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Objective:To investigate the effects of riboflavin-ultraviolet A scleral collagen cross-linking on the retina under different irradiation time, and to determine the safe irradiation time.Methods:Sixty healthy New Zealand white rabbits were randomly divided into control group (0 minute group), 10 minutes group, 20 minutes group, 30 minutes group and 40 minutes irradiation group according to the irradiation time, with 12 rabbits in each group.The left eye was irradiated with riboflavin-ultraviolet A scleral collagen (370 nm, 10 mW/cm 2). The histopathological change of retina was observed by light microscope and transmission electron microscope and compared among different groups.The concentration of MDA and the activities of SOD, CAT and GSH-Px in retinal tissue were detected by corresponding kits.The expression levels of SOD and CAT proteins in retinal tissue were detected by Western blot method.The study protocol was approved by the Binzhou Medical University Laboratory Animal Ethical Committee (No.2017-80). The use and care of animals complied with the statement of ARVO and the Regulation on the Management of Laboratory Animal Quality of China. Results:Under the light microscope, the structure of the retinas in the control group was orderly arranged.Under the transmission electron microscope, the lamellar structure in the inner segment and the mitochondrial structure in the outer segment of the photoreceptor cells were intact, and the mitochondrial ridge was continuous in the control group.There was no obvious difference in retinal morphology between the 10 minutes irradiation group and the control group under both the light microscope and the transmission electron microscope, and the retinal damage became more severe with the prolongation of irradiation time.The concentration of MDA in the retina of each group was elevated gradually with the increase of irradiation time, and the difference was statistically significant ( F=65.25, P<0.05). The concentration of MDA was (11.31±1.84), (14.94±1.04)and (18.25±1.42)nmol/mgprot in the 20 minutes, 30 minutes and 40 minutes irradiation groups respectively, which were significantly higher than (1.13±0.02)nmol/mgprot in the control group (all at P<0.05). The MDA concentration in 20 minutes, 30 minutes and 40 minutes irradiation groups was increased successively, showing statistically significant differences (all at P<0.05). With the prolongation of irradiation time, the activities of SOD, CAT and GSH-Px as well as the expression levels of SOD and CAT proteins were significantly decreased gradually ( F=44.09, 34.18, 35.60, 115.75, 78.86; all at P<0.05). The differences between the control group and 20 minutes, 30 minutes, 40 minutes irradiation groups, and the differences among 20 minutes, 30 minutes, 40 minutes irradiation groups were statistically significant (all at P<0.05). Conclusions:Riboflavin-ultraviolet A 10 mW/cm 2 scleral collagen cross-linking irradiation for 10 minutes is safe.Excessive irradiation time can cause damage to the retina of rabits.
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Objective:To optimize the matrix formulation of Erhuang analgesic gels. Methods:Central composite design-response surface methodology was adopted to optimize the best formulation of Erhuang analgesic gels by using carbomer 940, triethanolamine and glycerine as independent variables, the appearance, stability, viscosity and in vitro release of berberine hydrochloride as comprehensive evaluation indices. Results:The fitting regressing equation was Y= 82.25 + 4.95 A+ 5.19 B + 1.41 C+ 1.51 AB + 0.904 0 AC- 0.531 9 BC- 2.92 A2-1.80 B2-0.182 1 C2. P value of the model was less than 0.000 1, and the correlation coefficient r value was 0.977. The optimal formulation of Erhuang analgesic gels consisted of 1.84% carbomer 940, 1.30 times triethanolamine of carbomer 940 and 13.99 grams of glycerine. The average comprehensive scores of three verification experiments was 88.56, and the deviations from the predicted values were 2.93%, 2.85% and 1.55%. Conclusion:The formulation process by central composite design-response surface methodology was stable and the formulation of Erhuang analgesic gels has been optimized.
