1.The Influences of Pre-injection of Donor Apoptotic Cells on Survival of Islet Grafts and Function of T Lymphocytes
Shuangxi LI ; Yuan LIU ; Baolei LI ; Shu YANG ; Hong CHEN ; Dehong CAI ; Zhen ZHANG
Tianjin Medical Journal 2014;(2):160-163
Objective To study the influence of pre-injection of donor apoptotic cells in the survival of islet grafts and the function of T lymphocytes in the peripheral blood. Methods The donor apoptotic cells and necrotic cells were ob-tained respectively by X-irradiation from electron linear accelerator and a heat-shock procedure (water bath box 56℃, 1 h). The diabetic rats for islet transplantation (n=42) were induced by a single intraperitoneal injection of streptozotocin (STZ), then were randomly divided into four groups:rats were injected by physiological saline group (n=9), normal cells group (n=12), apoptotic donor cell group (n=12) and necrotic donor cell group (n=9). On the seventh day, each group received islet transplantation under the renal capsule. The blood glucose level was detected to reflect the survival of the islets. The periph-eral blood samples of three rats in each group were obtained at different observation times. The proliferative activity of T lym-phocytes was determined by MTT method. The levels of cytokines interferon (IFN)-γ, interleukin (IL)-10 in peripheral blood were measured by Luminex 100 Integrated System, and transforming growth factor (TGF)-β1 by ELISA respectively at 0 d, 1 week, 2 weeks and after rejection. Results The survival time of islets was significantly prolonged by the pre-intervention of apoptotic cells, and the proliferative activity of T lymphocytes stimulated by ConA was inhibited. Meanwhile, the extent of the increased level of IFN-γwas inhibited significantly at 1 week and 2 weeks after transplantation (P<0.05), the levels of IL-10 and TGF-β1 were significantly increased before transplantation, 1 week and 2 weeks after transplantation (P<0.05). Conclusion Our results demonstrated that the pre-treatment of donor apoptotic cells can regulate the recipient’s immune reactive state by inhibiting the proliferative activity of T lymphocytes and changing the levels of cytokines from different sub-sets of T lymphocytes, and finally resulted in the prolonging of the survival of islet grafts.
2.Genotype analysis of ESBLs-producing Klebsiella pneumoniae isolates
Shu-Zhen CHEN ; Rong-Lin SHI ; Fen YAO ; Ying-Mu CAI ; Yuan-Su QIAN ;
Chinese Journal of Infection and Chemotherapy 2006;0(04):-
Objective To identify the genotypes of ESBLs-producing Klebsiella pneumoniae isolates from the First Affiliated Hospital,Shantou University Medical College.Methods The MICs of 10 antibiotics were determined by agar-dilution against the clinical isolates of ESBLs-producing K.pneumoniae.PCR were performed with specific primers for blaTEM,blaSHV, blaCTX-M and blaOXA respectively.PCR products were cloned and sequenced.Results The results of PCR showed that a- mong the 83 strains of ESBLs-producing K.pneumoniae,75 were positive for blaTEM,41 positive for blaSHV,25 poitive for blaCTX-M,9 positive for hlaOXA.Three genotypes were found in 13 strains(15.7%),2 genotypes in 59 strains (71.1%) and single genotype in only 11 strains(13.2%).The genes of CTX-M-3,TEM-1 and SHV were found co-existent in 9 strains. The strains carrying 2 or 3 ESBL genes were more resistant to antibiotics than those carrying only 1 ESBL gene.Conclusions The genotypes of ESBLs-producing Klebsiella pneumoniae in this hospital are blaTEM,blaSHV,blaCTX-M and blaOXA. Most strains carry 2 or 3 ESBL genes.
3.Cloning and Preliminary Analysis of the Stem Specific Promoter of Sugarcane
Wen-Wei CAI ; Zheng-Peng WANG ; Shu-Zhen ZHANG ; Ben-Peng YANG ;
China Biotechnology 2006;0(08):-
Sugarcane stem is an ideal organ for producing foreign pharmaceutical proteins and chemicals by genetic engineering.A perfect promoter driving foreign gene to express strongly and specifically in sugarcane stem is necessary for this purpose.In order to isolate a Sugarcane stem-specific promoter,a fragment of 1968bp nucleotide sequence(Ppst2a)upstream 5' of sugarcane pst2a gene,which was demonstrated to express specifically in sugarcane stem previously was isolated by using chromosomal walking.Bioinformatical analysis of this sequence shows that the sequence contains some typical elements of a promoter.To identify the stem-specific of this promoter,a construct was derived from pCAMBIA1301,which original CaMV 35S promoter was replaced by the 1968bp nucleotide sequence,and named as pCAMBIA1900.Transformations of pCAMBIA1900 and pCAMBIA1301 to leaves and stem pieces of sugarcane were carried out by using particle bombardment.The transient expression of gus showed that the gus expressed specifically in sugarcane with a little higher level compared with CaMV 35S.It is the first report that pst2a promoter is a potential stem-specific promoter which can further be used in transgenes into sugarcane.
