocking the PI3K/Akt pathway and activating the p38 MAPK signaling pathway.

Aquaporin 3 ; metabolism ; Aquaporins ; metabolism ; Fatty Liver ; metabolism ; pathology ; Gene Expression Regulation ; drug effects ; Hep G2 Cells ; Humans ; Oleic Acid ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; metabolism