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Objective To establish a simple and efficient method for isolation and culture of adult rat cardiac micro-vascular endothelial cells. Methods Wistar rats were anesthetized and thoracotomy was performed. Cardiac micro-vascular endothelial cells were isolated by heart perfusion and enzyme digestion. The morphology and growth curve of the third generation cells were examined by inverted microscopy. Its molecular markers factor Ⅷ-related antigen and CD31 were detected by immunocytochemistry and immunofluoresence. The angiogenic ability was examined by matrigel. Results The primary cultured cardiac microvascular endothelial cells in vitro were polygonal or fusiform with typical paving stone-like growth characteristics. At the same time,the cells were positive stained by factorⅧ-related antigen and CD31 and showed angiogenic ability. Conclusions Cardiac microvascular endothelial cells cul-tured by this method possess high purity. The method is easy to operate and can be used for laboratory and clinical research of cardiovascular diseases.
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Oxytocin (OT) is a 9-amino-peptide hormone synthesized mainly in the Paraventricular Nucleus (PVN) and Supraoptic Nucleus (SON) of mammals'hypothalamus. Small part of OT is synthesized in peripheral organs. In addition to promoting the lactation and uterine contraction, OT has many other physiological functions, such as analgesia. It has been proved that neuropeptide can exert analgesic effect. OT is indeed a neuropeptide secreted naturally by mammals. Sdudies have shown that OT plays an important role in relieving pain and can avoid problems like addiction and other side effects.However, its underlying mechanism of analgia is still unclear. Therefore, further study of the analgesic mechanism of oxytocin has a great theoretic significance and very broad clinical prosperities.
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<p><b>OBJECTIVE</b>To examine serum adiponectin level in preterm infants and to evaluate the relationship between serum adiponectin and bone mineral density in preterm infants.</p><p><b>METHODS</b>Seventy-two appropriate-for-gestational-age neonates were classified into three groups according to their gestational ages: early preterm (31-33(+6) weeks, 13 cases), late preterm (34-36(+6) weeks, 16 cases), and full-term (37-42 weeks, 43 cases). Venous blood was collected at one week of their life to measure serum adiponectin concentration. During the period, omnisense ultrasound bone sonometer was applied to measure speed of sound (SOS) of the left tibia.</p><p><b>RESULTS</b>The median of tibia SOS in the early preterm group was significantly lower than in the late preterm and full term groups (P<0.05), and the median of tibia SOS in the late preterm group was lower than in the full-term group (P<0.05). Serum adiponectin level was lowest in the early preterm group, and the full-term group had the highest serum adiponectin level. Serum adiponectin level was positively correlated with tibia SOS in preterm infants (r=0.664, P<0.05). According to the result of multivariate linear stepwise regression analysis, serum adiponectin and birth weight were independent predictor of tibia SOS in preterm infants.</p><p><b>CONCLUSIONS</b>Serum adiponectin level is lower in preterm infants than that in full-term infants. There is a positive correlation between serum adiponectin and bone mineral density in preterm infants.</p>
Subject(s)
Female , Humans , Infant, Newborn , Male , Adiponectin , Blood , Birth Weight , Bone Density , Infant, Premature , Blood , Linear ModelsABSTRACT
Astragalus polysaccharide has been widely used in food and medicinal industry owing to its health-promoting properties. In order to characterize better the relationship among molecular weight, structure-activity and activities, a simple method was used different concentration of ethanol including 30% (PW30), 50% (PW50), 70% (PW70), 75% (PW75), 80% (PW80) and 90% (PW90) to precipitate Astragalus polysaccharides into different molecular weight. As a result, PW90 showed smooth surface and the strongest antioxidant activity among these six fractions (P < 0.05). In conclusion, graded ethanol precipitation was a simple method to separate Astragalus polysaccharides into different molecular weight with different antioxidant activity fractions.
