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OBJECTIVES@#To develop a multiplex PCR amplification system (EX20+30Y for short) of 19 autosomes, 30 Y-STR loci plus the gender indicator, and evaluate its forensic application value.@*METHODS@#With the six-color fluorescence labeling technology, a multiplex amplification system of 19 autosomal STR loci and 30 Y-STR loci plus the gender indicator was constructed. Blood samples from 210 unrelated individuals, 69 daily case samples and standard samples 9948 and 9947A were collected for loci detection and analysis. The EX20+30Y multiplex amplification system was evaluated by its sensitivity, mixed sample detection ability, species specificity, balance, direct amplification ability, sample applicability and anti-inhibition ability.@*RESULTS@#Multiplex amplification of blood samples from 210 unrelated individuals by the detection system obtained accurate genotyping results. The detection sensitivity of standard samples was 0.125 ng and the species specificity was high. The 69 samples from daily cases were genotyped correctly. Moreover, standard sample 9948 could be accurately genotyped even if the samples contained a certain concentration of inhibitors.@*CONCLUSIONS@#The multiplex amplification system established in this study can conduct combined examination of 19 autosomes, 30 Y-STR loci plus the gender indicator with accurate genotyping and high sensitivity. It has a good forensic application prospect.
Subject(s)
Humans , Chromosomes, Human, Y/genetics , DNA Fingerprinting/methods , Forensic Medicine/methods , Microsatellite Repeats , Multiplex Polymerase Chain Reaction , Species SpecificityABSTRACT
Objective To explore the therapeutic effect of astragalus polysaccharides (APS) on high fat-induced nonalcoholic fatty liver disease (NAFLD) in rats. Methods Totally 36 male SD rats were randomized into normal control group (n=8) and disease model group (n=28). The rats in the disease model group were fed with high-fat diet for 6 weeks to establish rat model of NAFLD. Twenty-five of 28 NAFLD model rats were randomized into NAFLD model group (n=7), simvastatin group (n=9, intragastric administration of simvastatin [6.7 mg · kg-1 · d-1]) and APS group (n=9, intragastric administration of APS [700 mg · kg-1 · d-1]). The rats in the normal control group and the NAFLD model group were intragastrically given the same volume of double distilled water. The rats in each group were continuously fed for 8 weeks. The dietary intake, energy intake, drinking water, excretion and serum biochemical parameters were compared between the four groups. Liver tissues were collected to calculate the liver index and measure the contents of total cholesterol (TC) and triglyceride (TG). The liver tissue structure and cell morphology were observed under the optical microscope. Results The energy intake of rats in the NAFLD model group was significantly higher than that in the normal control group (P0.05). Compared with the NAFLD model group, serum levels of TC, TG, lowdensity lipoprotein-cholesterol and malondialdehyde were significantly decreased, and the level of high-density lipoproteincholesterol and the activity of total superoxide dismutase were significantly increased in the APS and simvastatin groups (all P0.05). Conclusion APS can effectively regulate the levels of serum glucose, blood lipids and hepatic lipid droplets in NAFLD rats, which may be related to the antioxidant effect of APS.
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OBJECTIVE@#To establish a multiplex STR genotyping method for autosomal STR and Y-STR loci in forensic biological practice.@*METHODS@#Widely used autosomal STR loci and Y-STR loci were selected. A set of PCR primers was designed, and a 5-dye fluorescent labeled STR multiplex PCR reagent kit was developed.@*RESULTS@#A kit was developed which can simultaneously detect 15 autosomal STR loci, 10 Y-STR loci, and an Amelogenin.@*CONCLUSION@#The 15 autosomal STR plus 10 Y-STR kit in combination with capillary electrophoresis method was used to STR genotyping with accurate and reliable results. The new one-step testing kit can potentially be widely used in forensic cases and DNA databank in the future.
Subject(s)
Humans , Amelogenin , Chromosomes, Human, Y/genetics , DNA Primers , Databases, Nucleic Acid , Forensic Genetics/methods , Genotype , Genotyping Techniques/instrumentation , Indicators and Reagents , Microsatellite Repeats , Multiplex Polymerase Chain ReactionABSTRACT
To establish an analytical method for determination of four bisphenols (BPA, BPB, BPF, and BPS) and two alkylphenols (4-n-OP, 4-n-NP) in water by ultra performance liquid chromatography- tandem mass spectrometry (UPLC/MS/MS). The water samples were extracted and condensed with solid-phase extraction (SPE) using C18 cartridges and eluted by acetonitrile. Separation was carried out with Acquity BEH C8 column and detection were performed by UPLC/MS/MS. Quantification was calculated by using the internal standard BPA-d16 and 4-n-NP-d8. The linear correlation coefficients of these compounds in the range of 1.0-100.0 μg/L were all over 0.999. The minimum detectable concentrations were 0.75-1.0 ng/L, and the recoveries ranged from 87.0% to 106.9%. Relative standard deviations (RSDs) were between 1.26% and 3.67%. Applying this method to detect the source water of Chaohu Lake and drinking water of Hefei, six target compounds were detected in different levels. This method is simple with high sensitivity and selectivity, could be suitable for the determination of these compounds in source and drinking water.
