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The structure of fibrauretin made by our lab was modified. Fibrauretin was demethylated at 9-site under high temperature pyrolysis at 160℃-180℃ and was reacted with a series of acid chlorides. Twele derivatives of fibrauretin were obtained. The structure of each derivative was determined by1H-NMR and13C-NMR. The derivatives were 9-O-benzoyl-fibrauretin, 9-O-( 2-methylbenzoyl)-fibrauretin, 9-O-( 4-methylbenzoyl)-fibrauretin, 9-O-(3, 5-dimethylbenzoyl)-fibrauretin, 9-O-(4-(chloromethyl) benzoyl)-fibrauretin and other derivatives. The 12 derivatives are all new chemical compounds. Taking ATCI as substrate,the inhibitory activity on acetylcholinesterase (AChE) from the head of flies of the fibrauretin and its derivatives were screened. The results showed that most of the derivatives had improved their inhibitory activity on AChE through esterification reaction. Compounds 9-O-(4-methylbenzoyl)-fibrauretin, 9-O-(3,5-dimethylbenzoyl)-fibrauretinand 9-O-(4-(chloromethyl)benzoyl)-fibrauretin had significant inhibitory effect on AChE,and the inhibitory activity was stronger than the that of donepezil.
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OBJECTIVE: To evaluate the qualities of different processed products of Gastrodia elata Bl. f. glauca S. Chow, such as steamed, wine-processed, ginger-processed and tribulus-processed products, and determine the optimum processing method of Gastrodia elata. METHODS: HPLC was employed to determine gastrodin and 4-hydroxybenzyl alcohol, and phenol-sulfuric acid method was used to detect the content of gastrodia polysaccharide. RESULTS: Gastrodin and gastrodia polysaccharides had the highest contents(1.901 and 3.246 mg·g-1, respectively) in the tribulus-processed products; and the highest content of 4-hydroxybenzyl alcohol in ginger-processed Gastrodia elata was 0.80 mg·g-1; gastrodin in two kinds of wine-processed products was lower than the others, while the content of gastrodia polysaccharide in Gastrodia elata processed by wine-soaking for three days was obviously less than those in the other products. CONCLUSION: The traditional tribulus-processed method is better than the modern steaming method. This study can provide scientific basis for the mechanism of traditional processing method of Gastrodia elata.
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The study aims at screening the specific bands by PCR, quickly and accurately evaluating the quality of ginseng seeding, accelerating the process of ginseng breeding. Based on the correlation of genetic differences and saponin content between individuals, a pair of specific primer GC1 was screened by PCR. According to the experiment by L16 (45) orthogonal test, a PCR system most suitable for GC1 was established, which came out total 25 μL reaction system containing DNA 2.60 mg•L⁻¹, Mg²⁺ 1.44 mmol•L⁻¹, dNTP 0.19 mmol•L⁻¹, primer 0.32 μmol•L⁻¹ and Taq enzyme concentration 0.076 U•μL⁻¹. By comparing the saponin content and the GC1 PCR electrophoretogram of samples, the ginseng, with 1 200 bp specific band by PCR of GC1, the contents of 9 monosodium saponins and their additions were higher than others, which provided a reliable method for accelerating the process of ginseng breeding. The sequence was sequenced and 99% homologous to glycerol-3-phosphate dehydrogenase.
