1.Pathology of White Matter Tracts at Core of Severe Spinal Cord Injury in Adult Rats
Xiaojiao PEI ; Zhaoyang YANG ; Peng HAO ; Jiasheng RAO ; Can ZHAO ; Xiaoguang LI
Chinese Journal of Rehabilitation Theory and Practice 2015;21(4):373-377
Objective To explore pathological characteristics of different white matter tracts at core of the spinal cord injury (SCI) in adult rats. Methods 21 adult female Wistar rats were divided into normal group (n=3), sham group (n=3) and lesion group (n=15). The rats of the lesion group were severely injured at T7-8 using the NYU impactor of 10 g×50 mm. The pathology of spinal cord injury was detected using Luxol fast blue (LFB) staining and NF200 immunohistochemistry staining 1 day, 3 days, 7 days, 14 days and 28 days after injury, while the hindlimbs behavior of rats was rated with Basso-Beattie-Bresnahan (BBB) scores of open-field. Results The BBB scores recovered 3 days after injury and reached a platform from 14 to 28 days after injury. LFB showed that there were spared white matters on ventral white matter (VWM) and lateral white matter (LWM). The percentage of spared white matters area decreased to the lowest 3 days after injury, and reached a platform from 7 to 28 days after injury. The number of NF200-IR axons reduced significantly in white matter tracts after SCI. Conclusion Ventral spared white matters plays a key role in the recovery of motor function in rat with SCI, and the first 3 days was a time window to protect the white matters from injury.
2.Expression, purification and preliminary activity study of recombinant hepatocyte growth factor protein in E.coli
Xiaojiao ZHENG ; Zhou GAO ; Rongrong SHEN ; Hang ZHAO ; Dong CEN ; Jianping LUO ; Jianxin Lü ; Renzhi PEI ; Shixuan HUA
Chinese Journal of Microbiology and Immunology 2012;(11):967-971
Objective To prepare hepatocyte growth factor(HGF) recombinant protein and confirm its activity preliminarily according to building HGF gene prokaryotic expression vector and transforming into E.coli.Methods Clone HGF inserted into the vector pET-26b(+) to construct prokaryotic expression vector pET-26b(+)-HGF and transform into E.coli Rosseta(DE3).The transformed bacteria induced by IPTG was purified through Ni-NTA resin affinity chromatography frozen-drying after renaturation.Results HGF gene recombinant prokaryotic expression vector pET-26b(+)-HGF was constructed successfully.E.coli Rosseta(DE3) which was transformed into pET-26b(+)-HGF expresses the target protein as the form of inclusion bodies,accounting for 38% of the total bacterial proteins,and confirmed by Western blot.HGF protein which was purified by Ni-NTA resin affinity chromatography,has a purity of about 95%,and can promote proliferation,migration,and inhibition of apoptosis for human non-small cell lung cancer cell line A549 cells after interaction.Conclusion HGF gene recombinant prokaryotic expression vector pET-26b (+)-HGF was constructed and expressed in transformed E.coli Rosseta(DE3) successfully.They resumed their recombinant HGF protein structure after purification and renaturation,and had biological activity confirmed by in vitro studies.
