Objective To study the characteristics of event-related potentials P300 in patients with anxiety disorder(AD). Methods P300 tests were carried out in 30 patients with AD and 30 healthy adult controls. ResultsPatients with AD had significantly delayed P3 latency ([326?16] ms vs [339?19]ms, P

Vitexin, a naturally occurring flavone glycoside in plants, has many pharmacological effects, which is widely distributed in nature. This paper reviewed the research progress of the distribution of vitexin in the plant resources and its pharmacological effects, and summarized its application prospects, aiming to provide a useful reference for the development of vitexin-enriched plant resources.
Animals ; Antineoplastic Agents ; pharmacology ; Antioxidants ; pharmacology ; Apigenin ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Myocardial Infarction ; drug therapy ; Plant Dispersal

Objective To determine the influence of protein fusion on the biological characteris- tics of hymidine kinase(TK)and human immunodeficiency virus(HIV)Tat recombinant protein. Methods By utilizing polymerase chain reaction(PCR)technique,different fragments containing two,four or six glycines(Gly)were inserted between the HIV Tat gene and TK,and cloned into PBK vector.After testified by sequencing,the vectors were transfected into E coli.After induced by iso- propyl thiogalactose(IPTG),bacilli were collected and destructed by ultrasonic,the fusion proteins were determined by monoclonal antibody against HIV protein.HepG2 cells were incubated in DMEM supplement with 10?g/mL HIV-Gly(n)-TK(n=0,2,4,6)fusion protein,TK-HIV Tat and only HIV Tat.HepG2 cells in different groups were detected by immunofluorescence assay 24 hours after transduction with HIV Tat monoclonal antibody.The rate of apoptosis after cells were incubated with gencilovir(10?g/mL)for 3 days was determined by cell flow cytometry,while survival cell ratio was recorded by trypan blue.The data were analyzed by statistics(t-test).Results The Tat-Gly(n)-TK (n= 0,2,4,6)recombinant genes were constructed and inserted into PBK vectors,which were expressed in E coli and then purified.Cells in different groups,which were incubated with Tat-Gly (n)-TK(n=0,2,4,6)fusion proteins,Tat-TK fusion protein,TK-Tat fusion proteins or only Tat proteins respectively,were detected by immunofluorescence assay.The intensities of fluorescence in different groups were almost same,but the ratios of cell survival or apoptosis were different.The highest ratio of cells apoptosis(14.77%)was in the group that cellular culture medium was mixed with Tat-Gly(4)-TK fusion protein,followed by the groups containing 6,2 glycines or no TK gene in genes(4.30%,12.69% and 1.03%,respectively).There were significant differences between each 2 groups among the all groups(t-test,P

Objective To assesse the quality of life of patients after cardiac pacemaker implantation using the Chinese version of SF-36.Methods Ninety-eight patients with permanent cardiac pacemaker implantation were investigated before and after the operation in terms of quality of life by using the Chinese version SF-36.Results Successful surgery was performed on all the 98 patients.The previous symptoms of the patients were improved to vari- ous extend after the operation.The quality of life of the patients was significantly improved after operation as demon- strated by the significant difference of the scores in 9 domains of SF-36 when compared with those before the operation (P

OBJECTIVETo explore the role of CD(8)(+)CD(28)(-) T regulatory cells (Tr) in the immunological pathogenesis of acute infection with Epstein-Barr virus in children.

METHODSThe present study enrolled 25 children with infectious mononucleosis (IM) and 25 age-matched healthy children. Flow cytometric analysis was performed to detect the percentage of CD(3)(+), CD(3)(+)CD(4)(+), CD(3)(+)CD(8)(+), CD(8)(+)CD(28)(+) by determining the ratio of positive cells in lymphocytes. Reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR were used to analyze IL-6, IL-10, IFN-gamma expression in CD(8)(+)CD(28)(-) Tr cells and ILT-3, ILT-4 expression in monocytes/macrophages.

RESULTSThe proportions of CD(8)(+)CD(28)(-)T cells in children with acute-phase IM was significantly higher than those in the controls (P < 0.01). The expression level of IL-6, IL-10, IFN-gamma, ILT-3, ILT-4 mRNA significantly increased compared to those of the controls (P < 0.01).

CONCLUSIONThe CD(28) expressed on CD(8)(+) T cells in vivo is gradually lost with age and CD(8)(+)CD(28)(-) cells increase up 50% to adult. EBV can directly infect B cells, trigger CD(8)(+) CTL response and destroy the target cells to cause serious immunopathological lesion. Therefore we speculate that the expansion of CD(8)(+)CD(28)(-) Tr cells in children with IM may be an adaptive immune response to avoid serious inflammation and autoimmune reactions.

CD28 Antigens ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; Cytokines ; immunology ; Epstein-Barr Virus Infections ; immunology ; Female ; Flow Cytometry ; Herpesvirus 4, Human ; Humans ; Infectious Mononucleosis ; immunology ; virology ; Male ; Membrane Glycoproteins ; metabolism ; Receptors, Cell Surface ; metabolism ; Receptors, Immunologic ; metabolism ; T-Lymphocytes, Regulatory ; immunology

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.

