1.Effects of?1 adrenoceptor antisense on blood pressure and?1 adrenoceptor mRNA and protein in renal hypertension rats
Yuan-Hong HANG ; Shu-Guang LIN ; Yan ZHOU ; Jin-Ming WANG ; Fang WANG ; Xi-Yong YU ;
Chinese Journal of Emergency Medicine 2006;0(11):-
Objective To evaluate the effects of 131 adrenoceptor anfisense on blood pressure and?1 adrenoceptor mRNA and protein levels in 2 kidney 1 clip(2K1C)rats.Method 2KIC hypertensive rots were produced by clipping renal artery of SD rats.Liposome/AS-ODNs 2.0 were tested intravenously in rats with 2KIC hypertension.Animals were divided into 5 groups(n=18 in each group):?1-AS-ODN group,?1-IN-ODN group,2K1C group,Sham group and SD group.Blood pressure was measured by tail-cuff method,the levels of myocardial?adreneceptor mRNA and protein were tested by RT-PCR and binding assay.Results On the basis of the magnitude and duration of hypotension,?1-AS-ODN decreased blood pressure by 39 mmHg at the most for 4 weeks.Compared with the 2KIC group,?1-AS-ODN did not significantly change the levels of myocardial?1 adrenoceptor mRNA but significantly decreased the levels of myocardial?1 adrenoceptor protein at 2,7,30 days (P
2.Expression of CXCR4 in Acute Leukemic Cells of Children and Its Signific ance
ri-ling, CHEN ; kang-rong, CAI ; ming-zhen, CHEN ; hui-qiong, ZHANG ; xi-min, FANG
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To analyze the chemokine receptor CXCR4 expression in acute leukemic cells of children and its relationship with extramedullary infiltration.Methods The immunotypes of cases of acute leukemia in children and the expression of CXCR4 in marrow leukemic cells were studied by flow cytometry respectively. The relationship between CXCR4 expression and extramedullary infiltration of leukemic cells were analyzed by statistical method.Results The expression rates of CXCR4 in ALL children were higher than those in NALL children(P
3.Analysis of iodine nutritional status of pregnant women and level of neonatal heel blood thyroid stimulating hormone in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai
Ming-xi, FANG ; Ying-jiu, ZHAI ; Li, XUE ; Qi, FANG ; Jin-fang, WU ; Guo-li, TIAN ; Yi-hua, WU ; Jing, GAO ; Xian-hua, CAI ; Xiu-hua, HU
Chinese Journal of Endemiology 2012;31(1):74-77
ObjectiveTo investigate the iodine nutritional status of pregnant women,newborn heel blood thyroid stimulating hormone(TSH) level and their relationship with urinary iodine(UI) level during pregnancy in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai.Methods A total of 993 urinary samples(the first,second and third trimesters of pregnancy were 200 people,respectively),breast feeding(193 people) and non-pregnant women (200 people) in Zhoupu and Kangqiao districts of Pudong New area were collected from Apr 2009 to Dec 2010.Two hundred copies of neonatal heel blood samples were collected.Median of UI was measured by arsenic-cerium catalysis.TSH in neonatal heel blood was analyzed 72 h after birth by time resolved fluoroisnmunoassay(TRFIA).ResultsMedian UI of all pregnant women was 161.35 μg/L,and that in third trimesters of pregnancy( 126.35 μg/L) was lower than that of the first,the second,the breast feeding and non-pregnant women (178.80,180.50,167.90,163.40 μg/L,all P< 0.05).The percentage of UI level less than 150 μg/L in the third trimester[57.5%(115/200) ] was higher than that of the first[39.0%(78/200) ],the second[39.5%(79/200) ],the breast feeding [ 16.6% (32/193) ] and non-pregnant women [ 23.0% (46/200) ],respectively (all P < 0.05).The percentage of UI level higher than 300 μg/L in the first [9.0%(18/200)],the second[8.0%(16/200) ] and the third trimester [ 5.0% ( 10/200 ) ] of pregnancy was lower than that of the breast feeding [ 20.2% (39/193) ] and nonpregnant [20.5%(41/200) ] women,respectively(all P < 0.05).The level of neonatal heel blood TSH was(2.92 ± 1.83)mU/L,the range was 0.01 - 9.76 mU/L,11.0%(22/200) of the neonates heel blood TSH level(5 mU/L)exceeded the ratio of World Health Organization (WHO) standard ( < 3% ) suitable for iodine nutrition.Conclusions The overall level of iodine nutrition among pregnant women in Zhoupu and Kangqiao districts of Pudong New Area of Shanghai is in the appropriate range,but the pregnant women in the third trimester is in mild iodine deficiencies,and the neonates in these districts may be prone to iodine deficiency.Monitoring of iodine nutrition of pregnant women should be strengthened and iodine supplementation should be done scientifically.