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Objective@#To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters.@*Methods@#By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used.@*Results@#The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889.@*Conclusions@#The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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Objective:To set up a method to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract and to measure their physico-chemical parameters. Methods:By using HPLC analytical methods to measure the content of baicaline, wogonoside, and paeonol in Jiedu-Zhixue Decoction extract. The RP18 column was adopted, gradient eluted with mobile phase of acetonitrile-1% phosphoric acid water, flow rate was 1 ml/min, detection wavelength was 274 nm, and the column temperature was 30 ℃. The method quantitatively analyze the compound extracts of baicaline, wogonoside and paeonol were used. Results:The method isrepeatable, stable with good recorey rates. The calibration curves of baicaline was in good linearity over the range of 0.191 2-1.147 2 μg. The calibration curves of wogonoside was in good linearity over the range of 0.041 5-0.207 4 μg. The calibration curves of paeonol was in good linearity over the range of 0.019 5-0.156 3 μg. There was no significant difference among the component of 3 different batches of Jiedu-Zhixue Decoction extract, and the pH value range was 5.240-5.753, relative density range was 0.846-0.889. Conclusions:The method used in this stydy to measure the effective components of Jiedu-Zhixue Decoction extract was stable and feasible, which is suitable for quality control of Jiedu-Zhixue Decoction.
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Objective To investigate the changes of metamorphopsia between before and after surgery in the patients with idiopathic epiretinal membrane and its influence factors.Methods A series cases observitional study included 39 eyes of 39 patients with idiopathic epiretinal membrane.Follow-up was carried out at 1 week before surgery and 3,6 months after surgery respectively.M-chart was used to quantify the severity of metamorphopsia (M-score).EDTRS visual chart was used to quantify best corrected visual acuity (BCVA) (converted to LogMAR).Central subfield thickness (CST),central foveal volumn (CV),cube average thickness (CAT),central foveal thichness (CFT),ganglion cell layer (GCL) thickness,inner nuclear layer (INL) thickness,outer nuclear layer (ONL) and outer plexiform layer (OPL) thickness,the integrity of external limiting membrane,ellipsoid zone and interdigitation zone were analyzed by using spectral domain-optical coherence tomography (OCT).This study protocol was approved by Ethic Committee of Beijing Tongren Hospital (No.TRECKY-012).Written informed consent was obtained from each subject before surgery.Results Mean M-score was significantly decreased from 0.8 (0.3,1.1) before surgery to 0.5 (0.2,0.8) at 3 months after surgery,with a significant difference between the two time points (Z=-2.013,P=0.044).Mean M-score was 0.6(0.2,0.8) at 6 months after surgery,which was not significantly different in comparison with before surgery and 3 months after surgery (Z =-1.873,P =0.061;Z =-0.288,P =0.773).Compared with before surgery,the horizontal M-score was significantly decreased 3 months and 6 months after surgery (Z =-2.329,P =0.020;Z =-2.858,P =0.004).No significant difference was found in vertical M-score among before surgery and 3,6 months after surgery (all at P＞0.05).The BCVA was improved from 0.40 (0.30,0.66) before surgery to 0.20 (0.06,0.42) 3 months after surgery and declined to 0.30 (0.10,0.52) at 6 months after surgery,and significant differences were obtained between 3 months after surgery and before surgery or 6 months after surgery (Z =-4.087,P＜0.001;Z =-2.235,P =0.025).Compared with before surgery,the BCVA in cataract combined with vitrectomy operative group was significantly improved in 3 months and 6 months after surgery (Z=-2.613,P=0.009;Z=-2.466,P=0.014) and the BCVA in only vitrectomy group was significantly improved at 3 months after surgery but decreased 6 months after surgery,showing significant differences between 3 months after surgery and before surgery or 6 months after surgery (Z =-3.104,P =0.002;Z =-3.464,P =0.001).Preoperative M-score was positively correlated with preoperative BCVA,preoperative CST or preoperative CFT (rs =0.384,P =0.016;rs =0.585,P＜0.001;rs =0.601,P＜0.001).No correlation was found between BCVA with GCL,INL or ONL + OPL thickness.Horizontal M-score was positively correlated with CST,postoperative CV and postoperative CAT (rs=0.322,P=0.045;rs=0.340,P=0.034;rs =0.336,P=0.036),and no correlation was found between horizontal M-score and BCVA,CFT,GCL thickness,INL thickness,ONL+OPL thickness in 6 months after surgery.The vertical M-score and mean M-score were not correlated with OCT parameters in 6 months after surgery.The mean M-score was positively correlated with preoperative mean M-score,preoperative CST,preoperative CV,preoperative CAT in 6 months after surgery (rs =0.589,P＜0.001;rs =0.330,P =0.040;rs =0.404,P =0.011;rs =0.410,P =0.009).In addition,and no significant correlation between mean M-score and preoperative BCVA,CFT,GCL thickness,INL thickness,ONL+OPL thickness.Multivariate stepwise linear regression showed that preoperative M-score was a predictor of postoperative M-values (adjusted R2 =0.211,P =0.002).Conclusions Most metamorphopsia can be alleviated after idiopathic epiretinal membrane surgery.The residue metamorphopsia after surgery probably is correlated with preoperative metamorphopsia and CFT.
