2. Research on purification and activity characterization of AmPR-10 ribonuclease from Astragalus membranaceus
Chinese Traditional and Herbal Drugs 2017;48(19):3945-3953
Objective To establish a stable and rapid separation and purification method for Astragalus membranaceus (Am) pathogenesis-related protein-10 (AmPR-10) using an automatic intelligent protein purification system AKTA Avant 25, and analyze its physiochemical and biological activity. Methods Am was extracted by Tris-HCl buffer. The crude extract was captured by anion exchange chromatography, and finely separated by hydrophobic chromatography and gel filtration chromatography. The relative molecular weight of AmPR-10 was measured by MALDI-TOF/TOF mass spectrometry, the protein identification was determined by mass spectrometry and MS/MS Ion Search, the glycoprotein identification was estimated by periodic acid-Schiff method, and the ribonuclease activity and effect factors were analyzed by agarose gel electrophoresis. Results The electrophoretically pure AmPR-10 was obtained by three-step purification of Q Sepharose Fast Flow, Butyl Sepharose High Performance and SuperdexTM 75 10/300 GL from the crude extraction. The relative molecular weight of AmPR-10 was 16 801. AmPR-10 was highly homologous to PR-10 and has no carbohydrate chains. Incubated at 56 ℃ for 30 min, AmPR-10 exhibited significant ribonuclease activity to total RNA of mammalian cells. The activity was insensitive to NaCl, pH value and mental ions, and weekly inhibited by 0.5 mol/L NaCl, pH 9.0, Mg2+ and Co2+. The activity was the same at EDTA as high as 20 mmol/L. Conclusion The three-step method of exchange chromatography-hydrophobic chromatography-gel filtration chromatography, a stable and rapid separation and purification method of AmPR-10, can be applied for other Chinese herbs. AmPR-10 might play an important role in resistance against virus.
3.Extrapleural solitary fibrous tumor: report of a case.
Jin-hong MEI ; Hong-ping WAN ; Han LIU ; Ren-sheng CHEN ; Hui-chun GAO ; Shan XU
Chinese Journal of Pathology 2006;35(8):510-511
4.CLONING OF FULL CRY1C GENE FROM B.T.C.AND EXPRESSION IN CROPS BENEFICIAL BACTERIA BACILLUS CEREUS
Yue-Hua CHEN ; Hong-Xiu LI ; Jin-Hong WANG ; Jun CAI ; Gai-Xin REN ;
Microbiology 1992;0(01):-
Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua.
5.Expression of survivin E-cadherin and Ki-67 in pituitary adenoma and correlation with invasiveness.
Chun-Liang WANG ; Jin-Hong MEI ; Hong-Ping WAN ; Ren-Sheng CHEN ; Lin-Lin XU
Chinese Journal of Pathology 2008;37(5):335-336
Adolescent
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Adult
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Aged
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Cadherins
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genetics
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metabolism
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physiology
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Female
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Humans
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Inhibitor of Apoptosis Proteins
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Ki-67 Antigen
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genetics
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metabolism
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Male
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Microtubule-Associated Proteins
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genetics
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metabolism
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physiology
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Middle Aged
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Neoplasm Invasiveness
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physiopathology
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Pituitary Neoplasms
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pathology
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physiopathology
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Young Adult
6.Inhibitory effect of an small peptide that interferes with Fc?-receptor recognition on antineutrophil cytoplasmic antibodies induced activity of neutrophils
Xiang-Ling WANG ; Nan CHEN ; Hai-Jin YU ; Hong REN ; Wei-Ming WANG ; Li-Yan NI ;
Chinese Journal of Rheumatology 2001;0(05):-