4.Study on Yangxue Qingnao granule in treating chronic cerebrovascular insufficiency.
Xi-xi GU ; Ding-fang CAI ; Yun-ke YANG ; Ying TENG ; Yi-ping CHEN ; Ming WEN
Chinese journal of integrative medicine 2005;11(1):22-26
OBJECTIVETo observe the therapeutic effect of Yangxue Qingnao granule (YXQNG) in treating chronic cerebrovascular insufficiency (CCI) and its possible mechanism.
METHODSEighty-three patients with CCI were randomly divided into YXQNG and nimodipine (ND) groups, the score of vertigo and the change in cerebral blood velocity before and after treatment were observed. And in the animal experiment, the authors adopted bilateral ligation of cervical carotid communis artery to establish CCI rat models in order to observe the effect of YXQNG and ND on incubation period of vertigo in rats and on memory performance.
RESULTSAfter clinical treatment, the vertigo score of YXQNG group was 2.34, and that of the ND group was 4.18, the comparison between the two groups showed that the difference was significant (P < 0.05). After treatment, the middle cerebral artery mean velocity (MCA Vm) of YXQNG group was 64.78 cm/s, vertebral artery mean velocity (VA Vm) was 29.78 cm/s, while that of ND group was 60.34 cm/s and 23.23 cm/s respectively, the comparison between these two groups showing statistical significance and the difference being obvious (P < 0.05). Experimental study showed that the rats in the model group after 12 weeks learning and memory were markedly lowered, the vertigo incubation period significantly lengthened, and compared with that of the model group, learning and memory of the YXQNG group was markedly improved and vertigo incubation period shortened, with the difference from that of the ND group insignificant, P > 0.05.
CONCLUSIONYXQNG could effectively improve CCI patients' vertigo and other clinical symptoms and increase the cerebral blood flow, lessen the vertigo incubation of model group rats, elevate model group rats' memory performance.
Aged ; Aged, 80 and over ; Animals ; Blood Flow Velocity ; drug effects ; Cerebrovascular Circulation ; drug effects ; Cerebrovascular Disorders ; complications ; drug therapy ; Drugs, Chinese Herbal ; adverse effects ; therapeutic use ; Female ; Humans ; Learning ; drug effects ; Male ; Memory ; drug effects ; Middle Aged ; Nimodipine ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Vasodilator Agents ; therapeutic use ; Vertigo ; etiology ; physiopathology
5.Identification and expression analysis of a full-length cDNA encoding a Kandelia candel tonoplast intrinsic protein.
Wei HUANG ; Xiao-Dong FANG ; Qi-Fen LIN ; Guan-Yi LI ; Wen-Ming ZHAO
Chinese Journal of Biotechnology 2003;19(2):147-152
Soil salinity is an important issue, as most crop plants are low in salt tolerance. Salt tolerance, a complex, multifactorial, and multigenic process, has been known to be a quantitative trait. The identification of the salt stress responsive genes or salt tolerance genes is essential for the breeding programs. Most recent efforts have been focused on the products of structural genes (transport proteins, ion channels, enzymes of solute synthesis) while little attention were paid to the regulatory aspects of these proteins. Since the first aquaporin gene from plants was cloned and functionally expressed in 1993, there has been a growing interest in the molecular biology of MIPs (membrane intrinsic proteins) and their bearing on the biophysics of water flow across plant membranes. In the last decades, studies on Mangroves, a special kind of wood plants, grow in high-salt and flooding conditions have been concentrated almost exclusively on their physiological and ecological characteristics. Kandelia candel, one of the dominant species of mangroves along the Chinese coast, lacks salt glands or salt hairs used for removal of excess salt in other mangroves. This makes K. candel a perfect model to study the molecular mechanism of salt tolerance in mangrove plants. Using cDNA RDA, a cDNA-specific modification of genomic representational difference analysis, a series of salt responsive genes of Kandelia candel were cloned. Among these gene fragments, a 183 bp fragment (termed as SRGKC1) encoding a tonoplast intrinsic protein (TIP) in Kandelia candel (KCTIP1) was identified. Based on the sequence of SRGKC1, two gene specific primers were designed, and the 3' and 5' end of the KCTIP1 gene were obtained using the SMART RACE cDNA Amplification Kit. RACE products were purified from low-melting agarose, and sequenced directly with GSPs as the sequencing primers. A 500-bp fragment corresponding to the 3'end of this gene was obtained using the GSP1 primer, and a 690 bp fragment corresponding to the 5' end of this gene was obtained using the GSP2 primer. Two primers that flank the putative open reading frame (ORF) were designed to obtain the cDNA containing the complete ORF by RACE PCR reaction. The full-length cDNA of KCTIP1, containing a 756 bp open reading frame (ORF), was approximately 1.1 kb; the start codon was located at the nucleotides of 