1.Prokaryotic Expression of Trichinella spiralis Gene Ts21 and Identification of the Recombinant Protein
Zhongquan WANG ; Li LU ; Jing CUI ; Lai WANG ; Rui WANG
Chinese Journal of Parasitology and Parasitic Diseases 1997;0(06):-
Objective To express the antigen gene Ts21 of Trichinella spiralis, purify the recombinant protein and test its antigenicity. Methods T. spiralis gene Ts21 was sub-cloned into the prokaryotic expression vector pMAL-c2X and the recombinant pMAL-c2X-Ts21 was constructed. The recombinant plasmid was transformed into E. coli TB1 strain and induced by IPTG. The expression products were purified by MBP-binding affinity chromatography. The antigenicity of the recombinant protein was examined by SDS-PAGE and Western blotting. Mice were immunized with the recombinant protein, the titer of the immune sera was detected by ELISA. The distribution of Ts21 protein in muscle larvae was observed by IFA. Results The molecular weight of the expressed fusion protein was about Mr 63 500 and the expression level peaked at 4 h post-incubation. The portion of the fusion protein accounted for 18.2% of all the protein by thin-layer gel optical scanning. Western blotting demonstrated that the recombinant protein was recognized by sera from mice infected by T. spiralis (T1) and T. nelsoni (T7) as well as sera of patients with trichinellosis, but not by sera from mice in-fected with T. nativa (T2), T. britovi (T3) and T. pseudospiralis (T4). The recombinant protein did not react with sera from patients with ancylostomiasis, cysticercosis and schistosomiasis, but cross-reacted with sera from patients with parag-onimiasis, clonorchiasis and echinococcosis. High titers of antibodies were produced in mice immunized with the recom-binant protein. IFA showed that the Ts21 protein was mainly distributed in the cuticle of muscle larvae. Conclusion The Ts21 antigen gene of T. spiralis has been expressed and the recombinant protein shows antigenicity.
2.COMPARATIVE STUDY ON THE STRUCTURAL CHARACTERISTICS OF MICROFILAMENTS IN CELLS WITH DIFFERENT METASTATIC ABILITY CLONED FROM HUMAN GASTRIC CANCER CELL LINE BGC-823
Rui LU ; Yang KE ; Guirong LU ; Tao NING ; Weili LIU ; Jing LIAO ; Bing WANG ; Zheng EH
Acta Anatomica Sinica 1957;0(04):-
Ten cell strains were obtained from the human gastric cancer cell line BGC-823 by using the method of single cell cloning. Through inoculation of these cell strains in nude mice and subsequent observation of their metastatic capability, four cell lines, C_1, C_6 with high, and C_5, C_8 with low metastatic capability were selected out; and their biological and morphological properties were studied. It was demonstrated that all of them did not show apparent difference in growth ability, but morphologically, the mictovilli of C_5 and C_8 cells were scarce and short; and their microfilaments were thick, straight and well organized. Whereas the microvilli of C_1 and C_6 strains were long and abundant comparatively, but their microfilaments were poorly organized. On the basis of present observation,we suggested that the organization of microfilaments in cancer cells appeared to bear reversed relationship with their metastatic capability.
3.Effects of 8 Hz,90 dB/130 dB infrasound on expression of NMDAR1 in hippocampus of rat
Zhao-Hui LIU ; Jing-Zao CHEN ; Jing LIU ; Jian-Yong QIU ; Rui LU ; Xiao-Chao YUAN ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(11):-
Objective To study the effect of infrasound on the changes of expression of NMDAR1 in hipp- ocampal cells.Methods Eighty-eight male Sprague-Dawley rats were randomized into eleven groups:control group,90 dB/1 d,7 d,14 d,21 d and 28 d infrasound exposed groups;130 dB/1 d,7 d,14 d,21 d and 28 d infra- sound exposed groups.All the animals in the test groups were put in an infrasound field with 8 Hz,90 dB or 130 dB for 2 hours daily.Immunohistochemistry methods were used to detect the changes of intracellular expression of NMDARI in hippocampal cells.Methods The expression of NMDAR1 in hippocampus after the rats were exposed to infrasound of 8 Hz,90 dB SPL showed a procedure from reducing on the 1st day to rising on the 7th and peaked on the 14th day,then to descending on 21st day and returning to the standard level on the 28th day.Exposure to infra- sound of 8 Hz,130 dB SPL induced opposite effects on the expression of NMDAR1 compared with 90 dB SPL,which showed a process of increasing,descending,reaching to the lowest,then ascending and returning to the normal.The lowest expression of NMDAR1 occurred on the 14th day in every observed hippocampal area.Conclusion 8 Hz, 90 dB/130 dB infrasound induced certain reversible reaction in the expression of NMDAR1 of hippoeampal cells in rats,which may disturb their learning and memory function.