Objective Despite regular treatment,antineutrophil eytoplasmie antibodies(ANCA)asso- ciated systemic vasculitis(AASV),in which the role of Fc?Rs has been established,are still associated with significant long-term mortality and remain an important cause of end-stage renal failure.ANCA plays an im- portant role in the pathogenisis of primary systemic small vessel vasculitis(PSV)by their potential to activate neutrophils.Because the interaction between ANCA and its receptors on the Fc portion of immunoglobulins (Fc?R)on neutrophils is essential in the activation process,we investigate the inhibitory,effect of tg19320 on ANCA induced activation of neutrophils,which is a tetrameric tripeptide that interferes with IgG/Fe?Rs in- teraction.Methods We prepared tg19320 by solid-phase peptide syntbesis.The binding between tg19320 and human IgG was assessed by enzyme-linked immunosorbent assay.The biological activity of tg19320 to intefere with FcF?receptor recognition was identified by rosette formation assay.ANCA IgG was prepared from the sera of active Wegener's granulomatosis(WG)and microscopic polyangiitis(MPA)patients.Neu- trophils isolated from the blood of healthy volunteers were primed with TNF-?(2 ng/ml)and then incubated with ANCA IgG(200?g/ml),or pretreated with tg19320(2.5 mg/ml)and then added with ANCA IgG.Su- peroxide burst of neutrophils was determined by Ferri-cytochrome reduction assay.Results We found that tg19320 bound tightly to human IgG in a dose dependent manner and the inhibition of the rosette formation between SRBC-IgG and U937 cells was statistically significant(20.3% vs 53.2%,P
7.The detection and significance of AKAP12 methylation levels in peripheral blood in patients with lung cancer
Hehe LIAO ; Yunfeng ZHANG ; Shuqiang WU ; Hanchun LI ; Lei JIN ; Hong REN
Chongqing Medicine 2014;(15):1834-1836
Objective To detect the levels of AKAP12 methylation by methylation‐specific high‐resolution melting curve(MS‐HRM ) in peripheral blood in patients with colorectal cancer and investigate its clinical significance .Methods We used MS‐HRM technology to detect the levels of AKAP12 methylation in peripheral blood in 60 lung cancer patients ,and analyzed the relationship between the levels of AKAP12 methylation and pathological parameters of lung cancer patients .Results 34(56 .7% ) of the 60 lung cancer patients were found to be methylated at the AKAP12 promoter region by MS‐HRM ,the methylation levels of 18 cases ranged between 1% -20% ,14 cases ranged between 20% -60% ,2 cases ranged between 60% -100% .There was no significant differences between the levels of AKAP12 methylation and lung cancer patients′age and gender(P>0 .05) .However ,it was signifi‐cantly higher in the patients with high pathological stage and differentiation degree (P<0 .05) .Conclusion AKAP12 promoter re‐gion methylation was related to tumor progression and malignant degree .
8.Electrolyzed Oxidizing Water Disinfection in Disinfection Supply Center of Peking University First Hospital:An Application Study
Hong TIAN ; Jinxia NIU ; Keshen ZHANG ; Wuai REN ; Dong YU ; Jin WANG ; Min LI
Chinese Journal of Nosocomiology 2009;0(17):-
1.0,achieved the standard request of Disinfection Technology standard.RESULTS Through EOW disinfection,the bacteria were not found on the object surfac of transports and conformed to the object surface disinfection request of Disinfection Technology Standard.CONCLUSIONS Applying EOW to disinfection of polluted instrument recycling vehicle for clinical purpose can fulfill the State standards completely;EOW has strongly germicidal activity,also with characteristics as rapidity,safety and environment security,without any irritant odor.These results indicate a widely potential use of EOW as in hospital disinfection supply center,especially,with a positive significance in health care for pediatric patients.
9.Effect of moxibustion on vascular endothelial cell and expression of vascular endothelial growth factor in rats with cutaneous wound.
Qi SUN ; Zhong-Ren SUN ; Qin-Hong ZHANG ; Di WANG ; Jin-Huan YUE
Chinese Acupuncture & Moxibustion 2014;34(7):679-684
OBJECTIVETo explore the effect and action mechanism of moxibustion on healing of cutaneous wound in rats.
METHODSTwenty-four SD rats were selected and made into linear full-thickness skin injury model. With randomized digital table, rats were randomly divided into a treatment group and a model group, 12 cases in each one. Then according to treatment time, each group was again divided into a 1d group, a 3d group and a 7d group, 4 cases in each one. The moxibustion at injured skin was applied in the treatment group, 30 min per time, once a day. Hematoxylineosin (HE) staining method was adapted to measure growth status of capillary and number of vascular endothelial cell; immunohistochemical method was used to measure the expression of vascular endothelial growth factor (VEGF).
