BACKGROUND: Paraquat (PQ) intoxication causes lung oxidative stress damage. Saturated hydrogen saline, a newly explored antioxidant, has been documented to play a powerful antioxidant role in preventing oxidative stress damage. This study aimed to investigate the protective effects and the possible mechanisms of intoxication on rats with acute lung injury (ALI) caused by paraquat poisoning. METHODS: Thirty PQ poisoned rats were randomly divided into a PQ intoxication group (intoxication group), a saturated hydrogen saline intervention group (intervention group), and a control group, with 10 rats in each group. The first two groups accepted an intragastric administration of PQ at a dose of 50 mg/kg for every single rat, and the control group was fed with a same volume of normal saline. Five mL/kg of saturated hydrogen saline was given to the intervention group three times a day by peritoneal injection for three days after intoxication. Arterial blood gas was detected on the third day. The rats were executed and their lungs were taken for measurement of wet dry weight ratio, homogenate malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OhdG). Histological changes of the lungs were also observed. RESULTS: Compared with the control group, the intoxication group had more serious hypoxemia, greater wet/dry weight ratio, higher MDA level, higher expression of 8-OhdG and more severe lung damage (P<0.01 or P<0.05). However, after intervention with saturated hydrogen saline, poisoned animals turned to have lighter hypoxemia, smaller wet/dry weight ratio, lower MDA level, lower expression of 8-OhdG, and milder lung damage (P<0.01 or P<0.05). CONCLUSIONS: Saturated hydrogen saline is effective in preventing acute lung injury caused by PQ. Possibly, it can neutralize toxic oxygen radicals selectively and alleviate the oxidative stress injury induced by PQ.

OBJECTIVETo investigate the association of surgical skills, anhepatic time and preoperative hepatic function grading with bacteria infection after the liver transplantation and identify the common bacterial flora involved for effective prevention and treatment of the posttransplant bacterial infection. METHODS;The clinical records of 31 cases of liver transplantation from August 2004 to August 2005 were reviewed and the collected data were analyzed statistically. RESULTS; Among the 31 cases, posttransplant bacterial infection occurred in 16 cases accounting for a total incidence of 51.61%, with the incidence of multi-system (or multi-organ) infection of 22.58%. The earlier cases had longer average surgery time and anhepatic period than the later cases, with also higher incidence of infection. Among the 19 patients with hepatic function class A before surgery, 7 acquired bacterial infection involving one system or organ, 2 had infections compromising multiple system or organ. In the 8 patients of hepatic function class B before surgery, 2 had single-system or -organ infection and 1 multi-system or -organ infection. Four out of the 5 patients with hepatic function class C before surgery acquired posttransplant bacterial infections, all involving multiple systems or organs. Pseudomonas aeruginosa was the most common bacteria responsible for the infections in these cases.

CONCLUSIONImprovement of surgical skills can obviously reduce the incidence of bacterial infection after liver transplantation. No evidences suggest the correlation between the incidence of infections (including severe ones) and hepatic function class A or B before the operation, whereas patients with preoperative hepatic function class C seems to be at higher risk for infection involving multiple systems or organs. The anhepatic time does not significantly impact on the incidence or severity of the posttransplant infections, and Pseudomonas aeruginosa is the most common bacteria causing the infections.

Adult ; Aged ; Bacterial Infections ; epidemiology ; etiology ; China ; epidemiology ; Female ; Humans ; Incidence ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Pseudomonas Infections ; epidemiology ; etiology ; Risk Factors