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The experiment was aimed to investigate the difference of plasma concentration and pharmacokinetic parameters between liposome and aqueous solution of toatal ginsenoside of ginseng stems and leaves in rats, such as ginsenosides Rg₁, Re, Rf, Rb₁, Rg₂, Rc, Rb₂, Rb₃, Rd. After intravenous injection of liposome and aqueous solution in rats, the blood was taken from the femoral vein to detect the plasma concentration of the above 9 ginsenoside monomers in different time points by using HPLC. The concentration-time curve was obtained and 3p97 pharmacokinetic software was used to get the pharmacokinetic parameters. After the intravenous injection of ginsenosides to rats, nine ginsenosides were detected in plasma. In general, among these ginsenosides, the peak time of the aqueous solution was between 0.05 to 0.083 3 h, and the serum concentration peak of liposome usually appeared after 0.5 h. After software fitting, the aqueous solution of ginsenoside monomers Rg₁, Re, Rf, Rg₂, Rc, Rd, Rb₃ was two-compartment model, and the liposomes were one-compartment model; aqueous solution and liposome of ginsenoside monomers Rb₁ were three-compartment model; aqueous solution of ginsenoside monomers Rb₂ was three-compartment model, and its liposome was one-compartment model. Area under the drug time curve (AUC) of these 9 kinds of saponin liposomes was larger than that of aqueous solution, and the retention time of the liposomes was longer than that of the aqueous solution; the removal rate was slower than that of the aqueous solution, and the half-life was longer than that of the water solution. The results from the experiment showed that by intravenous administration, the pharmacokinetic parameters of two formulations were significantly different from each other; the liposomes could not only remain the drug for a longer time in vivo, but also reduce the elimination rate and increase the treatment efficacy. As compared with the traditional dosage forms, the total ginsenoside of ginseng stems and leaves can improve the sustained release of the drug, which is of great significance for the research and development of new dosage forms of ginsenosides in the future.
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In this study, the total alkaloids of Huangteng were given to the rats by the methods of intragastric administration and tail vein. After the concentration changes of palmatine and jatrorrhizine in the plasma of rats were determined by RP-HPLC, pharmacokinetic parameters and oral bioavailability were calculated by 3P97 software. After the rats were pre-treated with total alkaloid 60 mg•kg⁻¹ by the methods of intragastric administration and tail vein, the main pharmacokinetic parameters were determined as follows： in the intragastric administration group, the Cmax of palmatine and jatrorrhizine were (0.91±0.06), (0.70±0.08) mg•L⁻¹; tmax of palmatine and jatrorrhizine were (35.24±0.83), (47.76±1.24) min; t1/2 of palmatine and jatrorrhizine were (187.03±1.53), (105.64±16.99) min, AUC of palmatine and jatrorrhizine were (280.30±18.69), (144.36±1.06) mg•min•L⁻¹; in the intravenous injection group, the t1/2 of palmatine and jatrorrhizine were (172.18±12.38), (147.26±1.82) min; AUC of palmatine and jatrorrhizine were (2 553.14±214.91), (328.83±10.81) mg•min•L⁻¹. The oral bioavailability of palmatine was 10.98% and jatrorrhizine was 43.90%.
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BACKGROUND/AIMS: Gastric hypersensitivity contributes to abdominal pain in patients with functional dyspepsia. Recent studies showed that hormones induced by stress are correlated with visceral hypersensitivity. However, the precise mechanisms underlying gastric hypersensitivity remain largely unknown. The aim of the present study was designed to investigate the roles of corticosterone (CORT) on excitability of dorsal root ganglion (DRG) neurons innervating the stomach. METHODS: DRG neurons innervating the stomach were labeled by DiI injection into the stomach wall. Patch clamp recordings were employed to examine neural excitability and voltage-gated sodium channel currents. Electromyograph technique was used to determine the responses of neck muscles to gastric distension. RESULTS: Incubation of acutely isolated DRG neurons with CORT significantly depolarized action potential threshold and enhanced the number of action potentials induced by current stimulation of the neuron. Under voltage-clamp mode, incubation of CORT enhanced voltage-gated sodium current density of the recorded neurons. Pre-incubation of GF109203X, an inhibitor of protein kinase C, blocked the CORT-induced hyperexcitability and potentiation of sodium currents. However, pre-incubation of H-89, an inhibitor of protein kinase A, did not alter the sodium current density. More importantly, intraperitoneal injection of CORT produced gastric hypersensitivity of healthy rats, which was blocked by pre-administration of GF109203X but not H-89. CONCLUSIONS: Our data strongly suggest that CORT rapidly enhanced neuronal excitability and sodium channel functions, which is most likely mediated by protein kinase C but not protein kinase A signaling pathway in DRG neurons innervating the stomach, thus underlying the gastric hypersensitivity induced by CORT injection.
