The partial segment of Marek′s disease virus (MDV) glycoprotein B (gB) gene was amplified by PCR. The segment was cloned into pET-28a vector to obtain the recombinant pET-gB plasmid. The recombinant plasmid was transformed into E.coli BL21,and expressed in very high level as inclusion body after induced with 1.0mmol/L IPTG. The inclusion body was solubilized in urea (8mol/L) . The purified protein was obtained by use of His?Bind affinity chromatography. Mice were immunized i.p. by the purified protein to make the polyclonal antibody. The titer of the antibody by indirect ELISA was 1?10~ -5 . Moreover, the analysis by western blot proved that antibody was specific to the recombinant protein. These works lay a favorable foundation for the study of the immune response by MDV gB.

In recent years,fluorescent probes become more available for the progress of biology and gene technology,which has accelerated the development of fluorescence molecular imaging.With these fluorescent probes,target molecular,protein and gene can be specifically located and analyzed,which make possible the early detection and treatment of disease.This paper gives an introduction of the fluorescent probe technology and its application in the fields of biology and medicine.

OBJECTIVE To investigate the distribution and antibiotic resistance of nonfermenting gram-negative bacilli isolated from lower respiratory tract infection patients from 2003 to 2007 for rational antimicrobial therapy in practice.METHODS The data of distribution and antibiotic resistance of nonfermenting gram-negative bacilli isolated from sputum samples of lower respiratory tract infection in our hospital from 2003 to 2007 were retrospectively analysed.RESULTS The most predominant nonfermenting gram-negative bacilli isolates were Pseudomonas aeruginosa,Acinetobacter baumannii,Acinetobacter lwoffii and Stenotrophomonas maltophilia.From 2003 to 2007,the resistance rates of these isolates to 15 antibiotics increased,especially P.aeruginosa to ceftazidime.The resistance changed from 30.2% in 2003 to 55.4% in 2005,54.8% in 2006,and reduced to 41.5% in 2007.The resistance to gentamicin,imipenem,netilmicin,ticarcillin and tobramycin varied in the similar way.The resistance of A.baumannii to imipenem increased from 21.1% to 59.7% within the five years.The resistance rate of A.lwoffi was still relatively low.S.maltophilis isolates were highly multi-resistant to antimicrobial agents tested with a rate ranging from 61.9%-100.0% except ceftazidime,ticarcillin and trimethoprim-sulfamethoxazole.CONCLUSIONS P.aeruginosa,A.baumanii and S.maltophilia are the main nonfermentive pathogens of lower respiratory tract infections and not susceptive to many antibiotics.The tendency of susceptibility and distribution changes of those bacteria should be considered in practice for proper antimicrobial therapy.

Objective To explore the effect of endoplasmic reticulum stress (ER stress) in uric acid?induced phenotypic change in renal tubular epithelial cells (HK?2). Methods (1) HK?2 cells were cultured with 0, 75, 150, 225, 300 mg/L uric acid for 24 h in vitro. (2) The cells were divided into normal control group, ER stress inhibitor 4?PBA (5 μmol/L) group, uric acid (150 mg/L) group and 4?PBA+uric acid group for 24 h. Morphological changes of HK?2 cells were observed under inverted microscope. MTT assay was used to detect the proliferation of HK?2 cells treated with 150 mg/L uric acid for 24, 48 and 72 h. The protein expressions of α?smooth muscle actin (α?SMA), vimentin, snail, glucose regulated protein 78 (GRP78) and the phosphorylation of eukaryotic initiation factor 2α(p?eIF2α) in HK?2 cells were measured by Western blotting. Results Compared with the control group, HK?2 cells in uric acid groups (150, 225, 300 mg/L) showed fibroblast?like appearance. The protein expressions of α?SMA, vimentin, snail, GRP78 and p?eIF2α in 150 mg/L and 225 mg/L uric acid groups were higher than those in the control group (all P<0.05). The proliferation of HK?2 cells in 150 mg/L uric acid group was lower than that in control group at 48 and 72 h (all P<0.01). Compared with the uric acid group, the cell morphology in 4?PBA+uric acid group was improved, and the protein expressions ofα?SMA, vimentin, snail, GRP78 and p?eIF2α were decreased (all P<0.05). Conclusions Uric acid may induce the phenotype transformation of renal tubular epithelial cell, and ER stress is involved. 4?PBA may inhibit the uric acid?induced ER stress response and phenotypic transformation, and may be beneficial in attenuating uric acid?induced renal tubular damage.

