3.Study of Expression of Pneumococcal Surface Protein and Immunogenicity
Qian-Ying CAI ; Liang FANG ; Jin-Zhong HUANG ; Hai-Ying LIN ; Yang-Hao GUO ; Chun MENG ;
China Biotechnology 2006;0(04):-
The specific fragment of Pneumococcal surface protein A(PspA)and Pneumococcal Surface Adhesin A(PsaA)gene was amplified by PCR from Streptococcus pneumonia 5 and Streptococcus pneumonia 19.The amplified fragnent of PspA and PsaA gene was ligated into pET-27b(+)vector and transformed into BL 21 E.coli for expression and obtain the expressive production of PspA and PsaA.Induced by IPTG,the expression level was as high as 75 % of the total disolube protein.The result showed that the recombinant plasmid could express a specific 75 kDa and 37 kDa fusion protein in E.coli BL 21,which showed the good immunogenicity and a broadly cross reactivity with the other serotypes.
4.Growth activity of osteoblast on a novel strontium incorporated calcium sulfate.
Chun-Li ZHANG ; Yan-Tao ZHAO ; Shu-Xun HOU ; Hong-Bin ZHONG ; Zhong-Hai LI ; Yan LIU ; Ying ZHOU
China Journal of Orthopaedics and Traumatology 2014;27(5):415-418
OBJECTIVETo investigate the growth activity of osteoblast on a novel strontium incorporated calcium sulfate and make comparison with normal calcium sulfate material.
METHODSOsteoblast was inoculated on samples and cell proliferation was measured on the 1st, 3rd, 5th days, and the activities of ALP and osteocalcin were observed on the 5th day. And microcosmic morphology of osteoblast was observed by scanning electron microscopy(SEM).
RESULTSOsteoblast grows robustly on tested material. Cell quantity on the surface of novel material was obviously higher than normal calcium sulfate material (P < 0.05). The activity of ALP and osteocalcin on novel material was 57.8% and 40.2% higher than on normal calcium sulfate material respectively (P < 0.05). On strontium incorporated surface, osteoblast spread well. Cells were polygonal with abundant cytoplasm and the morphology was active.
CONCLUSIONStrontium incorporated calcium sulfate can sustain robust growth activity of osteoblast, which is promising to be used for bone substitute materials.
3T3 Cells ; Alkaline Phosphatase ; metabolism ; Animals ; Bone Substitutes ; chemistry ; pharmacology ; Calcium Sulfate ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Mice ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteocalcin ; metabolism ; Strontium ; chemistry
6.A comparison of four methods for extraction of human fecal DNA by using real time PCR
Zhong-Wen WU ; Ying HAN ; Hai-Feng LU ; Lan-Juan LI ; Ji-Fang SHENG ; Jian ZUO ;
Chinese Journal of Laboratory Medicine 2001;0(01):-
Objective To compare the relative efficacy and quality of extraction of human fecal DNA using four methods.Methods Real-time PCR were utilized for analysis both quantification and quality of the fecal targeted bacteria(including gut all eubaeterium,Bacteriodes-PrevoteUa group,Bifidobacterium spp Enterobacteriaceae and Enterococcus spp)by using 16s rRNA gene-targeted genus or group-specific primer sets.Results The negative rat of PCR product from method 3(phenol-chloroform plus bead-beating) was about 40%(4/10)by using universal primers,the PCR inhibition disappeared after fecal DNA purified with column.The total fecal 16s rRNA gene copy numbers(per gram of wet weight of feces)as well as the numbers of Bacteriodes-Prevotella group from method 1(QIAamp~DNA stool mini kit)and 4(QIAamp~ DNA stool mini kit combined with bead-beating)was higher significantly than that from method 2(FastDNA ~Kit,Biol01)and 3(P
7.Cloning of Endoglucanase I Gene from Trichoderma longi-brachiatum and Its Expression in Pichia pastoris
Hai-Ying LIU ; Juan WANG ; Zheng-Yu SHU ; Song-Gang WU ; Jian-Zhong HUANG ;
Microbiology 1992;0(03):-
A cellulase high-yield strain was identified and named as Trichoderma longibrachiatum SSL by ITS sequence identification. The endoglucanase1 gene (eg1) encoding endo-l,4-?-D-glucanase I was ampli-fied by RT-PCR method, which including 1386 bp and encoding 461 amino acid. Sequence analysis showed that: This gene has a more 90% homology with the T. longibrachiatum eg1 gene. The eg1 gene encoding the mature peptide was inserted into the Pichia pastoris expression vector pPIC9K, which resulted in construc-tion of the recombinant expression plasmid, pPIC9k-eg1. The pPIC9k-eg1 was then introduced into the host Pichia pastoris GS115. After the induction of methanol, extracellular recombinant endoglucanase I from the supernatant of the recombinant Pichia pastoris strain reached 73 U/mL. A clear strengthening of the protein bands, whose molecular weight is about 58 kD, appeared in the SDS-PAGE.