Subject(s)
Animals , Humans , Rats , Abdominal Pain , Action Potentials , Corticosterone , Cyclic AMP-Dependent Protein Kinases , Diagnosis-Related Groups , Dyspepsia , Ganglia , Ganglia, Spinal , Hypersensitivity , Injections, Intraperitoneal , Neck Muscles , Neurons , Protein Kinase C , Protein Kinases , Sodium , Sodium Channels , Spinal Nerve Roots , Stomach , Visceral PainABSTRACT
OBJECTIVE: To establish an HPLC method for simultaneous determination of ginsenoside Rg1, Re, Rb1 and jujuboside A, B in Renshen Jianpi Pellets (RSJPW). METHODS: The determination was conducted on an Eclipse XDB C18 column (4.6 mm × 250 mm, 5 μm) with the mobile phase of acetonitrile (A)-water (B) gradiently eluted at the following flow rates: 0-90 min, 1.0 mL · min-1; 90-130 min, 1.0-0.5 mL · min-1; 130-140 min, 0.5 mL · min-1. And the column temperature was maintained at 35℃; the detection wavelength was set at 203 nm. RESULTS: The calibration curves of ginsenoside Rg,, Re, Rb, and jujuboside A, B were in good linearity over 0.32-2.24 μg (r=0.9982), 0.16-1.12 μg (r=0.995 3), 0.32-2.24 μg (r=0.9996), 0.16-1.12 μg (r=0.9915), 0.08-0.56 μg (r=0.9996). The corresponding average recovery rates were 97.18%, 97.62%, 98.79%, 98.48%, 94.51%; the standard deviations were 1.10%, 0.98%, 0.34%, 1.09%, 1.88%, respectively. CONCLUSION: The established HPLC method is accurate, reliable and reproducible, which can be used as a reference for the quality control of RSJPW.
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<p><b>OBJECTIVE</b>To establish a new method for detection of red blood cell osmotic fragility by using flow cytometry.</p><p><b>METHODS</b>The hypotension salt solution of different concentrations (0.70 ml normal saline+0.3 ml deionized water, 0.60 ml normal saline+0.40 ml deionized water and 0.55 ml normal saline+0.45 ml deionized water) were prepared with normal saline and deionized water, in which the red blood cells were suspended, and the residual red blood cells were detected by flow cytometer.</p><p><b>RESULTS</b>There was no significant difference in percentage of residual red blood cells between different time points detected by flow cytometer in 3 different hypotonic salt solutions. The percentage of residual red blood cells in B+C+D+E+F+G detected time region was different among 3 NaCl dilution groups. The percentage of residual red blood cells in normal control was lower than that in hemoglobinopathy group. The percentage of residual red blood cells in hereditary spherocytosis (HS) group was obviously lower than that in hemoglobinopathy and normal control groups. The comparison of 3 different dilution concentrations found that the second concentration (0.60 ml normal saline+0.40 ml deionized water) is more suitable to screen HS by FC500 flow cytometer.</p><p><b>CONCLUSION</b>The detection of red cell osmotic fragility by using flow cytometry is a simple, rapid, objective and economic way that can be an effective screening method for diagnose the HS.</p>
Subject(s)
Humans , Erythrocytes , Cell Biology , Flow Cytometry , Osmotic Fragility , Spherocytosis, HereditaryABSTRACT
