Objective To study the application of a new magnetic beads made in China in DNA extraction of forensic biological samples with automation workstation.Methods DNA was extracted from common forensic biological samples by QIAGEN Bio-Robert Universal System and a new magnetic beads made in China,and then typed with Identifiler system in ABI3130XL Genetic Analyzer.210 of these samples were also quantitated by ABI7500 Real Time System.Results Total of 9100 genomic DNA was extracted from various forensic biological samples by the new magnetic beads made in China and automation workstation methods,and most of them were successfully typed for STR analysis.In these biological samples,oral swabs and muscles were of the highest Success rate of STR typing(100%),and the lowest was touched cell samples (50.0%).Conclusion The new magnetic beads made in China with automation workstation methods can be applied to DNA extraction of most forensic biological samples.

Actins ; metabolism ; Animals ; Collagen Type III ; metabolism ; Collagen Type IV ; metabolism ; Desmin ; metabolism ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; metabolism ; pathology ; HSP47 Heat-Shock Proteins ; metabolism ; Kidney Glomerulus ; metabolism ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vimentin ; metabolism

BACKGROUND:Erythromycin spreads widely in the body with a short period of effective concentrations and has a lot of adverse effects.Therefore,it is necessary to make erythromycin as targeted medicine.OBJECTIVE:To optimize the preparation conditions of erythromycin gelatin microspheres.DESIGN,TIME AND SETTING:An orthogonal controlled test was performed in the Department of Pharmacy,Guangdong Pharmaceutical University from June to December in 2005.MATERIALS:Erythromycin and gelatin.METHODS:According to the emulsion principle,erythromycin dispersed in the gelatin solution.In the process of preparing microspheres,the gelatin solution and oil should form W/O emulsion and then it turned into spheres by solidification.The formation and quality of microspheres were influenced by four factors,namely the concentration of gelatin,dosage of emulsifier,the solidification time and the speed of mixing.The arithmetic mean diameter of microspheres,the drug loading efficiency and the encapsulation efficiency were targets for the survey in this study on the basis of pretests.The best preparation conditions were optimized in accordance with the results of L9 (34) orthogonal tests.The optimized preparation conditions were obtained according to the results of orthogonal tests.MAIN OUTCOME MEASURES:The mean diameter of microspheres,the drug loading efficiency,the encapsulation efficiency,and the orthogonal tests were examined.RESULTS:The optimized preparation conditions of erythromycin gelatin microspheres included 15% gelatin,3.0 mL emulsifier,0.5 hour solidification and mixing at 1 000 r/rain.The erythromycin gelatin microspheres were regular in their morphology.Drug was enveloped in microspheres.The average particle size was (14.15±0.20) μm;the drug loading efficiency and the encapsulation efficiency were (5.83±0.38)% and (65.70±0.56)%,respectively.Over 90.16% of the microspheres was in the range of 7-25 μm;The reappearance of pharmaceutical technology was good.CONCLUSION:The optimized preparation conditions of erythromycin gelatin microspheres are obtained using L9 (34)orthogonal tests.The microspheres prepared meet the requirement of the size for lung targeting.

Adolescent ; Adult ; Aged ; Child ; Decision Trees ; Female ; Hepatitis B, Chronic ; diagnosis ; Humans ; Male ; Middle Aged ; Young Adult

PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8+T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8+CLA+T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8+CLA+T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8+CLA+T cells were evaluated. The proliferative responses of CD8+CLA+T cells were assessed by flow cytometry, and the levels of transforming growth factor-beta1 (TGF-beta1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8+CLA+T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8+CLA+T cells in AD. Meanwhile, the levels of TGF-beta1 produced by Tregs were significantly lower in AD, and anti-TGF-beta1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8+CLA+T cells, mediated by TGF-beta1, plays an important role in the pathogenesis of AD.
Adult ; Aged ; CD8-Positive T-Lymphocytes/drug effects/*immunology ; Case-Control Studies ; Cell Proliferation ; Cell Separation ; Dermatitis, Atopic/*immunology/pathology ; Female ; Granzymes/metabolism ; Humans ; Interleukin-10/metabolism ; Lymphocyte Count ; Male ; Perforin/metabolism ; Skin/*immunology/pathology ; T-Lymphocytes, Cytotoxic/drug effects/immunology ; T-Lymphocytes, Regulatory/drug effects/*immunology ; Transforming Growth Factor beta1/pharmacology