4.Study on interactions between targeted ultrasound contrast agents with anti-human interleukin-8 monoclone antibody and activated vascular endothelial cellls
Wei YANG ; Yongping LU ; Yuan YUAN ; Hong ZHOU ; Rui LIU ; Zhi JING ; Yueyue TANG
Chinese Journal of Ultrasonography 2012;21(8):715-718
Objective To observe the interactions between a anti-interleukin-8 monoclonal antibody (anti-IL-8 mAb) carried targeted ultrasound contrast agents and the injured vascular endothelial cells,as well as to explore the role of IL-8 in the formation of atherosclerotic plaques and a new assessing method of vascular endothelial functions.Methods The targeted ultrasound contrast agent was prepared by using a erosslinking agent to couple a anti-IL-8 mAb with SonoVue microbubbles.The interactions between SonoVue microbubbles/the targeted microbubbles and normal/injured endothelial cells were observed under an inverted microscope,respectively.The numbers of endothelial cells and adhered microbubbles were counted under high power magnification.The ratio of microbubbles to endothelial cells was calculated for the quantitative analysis of the interactions.Results In the control group,only a slight amount of original SonoVue microbubbles were bound to normal/injured endothelial cells.In contrast,it was visible under the microscope that the anti-IL-8 mAb carried SonoVue could be bound to endothelial cells,and the number of microbubbles bound to the surface of injured endothelial cells was significantly higher than that bound to the normal endothelial cells.Conclusions The anti-IL-8 mAb carried targeted ultrasound contrast agent could be readily bound to the surface of the injured cells specifically,and thus suggesting a new direction for ultrasonic detection of vascular endothelial injury and the ultrasonic assessment of vascular endothelial functions.
5.Levels of main platelet thrombin receptors in older chronic haemodialysis patients.
Yan LI ; Lin SHEN ; Rui CHEN ; Fu-rong LU ; Jing LI ; Jian-guo LIU
Chinese Medical Journal 2010;123(17):2495-2496
Aged
;
Aged, 80 and over
;
Blood Platelets
;
chemistry
;
Humans
;
Receptor, PAR-1
;
blood
;
Receptors, Thrombin
;
blood
;
Renal Dialysis
;
Uremia
;
blood
;
therapy
6.DNA barcoding the medicinal plants of the genus Paris.
Yingjie ZHU ; Shilin CHEN ; Hui YAO ; Rui TAN ; Jingyuan SONG ; Kun LUO ; Jing LU
Acta Pharmaceutica Sinica 2010;45(3):376-82
DNA barcoding is a technique in which species identification and discovery are performed by using short and standard fragments of DNA sequences. In this study, eleven species of Paris, including seven varieties, were sampled. Five chloroplast sequences, psbA-trnH, rpoB, rpoC1, rbcL, matK, and one nuclear marker, the second internal transcribed spacer (ITS2) of ribosomal DNA, were amplified and sequenced. The PCR amplification and sequencing efficiency, intra- and inter-specific divergence and barcoding gap were used to evaluate different loci, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The ITS2 sequences in the studied samples of Paris were amplified and sequenced successfully using primers designed by our group, while matK showed low level in the amplification and psbA-trnH was difficult for sequencing because of over 800 bp and poly (A) structure. Analysis of the intra- and inter-specific divergence and barcoding gap showed ITS2 was superior to other loci. The ITS2 showed a much higher percentage of success (100%) in identification than other five loci, none of which indicated more than 50% except matK (52.9%). The 2-locus combination of rbcL+matK didn't improve ability of authentication. In addition, the rate of successful identification with ITS2 kept 100% when the samples were expanded to 67 samples of 29 species. In conclusion, ITS2 can be used to correctly identify medicinal plants of Paris, and it will be a potential DNA barcode for identifying medicinal plants of other taxa.