Hydrogen sulfide (HS) contributes to visceral hyperalgesia in primary sensory neurons, but its role in central nervous system remains largely unknown. This study was to investigate the roles and underlying mechanisms of HS and its endogenous synthesis enzymes in the arcuate nucleus (ARC) in rat pancreatic hyperalgesia. Chronic pancreatitis (CP) was induced in male adult Sprague-Dawley rats by intra-pancreatic ductal injection of trinitrobenzene sulfonic acid (TNBS). Abdominal hyperalgesia was assessed by referred somatic behaviors to mechanical stimulation of rat abdomen. Western blot analysis was performed to detect protein expression in the ARC. CP markedly upregulated cystathionine β-synthetase (CBS) expression but did not alter cystathionine-γ-lyase level in the ARC at 4 weeks after TNBS injection. Although the expression of total GluN2B was not altered, CP greatly enhanced the phosphorylation level of GluN2B in the ARC when compared with age- and sex-matched control rats. CP also significantly increased expression of protein kinase Cγ (PKCγ) in the ARC. Arcuate microinjection of O-(Carboxymethyl) hydroxylamine hemihydrochloride (AOAA, an inhibitor of CBS) significantly attenuated abdominal pain in CP rats in a dose-dependent manner and reversed the CP-induced upregulation of p-GluN2B and PKCγ in the ARC. Furthermore, the GluN2B inhibitor or specific PKC inhibitor chelerythrine significantly attenuated abdominal hyperalgesia in CP rats. The p-GluN2B expression was also suppressed by PKC inhibitor. Taken together, our results suggest that the upregulation of CBS in the ARC leads to an activation of GluN2B via PKCγ, which may play an important role in generation of pain hypersensitivity of CP.
Subject(s)
Animals , Male , Rats , Acute Disease , Arcuate Nucleus of Hypothalamus , Cystathionine beta-Synthase , Hyperalgesia , Pain , Pancreatitis, Chronic , Phosphorylation , Protein Kinase C , Rats, Sprague-Dawley , Receptors, N-Methyl-D-Aspartate , Up-RegulationABSTRACT
BACKGROUND/AIMS: This study was to investigate whether transforming growth factor-β1 (TGF-β1) plays a role in hyperalgesia in chronic pancreatitis (CP) and the underlying mechanisms. METHODS: CP was induced in male adult rats by intraductal injection of trinitrobenzene sulfonic acid (TNBS). Abdominal hyperalgesia was assessed by referred somatic behaviors to mechanical stimulation of rat abdomen. Dil dye injected into the pancreas was used to label pancreas-specific dorsal root ganglion (DRG) neurons. Whole cell patch clamp recordings and calcium imaging were performed to examine the effect of TGF-β1 on acutely isolated pancreas-specific DRG neurons. Western blot analysis was carried out to measure the expression of TGF-β1 and its receptors. RESULTS: TNBS injection significantly upregulated expression of TGF-β1 in the pancreas and DRGs, and TGF-β1 receptors in DRGs (T9-T13) in CP rats. Intrathecal injection of TGF-β receptor I antagonist SB431542 attenuated abdominal hyperalgesia in CP rats. TGF-β1 application depolarized the membrane potential and caused firing activity of DRG neurons. TGF-β1 application also reduced rheobase, hyperpolarized action potential threshold, and increased numbers of action potentials evoked by current injection of pancreas-specific DRG neurons. TGF-β1 application also increased the concentration of intracellular calcium of DRG neurons, which was inhibited by SB431542. Furthermore, intrathecal injection of TGF-β1 produced abdominal hyperalgesia in healthy rats. CONCLUSIONS: These results suggest that TGF-β1 enhances neuronal excitability and increases the concentration of intracellular calcium. TGF-β1 and its receptors are involved in abdominal hyperalgesia in CP. This and future study might identify a potentially novel target for the treatment of abdominal pain in CP.