4.Effects of sodium tanshinone B on the protein expression of NMDAR1 in rat hippocampal subfields following focal ischemia/reperfusion injury.
Qing CAI ; Shu-yun HUANG ; Jun-zhen TAN
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(8):1073-1076
UNLABELLEDOBJECTIVE To observe the changing laws of the protein expression of N-methyl D-aspartate receptor (NMDAR) in rat hippocampal subfields following focal ischemia/reperfusion injury, and to study the effects of sodium tanshinone B (STB) on it, thus exploring the possible mechanism of STB for treating cerebral ischemia.
METHODSThe rat model of focal cerebral ischemia/reperfusion injury was established using middle cerebral artery occlusion (MCAO) by reversibly inserting a nylon thread. The Wistar rats were randomly divided into the sham-operation group, the I/R model group, and the low, middle, and high dose STB groups. The neural functional disturbance was scored referring to the 5-grade Zea Longa EL standard. The protein expression of NMDAR1 in the ischemic side was detected using immunohistochemical assay.
RESULTSThere was statistical difference in the scores of the neural functional disturbance in the middle and high dose STB groups when compared with the model group (P < 0.01). Results of the immunohistochemical assay showed the expression of NMDAR1 in CA1 region was obviously higher in the I/R model group, the low and middle dose STB groups than in the sham-operation group (P < 0.01). The expression of NMDAR1 in CA1 region was obviously lower in the high dose STB group than in the I/R model group (P < 0.01), the low (P < 0.01) and middle dose STB groups (P < 0.05). The expression of NMDAR1 in CA3 region was obviously higher in the low dose STB group and the I/R model group than in the sham-operation group, the middle and high dose STB groups (P < 0.01). The expression of NMDAR1 in CA3 region was obviously higher in the high and middle dose STB groups than in the sham-operation group (P < 0.05).
CONCLUSIONSSTB could promote the recovery of neural functions in cerebral ischemia/reperfusion injury rats. STB fought against cerebral ischemia/reperfusion injury by lowering excitable neurotransmitter glumatic acid and reducing the protein expression of NMDAR1.
Animals ; Brain Ischemia ; metabolism ; Diterpenes, Abietane ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; metabolism ; Reperfusion Injury ; metabolism
5.Iodine nutrition level of children aged 8 - 10 in low-coverage area of iodized salt of Yushu Qinghai province in 2009: an analysis of surveillance results
Lan-sheng, HU ; Hui-zhen, YU ; Li-lin, CHEN ; Ya-nan, LI ; Pei-zhen, YANG ; Sheng-hua, CAI ; Shu-bang, LI ; Fa-rong, ZHANG
Chinese Journal of Endemiology 2011;30(3):316-318
Objective An analysis was conducted to investigate the iodine nutrition level of children aged 8 - 10 in low-coverage area of iodized salt of Yushu Qinghai province for providing a scientific basis for the development of effective preventive measures. Methods Yushu, Chengduo, Nangqian and Zaduo counties with higher non-iodized salt coverage rate in Yushu Qinghai province were chosen as survey counties in 2009. Three townships were selected in each county, and 2 primary schools were selected in each township and 40 urine samples of children aged 8-10 were collected randomly in each primary school. The content of urinary iodine was analyzed by As-Ce catalytic spectrophotometery. Results Median urinary iodine of children aged 8 - 10 in Nangqian and Zaduo was < 100 μg/L. The percentage of median urinary iodine < 50 μg/L in Yushu was over 20%. Median urinary iodine of children aged 10 in Zaduo was 81.5 μg/L, the percentage of median urinary iodine < 50 μg/L of children aged 9 and 10 was over 20%. The percentage of median urinary iodine < 50 μg/L in children aged 9 and 10 of Yushu was over 20%. Median urinary iodine of girls in Zaduo was 87.1 μg/L, the percentage of median urinary iodine < 50 μg/L of boys in Zaduo was over 20%. The percentage of median urinary iodine < 50 μg/L of girls in Yushu was over 20%. Conclusions The iodine nutrition level of children aged 8 - 10 in Nangqian, Zaduo and Yushu counties were very low due to the impact of non-iodized salt. We propose salt market in the region to strengthen management and improve the coverage and consumption rates of iodized salt to improve the level of iodine nutrition for effective prevention of iodine deficiency disorders.
