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Objective:To investigate the correlation of serum pro-gastrin-releasing peptide (ProGRP), sugar chain antigen 242(CA242), procalcitonin(PCT) levels in patients with chronic atrophic gastritis (CAG) with intestinal metaplasia and Helicobacter pylori(Hp) infection and prognosis.Methods:One hundred patients with CAG intestinal metaplasia in Emergency General Hospital were selected as the research objects, and they were divided into infection group (75 cases) and non-infected group (25 cases) according to whether they had Hp infection. The clinical data, levels of serum ProGRP, CA242, and PCT were compared between the two groups, and the diagnostic value of the combination of serum indicators in the diagnosis of Hp infection in CAG intestinal metaplasia patients and their correlation with prognosis were analyzed.Results:The degree of atrophy and intestinal metaplasia in the infected group were higher than those in the non-infected group ( P<0.05). The levels of serum ProGRP, CA242 and PCT in the infected group were higher than those in the non-infected group: (159.41 ± 42.38) ng/L vs. (105.84 ± 18.29) ng/L, (7.24 ± 2.28) kU/L vs. (4.12 ± 1.30) kU/L, (3.84 ± 1.12)μg/L vs. (2.57 ± 0.82) μg/L, there were statistical differences ( P<0.05). The results of Spearman analysis showed that the levels of ProGRP, CA242, PCT had positive correlation with atrophy degree ( r = 0.614, 0.629, 0.672, P<0.05), and had positive correlation with intestinal metaplasia degree ( r = 0.574, 0.591, 0.603, P<0.05). The area under the curve (AUC) for the combined diagnosis of Hp infection in patients with CAG intestinal metaplasia by serum ProGRP, CA242, and PCT was 0.874 (95% CI 0.793 - 0.932), the diagnostic sensitivity and specificity were 76.00% and 92.00%, respectively. The incidence of gastric neoplasms in patients with Hp positive combined diagnosis of serum ProGRP, CA242, and PCT within 2 years (11.86%) was higher than that of negative patients (0), but the difference was not statistically significant ( P>0.05). Conclusions:The levels of serum ProGRP, CA242 and PCT in patients with CAG intestinal metaplasia are closely related to Hp infection. The combination of various indicators has high application value in the diagnosis of Hp infection.
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The National Medical Products Administration has authorized sodium oligomannate for treating mild-to-moderate Alzheimer's disease.In this study,an LC-MS/MS method was developed and validated to quantitate sodium oligomannate in different biomatrices.The plasma pharmacokinetics,tissue distri-bution,and excretion of sodium oligomannate in Sprague-Dawley rats and beagle dogs were system-atically investigated.Despite its complicated structural composition,the absorption,distribution,metabolism,and excretion profiles of the oligosaccharides in sodium oligomannate of different sizes and terminal derivatives were indiscriminate.Sodium oligomannate mainly crossed the gastrointestinal epithelium through paracellular transport following oral administration,with very low oral bioavail-ability in rats(0.6％-1.6％)and dogs(4.5％-9.3％).Absorbed sodium oligomannate mainly resided in circulating body fluids in free form with minimal distribution into erythrocytes and major tissues.So-dium oligomannate could penetrate the blood-cerebrospinal fluid(CSF)barrier of rats,showing a con-stant area under the concentration-time curve ratio(CSF/plasma)of approximately 5％.The cumulative urinary excretion of sodium oligomannate was commensurate with its oral bioavailability,supporting that excretion was predominantly renal,whereas no obvious biliary secretion was observed following a single oral dose to bile duct-cannulated rats.Moreover,only 33.7％(male)and 26.3％(female)of the oral dose were recovered in the rat excreta within 96 h following a single oral administration,suggesting that the intestinal flora may have ingested a portion of unabsorbed sodium oligomannate as a nutrient.