8.Modulatory Effect of 5-Hydroxytryptamine 2A Receptor on Discharge Activities of Inspiratory Neurons in the Medial Region of Nucleus Retrofacialis of Neonatal Rats in Vitro
Zhi-bin, QIAN ; Ming-li, JI ; Ying, QI ; Zhong-hai, WU
Journal of Applied Clinical Pediatrics 2008;23(12):965-967
Objective To explore the modulatory effect of 5-HT2A receptors on the discharge activities of inspiratory neurons in medial region of nucleus retrofacialis of neonatal rats. Methods Experiments were performed in vitro brainstem slice preparations from neonatal rats. These preparations included the medial region of nucleus retrofacialis with the hypoglossal nerve rootlets retained. The rhythmic discharges of the inspiratory neurons and activities of the hypoglossal nerve rootlets were simultaneously recorded by using microelectrodes and suction electrodes, respectively. Roles of 5-HT2A receptors in modulation of the discharge activities of inspiratory neurons were investigated by administration of the 5-HT2A receptor agonist 1-(2,5-dimethoxy-4-iodopbenyl)-2-aminopropane (DOI), and its specific antagonist ketanserine dissolved in modified Kreb's solution for perfused slices. Results In DOI group, the inspiratory time (TI) was (0.864±0.07)s, expiratory time (TE) was (10.78±1.06)s, respiratory cycle (RC) was (11.79±1.64)s, integral amplitude (IA) was (357.98±37.21)(μV·s) and the peak discharge frequency (PF) was (37.83±3.66)Hz. In control group, they were (0.68±0.06)s, (13.89±2.14)s, (14.77±1.92)s, (273.57±24.39)(μV·s), and (29.92±4.50)Hz, there were significant differences between the 2 groups (Pa<0.01). In ketanserine group, TI was (0.55±0.07)s, TE and RC were (18.43±3.28)s and (20.17+2.91)s respectively, IA and PF were (214.37±33.52)(μV·s) and (22.17±3.92)Hz, there were significant differences between ketanserine group and DOI, control group (Pa<0.01). Conclusion 5-HT2A receptors take part in modulate the discharge activities of inspiratory neurons in neonatal rat brainstem slices.
9.Clinical Features of Juvenile Dermatomyositis
hai-feng, CHU ; yu-qin, CHU ; ying-jie, ZHONG ; shan-rui, ZHAO
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To investigate the clinical manifestations and treatment regiment of children with juvenile dermatomyositis(JDM).Methods The clinical manifestation,changes of serum muscale enzyme,myopathic laboratory examination,treatment and prognosis of 15 children with JDM retrospectively admitted from Jan.1990 to Jan.2004 were analyzed.Results All of the children had symmetrical weakness of the proximal muscles.The most frequent features were heliotrope and Gottron's papules.Elevated muscle enzymes were noted in all cases.Electromyography revealed typical change of myopathic type and muscle biopsy was compalible with myositis in all cases.Most of patients achieved normal muscle enzymes within 1 month and had improved muscle strength with 2.5 monthes of the initiation of corticosteroid therapy.Conclusion It is very important to know the clinical features of JDM,and prompt diagnosis and treatment will result in an improved prognosis.
10.Modulatine Role of Gamma-Aminobutyric Acid A Receptor on Respiratory Rhythmical Discharge Activity in Brainstem Slice of Neonatal Rat
zhi-bin, QIAN ; ming-li, JI ; ying, QI ; zhong-hai, WU
Journal of Applied Clinical Pediatrics 1992;0(06):-
Objective To explore the modulation roles of gamma-aminobutyric acid A(GABAA) receptor on respiratory rhythmical discharge activity in brainstem slice of neonatal rats.Methods Experiments were performed on in vitro neonatal rats brainstem slices.The slices containing the medial region of the nucleus retrofacialis(mNRF)with the hypoglossal nerve rootlets retained was prepared in which the respira-tory-related rhythmic discharge activity(RRDA)were recorded from hypoglossal nerve rootlets by suction electrode.The possible role of GABAA receptor on RRDA was investigated by administration of GABA and its antagonist bicuculline in modified Kreb′s solution(MKS),for perfusion of the brainstem slices.Eighteen slices positive of RRDA were randomly divided into three equal groups:the first group of slices was perfused with different concentrations(10,20,40,60 ?mol/L) GABA and the most effective concentration was selected;the second group of slices was perfused with 10 ?mol/L bicuculline;the third group of slices was perfused with 10 ?mol/L bicuculline plus 40 ?mol/L GABA.The RRDA in 3 groups were observed.Results GABA decreased RRDA,40 ?mol/L was the most effective concentration.It shortened inspiratory time(TI),decreased integral amplitude(IA),prolonged respiratory cycle(RC) and expiratory time(TE) respectively.By contrast,10 ?mol/L bicuculline produced opposite effects on RRDA,it prolonged TI,increased IA,shortened RC and TE respectively.40 ?mol/L GABA plus 10 ?mol/L bicuculline had no significant effects on RRDA.Conclusion GABA plays an important role in modulate RRDA of neonatal rats via GABAA receptor.