Objective At present,there is still a lack of effective means for the treatment of diabetic cystopathy,and to find natural antioxidants for this purpose has become a hot spot in research. This study is to investigate the protective effect of inosine on the bladder of diabetic rats and its antioxidative stress mechanisms. Methods A total of 60 adult male Sprague-Dawley rats were ran-domly divided into three groups of equal number:normal control,diabetes mellitus(DM) model control,and inosine intervention. The DM model was made by intraperitoneal injection of streptozotocin at 60 mg/kg. The DM model controls were injected with saline while the model rats in the intervention group with inosine, all at 75 mg/kg, ip,bid. After 4 and 8 weeks of treatment, the bladder tissues were collected from the rats for examination of the structural changes by HE staining,determination of the expressions of c-kit and nerve growth factor (NGF) by immunofluorescence assay, and observation of the ultrastructure of the bladder tissue under the electron microscope,de-tection of the cell apoptosis by TUNEL,and measurement of the con-tents of malondialdehyde (MDA),superoxide dismutase (SOD),and glutathione (GSH). Results HE staining indicated signifi-cant mucosal hyperplasia, disordered arrangement, loose structure, fracture, expanded intervals and collagen fiber filling of muscle bundles,muscular atrophy,lymphocytes infiltration,vascular hyperplasia and congestion,and few muscle bundles,while electron mi-croscopy manifested disordered arrangement, interrupted connection, mitochondrial vacuolation in muscular and interstitial cells, shrinkage of nuclear membrane,disappearance of nucleoli,and irregular chromatin margination and condensation in the bladder tissues of the DM rat models. Immunofluorescence assay showed that the signals of c-kit and NGF were reduced in the DM models as compared with those in the normal controls. After 4 and 8 weeks of intervention,the cell apoptosis rate was significantly higher in the DM model control ([1.68±3.04]% and [10.51±0.90]%) and inosine-treated rats ([7.00±1.72]% and [7.24±1.66]%) than in the normal controls ([4.65±3.04]% and[5.48±2.00]%),but remarkably lower in the inosine-treated than in the DM model controls(P<0.01). The contents of SOD and GSH were increased(P<0.05) while that of MDA decreased markedly in the DM models(P<0.05),but the former decreased (P<0.05) while the latter increased significantly in the inosine intervention group as compared with the DM model control group (P<0.05). At 8 weeks,the contents of SOD and GSH were remarkably lower in the DM model than in the normal con-trols (P<0.01),while that of MDA markedly higher than in both the normal control and inosine intervention groups (P<0.01). The wet weight of the bladder was significantly increased in the DM model and inosine intervention groups in comparison with that of the nor-mal controls(P<0.01). Conclusion Oxidative stress plays an important role in the development and progression of diabetic cystopa-thy. Inosine can protect the bladder structure and function of the DM rat by reducing oxidative stress and injury to the bladder tissue.

OBJECTIVETo establish a qualitative and quantitative reversed-phase high-performance liquid chromatography (RP-HPLC) with fingerprinting technique for quality control of compound dandelion enema.

METHODSHPLC was utilized for quality assessment of 10 batches of samples. RP-HPLC analysis was performed on a Hypersil BDS C18 column (4.6 mm x 250 mm, 5 microm) with the mixture of acetonitrile (A) and potassium phosphate solution (B) (pH3.2) as the mobile phase in gradient mode. The concentrations of solvent A were 10%, 80% and 80% at 0, 38 and 40 min, respectively. The column temperature was set at 35 degrees C, the flow rate at 0.7 ml/min and the detection wavelength at 254 nm.

RESULTSHPLC fingerprinting was established from the 10 batches, and the data showed 23 characteristic peaks in the compound dandelion enema for use as index peaks for qualitative identification. Comparison of the retention time and the on-line UV spectra of the samples with the chemical standards identified peaks 3, 4 and 8 as protocatechualdehyde, caffeic acid and ferulic acid, respectively. The contents of caffeic acid in the compound dandelion enema ranged between 63.7 and 136.8 microg/ml.

CONCLUSIONHigh specific chromatographic fingerprinting and quantitative measurement of caffeic acid allows rigorous quality control of compound dandelion enema.

Caffeic Acids ; analysis ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Reference Standards ; Reproducibility of Results ; Taraxacum ; chemistry

OBJECTIVETo observe the changes in spinal cord pathophysiology, motor function and electrophysiology after spinal cord injuries induced by punctures with different needles, and explore a new means for studying spinal neurotoxicity of local anesthetics.

METHODSA total of 144 SD rats were randomly allocated into the sham-operated group (n=36) and 3 spinal cord injury groups (n=36) with the L4-5 segment of the dura mater of the spinal cord punctured using 29G, 25G, and 21G needles. The BBB scores before surgery were recorded, and at 8 h, 24 h, 72 h, 1 week, and 2 weeks after the surgery, the motor evoked potential (MEP), spinal cord pathology and the BBB scores were examined.

RESULTSIn the control group, the rats showed normal BBB score, spinal function and microstructure. Spinal cord puncture with 29G needle did not cause obvious pathologies of the spinal cord, whereas puncture with 21G needle resulted in marked changes in the motor function, electrophysiology and histology of the spinal cord, which showed significant improvements at 2 weeks postoperatively.

CONCLUSIONPuncture with a 29G needle causes less injuries and minimal functional changes of the spinal cord, which can serve as a new means for studying spinal neurotoxicity of local anesthetics.