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DNA polymerases are widely used in PCR and play important roles in life science research and related fields. Development of high-performance DNA polymerases is of great commercial interest as the current commercial DNA polymerases could not fully satisfy the requirements of scientific research. In this study, we cloned and expressed a family B DNA polymerase (NCBI accession number TEU_RS04875) from Thermococcus eurythermalis A501, characterized its enzymatic property and evaluated its application in PCR. The recombinant Teu-PolB was expressed in E. coli and purified with affinity chromatography and ion-exchange chromatography. The enzymatic properties of Teu-PolB were characterized using fluorescence-labeled oligonucleotides as substrates. The application potential of Teu-PolB in PCR was evaluated using the phage λ genomic DNA as a template. Teu-PolB has DNA polymerase and 3'→5' exonuclease activities, and is highly thermostable with a half-life of 2 h at 98 ℃. The most suitable PCR buffer is consisted of 50 mmol/L Tris-HCl pH 8.0, 2.5 mmol/L MgCl2, 60 mmol/L KCl, 10 mmol/L (NH4)2SO4, 0.015% Triton X-100 and 0.01% BSA, and the optimal extension temperature is 68 ℃. Under the optimized conditions, a 4 kb target fragment was successfully amplified with an extension rate of 2 kb/min. The yield of the Teu-PolB amplified-DNA was lower than that of Taq DNA polymerase, but its extension rate and fidelity was higher than that of Taq and Pfu DNA polymerases. The biochemical properties of Teu-PolB demonstrate that this enzyme can be used in PCR amplification with high thermostability, good salt tolerance, high extension rate and high fidelity.
Subject(s)
DNA-Directed DNA Polymerase/genetics , Escherichia coli/genetics , Polymerase Chain Reaction/methods , Temperature , Thermococcus/geneticsABSTRACT
Objective: To systemically investigate the volatile organic compounds (VOCs) of Viola yedoensis, and to compare the VOCs differences of V. yedoensis obtained by the needle trap, static headspace and hydrostillation methods. Method: The needle trap, static headspace and hydrostillation methods coupled with gas chromatography-mass spectrometer (GC-MS) were developed for isolation and identification of the VOCs in V. yedoensis. The relative content of each component was obtained by peak area normalization with a triple-bed needle packed with Tenax, Carbopack X and Carboxen 1000 sorbents. Result: The 112 compounds were trapped by using needle trap, mainly moderate volatile components, including aldehydes, ketones, alcohols, monoterpenes, sesquiterpenoids and aromatic compounds. The static headspace and hydrodistillation methods were allowed to obtain 37 (mainly the high-volatile components) and 78 compounds (mainly low-volatile components), respectively. Only 13 common volatile components were detected in all these three methods. Conclusion: The results clearly demonstrated that the needle trap method is an alternative method for sampling VOCs of herbs, characterized by fast analysis, simple operation, good enrichment effect and high sensitivity.These three methods for VOCs analysis are complementary for each other.
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Aim To establish human U87-MG glioma model in nude mice brain and to observe the characteristics of the tumor growth. Methods Human U87-MG glioma cells were cultured in vitro. 5 μL of cell suspension containing 3.0 ×1010·L-1, 4.0×1010·L-1and 5.0×1010·L-1respectively was inocula-ted into the right caudate nucleus of 18 male nude mice brain un-der the guidance of stereotaxic apparatus, separately, whereas another 6 nude mice as the control group, were inoculated into the same volume of Hanks solution. The moving and survival state of rats with gliomas were observed. The examinations of the tumors formation, volumes, metastasis and histopathology were performed and the obtained brain samples were stained with HE and immunohistochemistry. Results All the tested rats of dif-ferent inoculation doses developed brain tumors without extracra-nial metastasis. The mean survival time of three groups was (46.50 ± 3.27) d,(38.50 ± 3.28) d and (30.67 ± 3.51) d,respectively. The tumors showed the similar morphological fea-tures and immunophenotype to human glioma. There was positive expression of GFAP and S-100 in the tumors. Conclusions The orthotopic implantation model of human U87-MG glioma, by in-oculating quantitative U87-MG cells stereotaxically into the brains of the nude mice, is successfully established with 100 yield of intracranial tumor and no extracranial growth extension. It resembles the histopathological and morphological features of human glioma,which can be used as a reliable animal model for the study of the tumorigenesis, pathogenesis, biological charac-teristics and therapy of glioma.