6.Correlation between matrix metalloproteinases activities and myocardial injury in neonatal rats after asphyxia.
Shu-zhen XU ; Cai-xia WANG ; Wei ZHAO ; Jian-fang CHU ; Wen-dong LIU ; Ming LI
Chinese Journal of Pediatrics 2007;45(2):134-137
OBJECTIVETo study possible correlation between matrix metalloproteinases (MMPs) activities and myocardial injury after asphyxia in neonatal Wistar rats.
METHODSixty neonatal Wistar rats (7 to 10 days old) were randomly divided into four groups: control group (group D); asphyxia groups A, B and C (1 day, 7 days, 14 days after asphyxia), every group had 15 rats. In the asphyxia groups, animal model was produced by normobaric asphyxia. Groups A, B and C were sacrificed on days 1, 7 and 14 days after asphyxia, and group D rats were sacrificed on the 7 th day. Then the heart blood was taken to tested the serum cTnI. The myocardial MMPs-3 and 9 activity was measured by using immunohistochemical assay. Histological sections of the hearts were stained with hematoxylin-eosin and myocardial histopathological scores were determined under an optical microscope. The amount of myocardial collagen was measured by means of chloramines T.
RESULTScTnI was significantly higher in group A (0.3680 +/- 0.40 ng/ml) than group D (0.0783 +/- 0.06 ng/ml) (P < 0.05), and was lower in group B (0.1889 +/- 0.15 ng/ml) but still significantly different from that of group D (P < 0.05), and declined to the normal level in group C (0.1338 +/- 0.07 ng/ml), but the difference between groups C and D was not significant (P > 0.05). Myocardial tissue MMPs-3 activity was transiently high in group A (0.1847 +/- 0.04), higher in group B (0.2780 +/- 0.05) as compared to group D (0.1213 +/- 0.03) (P < 0.05 for all). The activity of MMPs-3 increased earlier than that of MMPs-9. The amount of myocardial collagen of group B (38.94 +/- 0.67) and C (40.69 +/- 0.75) was significantly greater than that of group D (P < 0.05). Myoardial tissue MMPs-3 and MMPs-9 positively correlated with myocardial histopathological scores (r = 0.669, 0.667, P < 0.05) and myocardial collagen (r = 0.482, 0.679, P < 0.05).
CONCLUSIONSIn rats with asphyxia, there was an excess activation of myocardial MMPs-3 and MMPs-9 activities and secondary to which, the quantity of myocardial collagen increased. The injuries of myocardium may be closely associated with myocardial tissue MMPs. MMPs may be used to evaluate the severity of myocardial interstitial damage.
Animals ; Asphyxia ; metabolism ; pathology ; Collagen ; metabolism ; Disease Models, Animal ; Matrix Metalloproteinase 3 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Wistar ; Troponin I ; blood
7.Cell membrane phospholipid variation and protein kinase C expression effects on hepatic metastasis of large intestinal carcinoma.
Shi-yong LI ; Bo YU ; Ping AN ; Zhen-jia LIANG ; Shu-jun YUAN ; Hui-yun CAI
Chinese Journal of Surgery 2004;42(2):84-87
OBJECTIVETo study cell membrane phospholipid variation and protein kinase C (PKC) isoenzyme expression and their effects on hepatic metastasis of large intestinal carcinoma.
METHODSHigh function liquid chromatography was used to separate and detect cell membrane phospholipids of phosphatidylinosital (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in primary foci, paratumor intestine mucosa and hepatic metastasis of large intestinal carcinomas. And mRNA expression levels of PKC-alpha, -beta II, -delta, -epsilon, -lambda, -zeta isoenzymes were detected using QRT-PCR technique.
RESULTSFifty-eight cases of colorectal cancer were examined.
CONTENTSof PI, PC and PE in primary foci and hepatic metastasis were higher than those in paratumor mucosa. PE content in hepatic metastasis was much higher than that in primary foci (t = 98.88, P < 0.01). But PI and PC contents had no significant differences between primary and hepatic metastasis (t = 1.73, 1.36, P > 0.05). PKC-beta II, -delta, -epsilon, -lambda, -zeta expression were enhanced in primary foci and hepatic metastasis, but PKC-alpha level decreased in comparison with paratumor mucosa. And PKC-delta, -epsilon, -lambda, -zeta levels in hepatic metastasis were higher than those in primary foci (t = 4.31, P < 0.05). PI and PC had positive correlations with PKC-beta II expression. PE had positive correlations with PKC-delta, -epsilon, -lambda, -zeta, but a negative correlation with PKC-alpha.