Subject(s)
Adult , Animals , Humans , Male , Rats , Abdomen , Abdominal Pain , Action Potentials , Blotting, Western , Calcium , Diagnosis-Related Groups , Fires , Ganglia, Spinal , Hyperalgesia , Injections, Spinal , Membrane Potentials , Neurons , Pancreas , Pancreatitis, ChronicABSTRACT
The polysaccharides from pumpkin fruit (PP) were obtained and purified by hot-water extraction, anion-exchange chromatography, and gel column chromatography. The physicochemical properties of PP were determined by gel filtration chromatography, gas chromatography, fourier transform infrared (FTIR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Results indicated that the molecular weight of PP was about 23 kDa and PP was composed of D-Arabinose, D-Mannose, D-Glucose, and D-Galactose with a molar ratio of 1 : 7.79 : 70.32 : 7.05. FTIR and NMR spectra indicated that PP was the polysaccharide containing pyranose ring. Additionally, PP protected islets cells from streptozotocin (STZ) injury in vitro via increasing the levels of super-oxide dismutase (SOD) and malondialdehyde (MDA) and reducing the production of NO. The experiment of reverse transcriptase-polymerase chain reaction further proved that PP inhibited apoptosis via modulating the expression of Bax/Bcl-2 in STZ-damaged islet cells. In conclusion, PP could be explored as a novel agent for the treatment of diabetes mellitus.
Subject(s)
Animals , Rats , Apoptosis , Chromatography, Gas , Chromatography, Gel , Cucurbita , Chemistry , Diabetes Mellitus, Experimental , Drug Therapy , Islets of Langerhans , Wounds and Injuries , Magnetic Resonance Spectroscopy , Malondialdehyde , Molecular Weight , Monosaccharides , Nitric Oxide , Polysaccharides , Chemistry , Pharmacology , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Spectroscopy, Fourier Transform Infrared , Superoxide Dismutase , bcl-2-Associated X ProteinABSTRACT
Objective: To investigate the suppression effect of mouse marrow-derived endothelial progenitor cells (EPCs) modified by cytosine deaminase (CD) gene and enzyme prodrug 5-fluorocytosine (5-Fc) on the growth of hepatocellar carcinoma in orthotopic H22 cell-transplanted mice. Methods: The EPCs were transfected by lentiviral vector carrying CD gene (plenti6.3-EGFP-CD) using Polybrene technique. The mouse hepatoma H22 cells were treated by the supernatant of EPCs carrying CD gene and 5-Fc, and then the number and morphology of the cells were observed under an inverted microscope. C57BL/6 mice bearing orthotopic transplanted H22 hepatocellar carcinoma were treated by EPCs carrying CD gene and 5-Fc. The volume of tumor in mice was monitored by magnetic resonance imaging, and the apoptosis in tumor was tested by terminal deoxynucleotidyl transferase-mediated dUTP nick and labeling (TUNEL) assay. The expression of CD protein in the transplanted tumor was identified by Western blotting. Results: The proliferation of H22 cells treated by the supernatant of EPCs carrying CD gene combined with 5-Fc was significantly reduced. Compared with the control, the tumor growth in the hepatocellular carcinoma orthotopic-transplantation mouse models treated by EPCs carrying CD gene and 5-Fc was inhibited to (47.29±5.81)% (P < 0.05), and the apoptosis index of tumor cells was markedly increased [(39.98±5.13)%]. Conclusion: The EPCs modified by CD gene combined with 5-Fc administration can effectively inhibit the proliferation of the mouse orthotopic transplanted hepatocellular carcinoma, and induce the apoptosis of tumor cells. Copyright © 2014 by TUMOR.
