Objective Alzheimer's disease(AD) is a common and devastating disease and there is no readily available biomarker to aid diagnosis or to monitor disease progression. This study is to further understand the pathogenesis of Alzheimer's disease and seek new biomarkers for AD. Methods AD rats made by Aβ1-40 were assessed using the Morris water maze method. Sera of two groups were collected and albumin, IgG and sodium chloride were removed from the serum before concentration. Two-dimensional electrophoresis(2-DE) was performed on serum protein. Altered proteins were identified by matrix assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results Compared with untreated samples, 10 altered proteins were found through 2-DE and all of them were down-regulated of which 2 were identified by MALDI-TOF-MS to be plasma retinol-binding protein precursor and complement component 4, gene 2 (C4b). Conclusion C4b may be the biomarker of AD and it may be helpful to understand the pathogenesis of AD.

Objective Predicting protein structural class is the basis for predicting protein spatial structure,so it is important to improve the prediction accuracy of protein structural class.Methods We proposed 3-state and 8-state Hidden Markov model (HMM),and applied these HMMs to the prediction of protein structural class,respectively.We evaluated their accuracy on two different datasets through the rigorous jackknife cross-validation test.Results Prediction ability of 8-state HMM and 3-state HMM to all α class were excellent,the prediction accuracy of 3-state HMM even reached above 95％.Compared with Chou data set,the prediction accuracy of Zhou data set for all β class and α/β class of was improved,while overall prediction accuracy increased by 2％.Conclusion HMM is an effective method to predict protein structural class.

AIM: To develop a simple and sensitive high-performance liquid chromatography (HPLC) for the quantification of zidovudine and to study the pharmacokinetics of two kinds of zidovudine capsules in Chinese healthy volunteers. METHODS :The concentrations of zidovudine in plasma were determined by a validated HPLC method with UV detection. A randomized two-way crossover study was conducted in 18 fasting volunteers to compare plasma pharmacokinetic profile and single-dose tolerability of a new zidovudine capsules. RESULTS: The main pharmacokinetic parameters of two formulations, reference and test capsules, were as follows: cmax were (2 252±s 837) μg·L-1 and (2 300±1 099) μg·L-1; tmax were (0.49±0.19) h and (0.5±0.3) h;t1/2 ke were (0.93±0.19) h and (0.99±0.24) h; AUC0-t were (2 530±452) μg·h·L-1 and (2 467±605) μg·h·L-1;AUC0-∞ were(2 689 ± 414) μg·h·L-1 and (2 583±575) μg·h·L-1. The results of ANOVA and two one-side t test statistical analysis for lg AUC0-t, lg AUC0-∞ and lg cmax showed that two formulations were bioequivalent. CONCLUSION:The method is convenient, sensitive, accurate and reproducible, and could be applied to determining the plasma zidovudine concentration and studying on pharmacokinetics. Two zidovudine capsules are bioequivalent in Chinese healthy volunteers.

Twelve populations of Inula lineariifolia were used as materials to measure morphological characteristics, photosynthetic parameters and chemical constituents. It aims to provide a theoretical basis for germplasm resources evaluation. The results showed that I. lineariifolia had relatively rich morphological diversity, there were significant differences of morphological characteristics, photosynthetic parameters and chemical constituents among populations. There was positive correlation on morphological characteristics and P(n). Twelve populations were divided into three-type. The three populations of Xuyi, Mingguang and Fengyang were of narrower-longer leaf, bigger biomass,better photosynthetic and higher chemical constituents. Then they were classified for a similar group. It proved that the three populations were more suitable for cultivation and promotion.
Biomass ; China ; Flowers ; chemistry ; growth & development ; metabolism ; Inula ; chemistry ; classification ; growth & development ; metabolism ; Photosynthesis ; Plant Leaves ; chemistry ; growth & development ; metabolism ; Plant Stems ; chemistry ; growth & development ; metabolism

OBJECTIVEStudies on DNA fingerprinting of eight species of Paris and application of restriction site amplification polymorphism (RSAP) to the identification of Paris.

METHODSequence-related amplified polymorphism (SRAP) molecular markers were used to detect the genetic diversity of 7 accessions of Paris collected from Tianquan and Baoxing in Sichuan, and one from Lijiang in Yunnan.

RESULTThe DNA fingerprinting of 8 species were generated by 18 primer combination screened from 45 primer combinations. Eight accessions were clustered into 4 groups by genetic distance.

CONCLUSIONBased on molecular biology methods of RSAP analysis, accurate molecular identification could be performed on traditional Chinese medicinal material plants in Paris, and provided molecular evidence for taxonomy and identification of different species in Paris.

DNA Fingerprinting ; Genetic Markers ; Genetic Variation ; Liliaceae ; genetics ; Polymorphism, Genetic

BACKGROUNDBecause of the pivotal role of the human leukocyte antigen (HLA) class II molecules in regulating the immune response and their extensive polymorphism, it is not surprising that particular HLA class II alleles have been implicated in susceptibility to allergic diseases and in restriction of the IgE responses to a variety of allergens. We investigated the relationship between HLA-DRB genotype and allergies to various penicillins and explored HLA-DRB restriction of IgE responses to these derivatives of penicillin.

METHODSRadioallergosorbent test was used to examine 8 kinds of specific IgE antibodies (4 major and 4 minor antigenic determinants) in the sera of 248 patients with an allergy to penicillins and 101 healthy subjects without any allergic reaction. Some (113 patients and 87 healthy control subjects) were chosen from all subjects to type for HLA-DRB alleles by sequence specific primer-polymerase chain reaction.