Subject(s)
Chromatography, Liquid , Endocrine Disruptors , Phenols , Solid Phase Extraction , Tandem Mass Spectrometry , Water Pollutants, ChemicalABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of laparoscopic total mesorectal excision (laparoscopic TME) versus open total mesorectal excision (open TME) in the treatment of middle and low rectal cancer using meta-analysis.</p><p><b>METHOD</b>From 1991 to 2012, the Chinese and English articles of randomized controlled trails (RTCs) about laparoscopic TME versus open TME in the treatment of middle and low rectal cancer were collected, and a meta-analysis was performed with RevMan 5.1 software.</p><p><b>RESULTS</b>Eight RCTs including 863 patients with middle and low rectal cancer (428 cases in laparoscopic TME group, 435 cases in open TME group) were enrolled in the meta-analysis. Laparoscopic TME was associated with significantly less intraoperative blood loss (P<0.01), earlier to pass first flatus (P<0.01), shorter hospital stay (P<0.05), less postoperative incision infections (P<0.01) and postoperative bleeding (P<0.05) compared to open TME. There were no significant differences between laparoscopic TME and open TME groups in operative time, number of resected lymph nodes, anastomotic leak, ileus and pelvic abscess (all P>0.05).</p><p><b>CONCLUSIONS</b>As compared to open TME, laparoscopic TME has similar efficacy in terms of lymph nodes harvest, and it can promote postoperative recovery, and reduce incision infection and postoperative bleeding.</p>
Subject(s)
Humans , Laparoscopy , Methods , Mesentery , General Surgery , Randomized Controlled Trials as Topic , Rectal Neoplasms , General Surgery , Rectum , General Surgery , Treatment OutcomeABSTRACT
Objective To explore the value of liver acquisition with volume acceleration (LAVA) multi-phase dynamic enhanced technique combined with magnetic resonance diffusion weighted imaging(DWI) in evaluating prostate lesions. Methods Totally 67 prostate disease patients confirmed by histopathology and follow-up were included in the present study. The patients included 19 with prostate cancer (PCa) and 48 with benign prostate hyperplasia (BPH). LAVA multi-temporal enhancement and DWI scanning were performed, and the values of apparent diffusion coefficient (ADC) were calculated. Two experienced radiologists evaluated the results in a double-blinded manner; the qualitative diagnosis accuracy rates of the T-SI curves alone or in combination with ADC for PCa and BPH were compared. Results The accuracy rates of T-SI curve alone for qualitative diagnosis of PCa, BPH were 57.9% and 72.8%, and those of T-SI curve combined with ADC value were 73.7% and 87.5%, respectively. Conclusion LAVA multi-phase dynamic enhanced technique combined with ADC is of great value for differential diagnosis of benign and malignant prostate lesions.
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Objective To explore the changes of plasma corticotrophin releasing factor (CRF) levels in the young rats with hypoxic-ischemic brain damage.Methods Two hundred and forty young rats were randomly divided into three groups:hypoxic-ischemic brain damage group ( model group,n =80),sham-operated group ( n =80),and normal control group ( n =80).The plasma CRF levels of rats in three groups were detected at 1 h,3 h,6 h,12 h,l d,3 d,5 d and 18 d after hypoxia-ischemia,per ten rats for each time point.Plasma CRF levels were measured by radioimmunoassay.Results Plasma CRF levels of model group,shamoperated group and normal control group showed no significant difference in the young rats after 1 h,3 h,6 h,12 h of hypoxia-ischemia ( P ＞ 0.05 ).But plasma CRF levels in the model group were respectively significantly lower than those of sham-operated group and normal control group after 1 d and 3 d of hypoxia-ischemia ( P ＜0.001 ),and then recovered to the control group levels after 5 d and 18 d of hypoxia-ischemia ( P ＞0.05 ).Conclusion Hypoxia-ischemia affects plasma CRF levels in the young rats,which is related with the duration after hypoxia-ischemia.
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BALB/c mice were immunized with purified White spot syndrome virus (WSSV).Six monoclonal antibody cell lines were selected by ELISA with VP28 protein expressed in E.coll in vitro neutralization experiments showed that 4 of them could inhibit the virus infection in crayfish.Westernblot suggested that all these monoclonal antibodies were against the conformational structure of VP28.The monoclonal antibody 7B4 was labeled with colloidal gold particles and used to locate the VP28 on virus envelope by immunogold labeling.These monoclonal antibodies could be used to develop immunological diagnosis methods for WSSV infection.
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The Infectious hypodermal and hematopoietic necrosis virus (IHHNV) and Taura syndrome virus (TSV) are two important shrimp viruses in cultured shrimp in America. These two viruses were transmitted to China at the beginning of the 21st century. In this study, 214 shrimp samples of Penaeus vannamei were collected from seven different areas of China and tested by PCR for IHHNV and TSV infection. The results showed that there were a high prevalence of IHHNV (65.42%) and low prevalence of TSV (3.27%) in the tested samples. Several samples were found to be co-infected with these two viruses. A 3 kb fragment of 7 positive IHHNV samples and a structure protein region (ORF2) of three TSV positive samples were amplified and sequenced. The sequence comparison indicated that both IHHNV and TSV sequenced in China have a low genetic variations compared with the prototype IHHNV and TSV from Hawaii. Phylogenetic analysis showed that TSV isolates were clustered into two groups, Asia and America group, which was genetically correlated to geographic distribution.