Objective To investigate the calcium mechanism involved in the A? 25-35-induced neurotoxicity.Methods PC12 cells viability was detected by using MTT assay. Relative change of intracellular calcium concentration was measured by laser confocal microscope.Results MTT assay showed that the cell viability of the three groups (10 ?mol/L A? 25-35?10 ?mol/L A? 25-35+5 ?mol/L nifedipine?10 ?mol/L A? 25-35+10 ?mol/L nifedipine) was decreased by 34.5%, 25.1% and 11.0%,as compared with the control group. 10 ?mol/L nifedipine could protect PC12 cells from A? 25-35-induced damage. A? 25-35 of different concentration (0.1 ?mol/L, 1 ?mol/L, 10 ?mol/L, 20 ?mol/L and 30 ?mol/L) resulted in the elevation of intracellular calcium (about 6.38%, 6.42%, 62.2%, 69.3% and 107.5%) with a dose-dependent manner. Potassium (5 mmol/L, 15 mmol/L and 30 mmol/L) could increase the intracellular calcium after one minute pretreatment of different concentration A? 25-35. Both above mentioned actions were sensitive to the medium calcium and were antagonized by nifedipine, a L-voltage-gated calcium channel antagonist.Conclusion Soluble A? 25-35 may damage the neuronal calcium homeostasis in early stage and make neurons more vulnerable to physiological or pathological stimuli.

Adult ; Female ; Glottis ; Humans ; Laryngeal Mucosa ; transplantation ; Laryngoscopy ; Laryngostenosis ; surgery ; Lasers, Gas ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Young Adult

The gene encoding thermostable lactate dehydrogenase (Tm-LDH) was cloned into the plasmid pHsh from Thermotoga maritima, and expressed in Escherichia coli JM 109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 33 kDa. The optimal temperature and pH of Tm-LDH were observed 95 degrees C and 7.0. The purified enzyme had a half-life of 2 h at 90 degrees C, and exhibited better stability over a pH range from 5.5 to 8.0. The K(m) and V(max) values were 1.7 mmol/L, 3.8 x 10(4) U/mg of protein for pyruvate, and 7.2 mmol/L and 1.1 x 10(5) U/mg for NADH, respectively. The expression of Tm-LDH in T7 system could not obtain high efficiency, but it has been soluble over-expression in pHsh system and reached 340 mg/L. The superior stability and productivity of Tm-LDH will lay the foundation of its industrial-scale fermentation and application in the NAD regeneration.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; metabolism ; L-Lactate Dehydrogenase ; biosynthesis ; Molecular Weight ; Recombinant Proteins ; biosynthesis ; Temperature ; Thermotoga maritima ; enzymology

Adolescent ; Adult ; Female ; Humans ; Mastitis ; therapy ; Middle Aged ; Moxibustion ; Young Adult

Adult ; China ; Humans ; Male ; Neisseria meningitidis, Serogroup C ; drug effects ; genetics ; isolation & purification

Aged ; Humans ; Kidney Neoplasms ; pathology ; Male ; Neoplasm Metastasis ; Paranasal Sinus Neoplasms ; secondary

To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

Objective To obtain scientific data for control of iodine deficiency disorders(IDD) by reviewing the surveillance information of school children intelligence quotient(IQ) after the implementation of universal salt iodization. Methods One thousand five hundred and eighty children were selected from 11 primary school in Dalian city of Liaoning province during 2006 to 2009. IQ was measured by Combined Raven Test-C2(CRT-C2) in China. Groups of IQ were classified as outstanding(≥ 130), excellent (120- 129), above average (110- 119), average (90 - 109), below average(80 - 89), margin(70 - 79), low(≤69). Urinary samples of children were collected randomly. Urinary iodine were determined by As-Ce catalytic spectrophotometry. The growth characteristics of IQ were analyzed according to surveillance year and born year. Results The average IQ of children aged 8-10 were 110.4 ± 14.0,112.5 ± 12.4,117.2 ± 11.4,116.2 ± 12.6, respectively, increased year by year from 2006 to 2009. Its average annual increase from 2007 to 2009 were 2.1,3.4,1,9 compared with the IQ in 2006 respectively. The medians of urinary iodine were 224.7,266.7,222.1 μg/L from the year 2007 to 2009, respectively, which were all between 200 - 300 μg/L and can be classified as more than adequate level. The average IQ of children born during the year of 1994 to 2000 were 106.7 ± 13.0,108.1 ± 13.9,108.5 ± 13.4,111.3 ± 14.3,113.6 ± 12.5,115.3 ± 12.3,119.8 ± 11.2, respectively. Its average annual increase from 1995 - 2000 were 1.4,0.9,1.5,1.7,1.7,2.2 compared with the IQ born in 1994 respectively. The ratio of IQ in margin group and low group were all below 2% ; the ratio was increasing in excellent group and outstanding group and decreasing in average group and below average group significantly year after year(x2 = 52.471,34.329,66.483,11.148, all P<0.01). Conclusions Universal salt iodization improves IQ scores. IQ index should be brought into the surveillance project and put in use in IDD control.

Objective To study the effect of shenfu injection on the neuron apoptosis in hippocampal CA1 region of newborn with hypoxic-ischemic brain damage(HIBD).Methods The experiment included 2 parts.One was to measure the apoptosis rate by the flow cytometry,and the other was to investigate the expression of Bcl-2 and Bax of neurons in left hippocampal CA1 region.Models of postnatal 7-day Sprague-Dawley(SD)rats with HIBD were established,and were equally divided into 4 groups:sham operation(group S),control group(group C),shenfu injection pretreatment(group P),and shenfu injection treatment(group SF).The neuron apoptosis rate in hippocampal CA1 region in every group was measured at 2 h before and 2,12,24 h,3,7,14 and 28 d after hypoxic ischemic(HI) insult.The expressions of Bcl-2 and Bax were performed by immunohisochemistry.Results The apoptosis of neuron in hippocampal CA1 region in group P and group SF after HI insult was significantly less than that of group C(P

Objective To explore the accuracy of Vitek 2 compact advanced expert system (AES) in indicating and analyze the carbapenemases-resisting Enterobacteriaceae phenotypes,and further investigate the methods to make up the AES.Methods 28 Enterobacteriaceae strains with Imipenem-Nonsusceptible by Vitek 2 compact,but AES suggested all production of carbapenemases were isolated.And imipenem susceptibility was determined by the disk diffusion method.Modified Hodge test (MHT) and the metallo-β-1actamase was detected by the double disk synergy method.Resistance genes were detected by the PCR amplification.Results ESBLs gene was amplified from all 28 selected strains,16 of which was detected KPC gene,and no strain of metallo-β-1actamases-producing bacteria.With carbapenemase gene detection as the gold standard,the accuracy of AES was 57.1％.Disc diffusion method detection accuracy rate of imipenem was 100％,and for 100％ of MHT accuracy.PCR amplification,MHT and the disk diffusion displayed the same result in detecting carbapenemases,but different with AES (x2 =10.08,P＜0.05).Conclusion The indications of the presence of carbapenemases using AES was not completely correct with a certain false-positive,and it is necessary to take other methods,such as disk diffusion or MHT methods,and improve the reliability of medicine-sensitivity tests.