Anesthetics, Local ; toxicity ; Animals ; Disease Models, Animal ; Electrophysiological Phenomena ; Female ; Male ; Needles ; Rats ; Rats, Sprague-Dawley ; Recovery of Function ; Spinal Cord Injuries ; etiology ; physiopathology

OBJECTIVETo establish an HPLC method for the determination of three kind of diester diterpenoid alkaloids (DDAs) in Aconitum carmichaeli and its processed products namely mesaconitine, aconitine and hypaconitine.

METHODA Zorbax Eclipse XDB C18 column was used, and ammonium acetate solution-acetonitrile as the mobile phase, with a flow rate of 1.0 mL x min(-1) and a detection wavelength of 230 nm.

RESULTMesaconitine, aconitine and hypaconitine, were separated, and the calibration curves of them were in good linearity over the range of 0.035 7-1.784 microg (r = 0.9999), 0.0126-0.632 microg (r = 0.9997) and 0.0334-1.672 microg (r = 0.9997) respectively.

CONCLUSIONThis method is simple and accurate. It can be used in the identification and quality control of A. carmichaeli.

Aconitine ; analogs & derivatives ; analysis ; Aconitum ; chemistry ; Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; methods ; Hot Temperature ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Technology, Pharmaceutical ; methods

Objective:To establish an in vivo diffusion model of Treponema pallidum (Tp) in New Zealand rabbits. Methods:A standard strain of Tp (Nichols strain) was recovered in the testes of New Zealand rabbits, and isolated and passaged continuously. The suspensions of the second-passage Tp were collected and inoculated onto the dorsal skin of New Zealand rabbits. After 21-day infection, the New Zealand rabbits were anesthetized and sacrificed, blood samples were collected, and skin tissues at the infection site as well as liver, spleen, testes and lymph nodes were aseptically resected. Real-time fluorescence-based quantitative PCR was performed to detect the spread of Tp in different tissues and organs.Results:On day 21 after infection with Tp, skin lesions such as indurations and ulcers were seen at all inoculated sites of New Zealand rabbits. Pathological examination showed a lot of inflammatory cells in the infected lesions, mainly including plasma cells, macrophages and lymphocytes. Real-time fluorescence-based quantitative PCR revealed a large number of Tp in tissues and organs, such as liver, spleen and testes.Conclusion:After inoculation with Tp in the dorsal skin of New Zealand rabbits, Tp could spread to the liver, spleen, testes and other tissues and organs through blood and lymph nodes, and the in vivo diffusion model of Tp strains in New Zealand rabbits was successfully constructed.

INTRODUCTION: Cardiovascular morbidity and mortality in end-stage renal failure (ESRF) patients are high. We examined the incidence and predictors of death and acute myocardial infarction (AMI) in ESRF patients on different modalities of dialysis. METHOD: Data were obtained from a population-based database (National Registry Disease Offices) in Singapore. The study cohort comprised all adult patients initiated on dialysis between 2007 and 2012 who were closely followed for the development of death and AMI until September 2014. Cox regression methods were used to identify predictors of death and AMI. RESULTS: Of 5,309 patients, 4,449 were on haemodialysis and 860 on peritoneal dialysis (PD). Mean age of the cohort was 61 (±13) years (44% women), of Chinese (67%), Malay (25%) and Indian (7%) ethnicities. By September 2014, the incidence of all-cause death was 34%; close to a third of the patients died from a cardiovascular cause. Age >60 years and the presence of ischaemic heart disease, diabetes, stroke, peripheral vascular disease and PD were identified as independent predictors of all-cause death. PD patients had lower odds of survival compared to patients on haemodialysis (hazard ratio 1.51, 95% confidence interval 1.35-1.70, P<0.0001). Predictors of AMI in this cohort were older age (>60 years) and the presence of ischaemic heart disease, diabetes, stroke, peripheral vascular disease and current/ex-smokers. There were no significant differences in the incidence of AMI between patients on PD and haemodialysis. CONCLUSION The short-term incidence of death and AMI remains high in Singapore. Future studies should investigate the benefits of a tighter control of cardiovascular risk factors among ESRF patients on dialysis.
Adult ; Aged ; Female ; Humans ; Incidence ; Kidney Failure, Chronic/therapy* ; Male ; Middle Aged ; Myocardial Infarction/epidemiology* ; Peritoneal Dialysis ; Renal Dialysis