The pol gene of HIV-1 encodes mainly three enzymes: reverse transcriptase (RT), protease (PR) and integrase (IN). Currently, FDA approved drugs targeting RT and PR are available and administered in various combinations, while no anti-IN drug was approved. HIV-1 integrase is an essential enzyme for the viral replication and an interesting target for the design of new pharmaceuticals for multi-drug therapy of AIDS. The 288 amino acids of IN (32kDa) recognizes specific sequences in the long terminal repeats (LTRs) of the retroviral DNA. The IN protein catalyzes the 3′-processing step and the 5′-strand transfer step reaction in vivo, which was called integration and this reaction could be analysed by ELISA Assay in vitro. It has been reported that F185K and C280S mutations of HIV-1 integrase would improve the enzyme solubility, and the catalytical activity of the enzyme was the same as that of the wild-type enzyme in vitro. In order to build the platform of screening inhibitor against integrase of HIV-1 virus, the IN enzyme was expressed and the function of integrase protein was assayed. The cDNA of clade B HIV-1 genome was used as a template, overlapped PCR was used to construct site mutagenesis of F185K/C280S and NdeI/Xho I restriction sites were brought in. The PCR product was cloned into the prokaryotic vector pET-28a(+) to form a recombined plasmid, transferred into the host cell E.coli(BL21 DE3). The recombined clones were identified by PCR and Nde I/Xho I digestion .The positive plasmid was sequenced, and the successfully recombined plasmid in the host cell was induced by IPTG. The expressed IN protein was puriied sy the Co+ affinity chromatography column and SDS-PAGE was used to analyze the molecular weight and specificity. In addition, ELISA assay was used to analyze the function of the recombined IN protein. The recombinant protein was soluble, and expressed highly and stably in E.coli. The molecular weight of the expression product was identical to the expectation. The IN protein was proved to be functional in 3′ processing and 5′strand transfer by ELISA. It will be helpful to build the platform of screening inhibitors against HIV-1 integrase.

OBJECTIVETo establish a method suitable to determine the purgative biopotency of rhubarb and construct a new quality evaluation pattern of rhubarb.

METHODA series of factors such as observation index (mass of feces in 10 hours), animal strain (ICR mice), sex (male) and the dose of diphenoxylate complex (50 mg x kg(-1)) was investigated and fixed. The purgative biopotency as well as anthraquinone determination was used to evaluate the quality of different rhubarb samples.

RESULTThere wasn't a good linear relationship between the purgative biopotency and content of anthraquinone. The quality difference of rhubarb samples could be well characterized by combination of purgative biopotency determination and anthraquinone determination.

CONCLUSIONThe purgative biopotency determination can be used in quality control and evaluation of rhubarb.

Animals ; Anthraquinones ; analysis ; Biological Assay ; Cathartics ; pharmacology ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Linear Models ; Male ; Mice ; Quality Control ; Reproducibility of Results ; Rheum ; chemistry ; Sex Factors ; Time Factors

The only difference of primary structure between single-chain prourokinase (pro-UK or scu-PA) and two-chain urokinase (UK or tcu-PA) is the cleavage of a single peptide bond (Lys158-Ile159) and transform scu-PA into its active two-chain form. A 13-peptide (Thr-Leu-Arg-Pro-Arg-Phe-Lys-Ile-Ile-Gly-Gly-Glu-Cys), which spans the cleavage peptide bond, was synthesized and linked to KLH (Keyhole limpet hemocyanin). The Balb/c mice were immunized by the conjugated protein with proper adjuvant. According to the Kohler and Milstein's methods, a hybridoma cell line G7 secreting monoclonal antibody specific for scu-PA was obtained. The anti-scu-PA McAb, purified from the supernatant of porous microcarrier hybridoma cell culture, was conjugated to CNBr-activated Sepharose 4B to prepare an immuno-affinity chromatography column. The u-PA was purified only by this affinity column from the supernatant of cultivating the u-PA-producing recombinant CHO cell, the u-PA recovery ratio is 90.4%, the purification factor was about 50, with the specific activity of 1.2 x 10(5) IU/mg, the scu-PA ratio in the u-PA product was 96.3%. Compared to immuno-affinity chromatography, the 3-step process for purifying u-PA (cation-exchange column, gel filtration column and benzamidine affinity column) has a u-PA recovery ratio of about 65%, with a specific activity of 1.0 x 10(5) IU/mg, and an scu-PA ratio of about 90%. These results showed that immuno-affinity chromatography is simple to recover u-PA and effective to separate scu-PA from tcu-PA.
Animals ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Chromatography, Affinity ; Enzyme-Linked Immunosorbent Assay ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; immunology ; isolation & purification ; Urokinase-Type Plasminogen Activator ; immunology ; isolation & purification

OBJECTIVETo explore the clinical application of the tubularized incised plate (TIP) in the surgical treatment of hypospadias.

METHODSThis study included 169 cases of hypospadias treated by TIP surgery from January 2007 to April 2009. The patients ranged in age from 1.5 to 12 years (mean 3.68 yr). The TIP technique was modified based on that described by Snodgrass, with the urethral plate longitudinally incised and a urethral stent kept in place. The patients were hospitalized for 10 days postoperatively, and followed up for an average of 2 years, ranging from 6 months to 3 years.

RESULTSComplications developed in 18 (10.6%) of the patients, most frequently meatal stenosis (9 cases, 5.3%) and urethrocutaneous fistula (8 cases, 4.7%).

CONCLUSIONThe TIP technique, as a surgical method, can be applied to most hypospadias cases. The accumulation of clinical experience and skills may help raise the success rate and reduce the complications of TIP surgery.

Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Treatment Outcome ; Urethra ; surgery ; Urologic Surgical Procedures ; methods