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Objective: Meta-Analysis was used to assess the association between serum high-density lipoprotein-cholesterol (HDL-C) level and the development of breast cancer. Methods: Studies on the relationship of high-density lipoprotein-cholesterol and the development of breast cancer were retrieved from PubMed, Ovid, EBSCO, Cochrane Library, China National Knowledge Internet, VIP Periodical Resource Integration Service Platform, Wanfang Database and China Biomedical Literature Database from inception to January 9, 2017. Data were extracted from studies which met the inclusion criteria. Standardized mean difference (SMD) was used as an indicator of effect. Literature quality evaluation was carried out according to Newcastle-Ottawa Scale. Stata 12.0 software was used for Meta-Analysis. Results: A total of 12 studies on the association of serum HDL-C level with breast cancer were included in this Meta-Analysis, including 8 204 patients with breast cancer and 230 835 controls. Meta-Analysis showed that the serum HDL-C level was associated with breast cancer [SMD =0.35 (95% confidence interval: 0.58 to 0.12), P <0.001]. Subgroup analysis revealed that serum HDL-C level of premenopausal breast cancer patients was lower than that of the controls [SMD =0.15 (95% confidence interval:0.29 to 0.01), P = 0.040], but the postmenopausal serum HDL-C level had no obvious relationship with breast cancer [SMD = 0.44 (95% confidence interval:0.98 to 0.11), P = 0.110]. In different study regions, the relationship between serum HDL-C level and the development of breast cancer was not clear. Conclusion: HDL-C is related to premenopausal breast cancer, and low serum HDL-C level may increase the risk of premenopausal breast cancer. The relationship between postmenopausal breast cancer and HDL-C is not clear. Given the limitations of the included studies, a larger prospective cohort study is needed to validate the above findings.
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It is one of key scientific problems to identify the effective compounds in the study on new TCM. Because of the integrity of the effects of TCM, the diversity of constituents, the complexity of the targets and the unpredictability of the interaction among the components, the efficiency of the traditional efficacy substance research method is generally not high. Based on the research thought of quantitative composition-activity relationship, there are inevitable quantitative correlation between chemical compositions and its biological activity. The quantitative composition-activity relationship research is an effective method to detect the effective compounds. This article reviewed the common methods and technologies used in the quantitative composition-activity relationship research, with a purpose to provide some references for the study on basic effective components in TCM.
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Objective To study the determination method of total flavonoids in Gansu Astragali Radix and Hedysarum Polybotrys. Methods Calycosin glycosides etc. was selected as reference substances, comparison on the difference of absorption curves was done by ultraviolet spectroscopy and colorimetric method (NaNO2-Al(NO3)3-NaOH, AlCl3, Mg(Ac)2, NaOH, phosphomolybdic acid, HCl-Mg power). Results With colorimetric method, the maximum absorption wavelength of referrence and the test was inconsistent. The absorption peak shape was also different. With UV method, Calycosin glycosides in band Ⅱ (260 nm) showed a shoulder absorption. Astragali Radix and Hedysarum Polybotrys also showed characteristic shoulder absorptions in band Ⅱ with absorption wavelength at 263 nm and 265 nm. So the sample absorption wavelength is basically the same as that of the control sample. Conclusion Colorimetries usually used for determination of total flavonoids are not suitable for the comparison determination of Gansu Astragali Radix and Hedysarum Polybotrys. It is suitable for determining the contents of total flavonoids in samples by UV spectrophotometry at the band Ⅱ, which is the characteristic absorption band of isoflavone compound.
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“Multi-component Chinese medicine” (MCC Medicine) is a new TCM concept in recent ten years. It is a new formed TCM product accepted and approved by the new mode (component compatibility of medicines) for TCM research and development, which originated from TCM research and development and TCM pharmaceutics. MCC Medicine contains massive historical accumulation of TCM and distinctive characteristics of the times, which is closely connected with the TCM theory, current trend of the TCM development, clinical treatment requirements, and the development of modern science and technology. In order to promote the development of MCC Medicine, this article reviewed its original background and future trend, with a purpose to make clear the direction for the development of MCC Medicine.
