Objective To establish a rapid assay for genotyping of IL-6-597G/A and-572G/C polymorphisms by duplex real-time PCR assay.Methods One pair of primers and two pairs of fluorescent hybridization probes had been used to genotype two mutation sites in the promoter region of IL-6 gene by fluorescent resonance energy transfer and melting curves.Results Duplex real-time PCR method for genotyping of two mutation sites simultaneously had been developed and 123 health people samples were analyzed by this new method.The results showed that three genotypes were found in IL-6 gene-572G/C polymorphisms.They were GG,GC and CC genotypes.IL-6 gene promoter-597G/A polymorphism analysis showed 4 cases displayed GA genotype and other possessed GG genotype.No AA genotype had been found.Conclusions Duplex real-time PCR method is simple and fast.It provided an accurate and economic method which is suitable for clinical gene genotyping.

China ; epidemiology ; Hexanes ; toxicity ; Humans ; Occupational Diseases ; epidemiology ; Trichloroethylene ; toxicity

Objective To evaluate the influence of grape seed proanthocyanidin extract (GSPE) on sunburn cell formation and p53 protein expression induced by acute ultraviolet injury. Methods Ten volunteers were enrolled in this study. The buttock region served as the exposed region. Four areas were randomized and delineated on the buttock: one area (control area) received no exposure or product, the other 3 areas were exposed to two minimal erythema doses (MED) of simulated solar radiation (SSR) for 3 days. Of the 3 exposed areas, one area (SSR) received no product before exposure, one area (SSR + Veh) was pretreated with vehicle, the third area (SSR + GSPE) with the samples of GSPE. GSPE or vehicle was applied 30 minutes before each exposure at 2 μL/cm2. Skin biopsy was performed 24 hours after the last exposure, and skin specimens were subjected to hematoxylin eosin (HE) staining and histochemical analysis for p53 protein. Results There was a statistical difference in the number of sunburn cells per high power field (×200) between SSR sites and SSR + GSPE sites (29.8±11.1 cells vs 2.2±0.2 cells, P<0.01). A significant decrease was noticed in the account of p53 protein-positive cells per high power field (×200) in SSR + GSPE sites com-pared with the SSR sites (4.6±0.7 cells vs 19.3±3.4 cells, P<0.05). Conclusion GSPE exerts a poten-tial protective effect against acute ultraviolet injury and can serve as a natural sunscreen.

BACKGROUND: Compared with bone marrow-derived mesenchymal stem cells (MSCs),human umbilical cord blood (hUCB)-MSCs possess many advantages,including easy acquirement,low immunogenicity,able to tolerance a higher degree of HLA-matching inconsistency,and with higher purity.OBJECTIVE: To analyze the method and condition for in vitro isolation,purification,proliferation,and osteoblast and lipoblast differentiation of hUCB-MSCs.DESIGN,TIME AND SETTING: The present observational experiment was performed in the Key National Laboratory of Biological Treatment,Huaxi Hospital,Sichuan University (i.e.,Institute of Stem Cells and Tissue EngIneering) between September 2004 and November 2005.MATERIALS: Neonatal cord blood at gestational age 37 to 40 weeks.METHODS: hUCB-MSCs were collected in aseptic condition,isolated by density gradient centrifugation,sedimenting red cells with methylcellulose followed by density gradient centrifugation,or immunomagnetic beads negative technique of CD34+.Theisolated MSCs were used to carry on plastic adherent culture in L-DMEM +10% fetal bovine serum (FBS) or MesencultTM medium +10% FBS.The third passage of cells were used for surface antigen determination by flow cytometry and induced to differentiate towards osteoblasts and lipoblasts.MAIN OUTCOME MEASURES: ①Identification of hUCB-MSCs.②Confirmation of hUCB-MSC differentiation by alizarin red and oil red staining.RESULTS: The mononuclear cells isolated by sedimented and centrifuged way cultured in MesencultTM medium +10% FBS were most available.Obvious colonies appeared in the third passage.The cells obtained by only centrifugation in density gradient were hard to form colony,and those isolated by immunomagnetic beads were hard to culture.The surface antigens of these colony cells presented CD29,CD59,and CD71,hut did not express CD34,CD45,and HLA-DR,and so on.After alizarin red staining,osteoblast-differentiating colony cell cytoplasm exhibited mineralized matrices.After old red staining,lipoblast-differentiating colony cell cytoplasm was full of lipid vacuoles.CONCLUSION: The MSCs can be successfully isolated by sedimenting red cells with methylcellulose followed by centrifugation and cultured in MesencultTM medium +10% FBS.Obvious colony growth appears in the third passage.After induction,the MSCs can differentiate into osteoblasts and lipoblasts.

BackgroundEndostatin (ES) is currently the strongest endogenous angiognesis inhibitor,and it can inhibit the occurrence of neovascularization.Various studies demonstrated that the poly RGD sequence can enhance the function of the ES gene.ObjectiveThis study was to evaluate the use of gene therapy of modified ES for alkaline burn-induced corneal neovascularization (CNV).MethodsOne hundred and two clean SD rats were randomly divided into the normal control group,the pCI empty vector group,the pCI-ES group,and the pCI-RGDRGDES group.Corneal neovascularization models were established by placing a piece of 3 mm filter paper with 1 mol/L NaOH at the central cornea for 40 seconds.3 μg of the pCI blank vector,ES-tranfected pCI blank vector,or RGDRGD-ES-transfected pCI vector was injected into the superior bulbar conjunctiva after the alkali burn twice at 1-week intervals.Area of CNV and edema of the cornea in the various groups of rats were examined daily under the slit lamp biomicroscope.1,4,7 and 14 days after operation,the rats were sacrificed by the excessive anesthesia method and corneal tissues were obtained to evaluate pathological changes.The expression of CD34 in vascular endothelial cells was detected by immunochemistry to calculate the corneal neovascular density.The expressions of VEGF mRNA and Flk-1 protein in the corneas were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The use and maintenance of animals followed the Statement of ARVO.Results Seven to fourteen days after corneal alkali-burning,the corneal neovascular area was smaller in the pCI-ES group and pCI-RGDRGD-ES group compared with the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and nevascular area in the pCI-RGDRGD-ES group was smaller than that in the pCI-ES group (P＜0.05).The expression level of CD34 was significantly lower in the pCI-ES group and pCI-RGDRGD-ES group than that in the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and the expression level of CD34 was further declined in the pCI-RGDRGD-ES group compared with the pCI-ES group (P＜0.05 ).Compared with the normal control group and pCI vector group,the expressions of the Flk-1 protein and VEGF mRNA were decreased in the pCI-ES group and pCI-RGDRGD-ES group on the fourth day after corneal alkali-burning (P＜0.01,P＜0.05 ),and those in the pCI-RGDRGD-ES group were less than the pCI-ES group (P＜ 0.05,P＜ 0.05 ).Conclusions Subconjunctival injection of both ES and modified RGDRGD-ES genes result in significant suppression of CNV in vivo,and modified RGDRGD-ES appears to be more effective than native ES.The main mechanism of ES in inhibiting neovascularization is to downregulate the expression of VEGF and Flk-1.

Background Uveal effusion syndrome is uncommon in clinic.To understand the clinical characteristics of uveal effusion syndrome is helpful for rescuing visual acuity of patient.Objective This study was to discuss the diagnosis,classification and surgical outcome of uveal effusion syndrome.Methods This was a descriptive study.The clinical data of 14 eys from 10 patients with uveal effusion syndrome,ineluding ophthalmologic examination,B-scan sonography,ultrasound biomicroscopy (UBM),fundus fluorescence angiography (FFA),indocyanine green angiography (ICGA),surgical treatment and prognosis,were retrospectively analyzed.The follow-up period was 6 months.Results The fundus findings of all impacted eyes showed bullous-shape retinal detachment (RD).B-scan sonography revealed retinal and choroidal detachment.A annular peripheral ciliochoroidal detachment was observed in the cases under the UBM.FFA exhibited leopard spots without any leakage from choroid into the subretinal space.ICGA demonstrated diffusely choroidal granular hyperfluorescence in the very early phase,which presented with an increasing intensity as time lapse until the late phase.Full-thickness sclerectomy was performed on 4 eyes of 2 patients and subscleral sclerectomy was performed in 1 eye of 1 patient,achieving a retinal anatomic reattachment after surgery.All of the patients finished the fellow-up.No recurrence of RD was seen during the followup duration.Conclusions Comprehensive preoperative evaluation,including ophthalmologic ultrasonography,MRI and CT,is crucial for accurate classification of uveal effusion syndrome and determine of proper management strategy.