CONCLUSIONSThe increases of PI and PC and PKC-alpha/PKC-beta II ratio change are related with colorectal cancer genesis. High content of PE and enhanced expression of PKC-delta, -epsilon, -lambda, -zeta isoenzymes and decreased PKC-alpha level improved hepatic metastasis of large intestinal carcinoma.
Adult ; Aged ; Aged, 80 and over ; Chromatography, Liquid ; Female ; Humans ; Intestinal Neoplasms ; pathology ; Isoenzymes ; genetics ; Liver Neoplasms ; secondary ; Male ; Membrane Lipids ; analysis ; Middle Aged ; Phosphatidylcholines ; analysis ; Phosphatidylethanolamines ; analysis ; Phosphatidylinositols ; analysis ; Phosphatidylserines ; analysis ; Protein Kinase C ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
8.Ultrastructure and crystallin mutant molecular modeling of hereditary coralliform cataract.
Wei-zhen XU ; Shu ZHENG ; Qi DONG ; Shan-rong CAI ; Ke YAO ; Su-zhan ZHANG
Journal of Zhejiang University. Medical sciences 2005;34(3):243-247
OBJECTIVETo observe the correlation of gammaD-crystallin P23T mutant with lens ultrastructure of the hereditary coralliform cataract.
METHODSComplete ophthalmologic examinations were performed before lens extraction and lens samples were studied by transmission and scanning electric microscope respectively. Protein molecular modeling was performed using SWISS-MODEL(version 2.0).
RESULTSProtein structure modeling demonstrated that the mutant caused a decrease in molecular final total energy and changes in the surface structure of gammaD-crystallin. Ultrastructure study revealed crystals deposited in lens, extensive granules dispersed in uncommon oval structure and the disorganization of lens epithelial cells.
CONCLUSIONIt is possible that the gammaD-crystallin P23T mutant is associated with abnormal crystals in lens and disorganization of lens epithelial cells.
Cataract ; congenital ; genetics ; pathology ; Female ; Humans ; Lens, Crystalline ; ultrastructure ; Male ; Pedigree ; Phenotype ; Point Mutation ; gamma-Crystallins ; genetics
9.Intertransverse approach microendoscopic discectomy for far lateral lumbar disc herniation.
Shu-Wen LI ; He-Ping YIN ; Zhen-Hua CAO ; Ming BAI ; Zhi-Cai DU ; Yu-Peng WANG
China Journal of Orthopaedics and Traumatology 2014;27(9):752-755
OBJECTIVETo explore the clinical effect of intertransverse approach microendoscopic discectomy for far lateral lumbar disc herniation.
METHODSFrom February 2005 to February 2010, 73 patients with far lateral lumbar disc herniation were treated with intertransverse approach microendoscopic discectomy. Their clinical data were retrospectively analyzed. There were 41 males and 32 females, aged from 19 to 80 years old with an average of 56.5 years; courses of disease ranged from 1 to 25 months with an average of 4.5 months. The main symptom was low back pain and sciatica, especially the sciatica was seriously. Herniation level was in L3,4 of 9 cases, L4,5 of 49 cases, L5S1 of 15 cases. Preoperative, 2 weeks after operation, final follow-up, conditions of pain relief were assessed by visual analogue scale (VAS); total life quality of patients were evaluated by Oswestry Disability Index (ODI) before operation and last follow-up.
RESULTSAll operations were performed successfully, operative time was from 40 to 115 min (mean of 50 min); and blood loss was from 50 to 150 ml (mean of 110 ml). Incision infection had 1 case and incomplete nerve root injury had 1 case. All patients were followed up from 3 to 8 years with an average of 4.5 years. Postoperative VAS and ODI had obviously improved (P < 0.01).
CONCLUSIONThe technique of intertransverse approach microendoscopic discectomy is a feasible and effective method for far lateral lumbar disc herniation.
Adult ; Aged ; Aged, 80 and over ; Diskectomy ; methods ; Endoscopy ; methods ; Female ; Humans ; Intervertebral Disc Displacement ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Young Adult
10.Effects of small intestinal bacterial overgrowth on bowel gas production in patients with liver cirrhosis.
Shu-guang CAO ; Hao WU ; Chang-long XU ; Zhen-zai CAI ; Jun YAN
Chinese Journal of Hepatology 2010;18(11):864-865
Adult
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Female
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Gases
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Humans
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Intestine, Small
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metabolism
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microbiology
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physiology
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Liver Cirrhosis
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metabolism
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microbiology
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Male
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Middle Aged