Objective To investigate the relationship between drinking-water type of endemic arsenicosis and adult carotid artery atherosclerosis. Methods In 2009, 285 participants aged over 40 from drinking-water type of endemic arsenism areas and 293 residents aged over 40 from control areas were investigated in Yingxian county,Shanxi province. Portable-type B mode color ultrasound was used to examine the carotid artery of all participants.The carotid atherosclerosis were diagnosed and graded through the ultrasonograms. Content of water arsenic and hair arsenic of 10 people randomly selected in every villages were detected. Results A total of 5 villages with drinkingwater type of endemic arsenicosis as observation group and 5 villages without drinking-water type of endemic arsenicosis as control group were investigated. The prevalence rates of adult carotid atherosclerosis within observation group were 35.09%(20/57), 55.74%(34/61), 38.46%(20/52), 36.51%(23/63) and 46.15%(24/52), respectively,and standardized prevalence rates were 32.5%, 33.8%, 34.9%, 46.2% and 47.3%, respectively and the prevalence rates of adult carotid atherosclerosis within control group were 18.18%(10/55), 30.77%(16/52), 20.00%(10/50),18.67% (14/75) and 21.31% ( 13/61 ), respectively; the standardize prevalence rates were 22.4%, 17.7%, 10.7%,24.6%, 18.9%, respectively. The standardize prevalence rates were higher in observation group [39.50%(113/285) ]than that in control group[39.50%(113/285), T = 26, P ＜ 0.01 ]. The severity of adult carotid atherosclerosis (composition of 4 - 7 scores ) was compared between observation group [ 17.70%(20/113 )] and control group [ 14.06% (9/64) ], and the difference was insignificant(x2 = 0.26, P ＞ 0.05). Conclusions The prevalence rate of carotid atherosclerosis in drinking-water type of endemic arsenicosis areas is higher than that of the control areas.The study provides evidence that arsenic poisoning can cause atherosclerosis.

Objective To explore the association between polymorphisms of interferon-gamma gene intron 1 at position+874 (IFN-γ+874) gene and the susceptibility of HBV and/or HCV infection with different clinical outcomes. Methods IFN-γ+874 gene SNP were detected in 277 subjects including 79 chronic HBV/HCV coinfections,69 individuals only with HBV infection,55 individuals only with HCV infection and 74 controls,by sequence specific primers-PCR (SSP-PCR). Hepatocellular injury as suggested by alanine aminotransferase (ALT) was detected by Beckman LX-20. The status of viral particles in serum was determined by RT-nPCR. The possible association of the polymorphism of IFN-γ+874 with the susceptibility of HBV and/or HCV infection and the outcome of these infections were analyzed. Results (1) IFN-γ+874 AA frequency in individuals with chronic HBV,HCV,HBV/HCV coinfections were significant higher than that in controls (X~2=16.15,P=0.01); OR (95% CI) of IFNγ+874 AA in chronic infection with HBV,HCV,HBV/HCV coinfections appeared to be 2.70 (1.24-5.92),3.22 (1.43-7.25) and 4.02 (1.88-8.55) compared with + 874 TA. No significant differences were found among HBV,HCV,HBV/HCV coinfections (X~2=1.97,P=0.73). There were no significant association of IFN-γ +874 A/T allele frequency with HBV and/or with HCV infection (X~2=4.87,P=0.18). (2)The clinical outcomes of mild chronic hepatitis (CH),moderate/severe CH and cirrhosis with HBV and/or HCV infection were associated with IFN-γ+874 AA [X~2＝14.17,P＝0.03;OR＝3.09(1.51-6.33),3.85 (1.70-8.70),3.14 (1.08-9.17)]. No significant relationships were found between IFN-γ+874 A/T allele frequency and the clinical outcome of HBV/HCV infection (X~2＝6.07,P＝0.11). (3)There were no significant associations of IFN-γ+874 genotype/allele frequency with HCV duplication (X~2=2.36,P=0.31). (4) There were no significant associations of IFN-γ+874 genotype/allele frequency with abnormal ALT (X~2=0.15,P=0.93). Conclusion These results suggested that polymorphisms in the IFN-γ +874 had some influence on chronic HCV and/or HBV infection,and on the outcome of HCV and/or HBV infections. IFN-γ+874 AA genotype and T allele were possible risk to chronic HBV and/or HCV infections and to the outcomes of HBV and/or HCV infection. However,IFN-γ+874 TA genotype might serve as possible protective factors to them.