7.Epidemiological analysis of 5 577 cases of primary liver cancer in Xinjiang
Pengfei LU ; Rui MAO ; Jing MA ; Huicai WANG ; Song'an ZHANG ; Ge WU ; Yongxing BAO
Chinese Journal of Health Management 2016;10(4):270-274
Objective To analyze the clinical epidemiology of primary hepatocellular carcinoma (PHC) in the Xinjiang region. Methods Clinical data of the patients with PHC were collected at First Affiliated Hospital of Xinjiang Medical University from 5 577 cases from January 2002 to December 2014, their gender, race, age, household distribution, hepatitis virus-positive rate were analyzed retrospectively. Results Among the 5 577 eases, the men/women gender ratio was 3.45∶1;the proportion of Han, Uighur, Kazakh, and other ethnic groups (Hui, Mongolian, Manchu, Xibo) was 79.67%, 9.86%, 4.55%, 3.31%and 2.61%, respectively. The Constituent ratio difference between Uighur and Han was significant (P<0.05);4 232 patients had hepatitis B surface antigen (HBsAg) detection, and 3 833 patients had HCV antibody (HCV-Ab) detection. HBsAg was positive in 2 560 cases (60.49%), HCV-Ab was positive in 490 cases (12.78%). Hepatitis B virus detection positive rate in Uygur was 35.52%, in Kazak was 40.00%, which was lower than the Han's (65.68%, P<0.05). Urban and rural population had 3589 cases (64.35%) and 1988 cases (35.65%). Conclusion An increased risk for PHC was found in hepatitis virus-positive patients, the Xinjiang Uygur and Kazak people had significantly lower prevalence of HBV infection than the Han's. Appropriate measures should be taken for clinical diagnosis, treatment and prevention of PHC.
8.Adherent-invasive Escherichia coli isolated from the patient with inflammatory bowel disease in China
Hui CAO ; Ruifeng LU ; Liping CHEN ; Rui ZHOU ; Jing LIU ; Qiu ZHAO
Chinese Journal of Digestion 2016;36(7):481-485
Objective To study adhesion and invasion of adherent-invasive Escherichia coli strains and its ability of survival and proliferation in macrophage cells.Methods Bacteria from the mucosa tissues of ileum,colon and rectum of inflammatory bowel disease (IBD)patients were isolated,cultured and identified.The toxic genes were confirmed by polymerase chain reaction.The adhesion and invasion ability of bacteria and its survival and proliferation ability in macrophage cells were observed after co-culture with cells.Results Five adherent-invasive Escherichia coli strains were isolated from the intestinal mucosa specimens of two patients with Crohn′s disease and one patient with ulcerative colitis.The adhesion rates of five bacteria stains (2A,2B,15A,15B,19B)to Caco-2 cells were (1 .4 ± 1 .3 )%,(1 .5 ± 0.6 )%, (0.6±0.1)%,(1 .4±0.4)% and (1 .1 ±0.8)%,respectively.The adhesion rates to Int-407 cells were (1 .0±0.8)%,(1 .5 ±0.8)%,(1 .0 ±0.8 )%,(1 .0 ±0.8 )% and (0.3 ±0.0 )%,respectively.The invasion rates to Hep-2 cells were (10.1 ±7.0)%,(0.7 ±0.4)%,(0.4 ±0.3 )%,(2.2 ±1 .0)% and (2.1 ±1 .8)%,respectively.The invasion rates to Int-407 cells were (0.7 ±0.5 )%,(0.5 ±0.3 )%, (2.8±1 .2 )%,(0.7 ±0.5 )% and (0.5 ±0.4)%,respectively.All five adherent-invasive Escherichia coli strains could survive and proliferate in macrophage cells.Conclusions Five adherent-invasive Escherichia coli strains are isolated for the first time in Chinese patients with IBD,and all of them could survive and proliferate in macrophage cells.