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Objective To establish a UV spectrophotometry method and an HPLC method respectively for the determination of the total content of coumarin and contents of four main constituents of coumarin in Fraxini Cortex extract.Methods UV spectrophotometry was used for the determination of the content of total coumarin in Fraxini Cortex extract. The reference substance was Aesculin, and the maximum ultraviolet absorption wavelength was 334 nm. The HPLC method was used to determine the contents of Aesculin, Fraxin, Aesculetin and Fraxetin in Fraxini Cortex extract, using gradient elution with acetonitrile-phosphate solution (0.01%) as mobile phase on Agilent ZORBAX SB-C18 chromatographic column (4.6 mm × 250 mm, 5μm) at room temperature.Results For the UV method, the linear range of the mass concentration of Aesculin was 5.76-23.04μg/mL (r=0.999 9), and the average recovery was 100.6% (RSD=1.8%). For the HPLC method, the linear ranges of the mass of Aesculin, Fraxin, Aesculetin and Fraxetin were 0.055 0-3.850 0μg (r=0.9997), 0.053 9-3.773 0μg (r=0.999 8), 0.060 0-0.660 0μg (r=0.999 9), and 0.056 2-0.618 2μg (r=0.999 9), respectively, and the average recoveries were 96.97% (RSD=1.26%), 100.80% (RSD=2.22%), 99.04% (RSD=2.47%), and 98.77% (RSD=1.94%), respectively.Conclusion Both of the two methods are simple, accurate and reliable, and can be used for the quality control of total coumarin and the main constituents of coumarin in Fraxini Cortex extract.
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“Multi-component Chinese medicine” was derived from effective clinical prescriptions, which is similar to natural medicines, and its form is similar to the fixed-dose compounds. The adaptable indications mainly cast chronic, intractable, and complex diseases. Its research targets of pharmacology mostly are diseases, symptoms or pathological process. Vivo and vitro experiments can be used in its researches. Its dose-effect relationship mostly shows non-linear relationship, and three elements (dose, efficacy and syndrome) are involved. However, the comprehensive evaluation method is still in the exploratory stag. In the safety evaluation, there were few general pharmacological researches, and the basis of toxicity and the relationship between amount and toxicity need further study. In combined optimization aspect, including identification of components and component ratio optimization, many methods have been developed, but it still needs to be developed in theory and systemic integration. Pharmacological mechanism researches focus on the multi-component, multi-target, multi-pathway, and multi-pharmacodynamics aspects, combined with new ideas and new technologies, such as systems biology, network pharmacology, etc. But as multi-component medicine, it should also be studied about the correspondence and network relationships of component and target, with a purpose to explain the contribution of each component in overall mechanism.
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Objective To study the effects of different extraction processes on extraction rate of cortex fraxini total coumarin and active constitute percentage of major coumarin;To establish a proper extraction process.Methods Orthogonal design method was applied to set comprehensive index cortex fraxini total coumarin extraction rate as inspecting index. Water and alcohol were used as solvent respectively to optimize the extraction process of cortex fraxini.Results Optimal water extraction process:cortex fraxini decoction pieces mixed with nine times of water, decocted for three times, 90 mins each time. The pasty fluid generating rate of cortex fraxini was 28.87%, total coumarin percentage was 19.26%, extraction rate was 5.56%, total percentage of Aesculin, Aesculetin, Fraxin, Fraxetin was 13.47%, when water was used as solvent. Optimal alcohol extraction process:cortex fraxini decoction pieces mixed with eight times of 75% ethyl alcohol, refluxed twice, two hours each time. The pasty fluid generating rate of cortex fraxini was 30.47%, total coumarin percentage was 21.72%, extraction rate was 6.62%, total percentage of Aesculin, Aesculetin, Fraxin, Fraxetin was 15.29%, when alcohol was used as solvent. It was found that using alcohol as solvent had a 5.54% higher pasty fluid generating rate, a 12.77% higher total coumarion percentage, a 19.06% higher total coumarin extraction rate, and a 13.51% higher percentage of total four coumarin constitutes than using water, with statistical significance. Conclusion Extraction process by using alcohol as solvent is better than using water. So the optimal and stable extraction process of cortex fraxini total coumarin is using 75% alcohol as solvent.