Objective To study the relationship between polymorphisms in interleukin-2gene at position-330 (IL-2-330) and the clinical outcome of hepatitis B virus (HBV) and/or hepatitis C virus (HCV) infection. Methods 277 subjects were recruited including 79 chronic HCV co-HBV infection, 55 chronic HCV infection, 69 chronic HBV infection and 74 controls. Single nucleotide polymorphisms of IL-2-330 was investigated by restricted fragment long polymorphism-PCR (RFLP-PCR). Hepatocellular injury, as revealed by alanine aminotransferase (ALT) was detected by Beckman LX-20 analyzer. The presence of hepatitis C viral particles in serum was determined by RT-nPCR. Results ( 1 ) IL-2-330 polymorphisms showed close association with persistent HBV and/or HCV infection. IL-2-330 TT was associated with an increased risk, but IL-2-330 GG with a reduced risk of persistent HBV and/or HCV infection (χ2=14.24, P=0.03 ) with ORs (95%CI) as 7.14(2.13-23.81 ), 3.46 (1.17-10.02) and 2.93 (1.15-7.46) respectively. However,IL-2-330 TT/GG did not significantly differ between patients with HBV and/or HCV infection (χ2=2.09, P=0.72). IL-2-330 T allele was associated with an increased risk, but the -330G allele was associated with a reduced risk of chronic HBV/HCV infection (χ2=12.33,P=0.01),with ORs (95% CI) as 2.26 (1.39-3.69) , 1.82 ( 1.09-3.03 ) and 1.73 ( 1.10-2.73 ) respectively. (2) IL-2-330polymorphisms showed significant association with the outcome of HBV and HCV infection ( χ2=13.52, P=0.04). IL-2-330 TT was associated with an increased risk, but-330 GG with a reduced risk of mild CH, moderate/severe CH, and cirrhosis. The ORs (95%CI) appeared to be 3.33(1.75-6.32), 3.31 (1.75-6.26), 11.23 (3.09-40.76) respectively. IL-2-330 T allele was associated with an increased risk, but the -330 G allele was associated with a reduced risk of mild CH, moderate/severe CH and cirrhosis (χ2= 12.32, P=0.01 ), with ORs as 1.86(1.32-2.63), 1.71 (1.27-2.31) and 2.77(1.57-4.89) respectively. (3) The polymorphisms of IL-2-330 showed no association with HCV RNA replication (χ2=0.83, P=0.66; χ2=0.20, P=0.66). The polymorphisms of IL-2-330 were not significantly associated with abnormal ALT ( χ2= 1.10, P=0.58; χ2=0.08, P=0.78). Conclusion These results suggested that IL-2-330 TT/T was associated with an increased risk, but IL-2-330GG/G was associated with reduced risk of persistent HBV and/or HCV infection, and with the development of mild CH,moderated/severe CH,and cirrhosis.

OBJECTIVE To study the effect of polygonatum polysaccharide on zebrafish with Alzheimer disease. METHODS Zebrafish were trained in T maze for 7 d. The 40 zebrafish successfully trained were divided into 4 groups:blank group, model group, positive group and polygonatum polysaccharide group. Model group, positive group and polygonatum polysaccharide group were put in AlCl3100μg·L-1 for 6 d. The positive group was exposed to Huperzine A solution 4μg·L-1, and the polygonatum polysaccharide group was exposed to polygonatum polysaccharide solution 6 g·L-1 for 6 d. The model group was not treated, and the blank group was not treated. Each stage of zebrafish was recorded by video, and the time of each group in the EC region was analyzed. After administration, the brain tissue was taken out and the expression of N-cadherin, P38 and p-P38 protein factors was determined by Western blotting. RESULTS In behavior, the analysis of the time spent in the EC area, the blank group, the positive group and the polygonatum polysac?charide group were compared with the model group, respectively, there were statistically significant differences (P<0.05). At the protein level, compared with the model group, the P38 and p-P38 proteins in the positive group and the polygonatum polysaccharide group were down-regulated, while the N-cadherin protein was up-regulated, with statistical difference (P<0.05). CONCLUSION Polygonatum polysaccharide can improve the learning and memory ability of zebrafish with Alzheimer disease by up regulating the protein level of N-cadherin and hindering P38 phosphorylation.

BACKGROUNDThe stage-specific expression of genes is one of the most characteristics of parasites. It has been found that a lot of genes of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocercoid of Spirtmetra erinceieuropaei.

METHODSRNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DNA contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T12MA, T12MC, T12MG and T12MT anchor-primers, PCR was done using the same T12MN and one random primer with alpha 35S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes.

RESULTSEleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA.

CONCLUSIONSThe gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique.