2.Soluble Expression and Purification of Snake Venoms Fihrino(geno)lytic Emzyme Alfimeprase in E.coli
Shou-Tao ZHANG ; Yan-Sheng ZHOU ; Xue-Hua LAI ; Xing-Feng BAO ; Ai-Guang GUO ;
China Biotechnology 2006;0(03):-
Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.
3.A Study on Cellular Matrix Via Cell Extraction from Bovine Pericardium
Jian-Ye ZHOU ; Sheng-Shou HU ; Bing REN ; Guo-Qi QI
Acta Academiae Medicinae Sinicae 2001;23(2):193-195
Objective To obtain an acellular matrix from bovine pericardial tissue, as a scaffold for tissue engineering of heart valves or cardiovascular patching applications. Methods A four step detergent and enzyme link extraction procedure was practiced in this study. The mechanic properties of the acellular matrix were evaluated, and the components were analyzed biochemically. Results HE staining confirmed the removal of cells and Von Gieson staining showed the integrality of collagen and elastin. Biochemical analysis demonstrated the retention of collagen and some glycosaminoglycans while the percentage of the soluble proteins reduced slightly. The tissue fracture strength and shrinkage temperature made no significant differ ence after extraction. Conclusions The extraction process is effective to remove cells from bovine pericardial tissue while maintains its mechanical strength. This approach may eventually lead to a scaffold for heart valves and cardiovascular patching applications in tissue engineering.
4.Long Term Results of Breast Conserving Operation Plus Radiotherapy on 63 Patients with Early Breast Cancer
Yun Shou HAN ; Xiong Guo CHEN ; Liuguo TU ; Chun Yang ZHOU ; Sheng Huo XIA
Chinese Journal of Cancer 2001;20(6):635-637
Objective:This study was designed to evaluate the efficacy of breast conserving operation plus radiotherapy. Methods: From January 1989 to march 1994, 63 cases with early breast cancer, among them, 22 cases were in StageⅠ , 41 cases in Stage Ⅱ were treated using breast conserving surgery plus radiotherapy. Tangential field irradiation was used with dose of 45 Gy- 50 Gy. Tumor bed was irradiated additional 15 gray- 20 gray with electron beam. Results: The 5 and 10 years survival rates were 92.1% and 87.3% respectively. In all of 63 patients, only 1 cases developed radiation induced pneumonitis. The cosmetic effects were satisfactory for all patients. Conclusion: In early stage of breast cancer, the effects of breast conserving operation plus radiotherapy were simitar to that of radical mastectomy. Thus for legible early stage breast cancer breast conserving therapy could be the first choice.
5.Protective effects of resveratrol on lipopolysaccharide-induced inflammatory toxicity in primary cortical astrocyte cultures.
Ben LIU ; Qing CHEN ; Jiang GUO ; Hao ZHOU ; Bin WANG ; Jin LI ; Shou-yi YU
Journal of Southern Medical University 2011;31(12):2052-2056
OBJECTIVETo study the anti-inflammatory effect of resveratrol in primary cortical astrocyte cultures stimulated by lipopolysaccharide (LPS) and explore the underlying mechanism of this protective effect.
METHODSThe astrocytes were cultured in the presence of resveratrol at different concentrations for 12 h followed by stimulation with lipopolysaccharide (LPS) for another 24 h. Lactate dehydrogenas (LDH) leakage volumes were detected, the cytotoxicity of resveratrol was examined using cell counting kit-8 (CCK-8), the release of NO was measured by Griess reaction, and the expression levels of TNF-α and iNOS were measured using ELISA and Western blotting respectively.
RESULTSThe purity of the astrocytes cultured in vitro was (95.49∓1.86)%. LPS treatment increased LDH leakage and reduced the survival rate of the astrocytes, resulting also in significantly increased NO and TNF-α release and iNOS protein expressions. Within the concentration range of 5-50 µmol/L, resveratrol effectively improved the survival rate of the astrocytes and decreased LDH leakage with a dose-response relationship. Only 25 and 50 µmol/L resveratrol produced obvious inhibitory effect on NO and TNF-α release and iNOS expression, while 5 µmol/L resveratrol had no such effects.
CONCLUSIONHigh concentration of resveratrol can inhibit the release of inflammatory mediators and improve the inflammation injury induced by LPS in astrocytes, the mechanism of which may involve the inhibition of iNOS/NO expression pathway.
Animals ; Animals, Newborn ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Astrocytes ; cytology ; Cerebral Cortex ; cytology ; Inflammation ; chemically induced ; Lipopolysaccharides ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Primary Cell Culture ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; pharmacology ; Tumor Necrosis Factor-alpha ; metabolism
6.Suppression subtractive hybridization for the identification of differentially expressed genes in the hippocampus of the offsprings of lead exposed female rats.