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Objective:To explore the effects of Bufalin on the growth and proliferation of human glioma cells U251. Methods:Methyl thiazolyl tetrazolium(MTT)assay was used to detect the effect of Bufalin on the proliferation of human glioma cells U251. An in-verted microscope was used to observe the changes of cell number,morphology and activity. AnnexinV/ PI was used to measure the in-duction of cell apoptosis caused by Bufalin. Results:Bufalin at different concentrations(0. 001 - 10. 0μmol·L - 1 )inhibited the pro-liferation of U251 cells in a dose and time-dependent manner. Compared with that of the control group,the apoptosis rate of Bufalin group was increased significantly(P < 0. 01). Conclusion:Bufalin can inhibit the growth and proliferation of U251 cells in a dose and time-dependent manner,and induce the apoptosis of U251 cells.
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<p><b>OBJECTIVE</b>To investigate the expression and clinical pathological significance of EB virus (Epstein-Barr virus, EBV), PTEN and VEGF in angioimmunoblastic T -cell lymphoma (AITL).</p><p><b>METHODS</b>The EBV -encoded small RNA (EBER) expression in 21 cases of AITL was detected by in situ hybridization. The expressions of PTEN and VEGF were detected in 21 cases of AITL and 20 cases of lymph node reactive hyperplasia by immunohistochemical EnVision two-steps method. The expression and clinicopathological significance of EBV, PTEN and VEGF in AITL were analyzed.</p><p><b>RESULTS</b>The positive expression rate of EBER in 21 cases of AITL was 61.9%; the expressions of PTEN and VEGF in AITL and lymph node reactive hyperplasia were significantly different (P<0.05). The expressions of EBER and PTEN negatively correlated (P<0.05). The EBER positive expression rates of male patients in AITL group and the progressed group was 80% and 78.6% respectively, which were significantly higher than that in female patients and patients in non- advanced group (P<0.05); the PTEN expression rates in the AITL group accompanying B symptoms and progressed group were 31.3% and 21.4%, respectively, which were significantly lower than those in patients without B symptoms and non-progressed group (P<0.05). Survival analysis showed that the PTEN expression negatively correlated with the overall survival rate of patients (P<0.05).</p><p><b>CONCLUSION</b>EBV infection and low expression of PTEN may indicate the deterioration of angioimmunoblastic T-cell lymphoma. Whether the EBV involved in the ocurring of T-cell angioimmunoblastic lymphoma by down-regulating PTEN expression is unclear, further research is needed.</p>
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Female , Humans , Male , Epstein-Barr Virus Infections , Herpesvirus 4, Human , Immunoblastic Lymphadenopathy , In Situ Hybridization , Lymphoma, T-Cell , PTEN Phosphohydrolase , Survival Analysis , Survival Rate , Vascular Endothelial Growth Factor AABSTRACT
<p><b>OBJECTIVE</b>To investigate the renal protective activity of Hsian-tsao Mesona procumbens Hemsl. water extracts in diabetic rats.</p><p><b>METHODS</b>Thirty Sprague-dawley female rats were randomly divided into three groups (n = 10 each), "control group" with intraperitoneal saline injection, "diabetic group" with 60 mg of intraperitoneal streptozotocin injection per kg of body weight and "Hsian-tsao group" with intragastric administration of Hsian-tsao extraction everyday for 4 weeks after intraperitoneal streptozotocin injection. The body weight and blood sugar were measured before and after model induction in the three groups. Thrombospondin-1 (TSP-1) expressions in the kidney were monitored by immunohistochemistry. Kidney ultrastructural changes were also analyzed by using transmission electron microscopy.