1.Analysis of the Relationship Between Era and YggG in E.coli by Double-promoter Expression Vector pDH2-YggG-P_(tac)-Era
Yong HUANG ; Xiao-Nan ZHANG ; Bin ZHANG ; Tao WANG ; Li WANG ; Lu-Yuan GUAN ; Nan-Chun CHEN ; Su-Min CHEN ;
China Biotechnology 2006;0(03):-
yggG, a Era-binding protein gene, was isolated and cloned from the E.coli genomic DNA library. Previous studies indicated that the product of yggG gene, YggG294(amino acids 1-294), strongly inhibited the growth of host bacteria and caused the death of bacteria cells. To elucidate whether Era is related to the death of bacterial cells expressed YggG294,A double promoter expression vector that can express YggG294 and Era proteins controllably in cells was constructed. Using this vector to express YggG294 and Era protein in the same E.coli cells, then analyzed the relation between YggG294 and Era. The results showed that the ratio of Era proteins to total proteins increased with the increase of induction time in E.coli cells without YggG294 expression and with little YggG294 expression;the ratio of Era proteins to total proteins seemed to be a constant level in E.coli cells overexpressing YggG294;but we could not detect any Era hydrolyzate in E.coli cells overexpressed YggG294 could not be detected. The results also showed that pre-expression of Era protein did not produce any effect on the growth inhibition of E.coli cells caused by YggG294. These results indicate that YggG294 can not hydrolyze Era protein in E.coli cells, and that YggG-Era interaction is not associated with the death of bacteria expressed YggG294. It is thus reasonable to draw a conclusion that Era is not associated with the growth inhibition of E.coli cells caused by YggG294. YggG294 inhibits the growth of bacteria by other way.
2.Advances on biological indicators of diabetic retinopathy and characteristic peculiarities
Wan-Feng YANG ; Wei XIE ; Xiao-Qiong CHEN ; Tian TIAN ; Chun-Nan TAO
International Eye Science 2018;18(7):1241-1244
·Diabetic Retinopathy ( DR ) has gradually worsening and lingering, which is the leading cause of global young people blindness. With the progression of the disease, patients with diabetes mellitus ( DM) will have different degrees of DR. If you can not prevent and give acute early intervention, once the visual acuity decreased significantly, DR would be difficult to reverse. DR progressively has worsening, the treatment status has no optimistic. Therefore, DR in the early prevention and treatment will be indispensable. This article summarizes some of the early warning of the occurrence and development of biological markers and characteristic indicators in order to provide a basis for the early prevention of DR.
3.Serum neuropeptide Y level in hypertensive patients with obstructive sleep apnea syndrome
Xiao-Guang YAO ; Zhi-Tao YAN ; Nan-Fang LI ; Ying-Chun WANG ; Tao LI ; Jia ZHU ; Ling ZHOU ; Liang SHAO
Chinese Journal of Cardiology 2010;38(6):510-513
Objective To explore the potential role of neuropeptide Y (NPY) in the pathphysiological process of hypertension caused by obstructive sleep apnea sydrome (OSAS). Methods The concentration of serum NPY were measured with radioimmunoassay (RIA) in 417 subjects (97 normotensive controls without OSAS, 113 cases of normotensive with OSAS, 73 cases of hypertensive without OSAS and 134 cases of hypertensive with OSAS. Futher, the mean NPY level were compared in four groups and the possible effective factors on NPY were discussed. Results ( 1 ) The concentration of NPY in four groups were (50. 5 ±37. 2) pmol/L in normal controls, (76. 0 ±39. 9) pmol/L in normotensive with OSAS group, (66. 9 ± 36. 2 ) pmol/L in hypertensive without OSAS group and (86. 8 ± 36. 8 ) pmol/L in hypertensive with OSAS group. Whether the patients with OSAS combined with hypertension or not, the concentration of NPY in the serum raised remarkably compared with those without OSAS and hypertension,the highest level of serum NPY was detected in OSAS combined with hypertension group. (2) Pearson corelation analysis indicated that both SBP and DBP related to the serum NPY significantly in non-OSAS group (AHI < 10), while the BMI, abdominal circumference, AHI as well as the lowest level of SaO_2 correlated to NPY besides SBP in OSAS group with ( AHI ≥ 10). (3) Multiple linear regression model showed that the abdominal circumference and AHI were contributing factors to SBP, while neck circumference and BMI were contributing factors to DBP. The level of NPY in the serum were significantly affected by AHI and BMI, in which the former one had greater influnce. Conclusion The increased level of serum NPY may play weakly potential roles in the pathphysiological process of hypertension caused by OSAS.
4.Inhibitory effect of Genipin on uncoupling protein-2 and energy metabolism of androgen-independent prostate cancer cells.