Shou-Ming LI ; Chang-Ying LIU ; Guo-Xiu XIE ; Ming ZHOU ; Zhi-Yong LIU
Chinese Journal of Contemporary Pediatrics 2010;12(10):820-824
OBJECTIVETo screen and identify differentially expressed genes in the hippocampus of the offsprings of lead exposed female rats in order to provide a theoretical basis for identifying learning and memory deficits related genes.
METHODSRNA was extracted from the hippocampus of young rats with learning and memory deficits due to maternal lead exposure. Suppression subtractive hybridization was used to identify the differentially expressed genes in the hippocampus.
RESULTSAn effective subtracted library was constructed which consisted of approximately 200 clones. Sequencing for the library identified 93 clones harboring insertion fragments which included 43 different genes and 4 unknown genes. These genes might be related to learning and memory deficits due to maternal lead exposure.
CONCLUSIONSThe up-regulated genes in the hippocampus of young rats from pregnant rats under lead exposure include some housekeeping genes and some proteins involved in cellular protein folding, signal transduction, stress response and DNA methylation. These proteins might be directly related to a significant reduction in learning and memory abilities in the young rats.
Animals ; Female ; Gene Expression Profiling ; Gene Library ; Hippocampus ; drug effects ; metabolism ; Lead ; toxicity ; Learning ; drug effects ; Memory ; drug effects ; Nucleic Acid Hybridization ; methods ; Polymerase Chain Reaction ; Pregnancy ; Rats
7.Impact of complicated intra-abdominal infection on albumin synthesis rate.
Bo ZHOU ; Jian-an REN ; Yu CHEN ; Guo-sheng GU ; Jun CHEN ; Jie-shou LI
Chinese Journal of Gastrointestinal Surgery 2011;14(7):496-499
OBJECTIVETo study the impact of complicated intra-abdominal infections on albumin synthesis rate.
METHODSEight patients with complicated intra-abdominal infections associated with intestinal fistula were admitted to the Research Institute of General Surgery at the Jinling Hospital between December 2009 and October 2010. Eight healthy volunteers matched for age, sex, and body mass index were enrolled as controls. All the subjects were given a primed, constant infusion of sterile L-[ring-(2)H(5)]-phenylalanine solution (priming dose: 4 μmol/kg, infusion rate: 6 μmol·kg(-1)·min(-1)) via peripheral venous lines in fast state. Arterial blood samples(3 ml) were drawn before and throughout the infusion at hourly intervals. The enrichment of L-[ring-(2)H(5)]-phenylalanine from the plasma free amino acid pool and from albumin were determined by gas chromatography/mass spectrometry analysis.
RESULTSBoth plasma total protein concentration(62.2±1.0) g/L and plasma albumin concentration (32.5±4.0) g/L in patients with complicated intra-abdominal infection were lower compared with controls[(74.2±1.7) g/L and (46.1±2.6) g/L, both P<0.05]. Body temperature, neutrophil count and plasma C-reactive protein concentration in patients with infection were significantly greater than the levels in control subjects(P<0.05). Albumin synthesis rate in patients with intra-abdominal infection was significantly lower than that in the control group [(5.3±1.6)%/d and (7.8±1.2)%/d respectively, P<0.05]. The measurement of plasma free amino acid concentration showed that plasma glutamic acid level was greater than that in control subjects, and that plasma phenylalanine and proline levels were lower than those in controls.
CONCLUSIONComplicated intra-abdominal infection inhibits albumin synthesis rate in patients with intestinal fistula, which may partially contribute to the decrease of plasma albumin concentration.
Adolescent ; Adult ; Case-Control Studies ; Female ; Humans ; Intraabdominal Infections ; blood ; Male ; Middle Aged ; Serum Albumin ; biosynthesis ; Young Adult
8.Amplitude Changes of Low Frequency Fluctuation in Brain Spontaneous Nervous Activities Induced by Needling at Hand Taiyin Lung Channel.
You-long ZHOU ; Cheng-guo SU ; Shou-fang LIU ; Xiang-yu JIN ; Yan-li DUAN ; Xiao-yan CHEN ; Shu-hua ZHAO ; Quan-liang WANG ; Chang-lin DANG
Chinese Journal of Integrated Traditional and Western Medicine 2016;36(5):553-558
OBJECTIVETo observe amplitude changes of low frequency fluctuation in brain spontaneous nervous activities induced by needling at Hand Taiyin Lung Channel, and to preliminarily explore the possible brain function network of Hand Taiyin Lung Channel.
METHODSBy using functional magnetic resonance imaging (fMRI), 16 healthy volunteers underwent resting-state scanning (R1) and scanning with retained acupuncture at Hand Taiyin Lung Channel (acupuncture, AP). Data of fMRI collected were statistically calculated using amplitude of low frequency fluctuations (ALFF).
