Identification and verification of α-11 giardin-interacting protein / 中国血吸虫病防治杂志

Objective To identify and verify the interacting protein of α-11 giardin, so as provide the experimental evidence for studies on the α-11 giardin function. Methods The yeast two-hybrid cDNA library of the Giardia lambia C2 strain and the bait plasmid of α-11 giardin were constructed. All proteins interacting with α-11 giardin were screened using the yeast two-hybrid system. α-11 giardin and all screened potential interacting protein genes were constructed into pBiFc-Vc-155 and pBiFc-Vn-173 plasmids, and co-transfected into the breast cancer cell line MDA-MB-231. The interactions between α-11 giardin and interacting proteins were verified using bimolecular fluorescence complementation (BiFC). Results The yeast two-hybrid G. lambia cDNA library which was quantified at 2.715 × 107 colony-forming units (CFU) and the bait plasmid containing α-11 giardin gene without an autoactivation activity were constructed. Following two-round positive screening with the yeast two-hybrid system, two potential proteins interacting with α-11 giardin were screened, including eukaryotic translation initiation factor 5A (EIF5A), calmodulin-dependent protein kinase (CAMKL) and nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH), hypothetical protein 1 (GL50803_95880), hypothetical protein 2 (GL50803_87261) and a protein from Giardia canis virus. The α-11 giardin and EIF5A genes were transfected into the pBiFc-Vc-155 and pBiFc-Vn-173 plasmids using BiFC, and the recombinant plasmids pBiFc-Vc-155-α-11 and pBiFc-Vn-173-EIF5A were co-tranfected into MDA-MB-231 cells, which displayed green fluorescence under a microscope, indicating the interaction between α-11 giardin and EIF5A protein in cells. Conclusion The yeast two-hybrid cDNA library of the G. lambia C2 strain has been successfully constructed, and six potential protein interacting with α-11 giardin have been identified, including EIF5A that interacts with α-11 giardin in cells.

Establishment of dual reverse phase chromatography combined parallel reaction monitoring for targeted quantitative proteomics / 生物工程学报

Steady improvement in mass spectrometers technology has transformed the targeted proteome analysis into a new stage. Parallel reaction monitoring (PRM) technology has evolved from the basic multiple reaction monitoring (MRM) targeted proteomics methods in recent years. PRM performs with a higher sensitivity, throughput and reproducibility in targeted quantification, however its limitations in effectiveness and accurate quantification of samples with higher complexity still remain unsolved. In this study through improving the chromatographic conditions of PRM we established a simple and robust platform for targeted proteomic quantification. The newly established PRM system is equipped with columns with increased inner diameter (150 μm) and decreased total length (8 cm); faster liquid phase elution rate (800 nL/min) and shortened elution gradient (35 min). These modifications enable PRM platform to combine with dual reverse phase chromatography, to quantify up to 400 low abundance peptides in human 293T cells whole cell extract. Our findings would benefit the promotion of PRM technology, especially providing a technical option for accurate quantification of low abundance proteins.

Cloning and bioinformatics analysis of Giardia lamblia C2 strain SENP gene and prokaryotic expression of SENP catalytic domain / 中国人兽共患病学报

SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .

Effects of dihydroartemisinin on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lamblia / 中国人兽共患病学报

Effects of dihydroartemisinin (DHA) on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lam-blia was investigated in this study to explore the damage to skeleton protein of C 2 Giardia lamblia .Giardia lamblia was culti-vated respectively for 2 ,4 ,8 ,and 12 hours with modified TYI-S-33 medium containing 100 μg/mL and 200 μg/mL DHA , while the control group performed in the same experimental conditions without DHA .The expressive quantity of Alpha-7 .3 gi-ardin mRNA was determined by using real-time reverse transcription PCR ,and then we found that the expressive quantities of Alpha-7 .3 giardin mRNA with DHA were significantly lower than those in the control group .It’s suggested that dihydroarte-misinin has obvious inhibitory effect on the expression level of Alpha-7 .3 giardin mRNA in C2 Giardia lamblia .The actions of dihydroartemisinin on skeleton protein of C2 Giardia lamblia are effective .