Mao-liang YAO ; Jiang GU ; Yong-chun ZHANG ; Nan WANG ; Zhi-hui ZHU ; Qing-tao YANG ; Miao LIU ; Jian-feng XIA
National Journal of Andrology 2015;21(11):973-976
OBJECTIVETo explore whether the inhibitory effect of Genipin on uncoupling protein-2 (UCP-2) in mitochondria is involved in energy metabolism of androgen-independent PC3 prostate cancer cells.
METHODSPC3 prostate cancer cells were cultured and treated with Genipin at the concentrations of 40, 80, and 160 μmol/L for 48 hours. Then the proliferation of the cells was detected by MTT assay, the expression of UCP-2 mRNA determined by RT-PCR, and the content of intracellular pyruvic acid (PA) and the activity of succinate dehydrogenase (SDH) in the mitochondria measured by visible spectrophotometry.
RESULTSWith the increased concentration of Genipin, the proliferative activity of the PC-3 cells, the expression level of UCP-2 mRNA, the content of intracellular PA and the activity of SDH in the cells were all decreased, namely, with the enhanced inhibitory effect of Genipin on UCP-2, a trend of reduction was observed in the proliferation of the cells, intracellular PA content, and SDH activity in the mitochondria.
CONCLUSIONGenipin is involved in the energy metabolism of androgen-independent PC3 prostate cancer cells by reducing the content of intracellular PA and the activity of SDH in the mitochondria, which may be associated with its inhibitory effect on UCP-2.
Cell Line, Tumor ; drug effects ; Energy Metabolism ; Humans ; Ion Channels ; metabolism ; Iridoids ; pharmacology ; Male ; Mitochondria ; metabolism ; Mitochondrial Proteins ; metabolism ; Prostatic Neoplasms ; metabolism ; Pyruvic Acid ; metabolism ; RNA, Messenger ; Succinate Dehydrogenase ; metabolism ; Uncoupling Protein 2
5.Effects of NMDA receptor expression in rat's hippocampus after exposure to 1800 MHz radiofrequency field.
Chun-tao HUANG ; Ping LIU ; Hui-xin WU ; Jing-lin WANG ; Xi-nan WU
Chinese Journal of Preventive Medicine 2006;40(1):21-24
OBJECTIVETo identify the effects of NMDA receptor subunits NR2A and NR2B expression in rat's hippocampus after exposure to 1800 MHz radiofrequency radiation.
METHODSFour-week old female Wistar rats were randomly divided into four groups, with 12 animals for each. The subjects in two experimental groups had been continuously exposed to 1800 MHz microwave radiation (CW) with respective power density of 0.5 mW/cm(2) and 1.0 mW/cm(2) 12 hours each day for 21 days. Meanwhile, sham-controls were carried out. The brain tissue sections were performed by immunohistochemistry to demonstrate both expressions of NR2A, NR2B immune-activity in the hippocampal CA1, CA3 and DG by using computer-assisted image analysis system.
RESULTSIn NR2A: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA3 [(8.5 +/- 1.5) vs (11.1 +/- 1.8), P < 0.01] and had not been significantly changed in CA1 and DG. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(7.9 +/- 1.6) vs (9.7 +/- 1.5); (8.4 +/- 1.7) vs (11.1 +/- 1.8), respective P < 0.05, P < 0.01] and had not been significantly changed in DG. In NR2B: the expression of 0.5 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1 and CA3 [(16.4 +/- 1.0) vs (17.8 +/- 1.6); (9.6 +/- 1.9) vs (11.2 +/- 2.1), respective P < 0.05]. The expression of 1.0 mW/cm(2) power density group was significantly lower than 0 mW/cm(2) power density group in CA1, CA3 and DG [(13.1 +/- 2.4) vs (17.8 +/- 1.6); (9.3 +/- 1.4) vs (11.2 +/- 2.1); (7.3 +/- 0.1) vs (8.5 +/- 1.0), respective P < 0.01, P < 0.05, P < 0.05].
CONCLUSIONThere were findings of the effects on NMDA receptor subunits in different hippocampus sections after exposure to 1800 MHz radiofrequency radiation.
Animals ; Female ; Hippocampus ; metabolism ; radiation effects ; Immunohistochemistry ; Radio Waves ; Rats ; Rats, Wistar ; Receptors, N-Methyl-D-Aspartate ; biosynthesis
6.Pilot study of differentiation of bone marrow derived mesenchymal stem cells into endothelial cells induced by B16 melanoma cells in vitro.
Chun-Sheng NI ; Nan ZHAO ; Tao SUN ; Xiu-Lan ZHAO ; Xing-Hui WANG ; Qiang GU ; Bao-Cun SUN
Chinese Journal of Pathology 2009;38(6):402-407
OBJECTIVEBone-marrow derived mesenchymal stem cells (BMSC) have the potential to differentiate into endothelial cells. The aim of the study was to investigate the induction process of BMSC by B16 melanoma cells in vitro and to analyze the role of VEGF-a in the process.
