It is still controversial whether total splenectomy on adult animals would affect their immune function. In this study, total splenectomy and different kinds of splenic preservation operations including splenorrhaphy, partial splenectomy and spleen autotransplantation were performed on adult rats after the porta of the spleen of the rats was artificially ruptured and the splenic artery ligated.In the 4th month after the operations, the hemolysin antibody and the de-layed-type hypersensitivity reaction were determined in the rats. It was found that no significant difference occurred in the hemolysin antibody and the delayed type hypersensitivity reaction among the rats of the control group, the group with total splenectomy, and the groups with different kinds of splenic preservation operations.

BACKGROUND: One of the key neuropathological changes in Alzheimer disease is that neurofibrils over phosphorylated cytoskeletal protein (such as r and neurofilaments) composed of entwist together, and the phosphorylation of τ protein can be catalyzed by cyclin dependent kinase 5 (CDK5),however whether the phosphorylation of neurofilaments can be catalyzed by CDK5, as well as its role in the pathogenesis of Alzheimer diseases is less acknowledged.OBJECTIVE: To explore the role of over-expression of intracellular CDK5 in the phosphorylation of neurofilamentsDESIGN: Randomized controlled study.SETTING: Biochemical and Molecular Biological Department of Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: This study was conduced at Biochemical and Molecular Biological Department of Tongji Medical College, Huazhong University of Science and Technology between February and May 2001. In vitro cultured rat neuroblastoma cell strain (N2a) was adopted as subjects.METHODS: In vitro cultured N2a cells were divided into 2 groups, namely transfection group and non-transfection group. In transfection group,CDK5 gene was transfected into N2a cell line by using liposome transfection technique so as to obtain N2a/CDK5 cell line stably expressing CDK5, immune-precipitation and enzyme activity assay was used to detect the CDK5 activity, meanwhile immunofluorescence technique and immuneblot assay was used to detect CDK5 expression and phosphorylation of neurofilaments.MAIN OUTCOME MEASURES: Phosphorylation of neurofilaments in both groups.RESULTS: In transfection group of N2a cell line, CDK5 expression increased presented by deep coloration of SMI31 antibody and weak coloration of SMI32 antibody, implying hyper-phosphorylation of neurofilaments. Meanwhile, the activity of CDK5 was 3.5 times higher than that in non-transfection group.CONCLUSION: Intracellular over-expressison of CDK5 would lead to hyperactivity of CDK5 and hyper-phosphorylation of neurofilaments, however the hyper-phosphorylation of neurofilamentsmight invlove in the pathological development of AD.

ObjectiveTo investigate the effects of using transforming growth factor-β3 (TGF-β3) combined with dental pulp stem cells (DPSCs) on the regeneration of rabbit facial nerve.MethodsSixteen New Zealand adult rabbits were selected randomly.These rabbits were divided into three groups (TGF-β3 and DPSCs treated group,TGF-β3 control group and PBS control group.treatment group (silicone guidance channel with collagen protein sponge were filled with TGF-β3 and DPSCs),TGF-β3 control group (silicone guidance channel with collagen protein sponge were filled with TGF-β3) and PBS control group(silicone guidance channel with collagen protein sponge were filled with PBS).After 3 months,a series of examinations were performed,including gross morphology,histological staining,neuroelectrophysiological tset.ResultsIn the experimental group,the diameter of the regenerating nerve and near distal nerve stem were almost the same.There were no formation of neuroma.The adventitial angiogenesis was rich and with tough texture.3 months after operation,in the experimental group,the facial nerve membrane integrity,nerve fibers arranged in neat rows,form a more complete,myelin swelling.The total number of regeneration of nerve fibers in the experimental group were more than of control groups,statistical analysis was significant (P＜0.05).Diameter of regenerating nerve fibers in the experimental group were greater than that of control groups,statistical analysis was significant (P＜0.05).The ultra-thin section showed that the regenerated fibers in the treatment group were mainly myelinated never fiber.The layer structure of myelin sheath was clear,and there were rich organells axoplasma.The neuroelectrophysiological examinations revealed that the latency of nerve and muscle action conduction in the treatment group was shorter than that of the control groups,the treatment group:(1.96±0.32) ms,the TGF-β3 control group:(2.35±0.41) ms,the PBS control group:(3.42±0.55) ms.The wave amplitude of nerve and muscle action conduction in the treatment group was obviously higher than that of control groups,the treatment group:(11.06±3.25) mV,the TGF-β3 control group:(8.40± 1.68) mV,the PBS control group:(4.62±0.77) mV.ConclusionThe combination of TGF-β3 and DPSCs can improve the effects on the repair of facial nerve injury.