Subject(s)
Antioxidants , Pharmacology , Astragalus Plant , Chemistry , Chemical Precipitation , Ethanol , Chemistry , Polysaccharides , Chemistry , PharmacologyABSTRACT
?AlM:To observe the clinical effects of thelacrimal fistula excision combined with double silicone intubation in the treatment of chronic dacryocystitis with lacrimal fistula.? METHODS: Totally 25 cases ( 25 eyes ) of chronic dacryocystitis with lacrimal fistula were allocated into two groups:the double silicone intubation group of 13 cases (13 eyes) received lacrimal fistula excision combined with double silicone intubation, and the routine group of 12 cases (12 eyes) received routine dacryocystorhinotomy, we analyzed the curative effect.?RESULTS: The double silicone intubation group had a cure rate of 92. 3% (12 eyes), a effective rate of 100%, compared with the routine group had a cure rate of 91. 7%( 11 eyes ) , a effective rate of 100%, there were no statistically significant difference between the two groups (P>0. 05).?CONCLUSlON:Lacrimal fistula excision combined with double silicone intubation has a same effect with routine dacryocystorhinotomy in the treatment of chronic dacryocystitis with lacrimal fistula, but less invasive, no scar, less pain, and meet the patients' esthetic needs.
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<p><b>OBJECTIVE</b>To investigate the correlation of gestational age (GA) with carboxyterminal propeptide of type I procollagen (PICP), deoxypyridinoline (DPD), and bone sound of speed (SOS) in appropriate-for-gestational-age (AGA) neonates, as well as the relationship between bone turnover markers and bone SOS.</p><p><b>METHODS</b>Sixty-five AGA neonates were included in the study. The neonates were divided into three groups: preterm infant (GA ≤3 4 weeks, 14 cases), late preterm infant (34 weeks<GA<37 weeks, 13 cases), and full-term infant (GA ≥ 37 weeks, 38 cases). Birth weight and length were measured in all cases, and Ponderal index (PI) was used to estimate their nutritional status. Venous blood was collected within 7 days after birth to measure blood PICP concentration. Urine was collected to measure urinary DPD and creatinine (Cr) levels. Omnisense 7000P ultrasound bone sonometer was applied to measure the SOS of the left tibia in all cases within 7 days after birth.</p><p><b>RESULTS</b>There were significant differences in GA (F=140.199, P<0.001), birth weight (F=47.042, P<0.001), birth length (F=46.877, P<0.001), and PI (F=11.898, P<0.001) between the three groups; the higher the GA, the higher the birth weigh, birth length, and PI. There were significant differences in PICP (F=30.384, P<0.001), DPD/Cr (F=21.761, P<0.001), and SOS (F=20.052, P<0.001) between the three groups; the higher the GA, the lower the PICP and DPD/Cr and the higher the bone SOS. PICP and DPD/Cr were negatively correlated with GA, birth weight and bone SOS (P<0.01), while bone SOS was positively correlated with GA and birth weight (P<0.01), which still held true after adjustment for GA and birth weight.</p><p><b>CONCLUSIONS</b>Among AGA neonates, bone turnover markers are negatively correlated with GA, birth weight and bone SOS. High bone turnover is bad for bone health in AGA neonates.</p>
Subject(s)
Female , Humans , Infant, Newborn , Male , Amino Acids , Blood , Birth Weight , Bone Density , Gestational Age , Peptide Fragments , Blood , Procollagen , Blood , Tibia , Diagnostic Imaging , UltrasonographyABSTRACT
Ornithine decarboxylase (ODC) is the rate-limiting enzyme in polyamine biosynthesis and a target for chemoprevention. Hydroxydibenzoylmethane (HDB), a derivative of dibenzoylmethane of licorice, is a promising chemopreventive agent. In this paper, we investigated whether HDB would inhibit the ODC pathway to enhance apoptosis in human promyelocytic leukemia HL-60 cells. We found ODC enzyme activity was reduced during HDB treatment. Overexpression of ODC in HL-60 parental cells could reduce HDB-induced apoptosis, which leads to loss of mitochondrial membrane potential (Deltapsim), through lessening intracellular ROS. Furthermore, ODC overexpression protected cytochrome c release and the activation of caspase-3 following HDB treatment. The results demonstrated HDB-induced apoptosis was through a mechanism of down-regulation of ODC and occurred along a ROS-dependent mitochondria-mediated pathway.
