1.Expression of human Streptococcus suis 2 muramidase released protein and preparation and application of its monoclonal antibodies
Gongyi XU ; Haili WANG ; Changcheng GE ; Changjun WANG ; Jiaqi TANG
Chinese Journal of Veterinary Science 2009;29(7):873-876
The mrp gene of SS2 human strain Habb was truncated and cloned into a prokaryotic expression vector pGEX4T-2,and a fusion-expressed protein MRP-GST of 61 000 was obtained in E.coli.The GST was cut from MRP-GST with thrombin protease to gain the purified MRP of 35 000 which showed a strong reaction to the SS2 positive sera in Western blotting.BALB/c mice were immunitied intraperitoneally with purified MRP protein.Murine myeloma cells were fused with the splenocytes of the immunized mice after the third immunization.An indirect ELISA coated with purified MRP was used to screen hybridomas for production of specific antibody.The six McAb recognized MRP specially.According to the results of 71 standard strains (48 SS and 34 SS2)by Sandwich ELISA,the coincidence rate was 97.2%,but for 34 SS standard serotype was 100 %.The ELISA method has a potential value for clinical and epidemiological applicationgs.
2.A MICROELECTROPHORETIC DELIVERY OF HORSERADISH PEROXIDASE FOR DEMONSTRATING THE AFFERENT PROJECTION TO NUCLEUS RAPHE MAGNUS
Zi GE ; Lixia ZHU ; Changcheng ZHANG ; Zhiying WANG ; Wancheng SONG
Acta Anatomica Sinica 1954;0(02):-
20 adult healthy rats were used for microelectropheretic delivery of horseradish peroxidase (HRP). The diameter of the tip of the glass electrode was around 20~50?. It was inserted into nucleus raphe magnus, 20% HRP was delivered microelectrophoretically(5 ?A for 30 min; 10 ?A for 10 min). In eight rats the microelectrode was withdrawn immediately after the delivery and in the rest it was removed 10~20 minutes after delivery. Afterward the animals were allowed to survive for 40 hrs before decapitation, except 2 for 20 and 2 for 60 hrs. The brains were dissected out for cryostat sections and DAB reaction for peroxidase.The tip of the electrode was located in the nucleus raphe magnus, at the level of nucleus facialis. The diameter of the brain tissues occupied by HRP at the points of microelectophoretic delivery was 0.2~0.4mm and the maximum reached 0.6~0.7mm. The exogenous HRP granules were not visible in the cases with electrodes withdrawn immeditely after delivery and in the survivors of 60 hrs. In most cases the electrodes were left inside for more than 10 minutes after the delivery. The exogenous HRP granules were large, coarse, brown, steroscopic and distributed evenly in the perikaryon of some neurons in several levels of the brain. Those in the axon and dentrite were distributed just like strings of pearls. There were much more neurons with exogenous HRP positive granules dispersed in the reticular formation of the medula oblongata, most of them were polydendritic. Small and fusiformed HRP-positive cells are observed occasionally in the gray substance of subventriculum, reticular formation pontis and ventrolateral side of locus coeruleus. Weaker reactions were found in a few cells in the periaquaductous gray substance of the midbrain, nucleus raphe centro-superior, substantia nigra pars compacta and in the area around the nucleus supraopticus. Besides no HRP-positive cells were discovered in thalamus and subcortex structure.There were two types of cells with HRP positive granules around some blood vessels in reticular formation and subventricular gray substance: One was a small protoplasmic astrocyte with broad cytoplasma, small nucleus, and thick, short and irregular processes and another kind was polydendritic neurons filling with brown positive granules in perikarya and cytoprocesses, in the latter, the HRP positive granules were arranged as strips of pearls.
3.Erythropoietin mobilizes renal progenitor cells to reduce ischemic reperfusion injury in rats with nephron-sparing surgery
Yunpeng ZHU ; Ruipeng JIA ; Changcheng ZHOU ; Zhongle XU ; Yuzheng GE ; Liuhua ZHOU ; Ran WU
Chinese Journal of Nephrology 2017;33(10):775-780
Objective To investigate the effects of the erythropoietin (EPO) on ischemia reperfusion injury (IRI) in rats with nephron-sparing surgery (NSS). Methods Fifty-four Sprague Dawley rats were divided into 3 groups randomly after right kidney nephrectomy: Sham group, NSS group (PBS+NSS) and EPO group (EPO+NSS). During NSS, renal artery was clamped for 40 min to induce IRI. Sham group just adopted exposure renal artery without vascular clamped. Rats in NSS group were injected intraperitoneally with PBS for 3 days before NSS. Rats in EPO group were injected intraperitoneally with EPO for 3 days before NSS. After 12 h, 24 h, 72 h, blood sample and renal tissues were collected. The serum creatinine (Scr) and urea nitrogen (BUN) were evaluated. The pathology injury was evaluated by HE staining. The CD24/CD133 double-positived renal progenitor cells (RPCs) were tested by flow cytometry. The CD133 and PCNA protein were quantified by immunohistochemical staining. The expressions of Wnt7b and β-catenin protein were detected by Western blotting. Results Rats in NSS group had more elevated Scr, BUN and pathology injury scores 12 h, 24 h and 72 h after operation than those in Sham group (all P<0.05). Compared with those in the NSS group, the Scr and BUN in the EPO group were significantly lower 24 h after the surgery (all P<0.05), and the pathology injury score also decreased (P<0.05). The proportion of RPCs, expressions of CD133 and PCNA, and expressions of Wnt7b and β-catenin protein were significantly higher after 24 h of the surgery in NSS group than those in the Sham group (all P<0.05). While compared with those in the NSS group, the proportion of RPCs and expressions of CD133, PCNA, Wnt7b and β-catenin increased at the EPO group (all P<0.05). Conclusions EPO can reduce the IRI after NSS, and its mechanism may be related to the mobilization of the RPCs by the Wnt7b/β-catenin signal pathway.
4.Effects of Age and Gender on Lumbar Proprioception
Xiaochen LIU ; Fang CUI ; Yulin DONG ; Yili ZHENG ; Haoyu HU ; Di GONG ; Juan ZHANG ; Changcheng CHEN ; Ge SONG ; Xueqiang WANG
Chinese Journal of Rehabilitation Theory and Practice 2018;24(12):1446-1450
Objective To explore the difference in lumbar proprioception between the youth and the elderly, and the male and the female. Methods From September to November, 2017, 58 elderly aged 65 to 80 years and 56 youth aged 20 to 30 years were randomly picked up from the community. Their lumbar proprioceptions were measured by Con-Trex isokinetic dynamometer. The absolute error angles of lumbar flexion and extension in passive replication test was used to assess lumbar proprioceptive function. Results The absolute error angles of passive replication test were less on youth proprioception than on elderly ones (t > 4.218, P < 0.001), either in men or women (t > 2.844, P < 0.001), and was not different between men and women neither in the elderly group nor in the youth group (t < 1.473, P > 0.05). Conclusion The lumbar proprioception in the elderly was obviously inferior to that of the youth, both men and women. There was no gender difference between men and women in lumbar proprioception, both elderly and youth.