In the modern biotechnology era,the important disease genetic resources become the most dependent factor.We expound the actuality of the collection,storage and use of the important disease genetic resources in China,and discuss the problems and challenges we meet.So the chief mission we should do is establishing the database and collecting,storing and using these valuable resources under standard procedure with aim,plan and organization.

To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To investigate the dynamics of the level of S100 in cerebrum, brainstem, and serum following the diffuse brain injury in rats and provide the experimental evidences for estimating injury time. Methods ELISA was used to determine whether S100 protein is changed after diffuse brain injury in rats. Forty rats were sacrificed at 0.5 hour, 2 hours, 4 hours, 12 hours, 24 hours, 3 d and 7 d after diffuse brain injury and normal rats as control. Results The level of S100 in cerebrum, brainstem, and serum increased, followed by a decrease, and then further increased. The level of S100 could be detected to increase at 30 minutes and reached the peak at 4 hours after DBI. The level decreased gradually to the normal at 1d and till 3 d formed the second peak. The level returned to the normal at 7d following injury again. In the postmortem injury groups, there were no significant changes compared to the control group. Conclusion The present study showed that the time-dependent expression of S100 is obvious following diffuse brain injury in rats and suggested that S100 will be a suitable marker for diffuse brain injury age determination.

AIM:To compare the difference between Shuxiong Tablet(Radix et Rhizoma notoginseng,Flos Carthami,Rhizoma Chuanxiong)(SXT) and its effective compounds group(SECG) in anti-inflammatory,relieving pain,anticoagulation,anti-thrombus and myocardial protection.METHODS:We adopted experiments of carrageenan-induced paw edema,hot water shrink trail,clotting time in mice,and thrombus in arteriovenous shut,and pituitrin induced acute myocardial ischemia in rats.RESULTS:Both of SXT and SECG could inhibit the tumefaction,decrease PGE_2 and SOD in inflammatory tissue;enhance the pain threshhold of mice;extend clotting time;inhibit the thrombosis;inhibit myocardial ischemia,significantly decrease CK,LDH,MDA and increase SOD in myocardial tissue.CONCLUSION:SXT and SECG have the same effects on anti-inflammatory,relieving pain,anticoagulation,anti-thrombus and myocardial protection.Further more,their dose-response relationship curves were similar,suggesting that SECG having the main effective components of SXT,could take place of SXT in clinical research.

Background Choroidal neovascularization (CNV) is a main cause of visual impairment in many retinal diseases.To create an ideal CNV animal model is very important for the experimental and clinical study of CNV.The assessment method of repeatable and reliable for CNV model is still seldom.Objective This experiment was to explore the label value of fluorescent antibody for visualizing and quantifying the morphologic changes associated with laser-induced CNV.Methods Laser-induced CNV models were created in 30 eyes of 15 male SPF C57BL/6J mice by Krypton red laser irradiating fundus 2 spots around the optical disc with the wavelength 647.1nm,power 260 mW,spot diameter 50μm and exposure time 0.05 seconds.The CNV was evaluated at 5 minutes,4,7,14 and 28 days after laser injury by using fundus photography and fundus fluorescein angiography (FFA),and the successful models were identified as the rupture of Bruch's membrane.The mice were then immediately sacrificed and the eyeballs were enucleated to prepare the choroidal flatmounts.The posterior eye cups were fluorescently labeled with markers of cell nuclei (DAPI,4',6'-diamino-2-phenylindole),endothelial cells (isolectin-B4),and filamentous actin (phalloidin).The CNV areas from specimens were measured by Image pro plus 6.0.Two eyes from one matched mouse without receiving photocoagulation were used as the controlls.This study followed the Standard of Association for Research in Vision and Ophthalmology.Results No any CNV was seen in photocoagulated eyes in 5 minutes and 4 days after laser irradiation.The first sign of CNV appeared at 7 days following photocoagulation.The incidence of fluorescein leakage was 76.47% (26/34),81.81% (18/22),50.00% (5/10) at 7,14 and 28 days,respectively.The fluomicroscope examination showed that in unphotocoagulated areas,retinal pigment epithelial (RPE) cells were visualized with a uniform hexagonal array.Immediately after laser exposure,a circular area devoid of fluorescent labeling was observed,indicating disruption of the choroid-Bruch membrane-RPE complex.On the fourth day,cellular debris and fragmented nuclei were presented and an autofluorescent ring was visible at the site of Bruch's membrane disruption.The number of CNV vessels increased exponentially during the next 3 days.At 7 days,a well-defined isolectin-B4 labeled CNV network was exhibited and lasted for 28 days.The CNV areas were (7.99±0.42)×103μm2,(16.89±3.77)×103μm2,(14.37±4.02)×103μm2 at 7,14 and 48 days after photocoagulation respectively,showing a significant difference among these three groups (F=17.340,P=0.000),and the CNV area was significantly increased in the photocoagulating eyes in 14 days and 28 days compared with 7 days (q=16.46,q=15.54,P＜0.01).Conclusion Fluorescent antibody labeling allows the well identification and measurement of laser-induced CNV lesions in mouse choroid/RPE flatmounts.This technique offers excellent morphologic detail and facilitates the study of critical early events in CNV.CNV complexes are labeled at an early stage,providing a more accurate preclinical evaluation of antiangiogenic molecule.

The clinical outcomes of five groups of infertility patients receiving frozenthawed,cleavage-stage embryo transfers with exogenous hormone protocols with or without a depot gonadotropin-releasing hormone (GnRH) agonist were assessed.A retrospective cohort analysis was performed on 1003 cycles undergoing frozen-thawed,cleavage-stage embryo transfers from January 1,2012 to June 31,2015 in the Reproductive Medicine Center of Wuhan General Hospital of Guangzhou Military Region.Based on the infertility etiologies of the patients,the 1003 cycles were divided into five groups:tubal infertility,polycystic ovary syndrome (PCOS),endometriosis,male infertility,and unexplained infertility.The main outcome was the live birth rate.Two groups were set up based on the intervention:group A was given a GnRH agonist with exogenous estrogen and progesterone,and group B (control group) was given exogenous estrogen and progesterone only.The results showed that the baseline serum hormone levels and basic characteristics of the patients were not significantly different between groups A and B.The live birth rates in groups A and B were 41.67％ and 29.29％,respectively (P＜0.05).The live birth rates in patients with PCOS in groups A and B were 56.25％ and 30.61％,respectively (P＜0.05).The clinical pregnancy,implantation and on-going pregnancy rates showed the same trends as the live birth rates between groups A and B.The ectopic pregnancy rate was significantly lower in group A than in group B.We concluded that the live birth rate was higher and other clinical outcomes were more satisfactory with GnRH agonist cotreatment than without GnRH agonist co-treatment for frozen-thawed embryo transfer.The GnRH agonist combined with exogenous estrogen and progesterone worked for all types of infertility tested,especially for women with PCOS.

S100beta is one kind of the calcium binding proteins. As growth factor of neuraxon, it is excreted by neuroglial cell, and distributing in nerve tissue extensively. Although S100beta has very important values neurophysiological, it also has neurotoxicity with excreting overmuch. Concentration of S100beta changes regularity in serum after the brain injury. In addition, it has a close relations with the degree of brain damage, which can be regarded as the neural new marker of biochemistry after brain damage. The advances of S100beta protein, in the research on neurophysiological values and its application for nerve tissue injury, disease were reviewed.
Alzheimer Disease/pathology* ; Biomarkers/blood* ; Brain Injuries/physiopathology* ; Cerebrovascular Disorders/pathology* ; Humans ; Nerve Growth Factors/blood* ; Neuroglia/metabolism* ; Postmortem Changes ; S100 Calcium Binding Protein beta Subunit ; S100 Proteins/blood* ; Severity of Illness Index ; Time Factors

OBJECTIVETo investigate the alleles and genotypes frequency of 10 short tandem repeat (STR) loci (DXS101, DXS6789, DXS6799, DXS6804, DXS7130, DXS7132, DXS7133, DXS7423, HPRTB, DXS8378) on X chromosome of Chinese Hui nationality population.

METHODSThe study of 10 STR loci was performed by using the techniques of PCR, polyacylamide gel electrophoresis and silver staining.

RESULTSAmong unrelated Hui individuals, the allele numbers of 10 STR loci DXS101, DXS6789, DXS6799, DXS6804, DXS7130, DXS7132, DXS7133, DXS7423, HPRTB, DXS8378 were 9, 8, 4, 6, 6, 6, 4, 4, 5 or 5 respectively; the numbers of genotypes were 17, 22, 7, 14, 14, 15, 6, 7, 12 or 8 respectively. The distribution of genotypes from these 10 STR systems fitted the Hardy-Weinberg equilibrium (P> 0.05). Polymorphisms information content of 10 STR loci, except for DXS7133 (0.48) and DXS7423 (0.48), ranged from 0.54 (DXS6799) to 0.80 (DXS6789); the power of discrimination were from 0.89 (DXS7133, DXS7423) to 0.99 (DXS6789, DXS7132, DXS101).

CONCLUSIONThe loci of 10 STR on chromosome X are appropriate for individual identification, paternity testing involving a female child and for study on related disease.

Alleles ; Child ; China ; Chromosomes, Human, X ; genetics ; Electrophoresis, Polyacrylamide Gel ; Female ; Gene Frequency ; Genotype ; Humans ; Linkage Disequilibrium ; Male ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

Objective:To use polyamidoamine(PAMAM)dendrimer as gene delivery system for survivin gene anti- sense oligodeoxynucleotide(asODN)transfection for inhibition of HepG2 cancer cell growth.Methods:The first to the fifth generation of PAMAM and asODN were used to prepare a complex:PAMAM-asODN.The morphology of PAMAM- asODN was observed using agrose electrophoresis and atomic force microscope(AFM).PAMAM-asODN was then used to transfect HepG2 cells and cells transfected with asODN served as control.The transfection efficacy of PAMAM-asODN into HepG2 cells was observed under confocol microscope,the surviving mRNA expression was analyzed by RT-PCR,and the inhibition of HepG2 cell growth was determined by MTT assay.Results:Agrose electrophoresis showed strong complexing action between PAMAM and asODN and they formed a complex with a diameter of 25 nm.Confocol microscope showed the transfection efficacy of PAMAM-asODN was higher than that of asODN.RT-PCR showed a decreased expression of sur- vivin mRNA in PAMAM-asODN transfected cells.MTF results demonstrated that the growth of HepG2 cell was obviously inhibited after transfection of PAMAM-asODN and the inhibition rate increased with culture time,concentration of com- plex,the generation of PAMAM.PAMAM-asODN at 6.0?mol/L G4.0 resulted in a 55% inhibition of HepG2 cells 96 h after culture.Conclusion:PAMAM dendrimers can efficiently mediate the entry of survivin asODN into HepG2 cells,re- sulting in inhibition of HepG2 cells.PAMAM might be a promising gene carrier for potential molecular therapy of cancer.

Estimation of the postmortem interval (PMI) is a practical task in daily forensic casework. Researches on PMI is an important practical project in forensic field. Estimation of the time since death is influenced by internal and external, antemortem and postmortem factors, thus the old methods have limitations. Fourier transform infrared (FTIR) spectroscopy has been applied to study the pure protein, nucleic acid and carbohydrate and to detect the changes in complex cells and tissues. At present because the powerful software has could be used to achieve the spectrum transformation, smoothing, baseline correction and normalization, it is possible to analyze the samples quantitatively with the FTIR which has been applied in the biology and clinical medicine. This paper has reviewed the mechanism of FTIR and its application in biomedicine. The postmortem FTIR spectral changes were also discussed, which showed its potential for estimating PMI.
Animals ; Forensic Pathology/methods* ; Glycogen/biosynthesis* ; Humans ; Kidney Cortex/metabolism* ; Lung/metabolism* ; Muscle, Skeletal/metabolism* ; Myocardium/metabolism* ; Nucleic Acids/metabolism* ; Postmortem Changes ; Rats ; Spectroscopy, Fourier Transform Infrared/methods* ; Spleen/metabolism* ; Temperature ; Time Factors