9.Femoral-deep femoral crossover bypass for unilateral iliofemoral arteriosclerosis obliterans
Tao MA ; Jie MA ; Qingsheng LU ; Zhiqing ZHAO ; Junmin BAO ; Xiang FENG ; Rui FENG ; Zaiping JING
Chinese Journal of General Surgery 2012;(11):893-895
Objective To evaluate the clinical results of femoral-deep femoral crossover bypass in the treatment of long-segment unilateral iliac artery occlusive disease.Methods From July 1995 to December 2010,40 patients (28 males,12 females,aged from 66 to 90,with mean age of 73) with comprehensive unilateral iliac-superficial femoral arteriosclerosis obliterans were enrolled in this procedure.All patients suffered from unilateral common iliac,external iliac,common femoral,and superficial femoral arteriosclerosis obliterans.These patients were treated with femoral-deep femoral crossover bypass.Postoperative ankle-brachial index,blood flow velocity and patency rates in 5,7 and 10 years and limb salvage rates in 5,7 and 10 years were evaluated.Results There was no perioperative mortality nor extremity amputation.35 (87.5% ) patients were followed-up from 1 to 13 years (mean 5.7 y).Anklebrachial index rose from preoperative 0.23 ± 0.10 to postoperative 0.55 ± 0.11 (t =15.91,P =0.000 ).Popliteal arterial velocity rose from preoperative ( 14 ±6) cm/s to postoperative (34 ± 10) cm/s (t =15.63,P =0.000) ; Tibial arterial velocity rose from ( 10 ±4) cm/s to (22 ±7) cm/s (t =15.71,P =0.000).The primary and secondary patency rates were 60.1%,44.3%,25.3%,and 93.5%,86.8%,57.9% at 5,7 and 10 years,respectively.Limb salvage rates were 97.5%,95%,and 90%,at 5,7 and 10 years,respectively.Conclusions Femoral-deep femoral crossover bypass is safe and reliable in treating certain unilateral iliofemoral occlusive disease,especially for high-risk old patients or those who are not indicated for endovascular therapies or direct aortic approaches.
10.Total knee arthroplasty:Goodness-of-fit analysis of intraoperative knee society score of Peking Union Medical College
Haiying JIA ; Ming LU ; Xiaoping WANG ; Rui ZHENG ; Jing ZHANG ; Huasong MA
Chinese Journal of Tissue Engineering Research 2013;(35):6228-6233
BACKGROUND:Knee society score of Peking Union Medical Col ege has been promoted and used in the 306th Hospital of PLA for 2 years and we have accumulated some clinical data.
OBJECTIVE:To analyze the stability and feasibility for suitable clinical medical practice of knee society score of Peking Union Medical Col ege.
METHODS:Fifty-five patients with osteoarthritis of the knee were included, and the patients were divided into
three groups:the preoperative group;3 months postoperative fol ow-up group;6 months postoperative fol ow-up group. The measurement results obtained by the application of the scale were compared to those of the Western Ontario and McMaster Universityies Ostroarthritis Index, visual analog scale, and hospital for special surgery
score, and then knee society score goodness-of-fit analysis was performed based on the structural equation model.
RESULTS AND CONCLUSION:Knee society score performance assessment of the signs and symptoms of
osteoarthritis of the knee patients was positively correlated with the pain severity of visual analog scale score;the overal knee society score of the patients after 6-month fol ow-up was significantly improved compared with that before treatment. The knee society score after 3-and 6-month fol ow-up was significantly higher than that before treatment, and the score was quite with the hospital for special surgery score. The overal assessment of the knee society score after treated for 3 and 6 months was improved for more than grade Ⅰ, the visual analog scale
score was decreased for more than 30%. The pain assessment items and physiological functional assessment items of Western Ontario and McMaster Universityies Ostroarthritis Index after 6-month fol ow-up were
significantly better than those before treatment (P<0.05);the visual analog scale score was poor after 3-month fol ow-up, and there was significant difference in visual analog scale score before and after treatment which was related with the pain level;the score accompanied by function and joint stiffness was significantly improved. The knee society score of Peking Union Medical Col ege obtained four dimensions of 22 assessment projects, and the
goodness-of-fit analysis showed that the scale had good goodness which had scientific nature and application value.