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N-acetyl-L-cysteine (NAC) capped quantum dots (QDs) were synthesized by a hydrothermal method and coated with 2-amino-2-deoxy-D-glucose (DG), polyethylene glycol (PEG), and 9-D-arginine (9R). The optical properties, morphology and structure of 9R/DG-coated CdTe QDs were characterized by ultraviolet-visible spectrometry, fluorescence spectrum, Fourier transform infrared (FTIR), proton nuclear magnetic resonance (1H NMR), liquid chromatography-mass spectrometer (LC-MS), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transmission electron micrographs (TEM). Furthermore, the biocompatibility, tumor targeted ability and transmembrane action of 9R/DG-coated CdTe QDs were studied. Results indicated that 9R/DG-coated CdTe QDs was constructed successfully by ligand exchange. The 9R/DG-coated CdTe QDs with the size of 8-10 nm had good dispersity and the absorbance and fluorescence peaks of CdTe QDs after modification were red shifted from 480 nm to 510 nm and 627 nm to 659 nm, respectively. In addition, the CdTe QDs modified by PEG, DG and 9R displayed good biocompatibility, high targeted ability to the cancer cells with glucose transporter type 1 (GLUT1) receptor high expression and obvious transmembrane ability.
Subject(s)
Humans , Acetylcysteine , Chemistry , Cadmium Compounds , Pharmacology , Neoplasms , Drug Therapy , Polymers , Chemistry , Quantum Dots , Chemistry , Spectrophotometry, Ultraviolet , Tellurium , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To determine the long-term testicular effect after neonatal exposure to 2,2', 4,4',5,5'-hexa-chlorobiphenyl (PCB153).</p><p><b>METHODS</b>On birth day (Postnatal day 0, PNDO), the Sprague-Dawley (SD) male rats were mixed together and divided into 12 pups/litter. At PND1, the rats were grouped randomly into control and treatment groups according to different litters, 24 pups/group. They were treated by oral gavage with PCB153 in corn oil at doses of 0, 0.025, 0.250 and 2.500 mg/kg BW-day from PNDI to PND7. The rats were sacrificed at PND8 and PND90 by anesthesia. The testes were collected and weighed for histological examination and daily sperm production at PND8 or/and PND90. The epididymidis and the epididymidis cauda also were collected and weighed for determination the sperm counts at PND90.</p><p><b>RESULTS</b>The body weight of 2.500 mg/kg dose group was decreased significantly from PND3 to PND8 compared with that of control (P < 0.05). At PND8, the loose structure in seminiferous cord and the spermatogonia with enlarged volume and detached from the cord were observed in 2.500 mg/kg dose group by light microscope and electronic microscopy. With the increase of exposure doses, the testicular daily sperm production (DSP) and the sperm counts of epididymidis cauda were decreased in dose-dependent manner at PND90. The DSP in 0.250 mg/kg [30 x 10(6)/testis(g)] and 2.500 mg/kg [18 x l0(6)/testis(g)] dose groups were significantly reduced compared with that of control [36 x 10(6)/testis(g)] (P < 0.05). And there was a significant reduction in the sperm counts of epididymidis cauda in 0.250 mg/kg [42 x 10(7)/epididymidis cauda (g)] and 2.500 mg/kg [18 x 10(7)/epididymidis cauda (g)] dose groups compared with that of control [51 x 10(7)/epididymidis cauda (g)] (P < 0.05).</p><p><b>CONCLUSIONS</b>The spermatogenesis of adult testis is disturbed, which causes the decrease in the testicular DSP and the sperm counts of epididymidis cauda after neonatal exposure to PCB153. The long-term damage in male reproductive function is caused by neonatal exposure to chemicals.</p>
Subject(s)
Animals , Male , Rats , Animals, Newborn , Environmental Exposure , Polychlorinated Biphenyls , Toxicity , Rats, Sprague-Dawley , Sperm Count , Spermatogenesis , Testis , PathologyABSTRACT
