1.Protective effect of Schisandrin B on myocardial cells with hypoxia/reoxygenation-induced inj ury in neonatal rats and its mechanism
Wei LIU ; Chengyi ZHANG ; Nan SHEN ; Hong JIN ; Rui YUAN ; Xiaoming WAN ; Ling QI
Journal of Jilin University(Medicine Edition) 2014;(5):977-980
Objective To investigate the influence of Schizandrain B(Sch B)preconditioning in myocardial cells of neonatal rats with hypoxia/reoxygenation (H/R)inj ury, and to explore its possible mechanism.Methods The cultured myocardial cells were divided into control group, model group and 10, 50, 250 mg · L-1 SchB preconditioning groups.All the cells in various groups except control group were cultured in 2 mmol·L-1 Na2 S2 O4 for 2 h,then cultured in normal medium for 18 h to induce myocardial H/R injury.The morphological changes of myocardial cells in various groups were observed under inverted microscope. The survival rates of the myocardial cells in each group were examined by MTT. The activities of lactic dehydrogenase (LDH ), creatine (CK ), superoxide ismutase(SOD),and the (MDA)levels in the cells in various groupswere examined by detection kits. Results Compared with control group,the cells in model group were retracted,arrest or float,the survival rate was decreased significantly (P<0.01),the activities of LDH,CK and MDA level were increased (P<0.01),and the SOD activity was decreased (P<0.01);compared with model group,the cells in Sch B preconditioning groups remained beating,retraction was light,the cell survival rates were significantly increased (P<0.01),the activities of LDH,CK and MDA levels were decreased (P<0.01),and the SOD activities were increased (P<0.05 or P<0.01).Conclusion Sch B has protective effect on myocardial cells with H/R injury in the neonatal rats,which may be associated with anti-oxidative damage.
2.Association of some SNP in TGFA,IRF6 and nonsyndromic cleft lip with or without cleft palate by DNA microarray technology
Ya SHEN ; Lu CHENG ; Wei-Dong WAN ; Zu-Hong LU ; Lian-Ming LIAO ; Yu-Gui CUI ; Jia-Yin LIU ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To explore the relationship between some single nucleotide polymorphisms (SNP)loci of interferon regulatory factor 6(IRF6)gene,transforming growth faetor-?(TGFA)gene and nonsyndromic cleft lip with or without cleft palate(NSCL/P)in nuclear families consisting of fathers, mothers and affected offspring with NSCL/P from southeast China.Methods Some SNloci of IRF6 and TGFA were detected by applying microarray technology in nuclear families,and then haplotype relative risk (HRR)and transmission disequilibrium test(TDT)were performed.Results There were no significant difference in genotypes and alleles distribution between patients and their parents.The SNP locus——V274I of IRF6 was associated with NSCL/P(HRR:?~2=4.5816,P
3.The expression of interferon-regulatory factor genes in patients with systemic lupus erythematosus
Jie QIAN ; Nan SHEN ; Gui-Mei GUO ; Nian-Hong WAN ; Yan LIN ; Xin-Fang HUANG ; Hui WU ; Shun-Le CHEN ;
Chinese Journal of Rheumatology 2003;0(09):-
Objective To observe whether the expression of interferon-regulatory factor genes are re- lated to systemic lupus erythematosus (SLE).Methods The clinical data of 45 SLE patients and 37 normal controls were collected.Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression (indicated as-??Ct value) of IRFI,IRF4,IRF8 in patients with SLE and those in the controls.Results The levels of IRF1,IRF4 and IRF8 mRNA were-3.90?0.19,-8.04?0.25 and 3.60?0.15 respectively in normal controls.In SLE patients, IRF4 mRNA expression was -8.82?0.18,higher than that in normal (P=0.011).But IRF8 mRNA expression was 3.09?0.13,lower than that in normal (P=0.012).Conclusion Abnormal IRF mRNA expression is found in the peripheral blood of SLE patients.IRFs may play roles in the pathogenesis of SLE by affecting the differen- tiation of Th cells.
4.Variation of serum thyroid transcription factor-1 protein and its value in the diagnosis of lung carcinoma.
Xiao LI ; Li-yan WAN ; Hong SHEN ; Xiao-yan BAI
Journal of Southern Medical University 2011;31(9):1592-1596
OBJECTIVETo investigate the variation of serum thyroid transcription factor-1 (TTF-1) in different patients and explore its significance in the diagnosis of lung carcinoma.
METHODSDot-enzyme linked immunosorbent assay (dot-ELISA) and Leica Q500 MC image analysis system were used to quantitatively detect TTF-1 protein in the serum samples from normal healthy adults and from patients with benign lung disease, lung cancer, thyroid carcinoma and non-thyroid carcinoma.