99-101 and stop codon at the nucleotides of 855-857 followed by a poly (A) tail. The KCTIP1 cDNA sequence in this research was released in GenBank with accession number AF521135. Using ExPASy Proteomics tools provided by EMBL, the isoelectric point and MWt of KCTIP1 are estimated as 5.77 and 26.3 kD respectively. Transmembrane prediction analysis revealed the deduced KCTIP1 protein sequence contains six transmembrane regions at amino acid residues of 20 - 42, 57 - 79, 86 - 108, 113 - 135, 142 - 164 and 217 - 239. Two highly conserved asparagine-proline-alanine (NPA) motifs were located at 85 - 87 and 199 - 201 amino acid residues respectively. KCTIP1 is also predicted to contain the Cys residue (Cys 118) that are shown to confer Hg-sensitivity in Arabidopsis gamma-TIP and delta-TIP. Similarity analysis showed that KCTIP1 shared 77% - 79% amino acid sequence identity with the TIPs from Vitis berlandieri, Brassica oleracea and Arabidopsis thaliana. Expression analyses indicated that KCTIP1 had different expression among species of Mangroves. Expressions of KCTIP1 in Kandelia candel, Rhizophora apoculata and Ceriops tagal were suppressed by salt, and were insensitive to salt stress in unknown species of Mangroves. Previous studied showed that salt conditions might result in large and rapid changes in extracellular water potential and serious disturbance to the cytoplasm. In order to compensate for this imbalance, the relative contribution of water channels to flow across the root could thus vary. K. candel is a species that is native to intertial zone of tropical and subtropical coast and is well-adapted to salt conditions. The coordinated down-regulation of aquaporins in this plant may decrease membrane water permeability and thus increase the cellular water conserva- tion during periods of salt stress. The results reported here are consistent with the postulated roles for tonoplast water channels in regulating the hydraulic permeability of the vacuolar membranes and in adjusting the water homeostasis of the protoplasm under various physiological conditions. The identification of KCTIP1 as one of salt-responsive genes implies that intracellular osmotic equilibration is a part of salt-tolerant mechanisms in Mangroves.
Amino Acid Sequence
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Base Sequence
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Blotting, Northern
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DNA, Complementary
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genetics
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Membrane Proteins
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chemistry
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genetics
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Molecular Sequence Data
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Nucleic Acid Amplification Techniques
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Plant Proteins
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chemistry
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genetics
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Rhizophoraceae
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genetics
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Sequence Alignment
6.The Interaction Between Dendritic Cells and Penicillium marneffei Yeasts
Li MA ; Li-Yan XI ; Wen-Jie ZHAO ; Jiu-Feng SUN ; Chang-Ming LU ; Jun-Min ZHANG ; Xi-Qing LI ; Hong-Fang LIU ;
Microbiology 1992;0(04):-
To assay the influence of dendritic cells(DCs)on the function of anti-infective immunity to Penicillium marneffei.DCs were generated from peripheral blood mononuclear cells(PBMC)and pulsed with Penicillium marneffei yeasts.DCs morphology was observed by the inverted microscope and cell surface markers of DCs were analyzed by flow cytometry.The concentrations of IL-12p70 were detected by ELISA. Mixed lymphocyte reaction was performed to assay the proliferation of T cells.The mRNA of CCR7 and CXCR4 were detected by the Real-time PCR quantifications.The acquired DCs exhibited irregular appearance and numerous long dendrites under light microscope.DCs and Penicillium marneffei yeasts were co-cultured for 24 h,numerous yeasts were observed inside the cells;an enhanced expression of the cell sur-face markers CD86、CD83、HLA-DR and CD40 were demonstrated;the expression of CCR7 and CXCR4 mRNA were also increased;the improved proliferation of T cells were observed in the mixed lymphocyte reaction.Yeasts-pulsed DCs secreted more IL-12p70 than that of non-pulsed,but less than that of LPS-pulsed DCs.DCs can engulf the Penicillium marneffei yeasts.When pulsed with Penicillium marneffei yeasts,DCs improved their expression of the co-stimulatory molecules and chemokine receptor CCR7、CXCR4,enhanced their capacity to process antigen.DCs play an important role in host defense against Penicillium marneffei infection.But the low level of the IL-12p70 production may lead to deficiency in the cell-mediated immunity against Penicillium marneffei.