<p><b>OBJECTIVE</b>To study the serrated lesions of colon and to compare the malignant potential between traditional serrated adenomas (TSA) and conventional adenomas (CAD).</p><p><b>METHODS</b>A total of 5347 cases of colorectal polyps encountered in five regional hospitals during a five-year period were retrospectively reviewed. The serrated lesions were classified on the basis of histologic examination. One hundred and eighty-seven cases of CAD (including 160 cases of tubular adenoma and 27 cases of villous adenoma) and 36 cases of invasive adenocarcinoma were randomly selected as the controls. The degree of dysplasia and expressions of Ki-67, p53 and beta-catenin in TSA and CAD were compared.</p><p><b>RESULTS</b>Amongst the 5347 colorectal polyps studied, 258 cases (4.8%) of serrated lesions were found, which included 112 cases (43.4%, 112/258) of hyperplastic polyp, 78 cases (30.2%, 78/258) of TSA and 26 cases (10.1%, 26/258) of sessile serrated adenoma. Sixty-two cases of TSA were identified from 3 hospitals, in which moderate dysplasia was found in 13 cases. High-grade intraepithelial neoplasia and ICA were found in 6 cases (9.6%). Compared with the 187 cases of CAD, moderate dysplasia were found in 27 cases and high-grade intraepithelial neoplasia and invasive adenocarcinoma were found in 25 cases (13.3%, χ(2) = 19.373, P = 0.000). There was statistically significant difference between TSA and CAD in the degree of dysphasia. The expression of Ki-67, p53 and beta-catenin in TSA and CAD showed no significant difference (P > 0.05).</p><p><b>CONCLUSIONS</b>The incidence of serrated lesions is lower in northern Chinese population than that in Caucasians. TSA has obvious malignant potential; but the rate associated with high-grade intraepithelial neoplasia and invasive adenocarcinoma is lower than that in CAD.</p>
Subject(s)
Humans , Adenocarcinoma , Metabolism , Pathology , Adenoma , Classification , Metabolism , Pathology , Adenoma, Villous , Classification , Metabolism , Pathology , Cell Transformation, Neoplastic , Pathology , Colonic Polyps , Metabolism , Pathology , Colorectal Neoplasms , Classification , Metabolism , Pathology , Intestinal Polyps , Metabolism , Pathology , Ki-67 Antigen , Metabolism , Precancerous Conditions , Metabolism , Pathology , Rectum , Pathology , Retrospective Studies , Tumor Suppressor Protein p53 , Metabolism , beta Catenin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To screen and identify differentially expressed genes in the dorsal root ganglion (DRG) in early experimental diabetic rats.</p><p><b>METHODS</b>Diabetic model rats were induced by single intraperitoneal injection of streptozotocin (STZ). At the second week after STZ injection, the sensory nerve conduction velocities (SNCV) of sciatic nerve were measured as an indicator of neuropathy. The technique of silver-staining mRNA differential display polymerase chain reaction (DD-PCR) was used to detect the levels of differentially expressed genes in rat DRG. The cDNA fragments that displayed differentially were identified by reverse-hybridization, cloned and sequenced subsequently, and then confirmed by Northern blot.</p><p><b>RESULTS</b>The SNCV in the diabetic model group [n = 9, (45.25+/-10.38) m/s] reduced obviously compared with the control group [n = 8, (60.10+/-11.92) m/s] (P < 0.05). Seven distinct cDNA clones, one was up-regulated gene and the others were down-regulated ones, were isolated by silver-staining mRNA differential display method and confirmed by Northern blot. According to the results of sequence alignment with GenBank data, majority of the clones had no significant sequence similarity to previously reported genes except only one that showed high homology to 6-pyruvoyl-tetrahydropterin synthase mRNA (accession No. BC059140), which had not been reported to relate to diabetic neuropathy.</p><p><b>CONCLUSION</b>These differentially expressed genes in the diabetic DRG may contribute to the pathogenesis of diabetic peripheral neuropathy.</p>
Subject(s)
Animals , Male , Rats , Blotting, Northern , Diabetes Mellitus, Experimental , Genetics , Diabetic Neuropathies , Genetics , Ganglia, Spinal , Gene Expression , Gene Expression Profiling , RNA, Messenger , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Sciatic NerveABSTRACT