RESULTSThe sensitivity, specificity, standardized positive predicative value, standardized negative predicative value, standardized accuracy and standardized wrong diagnostic rate of the method were 90.91%, 82.22%, 83.64%, 90.04%, 86.57% and 13.43%, respectively. The cutoff value of serum TTF-1 in healthy normal adults was 36.39, with a ROC value of 0.95. Serum TTF-1 PU was significantly higher in patients with lung adenocarcinoma, squamous cell lung carcinoma and thyroid carcinoma than in healthy adults and patients with benign lung diseases and non-thyroid carcinoma (P=0.000). Serum TTF-1 PU was similar in lung adenocarcinoma, squamous cell lung carcinoma, small cell lung carcinoma, large cell lung carcinoma and thyroid carcinoma (P=0.744, 0.677, and 0.333, respectively). Serum TTF-1 PU was greater than the PU in the corresponding homogenate of lung adenocarcinoma, squamous cell lung carcinoma, small cell lung carcinoma, large cell lung carcinoma and thyroid carcinoma (P=0.000). Serum and homogenate TTF-1 PU was correlated to TNM stage of lung cancer patients (P=0.000) but not to gender, tumor types, differentiation or lymph node metastasis.
CONCLUSIONSLung adenocarcinoma, squamous cell lung carcinoma and thyroid carcinoma are suspected when serum TTF-1 PU is higher than 36.39. Serum TTF-1 is not helpful in differentiating the types of lung carcinomas and thyroid carcinoma. After exclusion of thyroid carcinoma, detection of serum TTF-1 can be helpful in the diagnosis of lung cancer. In different lung carcinomas and thyroid carcinomas, the serum TTF-1 is higher than the corresponding homogenate TTF-1 level. Serum TTF-1 increases with the progression of TNM stage of lung carcinoma.
Case-Control Studies ; Humans ; Lung Neoplasms ; blood ; classification ; diagnosis ; Nuclear Proteins ; blood ; Predictive Value of Tests ; Sensitivity and Specificity ; Thyroid Nuclear Factor 1 ; Transcription Factors ; blood
5.Classification of Cimicifuga species based on 1H-NMR fingerprint combined with pattern recognition technique.
Li SHEN ; Yan-Yan ZHAO ; Hong-Ping XIE ; Wan-Hui LIU
China Journal of Chinese Materia Medica 2013;38(2):217-222
The metabolomic analysis of three Cimicifuga species was performed using H-NMR spectroscopy and pattern recognition (PR) techniques. A broad range of metabolites could be detected by 'H-NMR spectroscopy without any chromatographic separation. The analysis using principal component analysis (PCA) and discriminant partial least square (DPLS) of the 1H-NMR spectrum showed a clear discrimination between C. foetida and the other two species. The major metabolites responsible for the discrimination were triterpenoid saponins and saccharides. These results indicated that the combination of 1H-NMR and PR provides a useful tool for chemotaxonomic analysis and authentification of Cimicifuga species, and could used for the quality control of plant materials.
Cimicifuga
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classification
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Discriminant Analysis
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Drugs, Chinese Herbal
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classification
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standards
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Magnetic Resonance Spectroscopy
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methods
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Metabolomics
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methods
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Pattern Recognition, Automated
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Principal Component Analysis
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Protons
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Saponins
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isolation & purification
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Triterpenes
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isolation & purification
6.Expression of thyroid-stimulating hormone receptor in mammary gland tissue of lactating rats
Si-yuan, WAN ; Li-xiang, LIU ; Mao-fu, WU ; Hong-mei, SHEN ; Ke-xin, LIU ; Yu, SUN
Chinese Journal of Endemiology 2013;(3):254-257
Objective To observe the protein and mRNA expression of thyroid-stimulating hormone receptor (TSHR) in mammary gland tissue of lactating rats,and to explore iodine uptake mechanism.Methods Eighty adult Wistar rats (60 female and 20 male),weighting 210-250 g were selected.All female Wistar rats were randomly divided into 6 groups according to their body mass:normal non-pregnant group,lactating for 5-,10-,15-and 20-day groups and weaning for 5 days group,10 rats in each group.All rats were fed with conventional fodder and tap water freely.In addition to the normal non-pregnant group,other five groups of female and male rats were mated at 3 ∶ 1,respectively.Then the rats in all groups were killed on the 5th,10th,15th and 20th day after lactation and on the 5th day after weaning to get the mammary gland tissue.The protein and mRNA expression of TSHR were determined by immunohistochemical staining and real-time quantitative PCR.Results TSHR protein was expressed in mammary acinar and ductal epithelial cytoplasm.The expression of TSHR in mammary gland showed significant differences between groups (x2 =14.612,P < 0.05),the staining intensity of mammary gland tissue in normal non-pregnant rats(weak,n =4; moderate,n =6) was weaker than that of lactating for 5 days(weak,n =2; moderate,n =3; strong,n =5) and 10 days groups(barely detectable,n =1;moderate,n =4; strong,n =5; x2 =4.113,5.250,all P< 0.05).The expression of TSHR mRNA in mammary gland showed significant differences between groups(F=20.488,P < 0.05); the expression of TSHR mRNA in lactating for 10 days group(0.31 ± 0.06) was higher than that of lactating for 5 days group(0.22 ± 0.04,P < 0.01),and the expression of lactating for 15 days group (0.16 ± 0.08) was significantly lower than that of lactating for 5 days group (P < 0.05).Conclusions TSHR is widely expressed in mammary gland of lactating rats.The iodine uptake of mammary gland is enhanced in early lactation period when the body may be more susceptible to iodine deficiency,therefore iodine should be supplemented reasonably.