7.Resveratrol and puerarin on peak bone mass in young rats.
Hui-Rong XI ; Fang-Fang YANG ; Yu-Hai GAO ; Hui-Ping MA ; Ke-Ming CHEN
China Journal of Orthopaedics and Traumatology 2018;31(7):635-641
OBJECTIVETo compare effects of resveratrol, puerarin and the compounds on peak bone mass in rats.
METHODSForty SPF Wistar rats weighed 109.45 g to 119.44 g with an average of 115.87 g were selected. After 3 days' adaption, rats were divided into control group (the same volume of distilled water per day), puerarin group(15.4 mg/kg puerarin daily), resveratrol group (8.4 mg/kg resveratrol daily), compound drug group (daily dose of 8.4 resveratrol added 15.4 mg/kg of puerarin) and 10 in each group. The body weight of the rats was monitored at every 7 days and body bone density was measured at every month. All rats were sacrificed after 3 months. The bone mineral density of femur and vertebrae was detected by dual energy X-ray absorptiometry; bone biomechanics, VG staining was used to analyze bone histomorphometry;ELISA was used to detect serum bone metabolic index and microstructure of femur were scanned with Micro-CT scanner.
RESULTSThere were no significant differences in body weight among groups during exoeriment. Bone mineral density results showed BMD of femur and vertebrae in the other three groups were significantly increased, and R+P group was significantly higher than PR group and RES group(<0.05) by compared with CON group;three-point bending and compression test results showed compared with CON group, other three groups of femoral and vertebral maximum load values were significantly increased, and P+R group was higher than PR group and RES group, but elastic modulus was not statistically significant. Bone histomorphometry showed that number of trabecular bone in other three groups were significantly increased compared with CON group, separation of trabecular bone were significantly reduced, continuity was improved, and R+P group was significantly better than RES and PR group. The results of Micro-CT scan showed that separation of trabecular bone were significantly reduced, continuity were improved in other three groups, and R+P group was significantly better than RES and PR group. The numbers of trabecular bone (Tb.N), trabecular bone thickness (Tb.Th), volume of trabecular bone (BV/TV) in PR group, RES group and R+P group were significantly higher than CON group, but trabecular bone separation (Tb.Sp) was significantly reduced. Serum levels results showed, level of OC in the other three groups were higher than control group(<0.05), content of TRACP 5b decreased, and level of OC in P+R group was significantly higher than PR group and RES group, content of TRACP 5b was no significant change.
CONCLUSIONSCompound of puerarin and resveratrol assigned in a 1:1 ratio could improve bone mineral density and bone mass in young rats, enhance biomechanical properties of bone, promote mineralization and maturation of osteoblasts, inhibit osteoblastic bone resorption, and is better than the role of their respective monomers. The paper showed that traditional Chinese medicine compound medicine will be used as a new way to prevent and treat osteoporosis.
8.Effect of ceftriaxone on the intestinal epithelium and microbiota in neonatal mice.
Qun WAN ; Ru-Yue CHENG ; Jia-Wen GUO ; Ke WANG ; Xi SHEN ; Fang-Fang PU ; Ming LI ; Fang HE
Chinese Journal of Contemporary Pediatrics 2018;20(4):318-325
OBJECTIVETo investigate the effect of ceftriaxone on the intestinal epithelium and microbiota in mice in the early-life stage, as well as the recovery of the intestinal epithelium and reconstruction of intestinal microbiota in adult mice.
METHODSA total of 36 BALB/C neonatal mice were randomly divided into control group and experimental group, with 18 mice in each group. The mice in the experimental group were given ceftriaxone 100 mg/kg every day by gavage within 21 days after birth. Those in the control group were given an equal volume of normal saline by gavage. Immunohistochemistry was used to measure the expression of Ki67, Muc2, and ZO-1 in the intestinal epithelium. qPCR and next-generation sequencing were used to analyze the overall concentration and composition of fecal bacteria.