OBJECTIVE: To standardize the management on drug use in primary hospitals in Beijing Dongcheng district so as to enhance the efficiency of drug administration.METHODS: A "Drug Use Monitoring Information Platform in Primary Hospital" comprising management information system on drug use,management system on information of drug wholesalers,drug monitoring management information system was established and its problems in the try-out were analyzed.RESULTS & CONCLUSIONS: The "Drug Use Monitoring Information Platform in Primary Hospital" still faced with the problems of legal basis,psychological concern,and construction of standard basic drug information database etc.However,its operation standardized the drug purchasing channel and management on drug use in hospitals of Beijing Dongcheng district and enhanced the drug administration level and efficiency,which thus deserves to be popularized throughout the country.

A rapid analytical method for the determination of dezocine and pethidine in hair samples using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established.After cleaned hair was extracted by grinding with methanol and ultrasonic, the final solution was analyzed by UPLC-MS/MS.The targets were gradient eluted on a Waters Acquity BEH C18 (2.1 mm × 100 mm, 1.7 μm) column with 0.1% formic acid-water and methanol as mobile phase at a flow rate of 0.4 mL/min.The ESI+ ion source and multiple reaction monitoring (MRM) were used to select the qualitative and quantitative ion pairs of dezocine and pethidine.Dezocine and pethidine showed good linearity in the range of 0.01-8 ng/mg, with the limit of detection of 0.005 ng/mg and the LOQs of 0.01 ng/mg.The accuracy, precision, matrix effect, extraction recovery, and stability all met the requirements.The established method is simple, rapid, and accurate for the qualitative and quantitative determination of dezocine and pethidine in hair, which can be applied in the case analysis of dezocine and/or pethidine abuse.

Lipase of 54/923 cases increased in viral hepatitis group with an abnormal rate of 5.85％.Lipase level of viral hepatitis patients gradually increased with the rising total bilirubin levels,and the difference was statistically significant (P ＜0.0l).Among 54 viral hepatitis cases with elevated lipase,11 died.The sustained abnormal rate of lipase (6/11) of the death group was statistically significant vs.4.65％ (2/43) of the survival group(P ＜0.01).Thus viral hepatitis could be concomitant with elevated lipase and elevated lipase might promote the progression of liver disease.The continuing rise of lipase is correlated with the severity and prognosis of the patients.

In this study, we studied the effect of glycogen synthase kinase-3 (GSK-3) overactivation on neurofilament phosphorylation in cultured cells. After N2a cells were treated with the specific inhibitor (wortmannin) of phosphoinositol-3 kinase (PI-3K) or treated with wortmannin and the specific inhibitor (LiCl) of glycogen synthase kinase-3 (GSK-3), GSK-3 activity and neurofilament phosphorylation were detected by using GSK-3 activity assay, Western blots and immunofluoresence. Our results showed that after treatment of N2a cells with wortmannin for 1 h, overactivation of GSK-3 caused a reduced staining with antibody SMI32 and an enhanced staining with antibody SMI31. When N2a cells were treated with wortmannin and LiCl, the activity of GSK-3 was reduced substantially. At the same time, the phosphorylation of neurofilament was also reduced. The study demonstrated that overactivation of GSK-3 induced hyperphosphorylation of neurofilament and suggested that in vitro overactivation of GSK-3 resulted in neurofilament hyperphosphorylation and this may be the underlying mechanism for Alzheimer's disease.