RESULTSCompared with control subjects, a significantly increased frequency of DR9 was present in 77 patients with allergic reactions, with immediate hypersensitive reaction and with urticaria (P = 0.011; P = 0.019; P = 0.005 respectively). Conversely, a significantly decreased frequency of DR14.1 was found in 80 patients with positive IgE antibodies, with immediate reaction and with urticaria compared with control group (P = 0.024; P = 0.038; P = 0.038). A possible excess of HLA-DR17 was found in subjects who were responsive to benzylpenicilloyl compared with those were not (chi(2) = 5.134, P = 0.023), and of HLA-DR4 was found in subjects responsive to phenoxomethylpenicillanyl (PVA, chi(2) = 4.057, P = 0.044).

CONCLUSIONHLA-DRB gene may be involved in allergy to penicillins through modulating specific serum IgE to penicillins.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Alleles ; Child ; Child, Preschool ; Drug Hypersensitivity ; genetics ; immunology ; Female ; Genotype ; HLA-DR Antigens ; genetics ; Humans ; Immunoglobulin E ; blood ; Male ; Middle Aged ; Penicillins ; adverse effects ; immunology

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

The present study was aimed to examine if protein kinase C (PKC) activation is necessarily involved in both the c-fos protein expression in the nocuously-activated c-fos protein-like immunoreactive (Fos-LI) neurons and the concomitant opioid receptor-mediated modulation in the dorsal horn circuitry of the spinal cord. Formalin was injected into a hindpaw of rats 5 min after the rats were pretreated with intrathecal (i.t.) administration of chelerythrine (Chel), an inhibitor of PKC, naloxone (Nal), combined administration of these two (Chel + Nal), or vehicle (n=5 in each group),respectively. By using immunocytochemical techniques, the formalin-induced Fos-LI neurons in the lumbar dorsal horn were calculated 1 h after formalin injection. The results showed that: (1) i.t. Chel significantly reduced the number of Fos-LI neurons in the dorsal horn of the spinal cord on the side ipsilateral to the formalin injection, showing a decrease by 60.3% (P<0.001) as compared to that observed in the i.t.vehicle group; (2) i.t. Nal significantly increased the number of Fos-LI neurons in the ipsilateral dorsal horn, with an increase of 46.0% (P<0.01) as compared to that in the i.t.vehicle group, the highest percentage increase being found in the deeper laminae of the dorsal horn; and (3) i.t. Chel + Nal also exhibited a significant decrease in Fos-LI neurons in the ipsilateral dorsal horn as compared to i.t. Nal group, showing a reduction of 53.2%, a value similar to that in the i.t. Chel group. These results suggest that: (1) PKC plays a role in the c-fos protein expression only in nearly one half of the Fos-LI neurons in the dorsal horn; and (2) PKC is possibly not involved in the concomitant modulation on the nociception mediated by micro- (and also partly delta-) opioid receptors in the spinal cord.
Animals ; Formaldehyde ; pharmacology ; Immunohistochemistry ; Male ; Naloxone ; pharmacology ; Narcotic Antagonists ; pharmacology ; Nociceptors ; physiology ; Pain ; metabolism ; physiopathology ; Posterior Horn Cells ; physiology ; Protein Kinase C ; metabolism ; physiology ; Proto-Oncogene Proteins c-fos ; biosynthesis ; physiology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, delta ; agonists ; Spinal Cord ; physiology

The purpose of this research is to investigate the critical period of voltage-gated Na(+) channel development in hippocampal CA1 neurons. Changes of Na(+) currents in acutely isolated hippocampal CA1 neurons of rats at different ages (0-4 weeks after birth) were recorded using the whole-cell patch-clamp technique. The results indicated that the maximum current density of Na(+) channels was increasing with age, and the amplitudes in 1, 2, 3 and 4 weeks respectively grew by (42.76 ± 4.91)%, (146.80 ± 7.63)%, (208.79 ± 5.28)% and (253.72 ± 5.74)% (n = 10, P < 0.05) compared with that in 0 week. The current density in CA1 neurons of 1-2 weeks after birth increased more significantly than those of other groups. The activation curve of Na(+) channel shifted to the left. The half-activation voltages (mV) in 0-2 weeks were -39.06 ± 0.65, -43.41 ± 0.52, -48.29 ± 0.45 (n = 10, P < 0.05), respectively, showing significant age-dependent decrease, and there were no significant changes in other groups. The slope factors of activation curve for each group did not change significantly. There were no regular changes in inactivation curve and no significant changes in half-inactivation voltage. The slope factors of inactivation curve in 1-2 weeks were: 5.77 ± 0.56, 4.42 ± 0.43 (n = 10, P < 0.05). The inactivation rate of the second week after birth was faster than that of the first week, and there were no significant changes during 0-1 week and 2-4 weeks. The recovery from inactivation curve of Na(+) channel shifted to the left. The recovery time declined in 1-3 weeks. Changes of action potential properties were consistent with Na(+) current. These results suggest that the period of 1-2 weeks after birth may be the critical development period of voltage-gated Na(+) channel in hippocampal CA1 neurons. During this time, the distribution of Na(+) channel increases significantly; the activation curve of Na(+) channel shifts to the left; inactivation rate increases as well as recovery time shortens.
Action Potentials ; Animals ; CA1 Region, Hippocampal ; cytology ; Neurons ; physiology ; Patch-Clamp Techniques ; Rats ; Sodium Channels ; physiology

OBJECTIVETo observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.

METHODSForty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.

RESULTS(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).

CONCLUSIONThe moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.

Acupuncture Points ; Animals ; Arthritis, Experimental ; metabolism ; therapy ; Disease Models, Animal ; Female ; Humans ; Moxibustion ; Potassium ; metabolism ; Rats ; Rats, Sprague-Dawley