RESULTSAfter 21 days of ceftriaxone intervention, the experimental group had a significant reduction in body weight, a significant reduction in the expression of Ki67 and ZO-1 and a significant increase in the expression of Muc2 in intestinal epithelial cells, a significant reduction in the overall concentration of fecal bacteria, and a significant increase in the diversity of fecal bacteria compared with the control group (P<0.05). Firmicutes was the most common type of fecal bacteria in the experimental group, and there were large amounts of Staphylococcus and Enterococcus. The experimental group had a certain degree of recovery of the intestinal epithelium, but there were still significant differences in body weight and the structure of intestinal microbiota between the two groups at 56 days after birth (P<0.05).
CONCLUSIONSEarly ceftriaxone intervention significantly affects the development of the intestinal epithelium and the construction of intestinal microbiota in the early-life stage. The injury of the intestinal microbiota in the early-life stage may continue to the adult stage and affect growth and development and physiological metabolism.
Animals ; Animals, Newborn ; Anti-Bacterial Agents ; pharmacology ; Ceftriaxone ; pharmacology ; Female ; Gastrointestinal Microbiome ; drug effects ; Intestinal Mucosa ; drug effects ; Ki-67 Antigen ; analysis ; Mice ; Mice, Inbred BALB C ; Mucin-2 ; analysis ; Zonula Occludens-1 Protein ; analysis
9.Effects of substance P on granulation tissue fibroblasts proliferation and expression of basic fibroblast growth factor mRNA.
Wei JIANG ; Zheng-guo WANG ; Xi-nan LAI ; Jin-ming ZHU ; Pei-fang ZHU
Chinese Journal of Surgery 2004;42(6):366-368
OBJECTIVETo explore the proliferation-promoting effect of sensory neuropeptide substance P (SP) on the cultured granulation tissue fibroblasts in vitro and its regulative effect on the gene expression of basic fibroblast growth factor (bFGF) mRNA.
METHODSThe proliferation-promoting effect of cultured granulation tissue fibroblasts was observed by means of MTT; the regulative effect of SP on gene expression of fibroblast bFGF by RT-PCR. The time and dose-efficiency relations were also observed.
RESULTSThere was a significant proliferation-promoting effect of SP on the cultured granulation tissue fibroblasts in vitro in a remarkable dose-dependent fashion. However, bFGF antibody only partly exerted its inhibitive effect. SP could induce the bFGF mRNA expression of the fibroblasts at the 3rd and 6th hour (P < 0.01). SP could promote the bFGF mRNA expression of the fibroblasts in the concentration of 10(-9) - 10(-5) mol/L and peaked in the concentration of 10(-7) mol/L.
CONCLUSIONSSP has a significant proliferation-promoting effect on the granulation tissue fibroblasts, which is correlated with SP inducing bFGF mRNA expression of fibroblasts.
Animals ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Fibroblast Growth Factor 2 ; genetics ; Fibroblasts ; cytology ; drug effects ; metabolism ; Gene Expression ; drug effects ; Granulation Tissue ; cytology ; drug effects ; metabolism ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Substance P ; pharmacology
10.Preparation and characterization of Forms A and B of benazepril hydrochloride.
Hong FANG ; Xiu-rong HU ; Jian-ming GU ; Guan-xi CHEN ; Jian-yue FENG ; Gu-ping TANG
Journal of Zhejiang University. Medical sciences 2012;41(6):639-646
OBJECTIVETo prepare Form A and Form B of benazepril hydrochloride and to compare the differences in spectrums, thermodynamics and crystal structure between two polymorphic forms.
METHODSForm A and Form B of benazepril hydrochloride were characterized by Fourier transform infrared spectroscopy (IR), thermal gravimetric analysis (TG), differential scanning calorimetry (DSC), powder x-ray diffraction (PXRD) and single crystal x-ray diffraction (SCXRD).
RESULTSPreparation method, crystal structure and polymorphic stability of Form A and Form B of benazepril hydrochloride were obtained. Based on the analysis of crystal structure of both polymorphs, Form A belonged to monoclone space group P2(1) with a=7.8655(4)Å, b= 11.7700(6)Å, c= 13.5560(7)Å, β= 102.9470(10)°, V=1223.07 (11)Å(3) and Z=2, while Form B belonged to orthorhombic space group P212121, with a=7.9353(8)Å, b=11.6654(11)Å, c=26.6453(16)Å, V=2466.5(4)Å(3) and Z=4. From the DSC and XRD results, Form B of benazepril hydrochloride could be transformed into Form A after heating treatment.
CONCLUSIONForm A and Form B of benazepril hydrochloride are both anhydrous and displayed different polymorphs due to different molecular configuration. Furthermore, Form A exhibits more stable than Form B at high temperatures.
Benzazepines ; chemistry ; Crystallization ; Drug Stability ; Molecular Conformation