2.32 patients of acute fulminant paraquat poisoning.
Wen-Sheng WANG ; Qing-Long LU ; Zeng-Xiang MA
Chinese Journal of Industrial Hygiene and Occupational Diseases 2008;26(12):772-773
Acute Disease
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Adolescent
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Adult
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Female
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Humans
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Male
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Middle Aged
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Paraquat
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poisoning
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Retrospective Studies
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Young Adult
4.Construction of shSet7/9 vector and its function in HepG2
Kejun MA ; Xingchen SHI ; Ping LI ; Xiaoqiang LI ; Wen REN ; Long QIN ; Xinhe SHI
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):753-757
Objective To silence human gene Set7/9 and screen out stable transfection cell line in hepatocellular carcinoma cell line HepG2 so as to investigate the impact of down-regulation of Set7/9 in cell line HepG2 and provide experimental foundation for studies on the effect of set7/9 in HepG2.Methods The target oligo was designed and synthesized;shRNA interference vector and the control vector were constructed and transfected into HepG2 cells;the stable transfection cells were screened out.Then Real-time PCR and Western blot were performed to detect the silence of Set7/9 according to both gene expression and protein expression level. Results The shRNA interference vector was constructed and transfected into HepG2 cells successfully.Compared with that in the negative control group,the expression of Set7/9 was dramatically downregulated (P < 0.05 ). Meanwhile,the expression of related protein Sirt1 and Suv39h1 was upregulated 8.4 folds and 1.1 fold, respectively.Conclusion Downregulation of Set7/9 expression can upregulate Sirt1 and Suv39h1,suggesting that Set7/9 may affect the activity of HepG2 cell lines.
5.Expression of a Novo Human Osteoprotegerin-Mycobacterial Heat Shock 70 Fusion Protein with Baculovirus-Insect Expression System
Shu LIU ; Quan-Geng ZHANG ; Yue ZHANG ; Jing MA ; Zhen-Long LIU ; Wen-Ming ZHAO ;
China Biotechnology 2006;0(05):-
Objective:Osteoprotegerin (OPG) is a key molecule negatively regulating osteoclast differentiation and activation; and the conserved mycobacterial heat shock 70 (HSP70) peptide p111-125 has also been found to inhibit inflammation reactions in chronic arthritis. BaculoDirectTM baculovirus expression system was selected to express recombinant OPG-HSP70 in insect cells.Methods:The human functional fragment (p22-194) of OPG and functional fragment (p111-125) of mycobacterial HSP70 gene were cloned into the transfer vector pENTRTM/SD/D-TOPO. The recombinant plasmid was performed an LR reaction with the BaculoDirectTM Linear DNA to generate recombinant baculovirus DNA. The cultured Sf9 insect cells were directly transferred with the recombinant baculovirus DNA,and the pure recombinant baculovirus was obtained. Then recombinant baculovirus was infected Sf9 insect cells again to express the OPG-HSP70 gene.Results:The target protein was detected at the time of 48h post infection,reached at highest yield at the time of 72h post-infection. A 28kDa protein immunostaining band was detected by Western blotting from lysate of those cells.And the purified protein was obtained by using Ni-NTA system. Functional stuies on the fusion protein showed it significantly reduce osteoclast cell number[(3.10?0.640) cells under each microscope field in treatment group by comparing to (10.70?0.817)cells in the control group] in the osteoclast inhibition test,and reduce the inflammation reaction in a delayed type hypersensitivity (DTH) mice model (P
6.Protein compositions in the seminal plasma are similar between severe oligospermic and healthy fertile men.
Jie BAI ; Ling SUN ; Hong ZHU ; Li-Wen ZHANG ; Jun-Long MA ; Yu-Long CONG
National Journal of Andrology 2008;14(4):298-302
OBJECTIVETo analyse the variability of proteins in the seminal plasma of severe oligospermic and healthy fertile men.
METHODSSpermatic fluid samples were collected from 11 healthy fertile men and 6 severe oligospermic male volunteers and tested by SELDI-TOF-MS with the CM10 protein chip to get the protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc.
RESULTSThe mean peak heights of 2 lower-abundance proteins expressed in the seminal plasma of the severe oligospermic men were statistically different from the healthy fertile males (P<0.05). Fifteen different proteins existed between the nonobstructive azoospermic and the severe oligospermic group, 7 of which, with m/z of 7,196.058, 7,547.610, 5,780.493, 7,059.844, 7,409.589, 5,379.173 and 10,778.810, also between the non-obstructive azoospermic and the healthy fertile males (P<0.05). Except the latter two, the contents of the other 5 proteins were decreased in the non-obstructive azoospermic men (P<0.05).
CONCLUSIONThe finger prints of the seminal plasma proteins of the severe oligospermic group were similar to those of the healthy fertile males, both significantly different from the non-obstructive azoospermic men. It is suggested that pathogenesis mechanisms differ exist between non-obstructive azoospermia and severe oligospermia but are not the simple accumulation of genetic factors.
Adult ; Humans ; Male ; Oligospermia ; metabolism ; Semen ; metabolism ; Seminal Plasma Proteins ; analysis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
7.The alteration of MTS1 gene in precancerous lesions and squamous cell carcinoma of oral mucosa.
