1.The medical gas central supply system and its quality control.
Yin-bao ZHONG ; Yue-guo QI ; Chao TANG
Chinese Journal of Medical Instrumentation 2002;26(5):363-365
This paper introduces the principles of the medical gas central supply system and its structure, Classification, standards, flow chart and its advantages, and its quantity guarantee and quality control.
Equipment Design
;
Equipment Safety
;
Equipment and Supplies, Hospital
;
standards
;
Gases
;
Humans
;
Quality Control
2.Synthesis of DNA Encoding Botulinum Neurotoxin Receptor syt II and Its Fusion Expression in E.coli
Jing SHI ; Hui WANG ; Shi-Zhong BAO ; Xiao-Jun HOU ; Jun YIN ;
China Biotechnology 2006;0(07):-
In order to construct a prokaryotic expression vector of human receptor syt II N-fragment and to express recombinant MBP-Syt fusion protein in E.coli and to purify and identify its activity. According to codon preference of E.coli, a DNA fragment encoding human syt II N-fragment was synthesized, and then cloned into prokaryotic vector pMAL-c2x for sequencing. Then the recombinant plasmid pMAL-Syt was introduced into E.coli ER2566 by transformation for expression and the obtained engineered bacteria were induced by IPTG. The fusion protein was purified by amylose resin affinity chromatography and identified by SDS-PAGE and Western blot. The binding activity of the protein was determined by ELISA. It is concluded that MBP-Syt protein is of good binding activity.
3.The Recombinant Expression and Receptor-binding Activity of the B Subunit of Shiga-like Toxin Type Ⅱ
Shi-Zhong BAO ; Jing SHI ; Kun CAI ; Jun YIN ; Hui WANG ;
China Biotechnology 2006;0(10):-
Objective:To express the B subunit of Shiga-like toxin type Ⅱ,and analyze its expression form and receptor-binding activity.Methods:The slt2b gene was obtained from EHEC O157∶H7 by PCR,and cloned to the expression vector pET22b(+).The genetically engineered bacteria pET22b(+)-stx2B/BL21 expressed the recombinant StxB after induced with IPTG.The renatured inclusion bodies were purified by ion exchange chromatography.The expression form of rStx2B was investigated by denaturing and native electrophoresis.The receptor-binding activity was confirmed by fluorescence detection and flow cytometer.Result:The constructed genetically engineered bacteria expressed the rStx2B at a high level.The purified protein was obtained after denaturation,renaturation and ion exchange chromatography.According to the denaturing and native electrophoresis,the rStx2B was expressed in a dimmer form,which consists of two monomers cross linked with disulfide bridge.The rStx2B showed good receptor-binding activity by Hela-binding assay.Conclusion:The genetically engineered bacteria were constructed successfully.The receptor-binding activity of rStx2B was independent of the pentamers.
4.Effect of emodin in attenuating endoplasmic reticulum stress of pancreatic acinar AR42J cells.
Li WU ; Feng ZHANG ; Shi-zhong ZHENG ; Yin LU ; Bao-chang CAI
China Journal of Chinese Materia Medica 2015;40(3):501-505
OBJECTIVETo explore the effect of emodin on endoplasmic reticulum (ER) stress of pancreatic acinar AR42J cells.
METHODRat pancreatic acinar AR42J cells were cultured in 6-well plates, and divided into the normal control group, the model group (with the final concentration at 1 x 10(-7) mol · L(-1) for cerulean and lipopolysaccharide at 10 mg · L(-1)) and the emodin group (10, 20, 40 μmol · L(-1)). Cells in each group were cultured in three multiple pores for 24 h, and their supernate was removed after cell attachment. The normal control group was added with haploids, the model group was added with the modeling liquid for haploids, and the treatment groups were added with different concentrations of emodin at 15-20 min before the modeling liquid. The cells were continuously cultured for 3 h under 37 °C and 5% CO2. Their intracellular protease and lipase expressions were detected with kits. The cellular morphology was observed under optical microscope. The level of calcium in endoplasmic reticulum was measured under laser confocal microscopy. Western blot assay were used to determine the protein expression of ER-related signaling molecules.
RESULTEmodin could significantly inhibit levels of amylase, lipase and intracellular calcium and ER.
CONCLUSIONEmodin could reduce pancreatic acinar cell injury induced by the combination of cerulean and lipopolysaccharide. Its action mechanism is correlated with the inhibition of intracellular calcium overload and ER stress.