7.Improving thermostability of Aspergillus niger phytase by elongation mutation.
Hui CHEN ; Hong-Ning WANG ; Wan-Shen YANG ; Hai-Xia ZHAO ; Qi WU ; Zhi SHAN
Chinese Journal of Biotechnology 2005;21(6):983-987
The phytase gene phyA(m) from Aspergillus niger N25 was recombined into E. coli expression vector pET-30b(+). Recombined at expression vectors pET30b-FphyA(m) was served as a template to amplify phytase gene, and the PCR product named elongation mutation gene phyA(e) was expanded with a 13 amino acid sequence from pET-30b-FphyA(m) vector at C-terminal of phytase gene phyA(m). Furthermore, phyA(e) gene was recombined into expression vector pPIC9k and expressed in Pichia pastoris. The comparison experiment of mutant phytase PP-NP0 with wild-type phytase PP-NP(m)-8 showed that: the optimum temperature of PP-NPe was increased by 3 degrees C, and its thermostability was increased by 21% when it was exposed to 10 min at 75 degrees C. Its effective reaction pH range with catalysis efficiency above 70% was pH 4.6 - pH 6.6, and wider 0.4 pH value than that of wild-type phytase.
6-Phytase
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biosynthesis
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genetics
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Amino Acid Sequence
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Aspergillus niger
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enzymology
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genetics
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Enzyme Stability
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genetics
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Escherichia coli
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enzymology
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genetics
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Fungal Proteins
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biosynthesis
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genetics
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Hot Temperature
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Molecular Sequence Data
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Mutation
8.Research about schwann cells and PLGA implanted to rat transected spinal cord.
Hong WAN ; De-zhi LI ; Fei YANG ; Jun-hua LI ; Shen-guo WANG ; Zhong-cheng WANG
Chinese Journal of Surgery 2007;45(12):843-846
OBJECTIVETo investigate the recovery of rat transected spinal cord injury after implantation of Schwann cells combined with poly (lactide-co-glycolide) (PLGA).
METHODSSchwann cells were expanded, co-cultured with PLGA for 9 days in vitro, and then analyzed with scanning electron microscope (SEM). Rat spinal cord at the level of T(9) was transected. Schwann cells labeled with BrdU and PLGA scaffold were implanted to injury site. After 1, 3, 6 months, BrdU/MBP immunohistochemistry double staining, semi-thin sections stained thionin and ultra-thin section were performed to investigate myelin renew. BBB open field locomotion, motor evoked potential (MEP), compound muscle action potential (CMAP) and somatosensory evoked potential (SEP) were recorded.
RESULTSSchwann cells grew well on PLGA under SEM. BrdU/MBP double positive cells would been seen, remyelination was thin and formed by Schwann cells at 6 months later under electron microscope (EM). BBB behavioral tests revealed no significant difference in recovery comparing with experiment group and control group. The results of MEP, CMAP and SEP showed no significant improvement in the conduction of spinal cord.
CONCLUSIONSThere are the compatibility between Schwann cells and PLGA. Although remyelination was found in morphology, function conduction of spinal cord failed to be established.