CBLB502 Protects mice radiation pneumonitis and pulmonary fibrosis / 军事医学

Objective To investigate the protective effect of CBLB 502 on radiation pneumonitis and pulmonary fibrosis for confirming the feasibility of CBLB502 as a clinical anti-radiation drug release.Methods With a single 20 Gy irradia-tion, C57BL/6J mice was sacrificed on 24 h, 1 month, 3 months and 5 months and lung tissue was assayed by TUNEL method for apoptosis of alveolar epithelial cells and endothelial cells , HE staining showing fibrosis changes , immunohisto-chemistry detecting the expression of specific indicators , as well as pathological changes of the fur and skin radiated site . Results CBLB502 inhibits apoptosis in mice alveolar epithelial cells and vascular endothelial cells after irradiation , slowing the process of pulmonary fibrosis , while reducing the expression of laminin and maintaining the expression of surfac-tant protein B, and the skin inflammation also significantly reduced .Conclusion CBLB502 could alleviate the occurrence of radiation pneumonitis and pulmonary fibrosis as well as radiation-induced skin injury .

Small interfering RNA in silencing Bcl-2 expression and enhancing radiosensitivity of esophageal cancer cells / 中华放射肿瘤学杂志

ObjectiveTo explore the effects of small interfering RNA (siRNA) specific to Bcl-2gene on radiosensitivity of esophageal cancer cells. Methods Bcl-2 gene siRNA ( Bcl-2 siRNA ) was induced into esophageal cancer EC9706 cells by lipofectamine.Bcl-2 protein expression and apoptosis of EC9706 cells were detected by flowcytometer. Clone forming assay was used to determine the inhibitory effects of X-ray radiation combined with Bcl-2 siRNA interference. ResultsWhen Bcl-2 siRNA had been induced into EC9706 cells, Bcl-2 protein expression in EC9706 cells was inhibited, and cell apoptosis was increased. Bcl-2 protein expression rates of EC9706 cells induced with Bcl-2 siRNA1, A2, A3 (25.13％ ±2. 04％ ,38.87％ ± 3.34％ , 30.55％ ± 2. 73％ ) were lower than the control group ( 84.28％ ± 1. 47％ )(t =4. 01,3.04,3.64, P ＜ 0. 05 ). After interference, the apoptosis rate of EC9706 cells ( 33.86％ ±1.04％ ) was higher than the control group and siRNA negative group (5.51％ ±0. 14％ and 5.59％ ±0. 46％ ) (t =6. 55,6. 54,P ＜0. 01 ). Bcl-2 gene siRNA interference enhanced X-ray inducing apoptosis of EC9706 cells (56.76％ ± 1.24％ ), which was higher than the radiation alone group ( 24.51％ ± 0. 48％ )(t =3.59,P ＜ 0. 05 ). The D0, Dq, and SF2 of combined treatment group were much lower than those of irradiation alone group . The sensitization enhancing ratio was 1.32 ( ratio of D0 values ) . Conclusions Bcl-2 gene siRNA could enhance the radiosensitivity of esophageal cancer EC9706 cells and may has a good future in clinical practice.

An investigation of influencing factors for depressive disorder in 1 000 retired cadres and people / 中国组织工程研究