<p><b>OBJECTIVE</b>To investigate the influence of the materials commonly used in the assisted reproduction procedure on human germ cells and embryo development.</p><p><b>METHODS</b>We used human sperm survival assay to detect the influence of two brands of culture dishes, two brands of injection needles, washed and unwashed ovum aspiration needles, embryo transfer catheters and surgical gloves on sperm motility. All the data obtained went through variance analysis with SPSS 13.0.</p><p><b>RESULTS</b>Sperm motility differed significantly between Nunclons and Falcon's culture dishes (52.68 +/-16.21 vs 45.36 +/- 15.25, P < 0.01) but not between the BD and the Jie-rui injection needles (P > 0.05), nor among the washed and unwashed ovum aspiration needles, embryo transfer catheters and surgical gloves (P > 0.05).</p><p><b>CONCLUSION</b>Different brands of assisted reproduction materials of similar use had different or similar influences on germ cells and embryos, while no difference existed in the influences of the washed and unwashed ovum aspiration needles, embryo transfer catheters and surgical gloves.</p>
Subject(s)
Humans , Male , Culture Media, Conditioned , Embryonic Development , Reproductive Techniques, Assisted , Sperm MotilityABSTRACT
<p><b>OBJECTIVE</b>To investigate whether two polymorphism sites of the G protein-coupled receptor 154 gene (GPR154) are associated with asthma in Han nationality of Hubei province in China.</p><p><b>METHODS</b>The polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 145 cases of allergic asthma and 120 healthy controls.</p><p><b>RESULTS</b>(1)The genotype frequencies of SNP563704 were 0.324 for CC, 0.524 for CT, 0.152 for TT in allergic asthma patients in a Chinese population. The genotype frequencies of SNP522363 were 0.289 for CC, 0.521 for CG, 0.190 for GG in allergic asthma patients in a Chinese population. (2)No significant differences in the distributions of SNP563704 and SNP522363 polymorphisms were found between allergic asthma and healthy control subjects (chi square is 1.880, P> 0.05; chi square is 0.700, P> 0.05, respectively). (3)No significant difference in serum total IgE was found between patients with allergic asthma with different genotypes of SNP563704 and SNP522363 (F is 0.714, P> 0.05; F is 0.083, P> 0.05, respectively). (4) The distribution of frequencies of 4 haplotypes showed significant difference(chi square is 16.50,P< 0.01). The haplotype frequencies of CT and GT were remarkably higher in asthma subjects than those in controls.</p><p><b>CONCLUSION</b>The polymorphisms of SNP563704 and SNP522363 are not associated with allergic asthma in Han nationality in Hubei Chinese population. However, the haplotypes are associated with allergic asthma.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Asian People , Genetics , Asthma , Genetics , China , Gene Frequency , Genetic Predisposition to Disease , Genetics , Genotype , Haplotypes , Genetics , Polymerase Chain Reaction , Polymorphism, Single Nucleotide , Receptors, G-Protein-Coupled , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To compare the prior coronary angiography (CAG) psychological status in chest pain patients with and without coronary artery disease (CAD).</p><p><b>METHODS</b>Ninety-nine patients with chest pain and scheduled for CAG were selected by cluster sampling method. The mental status was measured by Hamilton Anxiety Rating Scale (HAMA) and Hamilton Depression Rating Scale-17 (HAMD-17) 24 hours before CAG, and the risk factors for CAD were also determined.</p><p><b>RESULTS</b>There were 43 patients with HAMA score > or = 14, 18 patients with HAMD-17 score > or = 14 and 16 patients with both scores > or = 14. CAD was diagnosed in 46 patients by CAG. HAMA score was significantly higher in patients without CAD than patients with CAD (14.1 +/- 7.1 vs. 11.1 +/- 6.7, P < 0.05).</p><p><b>CONCLUSIONS</b>Incidences of anxiety and depression were high in chest pain patients prior CAG and incidence of anxiety prior CAG was significantly higher in chest pain patients without CAD compared to chest pain patients with CAD.</p>