Animals ; Calcium ; metabolism ; Cell Line, Tumor ; Emodin ; pharmacology ; Endoplasmic Reticulum Stress ; drug effects ; Pancreatic Neoplasms ; metabolism ; pathology ; Rats ; Unfolded Protein Response ; drug effects
5.Construction and Screening of a Phage Display Library of Repertoire Single Chain Fv Antibody from Mouse Immunized with BoNTB/Hc
Xiu-Qing YANG ; Hui WANG ; Jing SHI ; Kun CAI ; Xiao-Jun HOU ; Shi-Zhong BAO ; Jun YIN ;
Microbiology 1992;0(06):-
To produce antibodies capable of neutralizing botulinum neurotoxin type B(BoNT/B),We cloned the carboxy-terminal end of Hc containing the major determinants responsible for specific toxin,induced and purifed.The heavy-chain and kappa light-chain variable region gene repertoire of immunoglobulin were amplified individually from the spleen cell mRNA by RT-PCR and joined as a single-chain Fv(scFv)DNA fragment.These fragment were cloned into the phagemid pCANTAB5E and the phage display library was constructed.Results showed that the high affinity scFv was obtained after 4 rounds of panning,with its DNA sequence conforming to that of mouse antibody.
6.Bcl-2/Bax gene expression in different types of carotid plaque.
Bao-zhong MEN ; Ding-biao ZHOU ; Xiao-ming ZHANG ; Huai-yin SHI
Acta Academiae Medicinae Sinicae 2005;27(2):241-244
OBJECTIVETo investigate Bcl-2/Bax gene expression in different types of carotid plaque, and examine the relationship between gene expression and atherosclerotic plaque instability and the main cause of brain ischemic events.
METHODSTotally 42 human carotid plaque specimens obtained during carotid endarterectomy were divided into stable group (n=19) and unstable group (n=23) based on histopathological studies (HE staining). Eight aortic arteries and their branches from hepatic transplantation donors were taken as control group. Bcl-2/Bax was detected by immunohistochemistry (IHC) staining (n=42) and in situ hybridization (ISH) (n=25, stable 13/unstable 12).
RESULTSBcl-2 gene expression, which was expressed in smooth muscle cells (SMC), endothelial cells (EC), macrophages (MP) and foam cells, was detected in 20 and 9 cases in unstable plaque while 11 and 4 cases in stable plaque by IHC and ISH, respectively (P < 0.05). Bax, which was expressed in SMC and MP, was detected in 18 and 11 cases in unstable plaque, while 8 and 5 cases in stable plaque by IHC and ISH, respectively (P < 0.05).
CONCLUSIONThe expression rate of Bcl-2/Bax in unstable plaques was higher than in stable plaques. Bcl-2 was one of the elements that maintain plaque stability whereas Bax was one element that facilitates plaque instability. Therefore, Bcl-2/Bax expression in different stage of atherosclerosis may be one of the molecule regulation mechanisms in carotid atherosclerosis.
Apoptosis ; genetics ; Carotid Arteries ; metabolism ; pathology ; Carotid Artery Diseases ; metabolism ; Carotid Stenosis ; metabolism ; pathology ; Endothelium, Vascular ; metabolism ; Humans ; Macrophages ; metabolism ; Muscle, Smooth, Vascular ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Up-Regulation ; bcl-2-Associated X Protein
7.Effect of preoperative transcatheter arterial chemo-embolization on activity of cell proliferation in Wilms; tumor.
Wei-guang LIU ; Wei-zhong GU ; Yin-bao ZHOU ; Hong-feng TANG ; Min-ju LI
Journal of Zhejiang University. Medical sciences 2008;37(1):83-87
OBJECTIVETo investigate the effect of preoperative transcatherter arterial chemo-embolization (TACE) on the cell proliferation in Wilms; tumor.
METHODSForty-one cases of Wilms; tumor diagnosed by histopathology were divided into two groups: in TACE group, 23 patients received TACE first and were operated 2 weeks later; in control group, 18 patients were operated alone. A comparative analysis of the pathological finding was made in two groups, and the expression of PCNA and VEGF in tumor tissue was detected by immunohistochemistry.
RESULTThe degeneration of tumor tissue such as tumor cell necrotic, broken, disappearance occurred in 17 cases of TACE group and in 4 cases of control group, respectively (P <0.01). The expression of PCNA in TACE group and in control group was 1/23 (4.3 %) and 9/18(50.0 %), respectively (P <0.01). VEGF was expressed in 7/23 (30.4 %) of TACE group and 9/18 (50.0 %) of control group (P=0.283).
CONCLUSIONTACE can significantly inhibit proliferation and enhance degeneration of Wilms; tumor cells.
Cell Proliferation ; Chemoembolization, Therapeutic ; adverse effects ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Kidney Neoplasms ; therapy ; Male ; Preoperative Care ; Proliferating Cell Nuclear Antigen ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Wilms Tumor ; complications ; pathology ; surgery ; therapy
8.Separation and cryopreservation of cord blood mononuclear cells.
Jiong-Cai LAN ; Zhong LIU ; Mao-Zhou GAN ; Qiang CHEN ; Yin-Zhe ZHANG ; Qing-Bao MENG
Journal of Experimental Hematology 2002;10(4):351-354
The influencing factors on cord blood storage after collection and mononuclear cell separation as well as cryopreservation were studied. The mononuclear cell are separated from blood after blood collection, then cryopreserved and washed after thawed. Results showed that the cord blood kept at 4 degrees C or room temperature less than 24 hours after blood collection, mononuclear cell separated by hydroxyethylstarch and 2 centrifugations, mononuclear cell cryopreserved with 50% DMSO and autoplasma from cord blood as protectives and washing the cells after thawing. In conclusion, the optimal project in this study can effectively preserve cord blood mononuclear cells.