METHODSA co-culture system containing BMSC and B16 melanoma cells based on transwell indirect model was established, and the induction process of BMSC by B16 melanoma cells was studied in vitro.
RESULTSBMSC were isolated from the bone marrow of C57 mice. BMSC expressed CD105, CD90, CD73, CD44 and CD166, and acquired expressin of endothelial phenotype markers including VEGFR-1, VEGFR-2 and Factor VIII after co-culture with B16 melanoma cells for 48 hours. The expression level of VEGFR-2 would be double and Factor VIII threefold more by extending the co-culture time to 72 hours. In the co-culture system, B16 melanoma cells also up-regulated the expression of VEGF-a.
CONCLUSIONSVEGF-a plays a significant role in the differentiation of BMSC into cells of endothelial phenotype, therefore, is important to tumor angiogenesis.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cell Line, Tumor ; Coculture Techniques ; Endothelial Cells ; cytology ; metabolism ; Factor VIII ; metabolism ; Male ; Melanoma, Experimental ; metabolism ; pathology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Pilot Projects ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism
7.Determination of Furazolidone and Florfenicol in Soil by Ultra Performance Liquid Chromatography-Tandem Mass Spectrometry
Qiong NAN ; Chun Jing TANG ; Zhu Ruo HE ; Cheng Yu HU ; Tao WU
Chinese Journal of Analytical Chemistry 2017;45(11):1600-1605
A detection method for furazolidone and florfenicol in soil with various environmental matrices was established using ultra performance liquid chromatography-tandem spectrometry (UPLC / MS / MS) technique. Extracting solution of a mixture of phosphate buffer (pH = 3) and acetonitrile (3 : 7, V/ V) was used in this experiment. The extracted water samples were enriched by SAX-HLB solid phase extraction column before the process of nitrogen blowing ( high purity nitrogen). The enriched antibiotics were desalted with 8 mL of methanol. Waters BEH C18(2. 1 × 100 mm) column was used for the sample separation. UPLC / MS / MS was carried out for qualitative and quantitative analysis under multi-reaction monitoring mode. The detection limit of the method was determined by 3 times of signal-to-noise ratio, and the limit of determination of the method was determined by 10 times of signal-to-noise ratio. The results showed that the detection limits of furazolidone and florfenicol were 1. 19 and 0. 41 μg / kg, respectively, and the limits of quantitation of furazolidone and florfenicol were 3. 40 and 1. 37 μg / kg, respectively. Besides, recovery experiment showed that, for the soil samples spiked with 50 μg / L furazolidone and florfenicol, the recoveries were 79% for florfenicol and 92% for furazolidone. Similarly, for the soil samples spiked with 200 μg / L furazolidone and florfenicol, the recoveries were 96% for furazolidone and 86% for florfenicol.
8.High Survivin and Low Zinc Finger of the Cerebellum 1 Expression Indicates Poor Prognosis in Triple-negative Breast Carcinoma
Chun Tao SHI ; Jun MA ; Qi Feng SHI ; Ye ZHANG ; Hao Nan WANG
Journal of Breast Cancer 2019;22(2):248-259
PURPOSE: Triple-negative breast carcinoma (TNBC) is accompanied with high risk of metastasis and recurrence. The present study aimed to explore the clinicopathological and prognostic roles of putative tumor-related genes in patients with TNBC. METHODS: Thirty pairs of frozen-thawed tumors were used to select reliable indicators via real-time quantitative polymerase chain reaction (RT-qPCR). Then, 150 pathology specimens were used to evaluate the expression of proteins in TNBC through immunohistochemistry. In addition, Kaplan-Meier curves and Cox regression analysis were also performed to analyze the overall survival and disease-free survival. RESULTS: RT-qPCR results indicated that among all the proteins analyzed using fresh-frozen TNBC samples, the expression levels of only Survivin and zinc finger of the cerebellum 1 (ZIC1) were obviously different from those in the corresponding normal tissues. Survivin and ZIC1 expression had opposite effects on the clinicopathological diagnosis and prognostic assessment in TNBC patients. Further, there was a negative correlation between Survivin and ZIC1 expression. In addition, the “Survivin-positive ZIC1-negative group” was associated with histologic grade, lymph node metastasis, and TNM staging (p < 0.001) and this was also an independent factor for evaluating the prognosis of TNBC in patients. CONCLUSION: In summary, the expression levels of Survivin and ZIC1 in TNBC are different from those in normal tissues and are negatively correlated mutually. The combined detection of Survivin and ZIC1 expression levels could allow better comprehensive diagnosis and prognostic evaluation for TNBC patients.