BACKGROUND: Geriatric depressive disorder is a common psychological disorder in elderly people, and it can lead to the decline of various organic functions, and it also does great harm to elderly people and their family.OBJECTIVE: To investigate the influencing factors of depressive disorder in retired cadres and people, so as to provide basic evidence for enhancing the mental nursing in retired cadres and people and reducing the prevalence of geriatric depressive disorder.DESIGN: A sampling survey.SETTING: Department of Nursing, North China Coal Medical College.PARTICIPANTS: Between August and December 2002, 1 000 retired cadres and people living in the urban area of Tangshan city were investigated. There were 574 males and 426 females, aged from 60-88 years, the average age was 67.04 years, 851 of them were married, 10 were divorced and 139 had lost their spouses: the educational level was primary school and below in 159 cases, junior middle school in 362 cases, senior middle school in 280 cases, university and above in 199 cases.METHODS: The elderly people were investigated with the geriatric depression scale (GDS) and individual general information inventory by means of family visit. The investigative purpose and methods were introduced to the subjects before the investigation, and they all signed the informed consent, the form of without recording name was used. The strictly trained investigators were qualified by examination, which ensured the concordance of the investigative results. The pretesting was performed before the investigation to detect whether there were words,sentencesand items hard to understand, so as to avoid the information bias. The questionnaires were filled by the elderly people themselves or by the investigator if there was any difficulty for the elderly people in writing according to the their choice after reading the questions for them.in retired cadres and people of different age, gender and educational level:ferent marriage status.RESULTS: Totally 1 000 questionnaires were sent out, and 1 000 valid der was obviously higher in females than in males (27.0%, 18.6%, P ＜ 0.05).ple with the educational level of primary school and below, junior middle school, senior middle school, university and above were 33.9%, 22.1%,20.0%, 16.1% respectively, and it was decreased with the elevation of emarkedly higher in retired people than in retired cadres (25.4%, 13.6%,higher than in those having or had lost spouse (50.0%, 21.2%, 26.6%, P ＜ 0.05).CONCLUSION: The prevalence of depressive disorder is higher in retired cadres and people. More attentions should be given to the mental health care and social support of retired cadres and people, especially for the divorced ones.

Depressive status in retired elderly people and community nursing intervention / 中国组织工程研究

BACKGROUND: With the increase of senile population, it needs to further improve their mental health when improving their physical health. To study the depressive status of retired elderly people in order to provide basis for effective community nursing intervention.OBJECTIVE: To investigate and analyze the depression incidence among retired elderly people and conducted correlative factors analysis in order to improve the nursing service to aged people's mental health.DESIGN: Cross-sectional sampling study towards retired senior people.SETTING: Nursing department of a medical college.PARTICIPANTS: This study was completed in Nursing Department of North China Coal Medical College during September to December 2002. It conducted questionnaire survey to 1 000 retired people in Tangshan city by family visit. There were 574 male and 426 female who aged from 60 to 88years old with mean age of 67 years.METHODS: To adopt geriatric depression scale and general condition questionnaire investigation to individuals.MAIN OUTCOME MEASURES: Depression scale score, depression incidence and frequency of correlative factors RESULTS: Mild depression accounts for 17.2% while moderate and severe depression account for 5% among retired elderly people. There is close relationship between factors such as retired veterans, female, low education,lack of social support, empty life, lack of exercise and poor self-assessed health and depression incidence(x2 = 9.80 - 175.93, P ＜ 0. 05).CONCLUSION: The incidence of senile depression among retired elderly people is pretty high so that it is important to pay attention to mental care towards retried people when doing community nursing, especially to divorced, widowed, female and low educated retired people.

The Injury of Metronidazole on Morphology of Giardia lamblia in Vitro / 中国寄生虫学与寄生虫病杂志

Trophozoites of Giardia lamblia were axenically cultivated with modified TYI-S-33 medium contained 500 ?g/ml metronidazole(12h LC50).The morphology of drug-treated trophozoites was observed with light and electron micro-scopes at 2,4,8,12 h respectively.The light microscopy revealed that the trophozoites treated with MTZ showed swollen,detached from the wall of the culture tube,and were with vacuoles in the cytoplasm.Movement of the flagella become slowly or stopped.Electronic microscopy showed that the trophozoites were swollen and deformed;lots of vacuoles were seen in the cytoplasm;the contents of cytoplasm were depleted and the nuclei deformed.This study indicated that MTZ has injured the morphology of G.lamblia.