BACKGROUND:Titanium mesh is a new type of bone graft supports used for the treatment of spine, especial y the cervical spine disorders at home and abroad in recent years. But, at present, how to improve the cervical lordosis curvature during anterior surgery is the difficulty for the treatment of cervical spondylotic myelopathy.
OBJECTIVE:To observe the effect of straight titanium mesh and individual designed titanium mesh used in the bone graft fusion and internal fixation on the recovery of cervical curvature and Japanese Orthopaedic Association score during anterior cervical subtotal vertebrectomy in the patients with cervical spondylotic myelopathy.
METHODS:Forty-three patients with cervical spondylotic myelopathy treated with surgical indications of subtotal vertebrectomy and decompression internal fixation were included, and the patients were divided into two groups according to different internal fixation methods. There were 15 cases in the individual designed titanium mesh group and 28 cases in the straight titanium mesh group. The preoperative anteroposterior X-ray films of the cervical spine of al the patients were as the blank control group. The clinical data of the patients in two groups were retrospectively analyzed, and the postoperative Japanese Orthopaedic Association score, cervical curvature, intervertebral angles and intervertebral height were compared.
RESULTS AND CONCLUSION:The cervical lordosis angle and the intervertebral angle of surgery segment in the individual designed titanium mesh group were improved when compared with those in the straight titanium mesh group and the blank control group (P<0.01), and the straight titanium mesh group was better than the blank control group (P<0.05). The intervertebral height of surgery segment in the individual designed titanium mesh group and the straight titanium mesh group was increased for 3.69 mm and 3.22 mm respectively when compared with that in the blank control group, and there was significant difference between individual designed titanium mesh group and the straight titanium mesh group (P>0.05). There was no significant difference in Japanese Orthopaedic Association score between individual designed titanium mesh group and the straight titanium mesh group (P>0.05). During subtotal vertebrectomy for the treatment of cervical spondylotic myelopathy, cutting titanium mesh into trapezoidal shape to simulate the normal disc shape of former high to low can effectively restore the physiological curvature of cervical spine and the intervertebral height of surgery segment.

OBJECTIVE:To determine the concentration of dehydroisoandrosterone sulfate (DHEAS) in human plasma by LC-MS.METHODS:With estrogen sulfate (ES) served as an internal standard,the plasma samples were deproteinized with acetonitrile,extracted by solid phase,hydrolyzed and derivatized.Then the concentration of DHEAS was determined by HPLC-MSD on Agilent SB C18 with column temperature kept at 40℃.The mobile phase consisted of acetonitrile-water (in gradient elution).Atmosphere pressure chemical ion source in negative ion detection model was employed.The ions selected for SIM (selected ion monitoring) quantitative analysis included m/z 490.0 (DHEAS ) and m/z 472.1(ES)[M-H]-.RESULTS:The linear range of DHEAS was 250.0～320.0 ng?mL-1(r=0.999 4).The extraction recovery of the simulated human albumin samples ranged from 71.1%～78.9% and its relative recovery ranged from 98.3%～101.4%.Both the intra-day RSD and inter-day RSD were less than 10%.The mean concentration of DHEAS in 15 health aged male volunteers was (981.6?353.4) ng?mL-1.CONCLUSION:The method is simple,practical,accurate and sensitive,and it is applicable for the determination of plasma concentration of DHEAS.

Objective To investigate the iodine nutritional status of children aged 8 to 10 in both costal and non-costal areas of Rizhao city Shandong province,and provide a basis for scientific supplementation of iodine.Methods In 2009 and 2010,townships of Lanshan and Donggang were chosen as survey points which situated close to the coast,the counties of Wulian and Juxian that were more than 50 kilometers away from the coast were selected as controls.The iodine concentration of drinking water in every village and water supply point was determined.At the county level,5 to 9 towns were selected according to their sub-area positions of east,west,south,north and center,4 villages were selected in each chosen township,8 - 15 households were selected in each chosen village,edible salt from the households was collected; 5 primary schools were selected in each chosen township,60 students aged 8 - 10 were selected to take thyroid examination in each chosen school.Twenty copies of urine samples were collected from the 60 students to detect the iodine concentration.Iodine in drinking water was tested by cerous sulfate catalytic spectrophotometry,in iodized salt by direct titration,in urine by arsenic-cerium contact method,and thyroids were examined by palpation.Results Totally 3483 copies of drinking water samples,1164 copies of edible salt samples,and 476 copies of urine samples were tested,and a total of 1200 children aged 8 to 10 were investigated; and the ratio of water iodine frequency distribution ≤ 10 μg/L (in costal and non-costal area) accounted for 90.03％ ( 1011/1123 ) and 91.10％ (2150/2360),respectively,the medians of water iodine were 5.6 and 4.2 μg/L,respectively; the iodine medians of edible salt were 29.03 and 29.99 mg/kg,respectively; the consumption rates of qualified iodized salt were 96.77％ (569/588) and 97.05％ (559/576),respectively; total goiter rates were 1.17％(7/600) and 1.33％(8/600),respectively; the medians of urinary iodine were 144.05 and 159.15 μg/L,respectively; the percentages of urinary iodine that bellow 100 μg/L were 26.58％ (63/237) and 22.59％ (54/239),respectively; between 100 - 300 μg/L were 66.75％ (158/237) and 64.02％ (153/239),respectively;above 300μg/L were 6.75％ (16/237)and13.39％ (32/239),respectively.ConclusionsThe costal of Rizhao is an iodine deficient area.The current popularity of household's consumption of qualified iodized salt is suitable.The levels of iodine nutrition of children aged 8 to 10 is appropriate,which met the national standard of elimination of iodine deficiency disorders (IDD).There is no iodine excess.There are no significant differences in levels of iodine nutrition between costal and non-costal areas on the premise that the same iodized salt is supplied in both the areas,and there is no need to make differentiated supply of iodized salt.