A quantitative analysis method based on solid phase extraction-ultra performance liquid chromatography-tandem mass spectrometry (SPE-UPLC-MS/MS) for simultaneous determination of illicit drugs and their metabolites in wastewater was established. Samples filtered at pH of 2 and spiked with internal standard were loaded to Oasis Prime MCX cartridges for solid-phase extraction. The samples were washed with 4 mL of methanol and eluted with 4 mL of 5% ammonia in acetonitrile before reconstituting with 0.1% formic acid/water solution. ZORBAX Eclipse Plus C18 column was used for chromatography, and gradient elution was performed with 0.1% formic acid/water solution and acetonitrile as mobile phase. The samples were then detected by electrospray ionization (ESI) in positive ion mode, and multiple reaction monitoring mode (MRM) was adopted for quantitative analysis. All analytes had a good linear relationship (r ≥ 0.993 2) within the range of their respective standard curve; the limit of quantification was 1 ng/L (except amphetamine at 2.5 ng/L); the extraction recovery ranged from 82.13% to 99.96%; and the intra- and inter-day precisions were less than 9.43%. The method is accurate, reliable and reproducible, and is suitable for the quantitative determination of illicit drugs and their metabolites in wastewater and can provide an analytical method for real-time monitoring of drug abuse.

The waste water-based epidemiology is an important technique to fight against drug abuse by analyzing the concentration of illicit drugs in urban sewage, which can monitor the abuse of drugs.An SPE-UPLC-MS/MS method was developed for the analysis of 12 common drugs and their metabolites involving amphetamine and morphine.It was shown that the best result was achieved when hydrochloric acid/ acetonitrile (5∶95) was added to acidify the sample during the concentration process, guaranteeing the anti-across contamination of the analysis of organic nitrogen basic trace components, and improve the stability, specificity, and accuracy of the method.The optimized method meets the analytical requirements of complex sewage samples, and has been successfully applied to the assessment of urban drug abuse through sewage analysis.

To explore the intracellular signal pathways for beta-VLDL induced very low density lipoprotein receptor (VLDL-R) transcription up-regulation and their effects on lipid accumulation in macrophages, Western Blot was used to examine phosphorylated ERK1/2 protein and regulated effects by different singal kinase inhibitants. It was found that beta-VLDL induced an increase in ERK1/2 activity in a protein kinase C (PKC)-dependent manner in murine RAW264.7 macrophages. By using different protein kinases inhibitors or activators, it was observed that the effect of beta-VLDL induced VLDL receptor transcription, which was monitored by RT-PCR analysis of VLDL receptor mRNA, was not affected by the inhibitor of p38 kinase and cAMP analog, but extremely abolished by pretreating cells with PD98059, an inhibitor of ERK and GF 109203X, an inhibitor of PKC. These results demonstrated that the PKC-ERK1/2 cascade is the essential signaling pathway by which beta-VLDL activated VLDL-R mRNA expression. Inhibition of the ERK1/2 signaling cascade resulted in suppression of the cellular lipid accumulation induced by beta-VLDL in macrophages.
Cells, Cultured ; Lipoproteins, VLDL ; metabolism ; Macrophages ; cytology ; metabolism ; Mitogen-Activated Protein Kinase 1 ; metabolism ; physiology ; Mitogen-Activated Protein Kinase 3 ; metabolism ; physiology ; Protein Kinase C ; antagonists & inhibitors ; metabolism ; Receptors, LDL ; biosynthesis ; genetics ; Signal Transduction ; Transcription Factors ; metabolism ; Transcription, Genetic ; Up-Regulation