RESULTSUnder R1 significantly enhanced ALFF occurred in right precuneus, left inferior parietal lobule, bilateral superior temporal gyrus, bilateral middle frontal gyrus, left superior frontal gyrus, left inferior frontal gyrus, left medial frontal gyrus. Under AP significantly enhanced ALFF occurred in right precuneus, bilateral superior frontal gyrus, cerebellum, bilateral middle frontal gyrus, right medial frontal gyrus, and so on. Compared with R1, needing at Hand Taiyin Lung Channel could significantly enhance ALFF in right gyrus subcallosum and right inferior frontal gyrus. Significant decreased ALFF appeared in right postcentral gyrus, left precuneus, left superior temporal gyrus, left middle temporal gyrus, and so on.
CONCLUSIONNeeding at Hand Taiyin Lung Channel could significantly change fixed activities of cerebral cortex, especially in right subcallosal gyrus, right inferior frontal gyrus, and so on.
Acupuncture Therapy ; Brain ; physiology ; Brain Mapping ; Humans ; Magnetic Resonance Imaging
9.Risk factors for leukopenia in patients with gastrointestinal fistula.
Zheng ZHOU ; Jian-An REN ; Hai-Yan LIU ; Guo-Sheng GU ; Jie-Shou LI
Chinese Medical Journal 2010;123(23):3433-3437
BACKGROUNDWhite blood cell count is an important index to the outcome of patients. In hospital, leukopenia is accompanied by high mortality, morbidity and treatment costs. However, in infectious diseases, the reasons responsible for leucopenia was not well elucidated. We investigated patients with gastrointestinal fistula to find risk factors for leukopenia.
METHODSA prospective case control investigation was carried out in the Gastrointestinal Fistula Center, General Surgical Institute of Jinling Hospital. Cases included gastrointestinal fistula patients with leukopenia (n = 98) and controls composed of gastrointestinal fistula patients with normal white blood cell count (n = 78). The two groups were compared for risk factors of leucopenia by statistical analysis.
RESULTSFactors associated with an increased risk for leukopenia included bacterial infection (25.5%) and hypoalbuminaemia (61.2%). Multivariable Logistic regression analysis identified bacterial infection (80%), urinary catheter (70%) and central vein catheter (60%) as the independent determinants for mortality in cases.
CONCLUSIONSIn patients with gastrointestinal fistula, two independent factors for leukopenia and three significant predictors of mortality were elucidated. We suggest that clinicians give patients more supportive management and apply prevention strategies to treat and prevent leukopenia.
Adult ; Aged ; Bacterial Infections ; complications ; Case-Control Studies ; Catheterization, Central Venous ; adverse effects ; Female ; Gastric Fistula ; complications ; Humans ; Intestinal Fistula ; complications ; Leukopenia ; etiology ; mortality ; Logistic Models ; Male ; Middle Aged ; Prospective Studies ; Risk Factors ; Urinary Catheterization ; adverse effects
10.Epidemiology investigation on arsenism from drinking water along Huai'he River and the surrounding area of Hong'ze lake in Huai'an city of Jinngsu province in 2008
Cong-ying, JIA ; Wen-zhou, YANG ; Huai-rong, ZHAO ; Wei, HU ; Yi, WANG ; En-chun, PAN ; Shou-guo, YUAN ; Dao-kuan, SHUN ; Si-hong, CHEN ; Yong, TANG
Chinese Journal of Endemiology 2010;29(1):74-76
Objective To explore the distribution of water with high level arsenic and prevalence of arsenism along Huai'he River and the surrounding area of Hong'ze lake in Huai'an of Jiangsu. Methods Wate rsamples were collected and tested in 2008 from 18 villages of 6 towns according to history data in 3 counties like Xuyi,Jinhu and Hongze. Samples having arsenic level higher than 0.05 mg/L were investigated by epidemiological method and the patients were diagnosed by Standard of Diagnosis for Endemic Arsenism. Results All 5199 water samples were determined,and 260 water samples were exceeding the national drinking water quality level (0.05 mg/L) in 3 counties,the rates of exceeding diagnosis were 5.6%(247/4454),0.7%(4/597),6.0%(9/148) respectively. Total detected rate of endemic arsenic disease was 5.94%(128/2155). The detected rates of age group of 0 ~ ,20 ~,30 ~ ,40 ~ ,50 ~ ,60 ~ ,70 ~ ,80 ~ were 2.86%(1/35),2.11%(2/95),1.26%(3/239),3.10%(16/516),5.53% (32/579),10.07%(41/407),11.84%(27/228),10.71%(6/56) respectively. The detected rate of male (9.10%,78/857) was higher than that of female(3.85%,50/1298,χ~2 = 25.46,P < 0.01). Conclusions Huai'he River and the surrounding areas of Hong'ze lake like Xuyi,Jinhu and Hongze are identified existing endemic arsenic disease area. The prevention of arsenism should be strengthened in these areas.