Wen-xia WANG ; Shan-zhen SUN ; Pi-shan YANG ; Bo-long MA ; Yan LONG
Chinese Journal of Stomatology 2003;38(5):361-363
OBJECTIVETo investigate the expression and alteration (including homozygous deletion and mutation) of MTS1 gene in precancerous lesions and squamous cell carcinomas (SCC) of oral mucosa, and to analyse the function of MTS1 gene alteration in oral mucosal carcinogenesis.
METHODSThe expression of p16 protein produced by MTS1 gene was examined with immunohistochemical SP method in 10 normal oral mucosas, 30 precancerous lesions (10 mild, 10 moderate and 10 severe dysplasia respectively) and 45 squamous cell carcinomas (SCCI18, SCCII 19, SCCIII 8). The deletion and mutation of exon1 and exon2 of MTS1 gene were examined with methods of PCR and SSCP in these same samples.
RESULTSAll the precancerous lesions had p16 protein expression and no alteration of MTS1 gene. In SCC, the positive rate of p16 protein was 60.0% with 72.2% in SCCI, 57.9% in SCCII, 37.5% in SCC III, and there were no significant difference among the three groups by chi2 test (P>0.05). Gene homozygous deletion of exon1 and/or exon2 was detected in 10 cases, and gene mutation in 4 cases. The whole rate of gene alteration was 31.1% (14/45). The MTS1 gene alteration rate was 27.8% in SCCI, 31.6% in SCCII, 37.5% in SCC III and there was also no significant difference among the three groups by chi2 test (P>0.05). In SCC with local lymph nodes metastasis, MTS1 alteration rate was 57.1%, while in SCC with no lymph nodes metastasis was 8.3%, and there was significant difference by chi2 test (P<0.05).
CONCLUSIONSMTS1 gene alteration is not an early event in the carcinogenesis of oral mucosa and can not be used as a biology mark to examine oral precancerous lesions. MTS1 gene may play a certain role in the progression of oral squamous cell carcinomas.
Carcinoma, Squamous Cell ; chemistry ; genetics ; pathology ; Cyclin-Dependent Kinase Inhibitor p16 ; analysis ; Genes, p16 ; Humans ; Lymphatic Metastasis ; Mouth Neoplasms ; chemistry ; genetics ; pathology ; Mutation ; Precancerous Conditions ; genetics
8.Comparative analysis of proteins in seminal plasma of non-obstructive azoospermia patients and healthy fertile males.
Jie BAI ; Ling SUN ; Shi-Ling CHEN ; Li-Wen ZHANG ; Jun-Long MA ; Yu-Long CONG
National Journal of Andrology 2007;13(7):579-583
OBJECTIVETo analyse protein alterations in the seminal plasma of non-obstructive azoospermia patients.
METHODSSemen samples were collected from 11 healthy fertile and 6 azoospermia male volunteers respectively and tested by SELDI-TOF-MS with CM10 protein chip to get protein spectra maps, which were automatically treated with the special softwares of Ciphergen Inc.
RESULTSThe mean peak heights of 28 proteins expressed in the seminal plasma of the azoospermia patients were statistically different from those of the healthy fertile males (P < 0.05 ), of which 24 were of lower contents than in the normal controls, 4 with remarkably significant difference, M/Z 7 196.058, 7 630.573, 7 547.610 and 7 709.833 (P < 0.01).
CONCLUSIONThe seminal plasma proteins of the azoospermia patients were significantly different from those of the healthy fertile males, with decreased contents of most of the different proteins, which might be significantly correlated with the development of azoospermia.
Adult ; Azoospermia ; metabolism ; Humans ; Male ; Proteins ; analysis ; Proteomics ; methods ; Semen ; chemistry ; cytology ; Spectrometry, Mass, Electrospray Ionization ; Sperm Count ; Sperm Motility
10.Perioperative intervention of tetramethylpyrazine on free skin flap transplantation.
Wen-long MA ; Chun-sheng CHENG ; Zhi-wei ZHAO
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(7):714-716
OBJECTIVETo study the impacts of tetramethylpyrazine (TMP) intervention on hemorheology and blood coagulation before and after free skin flap transplantation, in order to provide new clues of applying Chinese drugs for activating blood circulation and removing stasis in microsurgery.
METHODSSixty patients undergoing free skin flap transplantation were randomly and equally assigned to two groups, the treatment group administered with tetramethylpyrazine, the control group with low molecular dextran, both were treated for 7 days. The survival rate of transplanted flap, hemorrheologic parameters and four blood coagulation associated indices (prothrombin time, thrombin time, activated prothrombin time and fibrinogen) were detected before and after treatment.
RESULTSComparisons of clinical efficacy and hemorrheologic indices showed no significant difference between the two groups (P > 0.05), but TMP showed less impacts on the four blood coagulation associated indices at the 24, 48 and 72 h after operation (P < 0.05).
CONCLUSIONTMP could prevent blood vessel crisis after free skin flap transplantation, so it is valuable in microsurgical clinical application.
Adult ; Female ; Hand Injuries ; drug therapy ; surgery ; Hemorheology ; drug effects ; Humans ; Male ; Microsurgery ; methods ; Middle Aged ; Perioperative Period ; Phytotherapy ; Pyrazines ; therapeutic use ; Surgical Flaps ; transplantation ; Young Adult