BACKGROUND:Co-culture technique makes different kinds of ceils cultured in the same system.The trans-differentiation from osteoblasts to adipocytes is usually analyzed under the action of adipogenesis inducers in vitro,but the cellular interactions in vivo are neglected.OBJECTIVE:To investigate the trans-differentiation from osteoblasts to adipocytes in indirect co-culture using Transwell system.METHODS:Mouse preadipocytes 3t3-l1 were induced to adipocytes.There were three groups:group A:mature adipocytes in the lower chamber of Transwell system;group B:mouse osteoblasts MC3T3-E1 in the upper chamber of Transwell system according to a ratio of 1:4 (MC3T3-E1:3t3-l1);group C:mouse osteoblasts MC3T3-E1 alone.At 7,14,and 21 days the cell morphology was observed under inverted phase contrast microscope,the relative level of triglyceride,expression of peroxisome proliferator-activated recoptor y2 in each group were detected,and red oil O staining and alizarin red staining were performed.The cell proliferation inhibition rate in the groups B and C were detected at 0,24 and 36 hours.RESULTS AND CONCLUSION:At 2 weeks after culture,spindle-shaped 3t3-l1 coils changed into round,the light and round lipid droplets in the cytoplasm were increased and were reserved after identified by oil red O staining.In the group B,the cells presented with spindle shape with no transparent lipid droplets after 7-day co-culture until black granules and small round lipid droplets appeared on day 14;and the cells changed from spindle shape to oval or round,and larger lipid droplets were found on day 21.Alizarin red staining results:the staining region in the group B was on a decline with time,while the group C showed no significant changes at each time point and all appeared with large staining region.Oil red O staining results:the staining region in the group B increased gradually in a time-dependant manner,while the group C was negative for oil red O staining and showed no significant changes at different time points.The relative level of triacylglycerol in the group B was increased with time,and there was significant difference between groups A and B (P ＜0.05),and the group C showed no significant differences (P ＞ 0.05).The coll proliferation inhibiting rate in the group B was increased with time,which showed significant difference from the group C (P ＜ 0.05).The expression level of peroxisome proliferator-activated recoptor y2 in the group B was on a rise with time,which had significant difference compared with the groups A and C (P ＜ 0.05).These results indicate that the trans-differentiation from osteoblasts to adipocytes appears in the Transwell system,and metabolic products and cytokines of adipocytes obviously inhibit the proliferation of osteoblasts,but all above conclusions need to be studied in depth.

OBJECTIVETo investigate potential contributions of genetic variants of cytochrome P-450 2C9 (CYP2C9) and vitamin K expoxide reductase (VKORC1) to the anticoagulation response during the initiation of warfarin therapy in the Han Chinese population.

METHODSA total of 798 Han Chinese patients received long-term warfarin anticoagulant therapy orally after valve replacement in our hospital between 2000 and 2008 were included in this study. Nine single nucleotide polymorphism (SNP) loci [rs12572351 G > A, rs9332146 G > A, rs4917639 G > T, rs1057910 A > C (CYP2C9(*)3), rs1934967 G > T, rs1934968 G > A, rs9923231 C > T (VKORC1-1639 G > A), rs2359612 G > A and rs10871454 C > T] in 2 genes including CYP2C9 and VKORC1, which were possibly correlated with warfarin pharmacokinetics and pharmacodynamics through literature retrieval, were selected and analyzed. Warfarin steady-state dose requirement, time to the INR (the international normalized ratio) within the therapeutic range and percent of the INR of more than 3.5 were compared among genotype subgroups. SNaPshot technique was used to detect gene SNPs; Hardy-Weinberg genetic equilibrium test was used to test population representativeness.

RESULTSCYP2C9(*)3 genotype did not affect the required warfarin dose while it was associated with increased risk of bleeding when treated with routine dosage regimen during the initiation of treatment. The allelic mutation frequency at VKORC1 gene rs10871454G > A and VKORC1-1639G > A SNP loci was 92.04% and 88.03%, respectively and rs10871454 was in perfect linkage disequilibrium with-1639. Patients with VKORC1 rs10871454 genetic mutation required lower warfarin dose in the first 28 days of therapy. VKORC1-1639 genetic polymorphism was also associated with shorter time to the INR within the therapeutic range and increased risk of over-anticoagulation.

CONCLUSIONDetecting genetic polymorphism of CYP2C9 and VKORC1 could guide clinical use of warfarin to reduce the risk of adverse reactions including bleeding in patients receiving chronic anticoagulation therapy.

Aged ; Anticoagulants ; pharmacology ; therapeutic use ; Aryl Hydrocarbon Hydroxylases ; Cytochrome P-450 CYP2C9 ; genetics ; Gene Frequency ; Genes ; Genetic Variation ; Genotype ; Hemorrhage ; Humans ; International Normalized Ratio ; Linkage Disequilibrium ; Mixed Function Oxygenases ; Polymorphism, Single Nucleotide ; Vitamin K Epoxide Reductases ; genetics ; Warfarin ; pharmacology ; therapeutic use