</p><p><b>RESULTS</b>Before diabetic model induction, there were no significant differences among the three groups in body weight and blood sugar. Four weeks after the induction of diabetes, the differences became statistically significant. Electron microscopy also revealed disruption of the foot processes of the podocytes and other damages in diabetic group. These damages were significantly less severe in Hsian-tsao group when compared with the diabetic group. TSP-1 expressions in the kidney were significantly increased in both the diabetic group and Hsian-tsao group, but it was relatively lower in Hsian-tsao group than in diabetic group.</p><p><b>CONCLUSION</b>Our results showed that Hsian-tsao treatment in the diabetic rats effectively prevented the pathological alterations in the kidney and decreased the TSP-1 expression. It was suggested that Hsian-tsao had protective effect on the kidneys of the diabetic rats.</p>
Subject(s)
Animals , Female , Rats , Diabetes Mellitus, Experimental , Drug Therapy , Metabolism , Pathology , Diabetic Nephropathies , Metabolism , Pathology , Drugs, Chinese Herbal , Therapeutic Uses , Kidney , Metabolism , Pathology , Lamiaceae , Chemistry , Rats, Sprague-Dawley , Thrombospondin 1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To describe a HPLC method for assessing betaine in Fufang Guilu granule.</p><p><b>METHOD</b>The content of betainephenaxcyl bromide in Fufang Guilu granule was determined by HPLC. The analytical column was a shim-pack CLC-ODS (6.0 mm x 150 mm) filling a 5 microm stationary phase; The mobile phase consisted of acetonitrile-water(35:65) with 0.1 mol x L(-1) NaClO4; The flow-rate was 1 mL x min (-1); The detector was set at 254 nm.</p><p><b>RESULT</b>The calibration curve was linear over the range of 0.09-0.585 microg (r = 0.9997). The average recovery of the method was 98.4%, RSD 2.5% (n = 5).</p><p><b>CONCLUSION</b>The results showed that this method was reliable and accurate, and can be used for quality control of Fufang Guilu granule.</p>
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Betaine , Chromatography, High Pressure Liquid , Methods , Codonopsis , Chemistry , Drug Combinations , Drugs, Chinese Herbal , Chemistry , Lycium , Chemistry , Plants, Medicinal , Chemistry , Quality Control , Reproducibility of ResultsABSTRACT
Objective To investigate the correlation between ankle-brachial index and ultrasonography in lower ex- tremities vascular in type 2 diabetic patients.Methods 218 type 2 diabetic patients were enrolled,and ankle-brachial index(ABI)measurements and uhrasonography in lower extremities vascular were performed in all patients.Lower extremi- ties vascular disease(LEVD)were divided into six species according to the uhrasonography results and scored in accord- ante with its serious degree,then investigate the correlation between ABI and uhrasonography results score.All patients were divided into deferent groups according to uhrasonography results and ABI range,and then compared their deference. Results The lower the ABI,the more serious of the LEVD showed by ultrasonography.ABI and ultrasonic inspection score was significantly negatively correlated(r=-0.65,P
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Objective To study a new clinicopathological subtype of primary cutaneous T-cell lym phoma (PCTCL). Methods A case of T-cell lymphoma was systematically evaluated clinically and by using H-E staining, special staining,immunohistochemical staining,gene rearrangement and PCR.Results The skin lesion presented as tender nodules with mucocele. Skin biopsies showed that in the dermis and subcutaneous tissue,most of the angiotropic tumor cells were small T cells; no obvious epidermotropic phenomenon was detected.A few vessels were observed with obvious fibromucinous matrix formation. Immunohislochemical studies showed the following:CD3(+),CD43(+),CD45RO(+),CD56(a few),CD68(-), CD79?(-),CD20 (-), CD30(-), CD117(-), ALK(-), S-100(-),CD45R(-),EMA(-),SMA(-).The mucoid matrix was positive for Alcian blue staining.The rearrangement of T-cell ? receptor gene was detected.EBV was not detected with PCR.Conclusion Fibromucinous T-cell lymphoma rich in blood vessels is a new and distinct variant of PCTCL; it is not a subtype of mycosis fungoides.