Blood Preservation
;
Cell Separation
;
methods
;
Cryopreservation
;
Fetal Blood
;
cytology
;
Humans
;
Leukocytes, Mononuclear
;
physiology
9.Expression of PTEN in athymic mice with HCC treated by complex prescription of Chinese crude drug.
Bao-guo SUN ; Ze-xiong CHEN ; Shi-jun ZHANG ; Yong-dong LIU ; Hong-zhong HUANG ; Li-rong YIN
China Journal of Chinese Materia Medica 2007;32(11):1057-1060
OBJECTIVETo research the treatment effect of complex prescription of Chinese crude drug in BALB/c athymic mice with human liver cancer, which were built by Bel-7402.
METHOD48 male BALB/c athymic mouse models were built by Bel-7402 with an indirect method. After 24 hours of postoperation, the 48 athymic mice were distributed randomly into 4 groups which were treated by intragastric administration with complex prescription of Chinese crude drug that had been deliquated into 3 groups by the different density as the low, middle, and high and FT207 (Tegafur) for 4 weeks. At last, athymic mice were put to death and PTEN was detected in hepatic tissue, latero-cancer tissue and cancer tissue by immunohistochemistry (PowerVision Two-Step Histostaining Reagent).
RESULTAll of the 48 athymic mice survived 12 to 28 days (Ms 24 days) and every mouse with liver cancer demonstrated by dissection. The result of immunohistochemistry represents that the intensity of PTEN in latero-cancer tissue is the highest, and then the hepatic tissue, the lowest is cancer tissue, P < 0.01. It also represents that the intensity of PTEN in treatment groups (A, B, C) is more higher than the control group (D), P < 0.05 or P < 0.01, and group B is the highest in the treatment groups, P < 0.05 or P < 0.01. However, there is no significant statistic difference between group A and group C.
CONCLUSIONThe higher expression of PTEN in the laterocancer tissue can represent the protective reaction of stress of the organism. And anticancer effect of this complex prescription of Chinese crude drug relates to an eligible density of it. Mechanisms of this complex prescription of Chinese crude drug healing HCC may partially be explained by enhancing the expression of PTEN in liver.
Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; enzymology ; pathology ; Cell Line, Tumor ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Immunohistochemistry ; Liver ; drug effects ; enzymology ; pathology ; Liver Neoplasms ; drug therapy ; enzymology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; PTEN Phosphohydrolase ; metabolism ; Phytotherapy ; Plants, Medicinal ; chemistry ; Random Allocation ; Xenograft Model Antitumor Assays
10.Study on the compatibility of yttria-stabilized zirconia framework bonded to the corresponding veneering ceramic.
Jia-yue YIN ; Zhong-ti ZHANG ; Hong-jun AI ; Wen-jie SI ; Yang BAO
West China Journal of Stomatology 2009;27(6):669-672
OBJECTIVETo investigate the bonding properties and interface characterization of a domestic 3mol yttrium-stabilized tetragonal zirconium polycrystal (3Y-TZP) framework fired on with 4 different veneering ceramics for zirconia.
METHODS4 different commercial veneering ceramics for zirconia (VITA VM9, SHOFU VINTAGE ZR, IPS e.max Ceram, Cercon ceram kiss) were sintered on 3Y-TZP rectangulars (15 mmx5 mmx5 mm) according to the manufacturers' instructions for shear bond strength test, a metal-ceramic system(Ni-Cr alloy/VITA VMK95) was fabricated in the same type as a control group. Two bilayered specimens (3Y-TZP/VITA VM9, Ni-Cr/VMK95) were prepared for scanning electron microscope (SEM) and energy distribution spectrum (EDS).
RESULTSThe values of shear bond strength test were (18.83 +/- 1.77) MPa for 3Y-TZP/VITA VM9, (23.83 +/- 7.05) MPa for 3Y-TZP/SHOFU VINTAGE ZR, (17.87 +/- 2.30) MPa for 3Y-TZP/IPS e.max Ceram, (22.26 +/- 7.45) MPa for 3Y-TZP/Cercon ceram kiss, (20.55 +/- 5.13) MPa for Ni-Cr alloy/VITA VMK95. There was no statistically significant between all-ceramic groups and the control group (P > 0.05). The failure modes in all-ceramic groups showed predominately adhesive at the interface. SEM showed the 3Y-TZP/VITA VM9 contacted tightly at the interface, while EDS detected Si element diffused into 3Y-TZP material.
CONCLUSIONThe results indicate that domestic 3Y-TZP has a good interface compatibility with 4 commercial veneering ceramics, as a dental framework material, it can satisfy the clinical requirements.
Ceramics ; Dental Bonding ; Dental Porcelain ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Yttrium ; Zirconium