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Objective To establish a method to analyze the composition and content of monoses in the garlic polysaccharide by ion chromatography. Methods The monoses in the solution of garlic polysaccharide hydrolysised by trifluoroacetic acid were determined by amino PAC column, eluted by gradient hydroxide sodium and identified by ampere detector (AU as working electrode, Ag/AgCl as reference electrode). Results The garlic polysaccharide was mainly composed by arabinose, galactose, glucose, xylose and fructose. Good resolution could be achieved among the above mentioned five kinds of monoses. Their linear correlations over the investigated concentration were above 0.998, while the average recovery rates were in the range of 95%-105%. Conclusion The ion chromatography method is fast, accurate, simple and reliable, and can be applied in the content determination and quality control of garlic polysaccharide.
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This study was aimed to optimize the extraction technology of Wei-Kang (WK) Capsule. Orthogonal test and comprehensive evaluation were used to optimize the extraction process of compound preparation. The icarrin, to-tal ginkgo flavone glycosides and the dried decocting rates were used as index components for optimizing the effect of ethanol concentration, ethanol volume, extraction duration and extraction frequency. The results showed that the opti-mal ethanol extraction technology was adding 12 times of 60% ethanol and extract for 3 times with 1 h for each ex-traction time. It was concluded that the optimized extraction technology was stable, practical, scientific and reason-able, which can be used in the large-scale industrial production.
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<p><b>OBJECTIVE</b>To establish a new quantitative method for simultaneous determination of multi-coumarins in Fraxini Cortex by using one chemical reference substance, and validate its feasibilities.</p><p><b>METHOD</b>The new quality evaluation method, quantitative analysis of multi-components by singer-marker (QAMS), was established and validated with Fraxini Cortex. Four main coumarins were selected as analytes to evaluate the quality and their relative correlation factors (RCF) were determined by HPLC-DAD. Within the linear range, the values of RCF at 340 nm of aesculin to asculetin, fraxin and fraxetin were 1.771, 0.799, 1.409, respectively. And the contents of aesculin in samples of Fraxini Cortex were authentically determined by the external standard method, and the contents of the three other coumarins were calculated by their RCF. The contents of these four coumarins in all samples were also determined by the external standard method.</p><p><b>RESULT</b>Within a certain range, the RCF had a good reproducibility (RSD 2.5%-3.9%). Significant differences were not observed between the quantitative results of two methods.</p><p><b>CONCLUSION</b>It is feasible and suitable to evaluate the quality of Fraxini Cortex and its Yinpian by QAMS.</p>
Subject(s)
Anticoagulants , Biomarkers , Coumarins , Drug Stability , Drugs, Chinese Herbal , Chemistry , Esculin , Feasibility Studies , Plant Extracts , Chemistry , Plants, Medicinal , Chemistry , Reproducibility of ResultsABSTRACT
BACKGROUND: Recently, with the increasing popularity of dental implant and orthognathic surgery, more and more people pay attention to the structure of mandibular canal and adjacent tissues. However, there are few reports addressing specimen measurements at home and abroad. OBJECTIVE: To measure the anatomical data of mandibular canal and adjacent tissue of mandible specimen to provide reliable reference data and anatomical evidence for alveolus surgery, tooth implantation, and orthognathic surgery. DESIGN, TIME AND SETTING: A controlled observation was performed at the Laboratory of Anatomy, Jilin Medical College between March 2007 and September 2007. MATERIALS: Twenty complete mandible specimens. METHODS: Sixteen adult, ex vivo mandibles with complete dentition and 4 edentulous mandibles were measured in every dental position of sagittal profile and posterior mental foramen in terms of molar, alveolar crest, and mandibular canal. Data were statistically analyzed. MAIN OUTCOME MEASURES: Mental foramen location, horizontal distance from anterior border of mental foramen to anterior border of mental canal, mandibular foramen location, distances from mandibular canal to molar root tip, lingual bone plate of mandible, inferior border of mandible, mandibular buccal bone plate, and alveolar ridge crest. RESULTS: Mandibular canal, located in the inferior region of the mandible body, ran towards the lingual side and was close to the inferior border of mandible and most close to the mandibular molar root tip. It turned towards posterolateral and buccal side in front of mental foramen and then passed through mental foramen. The distance of mandibular canal between edentulous mandible and alveolar ridge crest was obviously shortened. CONCLUSION: Multi-section observation and measurement of ex vivo mandible specimens provide precise, reliable evidence for preoperative design of orthognathic surgery, facilitate surgeons to better formulate surgical proposals, and prevent some complications, including intraoperative massive hemorrhage, interior alveolar nerve injury, mandibular angle fracture, and mental bone lateral wall perforation.