RESULTSThe wound healing indices in the treatment 7d group were higher than those in the model 7d group on both the 4th day and 8th day after treatment (both P < 0.05). The number of capillary in the treatment 1d group and 3d group was higher than that in the model 1 d and 3 d groups (both 1 < 0.05). The number of capillary in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The number of vascular endothelial cell in the treatment 3d group was higher than that in the model 3d group (P < 0.05). The number of vascular endothelial cell in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of number of vascular endothelial cell between the treatment 1d group and model 1d group was not significant (P > 0.05). Positive cells accumulated score of V EGF expression in the treatment 3d group was higher than that in the model 3d group (P < 0.05). Positive cells accumulated score of VEGF expression in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of positive cells accumulated score of VEGF expression between the treatment 1d group and model 1d group was not significant (P > 0.05).
CONCLUSIONMoxibustion could improve the healing of skin wound in rats, which could be related with regulating vascular endothelial cell and VEGF in wound tissue at different time.
Animals ; Endothelial Cells ; metabolism ; Female ; Humans ; Male ; Moxibustion ; Rats ; Rats, Sprague-Dawley ; Skin ; injuries ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wounds and Injuries ; genetics ; metabolism ; therapy
10.Effect of phacoemulsification with different corneal micro-incision sizes on postoperative corneal healing
Ning REN ; Lixia SUN ; Hong CUI ; Zhengri LI ; Chenglin LI ; Haiyan JIN ; Yingjun LI
Chinese Journal of Experimental Ophthalmology 2021;39(4):319-326
Objective:To investigate the effect of phacoemulsification using different corneal micro-incision sizes on the postoperative corneal healing.Methods:A non-random controlled study was performed.Seventy-six patients (76 eyes) with age-related cataract who underwent phacoemulsification and cataract extraction combined with intraocular lens (IOL) implantation in Yanbian University Hospital from May 2016 to May 2017 were enrolled.The subjects were divided into 2.2 mm incision group (37 eyes) and 1.8 mm incision group (39 eyes) according to corneal incision size.The intraoperative cumulative dissipated energy (CDE) and ultrasound time of the two groups were measured and compared.The corneal incision structure and corneal thickness of operative eyes were measured by anterior segment optical coherence tomography before surgery and at postoperative 1 day, 1 week and 1 month.The corneal endothelial cells count, the central corneal thickness (CCT), and the corneal volume in the central corneal diameters of 3.0 mm (CV3) and 10.0 mm (CV10) were measured by Pentacam.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the Yanbian University Hospital (No.2015153), and written informed consent was obtained from all subjects before surgery.Results:There were statistically significant differences in the corneal endothelial cells count, CCT, CV3, CV10, and corneal thickness at epithelial side of the incision between pre-operation and post-operation in each group ( Ftime=17.717, 67.356, 17.577, 13.559, 80.076; all at P<0.01).But there was no statistically significant difference in the indexes between the two groups ( Fgroup=0.788, 0.706, 3.692, 4.341, 4.182; all at P>0.05).The number of corneal endothelial cells in the two groups was gradually decreased after surgery, and was significantly reduced at one month after surgery in comparison with the pre-operation, and the differences were statistically significant (both at P<0.05).At 1 day after surgery, the CCT, CV3, CV10, and corneal thickness at epithelial side of the incision in the two groups were increased significantly in comparison with the pre-operation and the differences were statistically significant (all at P<0.05), and the differences were not statistically significant between preoperative and postoperative 1 week or 1 month (all at P>0.05).With time going by, the indexes were back to normal gradually.The incidence of endothelial gaping and dislocation at the corneal incision in the 1.8 mm incision group was 12.8% (5/39) and 5.1% (2/39), which were significantly higher than 0.0% (0/37) and 2.7% (1/37) in the 2.2 mm incision group, and the differences were statistically significant ( χ2=5.078, P=0.024; χ2=0.295, P=0.590).At 1 day after surgery, corneal thickness at the endothelial side in the 1.8 mm incision group was significantly thicker than that in the 2.2 mm incision group, and the difference was statistically significant ( P=0.042), and the corneal thickness at endothelial side of the incision was positively correlated with CDE in the two groups ( r=0.231, P=0.025; r=0.347, P=0.003). Conclusions:Compared with 2.2 mm incision, 1.8 mm corneal incision results in higher incidence of endothelial gaping and dislocation at the corneal incision, greater corneal thickness at endothelial side of the incision and slower recovery.