3.Research of cedilanid in the prevention of severe pneumonia complicated with heart failure in infants
Miaomiao XUE ; Yanyan DING ; Xiaojiao PAN ; Pei YUAN ; Shuyan CHI ; Li LI
Chinese Journal of Postgraduates of Medicine 2022;45(10):942-945
Objective:To investigate the efficacy and safety of cedilanid in the treatment of severe pneumonia in infants and the value of preventing heart failure.Methods:A total of 80 children with severe pneumonia admitted to Dezhou Maternal and Child Health Hospital from January 2019 to December 2020 were selected and randomly divided into the control group and the observation group, with 40 cases in each group. The control group received comprehensive treatment, while the observation group was treated with cedilanid (0.01 mg/kg, one-time intravenous injection) on the basis of the control group. The efficacy of both groups was observed after 5 d of treatment. The incidence of heart failure, correction time of heart failure, improvement time of symptoms and signs, and length of hospitalization time were compared between the two groups; the inflammatory markers, myocardial markers and arterial blood gas indexes were compared between the two groups before and after the treatment.Results:The total effective rate in the observation group was higher than that in the control group, and the incidence of heart failure in the observation group was lower than that in the control group: 90.0% (36/40) vs. 72.5% (29/40), 32.5%(13/40) vs. 10.0%(4/40), the differences were statistically significant ( χ2 = 4.02, 4.10, P<0.05). The improvement time of symptoms and signs (restlessness elimination, respiratory improvement, heart rate improvement and disappearance of rhonchus in lung) in the observation group were less than those in the control group ( P<0.05). The levels of procalcitonin (PCT) and N-terminal pro-brain natriuretic peptide (NT-ProBNP), myocardial troponin I(cTnI), and creatine kinase isoenzyme (CK-MB) in the observation group after treatment were lower than those in the control group: (6.15 ± 1.03) μg/L vs. (10.85 ± 2.12) μg/L, (112.02 ± 30.09) ng/L vs. (215.39 ± 55.08) ng/L, (0.68 ± 0.17) μg/L vs. (1.12 ± 0.34) μg/L, (19.05 ± 6.11) U/L vs. (28.97 ± 7.82) U/L, P<0.05. The levels of oxygen partial pressure (PaO 2), blood oxygen saturation (SaO 2) and oxygenation index (PaO 2/FiO 2) in the observation group after treatment were higher than those in the control group: (6.15 ± 1.03) μg/L vs. (10.85 ± 2.12) μg/L, (112.02 ± 30.09) ng/L vs. (215.39 ± 55.08) ng/L, (0.68 ± 0.17) μg/L vs. (1.12 ± 0.34) μg/L, (19.05 ± 6.11) U/L vs. (28.97 ± 7.82) U/L, P<0.05. Conclusions:Early application of small dose of cedilanid in infants with severe pneumonia can effectively reduce the occurrence of heart failure, improve the clinical symptoms and blood gas indicators, with significant curative effect, which is worthy of promotion.
4.NK4 growth inhibition of human Raji lymphoma xenografts by competitive interrupting HGF/Met signal pathway.
Zhou GAO ; Xiaojiao ZHENG ; Rongrong SHEN ; Hang ZHAO ; Dong CEN ; Jianping LUO ; Shixuan HUA ; Renzhi PEI ; Jianxin LYU
Chinese Journal of Pathology 2014;43(8):551-555
OBJECTIVETo observe the inhibition of NK4 protein in the proliferation of human Raji lymphoma xenografts in nude mice, and to explore its molecular mechanism.
METHODSModels of human Raji lymphoma xenograft transfected with HGF gene were established by subcutaneous inoculation in nude mice. After establishment of the models, the mice received continuous NK4 protein via tail vein for 4 weeks, and the weight and tumor growth were monitored every week. After 8 weeks, the expression of HGF mRNA and c-Met mRNA of tumor tissues was measured by real-time fluorescent quantitation PCR. The apoptotic index (AI) and microvessel density (MVD) were evaluated by terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) and immunohistochemistry, respectively.
RESULTSThe models of human Raji lymphoma xenograft were successfully established. Although the animal weights of all groups declined, especially in the groups with NK4 protein injection, there was no statistical significance (P > 0.05). The tumor volume in HGF gene transfected group was larger than those of the control groups (P < 0.01), and there was no statistical significance among the control groups (P > 0.05). However, the tumor volume of the NK4 protein injection group decreased significantly (P < 0.01). Expression of HGF mRNA and c-Met mRNA in HGF gene transfected group increased significantly after injection of NK4 protein (P < 0.01). AI in HGF gene transfected group (33.5% ± 12.3%) was significantly lower than that of control groups (89.1% ± 22.3% vs. 81.9% ± 27.0%, P < 0.05), but became significantly higher (119.1% ± 18.9%) after NK4 protein injection (P < 0.01). MVD in HGF gene transfected group (28.5 ± 2.0) was higher than that of control groups (12.2 ± 1.4, 13.8 ± 1.3, P < 0.01), although declined (15.5 ± 2.5) after NK4 protein injection (P < 0.01).
CONCLUSIONSNK4 protein suppresses significantly the growth of human Raji lymphoma xenografts transfected with HGF gene. The pathogenesis may be involved in promoting tumor cell apoptosis and restraining tumor angiogenesis through competitive interrupting HGF/Met signal pathway.
Animals ; Apoptosis ; Hepatocyte Growth Factor ; genetics ; metabolism ; Heterografts ; Humans ; Lymphoma ; genetics ; metabolism ; therapy ; Mice ; Mice, Nude ; Microvessels ; pathology ; Neovascularization, Pathologic ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; T-Box Domain Proteins ; administration & dosage ; Transfection ; Transplantation, Heterologous