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We have designed and carried out problem based learning (PBL) pedagogy since 2004. According to clinical eases, students learnt the pathophysiology of heart failure knowledge by themselves. Each group recommen-ded one student to make an oral presentation and wrote a review about heart failure. Preparing clinical cases and group discussions are very important. At the same time we should pay attention to the change in role of the teacher in PBL and cooperation with other disciplines.
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Objective To analyze the clinicopathological characteristics, immunophenotypes and immu-noglobulin heavy chain gene rearrangement of gastric mucosa-associated lymphoid tissue (MALT) lymphoma. Methods The clinical data of 35 patients with gastric MALT lymphoma who had been admitted to Southwest Hospital from January 1984 to June 2003 were retrospectively analyzed. The specimens of the gastric MALT lymphoma were obtained and their pathomorphological classifications were studied. The immunophenotypes and immunoglobulin heavy chain gene rearrangement were detected by immunohistochemistry and polymerase chain reaction, respectively. All the data were analyzed by chi-square test and Mann-Whitney U test. Results The main manifestations of patients with gastric MALT lymphoma were epigastric discomfort and abdominalgia. The percentage of patients with age≥40 accounted for 74% (26/35). The gastric MALT lyrnphoma has low differentia-tion in 32 patients, and high differentiation in 3 patients. There were 31 patients in Ⅰ E stage, 2 in Ⅱ E1 stage and 2 in Ⅳ stage. Pathological examination showed that gastric MALT lymphoma was composed of centrocyte-like cells and mononuclear-like B cells or lymphoplasma cells. Twenty-nine specimens were with lymphoepithelial lesions (83%), 2 with follicular colonization (6%), and 11 with plasma cells segregated beneath the surface epithelium (33%). All the 35 specimens were B cell immunotype. Restrictive expression of κ and λ light chains of the immunoglobulin was found in 19 cases, and Bcl-2 positive expression was found in 16 cases. High expres-sion of proliferating cell nuclear antigen was found in gastric MALT lymphoma with transformed blasts, which indicated that gastric MALT lymphoma with transformed blasts was more active in proliferation than those of low grade. A monoclonal pattern of immunoglobulin heavy chain gene rearrangement was detected in 33 patients (94%). Conclusions Digestive symptoms are the main manifestations of patients with gastric MALT lymphoma. The incidence of gastric MALT lymphoma increases with age. The final diagnosis of gastric MALT lymphoma depends on the pathological, immunohistochemical examination and laboratory test.
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Objective To screen the variation in NeuroD1 gene and to study its function in vitro and its clinical phenotypes and genetic characteristics in Chinese early-onset type 2 diabetic probands. Methods PCR-direct sequencing of NeuroD1 gene was performed in 85 early-onset type 2 diabetic probands, 95 late-onset type 2 diabetics with strong diabetic history and 87 non-diabetic control subjects. Distributions of the identified variation were calculated and compared among the three groups. Expression vectors with mouse NeuroD1 (mND1)cDNA wild type or mutant type and reporter vectors with human insulin promotor-linked luciferase were constructed. Then the above vectors were co-transfected into rat INS-1 cells. Relative luciferase activities were measured to compare transcriptional activities of insulin gene between WT and MT. Results S159P (T→C), a new mutation was identified in a proband, which was co-segregated with diabetes in 4 carriers from the paternal side. The functional study showed that the S159P mutant exhibited a 25% reduction in transcriptional activity of insulin gene as compared with the wild type. A45T (G→A), a common variation was identified. The AA + GA genotypic frequencies were markedly increased in early-onset type 2 diabetic probands as compared with late-onset type 2 diabetic probands and non-diabetic control subjects (P=0.006 and P=0.014, respectively). Conclusion The novel S159P mutation in the NeuroDl gene seems to contribute to the development of diabetes in the Chinese early-onset type 2 diabetic family. The A45T variation may increase susceptibility to or be in disequilibrium with early-onset type 2 diabetes mellitus in Chinese population. In addition, the A45T variation may affect the onset pattern of type 2 diabetes mellitns, such as early-onset but not late-onset type 2 diabetes mellitus.
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10 mm, and treat the primar disease in host. Drug-eluting stent is beneficial to reduce incidence of restenosis.