The expressing cassette, LoxP-CMV-gpt-IRES-LoxP( about 2.9kb), was amplified by PCR from a plasmid, pIRES-gpt, by use of the primers , which contained the loxP sites in 5' terminals, respectively. The loxP sites were designed into primers by the software of Primer primer 5.0. Then the cassette was cloned into the site of BalI in pBUS10 to obtain pUS-gptIRES(L). The sequencing analysis for pUS-gptIRES(L) indicated that two loxP sites with the same direction were correctly inserted into pUS-gptIRES(L).The gpt gene in pUS-gptIRES(L) was replaced by a fragment including the full length GFP gene as well as SV40 poly A sequence to get pUS-GFPIRES(L). pUS-GFPIRES(L) was transiently transfected into CHO cell lines, and then the green fluorescence could be seen, the results showed that GFP gene could be expressed correctly. Moreover, pUS-GFPIRES(L) was transfected into the CEF infected MDV CVI988 strain and recombinant virus was selected by the green fluorescence. The growth curve of virus showed the characteristic of recombinant virus was the same as that of CVI988 in vitro. These results give the basis for further studying the characteristic of MDV in vivo and the application of the Cre/LoxP system to MDV genome.

Objective: To investigate the effect of oleuropein (OE) on long term potentiation (LTP) at hippocampal perforant path-dentate gyrus synapses in vivo. Methods: An outer guide cannula, a monopolar recording electrode, and a bipolar stimulating electrode were implanted in the skull and extracellular recording technique was used to record the population spike in the dentate gyrus of anesthetized rats. Results: Oleuropein significantly increased the basal synaptic transmission and the amplitude of population spike was increased from (117.6 ± 2.3)% to (134.9 ± 3.7)% after administration with OE. OE also accelerated LTP induction and maintenance, the population spike amplitude after high frequency stimulation was increased from (167.2 ± 12.8)% to (225.5 ± 15.5)% and the maintenance phase of LTP was from (182.1 ± 15.1)% to (210.5 ± 9.0)% respectively after administration with OE. Conclusion: Present study showed that OE significantly improved different stages of LTP, which could be the molecular mechanism of its efficacy on attenuating AD-like pathology and delaying cognitive decline. OE can be a promising drug for AD and dementia.

This study is to investigate the anti-tumor activities of a novel cyclophosphamide derivate 4, 6-diphenyl cyclophosphamide (9b) in vivo and in vitro, and its possible mechanism of action. The inhibitory effects of 9b on human hepatoma cell line HepG2, human breast carcinoma cell line MCF-7 and human myeloid leukemia cell line K562 were measured by MTT assay in vitro. Cell cycle distribution and apoptotic rate were evaluated by flow cytometry. To evaluate the anti-tumor effect of 9b in vivo, mouse model bearing inoculated H22 tumor was established. The results indicated that 9b could inhibit the proliferation of HepG2, MCF-7 and K562 cells in a dose and time dependent manner. The ICo50 values of 9b were 32.34 micromol.L-1 to HepG2 cells, 87.07 micromol.L-1 to MCF-7 cells and 149.10 micromol.L-1 to K562 cells after incubation for 48 h. The results of flow cytometry indicated that after being treated for 48 h with different concentrations of 9b, the ratios of HepG2, MCF-7 cells at the Go/G1 phase and K562 cells at the G0/Gl phase and G2/M phase increased significantly compared with control group, and the apoptotic rate increased with the increase of the concentration of 9b. 9b could significantly reduce tumor weight of H22 solid tumor mouse model in vivo. To summarize, 9b showed significantly anti-tumor activity in vivo and in vitro, of which the mechanism might be associated with the change of cell cycle distribution and induction of tumor cell apoptosis.
Animals ; Antineoplastic Agents, Alkylating ; chemistry ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclophosphamide ; analogs & derivatives ; chemistry ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Inhibitory Concentration 50 ; Liver Neoplasms, Experimental ; pathology ; Male ; Mice ; Molecular Structure ; Random Allocation ; Tumor Burden ; drug effects