Subject(s)
Humans , Apoptosis/drug effects , Caspase 3/metabolism , Chalcones/metabolism , Chemoprevention , Cytochromes c/biosynthesis , Down-Regulation , Gene Expression , HL-60 Cells , Immunoblotting , Leukemia, Myeloid/enzymology , Membrane Potential, Mitochondrial/drug effects , Mitochondria/enzymology , Ornithine Decarboxylase/antagonists & inhibitors , Reactive Oxygen Species/analysis , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
A new method of automatic detection of brain lesion based on wavelet feature vector of CT images has been proposed in the present paper. Firstly, we created training samples by manually segmenting normal CT images into gray matter, white matter and cerebrospinal fluid sub images. Then, we obtained the cluster centers using FCM clustering algorithm. When detecting lesions, the CT images to be detected was automatically segmented into sub images, with a certain degree of over-segmenting allowed under the premise of ensuring accuracy as much as possible. Then we extended these sub images and extracted the features to compute the distances with the cluster centers and to determine whether they belonged to the three kinds of normal samples, or, otherwise, belonged to lesions. The proposed method was verified by experiments.
Subject(s)
Humans , Electronic Data Processing , Methods , Brain , Diagnostic Imaging , Brain Neoplasms , Diagnostic Imaging , Image Interpretation, Computer-Assisted , Methods , Intracranial Hemorrhages , Diagnostic Imaging , Tomography, X-Ray Computed , Methods , Wavelet AnalysisABSTRACT
<p><b>OBJECTIVE</b>To express, purify and refold recombinant luteinizing hormone releasing hormone-angiogenin (LHRH-Ang) toxin using E. coli. expression system.</p><p><b>METHODS</b>Recombinant LHRH-Ang expression vector was constructed by replacing of EGF fragment in plasmid pET28a/EGF-Ang with LHRH-PII fragment amplified from plasmid pET28/MSH-PE40. DNA sequencing would be used to verify the correction of fused LHRH-PII-Ang gene. Then, E. coli strain BL21 (DE3) was transformed by pET28a/LHRH-Ang vector. Expression of recombinant LHRH-Ang toxin was induced by Isopropyl-β-D-Thiogalactoside (IPTG). Refolding effects of gradient dialysis was evaluated by SDS-PAGE.</p><p><b>RESULTS</b>Prokaryotic expression vector pET28a/LHRH-Ang, containing LHRH-PII-Ang fusion gene, was constructed by PCR amplification, restriction enzyme digestion and ligation method. Sequence correction of fusion gene was confirmed by DNA sequencing. After IPGT induction, recombinant LHRH-Ang protein was expressed in BL21 (DE3) as inclusion body, it took 18.43% of total protein. Inclusion body was resolved in 8 mol/L urea and purified by DEAE-Sepharose FF column, the purity was 85%. Recombinant LHRH-Ang toxin was refolded and concentrated by gradient dialysis and PEG 20000, respectively.</p><p><b>CONCLUSIONS</b>Recombinant LHRH-Ang protein was expressed in E. coli and refolded successfully.</p>
Subject(s)