Breast Neoplasms
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Breast
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Cerebellum
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Diagnosis
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Disease-Free Survival
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Humans
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Immunohistochemistry
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Lymph Nodes
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Neoplasm Metastasis
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Neoplasm Staging
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Pathology
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Polymerase Chain Reaction
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Prognosis
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Recurrence
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Triple Negative Breast Neoplasms
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Zinc Fingers
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Zinc
9.Phosphorylcholine Oligomer-Grafted Graphene Oxide for the Transcatheter Arterial Chemoembolization of Liver Cancer
Hai-Tao DAI ; Yong-Hui HUANG ; Run LIN ; Su ZHOU ; Bin CHEN ; Tao ZHANG ; Nan LI ; Ke-Yu TANG ; Chun-Yong WEN ; Jian-Yong YANG
Journal of Sun Yat-sen University(Medical Sciences) 2018;39(1):18-25
[Objective]To investigate the safety and efficacy of phosphorylcholine oligomer grafted graphene oxide as a drug carrier for transcatheter arterial chemoembolization in the treatment of liver cancer.[Methods]Doxorubicin loaded folic acid labeled phosphorylcholine oligomer grafted graphene oxide(DOX@GO-PCn-FA)was prepared. Graphene ox-ide(GO)and DOX@GO-PCn-FA were injected intravenously via marginal ear vein in New Zealand white rabbits respec-tively to assess their safety and biodistribution for intravenous administration.Ten male New Zealand rabbits were used to establishe the VX2 liver cancer model and the tumor characteristics were confirmed by dynamic contrast enhanced CT scan.Catheter was inserted via femoral artery and advanced into hepatic lobar or segmental artery.Digital subtraction angi-ography(DSA)was performed to validate the tumor feeding vessels.DOX@GO-PCn-FA was injected through the cathe-ter to carry out selective transcatheter arterial chemoembolization(TACE). Dynamic enhanced CT scan and pathological examinations of major tissues and organs were implemented 7 days post TACE to evaluate the efficacy of embolization effect of DOX@GO-PCn-FA against liver tumor as well as the biodistribution and safety.[Results]Intravenous injection of GO resulted in significant thrombosis and pulmonary embolism whereas DOX@GO-PCn-FA of same dosage did not. DOX@GO-PCn-FA was capable of effectively diminishing the blood supply of liver tumors when applied in TACE. Pathologic exploration revealed that DOX@ GO-PCn-FA mainly deposited in the tumor,and no obvious complications were observed.[Conclusions]GO-PCn presented superior biocompatibility and exerted effective chemoembolization against liver cancer.
10.Pulmonary artery perfusion with hypothermic solution inhibits the apoptosis of lung parenchymal cells during cardiopulmonary bypass.
Bo WEI ; Ying-long LIU ; Cun-tao YU ; Yong-nan CHANG ; Chun-hua LI
Chinese Journal of Surgery 2004;42(4):227-229
OBJECTIVETo study effects of pulmonary artery perfusion with hypothermic solution on the apoptosis of lung parenchymal cells during cardiopulmonary bypass.
METHODSForty children with tetralogy of Fallot were divided into control group (n = 20) and protective group (n = 20). The patients in control group were performed using routine approaches. Patients' pulmonary artery were infused with 4 degrees C protective solution during cardiopulmonary bypass in protective group. Lung biopsy specimens were obtained after operations in order to study the apoptosis of lung parenchymal cells using tunnel techniques. At same time, patients' pulmonary functions and clinic index were monitored.
RESULTSThe rate of apoptosis cells of lung parenchymal cells was (18 +/- 7)% in control group, whereas (10 +/- 2)% in protective group. There was significant difference between both groups (t = -2.95, P < 0.05). Index O(2) in protective group was higher than that in control group at 0, 6 and 12 hours after operations [(492 +/- 172), (444 +/- 104), (489 +/- 58) mm Hg versus (369 +/- 126), (347 +/- 107), (340 +/- 119) mm Hg, t = 2.59, P < 0.05; t = 2.88, P < 0.01; t = 5.06, P < 0.01, respectively)]. The time of mechanical ventilation was significantly shorter in protective group than in control group [(15 +/- 11) hours versus (26 +/- 15) hours, t = -2.76, P < 0.01].
CONCLUSIONPulmonary artery perfusion with hypothermic solution can inhibit the apoptosis of lung parenchymal cells and relieve cardiopulmonary bypass-induced lung injury.
Apoptosis ; Cardiopulmonary Bypass ; Case-Control Studies ; Child, Preschool ; Female ; Humans ; Hypothermia, Induced ; methods ; Infant ; Lung ; blood supply ; pathology ; Male ; Perfusion ; Pulmonary Artery ; Tetralogy of Fallot ; surgery