Animals ; Cells, Cultured ; Disease Models, Animal ; Evoked Potentials, Motor ; Female ; Immunohistochemistry ; Lactic Acid ; chemistry ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Nerve Regeneration ; Polyglycolic Acid ; chemistry ; Prostheses and Implants ; Rats ; Rats, Wistar ; Schwann Cells ; chemistry ; transplantation ; ultrastructure ; Spinal Cord Injuries ; physiopathology ; surgery ; Tissue Engineering ; methods
9.Effect of different levels of iodine nutrition on secretion of placental hormone in pregnant rats
Xue-jiao, WANG ; Rui-qiang, DONG ; Hong-mei, SHEN ; Li-xiang, LIU ; Ke-xin, LIU ; Si-yuan, WAN ; Mao-fu, WU
Chinese Journal of Endemiology 2011;30(6):616-619
Objective To study the effect of different levels of iodine nutrition on secretion of placental hormone in pregnant rats.Methods Two hundred and twenty five Wistar rats (165 female,60 male),weighing about 80 - 100 g were used in the study.Female rats were randomly divided into five groups according to their body weights:low iodine group Ⅰ(LⅠ),low iodine group Ⅱ (LⅡ),adequate iodine(control) group(Al),high iodine group Ⅰ ( HⅠ ),and high iodine group Ⅱ (H Ⅱ ),and 33 rats in each group.Animals in the low iodine groups were fed low-iodine diet,the iodine content was 13.46 μg/kg,in addition,these rats drank deionized water which containing potassium iodated,the dose was 0 and 5 μg/L,respectively.The rats of adequate and the two high iodine groups were fed normal diet,the iodine content was 22.00 μg/kg,they also drank deionized water,containing potassium iodated 50,3000,and 10000 μg/L,respectively.The rats mated after 3 months of feeding,and were respectively sacrificed at early pregnancy(5 ± 2)d,second trimester( 12 ± 2)d,and third trimester of pregnancy(17 ± 2)d,and then their serum was taken.Serum human chorionic gonadotropin(HCG),human chorionic thyrotropin(HCT),and progesterone were measured by enzyme-linked immunosorbent assay (ELISA).Results In the third trimester,the serum levels of rat HCG were significantly different between groups(F =4.16,P < 0.05).The means of rats serum HCG of the two low iodine groups [ (16.08 ± 4.45),(17.43 ± 2.70)U/L] were significantly higher compared with that of AI group[ (13.68 ± 3.52)U/L] in the third trimester(all P < 0.01 ).In the second and third trimester,the levels of rats serum HCT were significantly different between groups(F =3.59,3.40,all P < 0.05).The means of rats serum HCT of HI group [(70.11 ± 10.97)μU/L] in the second trimester and HII group[(74.93 ± 13.22)μU/L] in the third trimester were higher than those of AI group[ (57.14 ± 12.56),(58.17 ± 8.54)μU/L] significantly(all P < 0.01 ).There were statistical differences of the means of serum progesterone among trimester of pregnancy(F =4.06,4.43,all P < 0.05).The level of serum progesterone of the third trimester[ ( 1462.80 ± 286.48 )pmoL/L] compared to those of the first[ (1929.93 ± 158.37) pmol/L] and the second trimester[ (1856.44 ± 542.08)pmol/L] was decreased significantly(all P < 0.05) in LI group.In the control group,the level of serum progesterone of the second trimester [ (2046.45 ± 475.67)pmol/L ] was significantly higher than the first trimester[ (1714.39 ± 461.71 )pmol/L,P < 0.05 ].Conclusions During pregnancy,placenta could promote HCG secretion under iodine-deficient conditions.In addition,the placenta increases the secretion of HCT under conditions of excess iodine.In the condition of severe iodine deficiency,the secretion of serum progesterone decreases,and further decreases with prolongation of pregnancy,but it is opposite to the change of HCG during pregnancy.This phenomenon could lead to harmful pregnant outcomes easily.
10.Effect of Intravenous Nutrition on Immune Function in Premature Infants
xiao-fang, ZHU ; li-ping, MAO ; shen-hong, WAN ; lin-yun, LI ; hua-ling, ZHANG ; yan-ling, ZHOU ; li-wen, CHANG
Journal of Applied Clinical Pediatrics 2006;0(13):-
Objective To observe the effect of intravenous nutrition on blood serum triglyceride (TG),humoral immunity and cellular immunity function in premature infants.Methods Sixty premature infants were randomly divided into 3 groups:amino acid group (group A),amino acid plus medium-long chain fatty group (group B) and amino acid plus long chain fatty group (group C).Amino acid and the fatty were used on them from 24 hours after their birth,started from 1.0 g/(kg?d),increased progressively 0.5 g/(kg?d) until 3.0 g/(kg?d),totally 7 days.TG and immunoglobulin IgA,IgM,IgG,complement C3,C4 and T-lymphocyte subsets CD3,CD4,CD4/CD8 and NK cell were checked in the first day before treatment and the ninth day after treatment.Results Compared with before treatment,TG of 3 groups were elevated(Pa0.05).Compared with group A and B,NK cell in group C were decreased obviously(Pa