Escherichia coli , Metabolism , Gene Expression , Genetic Vectors , Gonadotropin-Releasing Hormone , Genetics , Plasmids , Recombinant Fusion Proteins , Genetics , Ribonuclease, Pancreatic , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To evaluate the safety and immunological effect of domestic split influenza virus vaccine.</p><p><b>METHODS</b>All 606 subjects were divided into three groups by under 6, 16-60 and above 60 years old. Each age group was divided as study group (n = 213), control group 1 (n = 195) and control group 2 (n= 198) by Table of Random Number, one domestic vaccine and two imported vaccines were respectively inoculated in three group people. The differences of clinical side effect rate, antibody positive rate, protective rate and geometric mean titer (GMT) of these three vaccines were compared by using the statistical software with statistical significance of P < 0.05.</p><p><b>RESULTS</b>The side effect rate of study group, control group 1 and control group 2 was 3.76% (8/213), 4.10% (8/195), and 3.54% (7/198), respectively without statistical significance(chi2 = 0.87, P =0.93). The positive seroconversion rates of H1N1, H3N2 and B in these three groups were respectively 89.2% (190/213), 63.4% (135/213), 86.4% (184/213), 88.7% (173/195), 61.5% (120/195), 87.2% (170/195), 87.9% (174/198), 61.6% (122/198) and 84.8% (168/198). There were no statistical significance in the total positive seroconversion rate of each antibody type (chi2(H1N1) = 0.94, P(H1N1) = 0.63; chi2(H3N2) = 0.94, P(H3N2) = 0.63; chi2(B) = 0.75, P(B) = 0.69). The average growth multiple of H1N1, H3N2 and B in these three groups were 10.7, 7.3, 8.4, 10.5, 6.3, 8.3, 10.2, 7.1, 8.8 times. There were no statistical significances in the GMT growth multiple of each antibody type (F(H1N1) = 0.35, P(H1N1) = 0.70; F(H3N2) = 2.22, P(H3N2) = 0.11; F(B) = 1.51, P(B) = 0.35). The antibody protective rates of H1N1, H3N2 and B were 100% (213/213), 70.0% (149/213), 95.3% (203/213), 100% (195/195), 66.7% (130/195), 97.9% (191/195), 99.5% (197/198), 66.2% (131/198), 96.5% (191/198) respectively. There was no statistical difference among the three vaccines (chi2(H1N1) = 2.04, P(H1N1) = 0.36; chi2(H3N2) = 0.74, P(H3N2) = 0.69; chi2(B) = 0.42, P(B) = 0.82).</p><p><b>CONCLUSION</b>The domestic influenza split vaccine might be suitable for colony vaccination for its having clinical safety and immunological effect.</p>
Subject(s)
Adolescent , Adult , Child , Humans , Middle Aged , Young Adult , Influenza A Virus, H1N1 Subtype , Allergy and Immunology , Influenza A Virus, H3N2 Subtype , Allergy and Immunology , Influenza Vaccines , Allergy and Immunology , Influenza, HumanABSTRACT
In order to explore the activity of a peptide containing rat sodium pump α2 subunit M1-M2 extramembrane fragment (RES2 derivative) in vitro, the peptide (Leu-Ala-Ala-Met-Glu-Asp-Glu-Pro-Ser-Asn-Asp-Asn-Gly-Gly-Gly-Ser) was synthesized by peptide synthesizer with Fmoc method and purified by high performance liquid chromatography (HPLC). Its binding activity was identified by radioligand-receptor binding assay (RRA) and its bioactivity was measured by erythrocyte (86)Rb uptake. The results of saturation binding experiment and competitive binding experiment showed that the synthesized RES2 derivative had the capability to bind to (3)H-ouabain. The dissociation constant (K(d)) was 38.46 nmol/L and IC(50) was 6.353 nmol/L. Erythrocyte (86)Rb uptake experiment showed that the RES2 derivative blocked the inhibitory effect of ouabain on the sodium pump on erythrocyte membrane in a dose-dependent manner. The results showed that the RES2 derivative is capable of binding to ouabain and improving the activity of sodium pump on erythrocyte membrane, suggesting that the RES2 derivative might become an effective antihypertensive drug in the future.