The secondary metabolites of plants are important sources of natural drugs. Betula plants have abundant pharmacological value, complex mechanism and wide applications, which are closely related to the triterpenoids of theirs. Triterpenoids in Betula species are mainly divided into dammarane-type, ocotillol-type, oleanane-type, lupane-type and cycloaltunane-type. The extracts of Betula species have varieties of activities such as anti-tumor, anti-inflammatory, anti-oxidant, anti-bacterial, etc. And the biosynthetic pathways of triterpenoids after 2,3-oxidosqualene are split into four branches of dammarenediol-II, lupeol, cycloartenol and amyrin according to the different oxidosqualene cyclases. This review summarizes the chemical constituents, pharmacological activities and biosynthetic pathways of triterpenoids in Betula plants. It provides a reference for the research and development of new drugs and the production of these triterpenoids in microbial cell factories by synthetic biology methods.

BACKGROUNDElectrocardiographic (ECG) changes occurring during the course of acute brain injury (ABI) have been described frequently, but their significances remain uncertain. The present study was designed to investigate the relation of ECG abnormalities to outcome in the patients with ABI.

METHODSWe performed a retrospective, observational study on the ABI patients admitted to the Department of Neurosurgery of the Beijing Tiantan Hospital between December 2005 and December 2007. All the patients accepted 12-lead electrocardiographic examination within 24 hours after injury, then divided into three groups according to the Glasgow coma score (GCS). In-hospital mortality and one-month outcome assessed by the Glasgow outcome score (GOS) were investigated.

RESULTSOf 335 ABI patients (mean ages 32.4 years), 246 patients (73.4%) had abnormal ECGs. The most common abnormality was ST-T changes (41.5%), followed by sinus tachycardia (23.6%). ECG changes had a significant association with the severity and outcome. Logistic regression analysis showed the presence of ST-T changes (OR 2.587, 95%CI 1.009 to 6.629, P = 0.048) and QT dispersion prolongation (OR 4.656, 95%CI 1.956 to 11.082, P = 0.001) significantly associated with short outcomes.

CONCLUSIONSABI can lead to myocardial damage and ECG changes had a significant association with the severity. ST-T changes and QT dispersion prolongation were the independent prognosis factors for the negative outcome of ABI patients.

In this study, based on the transcriptome data, we cloned the full-length cDNAs of TwAACT gene from Tripterygium wilfordii suspension cells, and then analyzed the bioinformation of the sequence, detected the genetic differential expression after being induced by methyl jasmonate (MeJA) by RT-PCR. The full-length cDNA of the TwAACT was 1 704 bp containing a 1 218 bp open reading frame (ORF) encoding a polypeptide of 405 amino acids (GeneBank accession No. KP297934). The deduced isoelectric point (pI) was 6.10, a calculated molecular weight was about 41.20 kDa, and online prediction showed that TwAACT had two catalytic active sites. After the induction of MeJA, the relative expression level of TwAACT increased rapidly. The expression level of TwAACT was highest at 24 h. TwAACT was cloned firstly, that laid the foundation for identifying thegene and illustrating thebiosynthesis mechanism and its synthetic biology.
Acetyl-CoA C-Acetyltransferase ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Cloning, Molecular ; Gene Expression Regulation, Plant ; Models, Molecular ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; chemistry ; genetics ; metabolism ; Sequence Alignment ; Tripterygium ; chemistry ; classification ; enzymology ; genetics

Objective To learn the present status of defluoridation water improvement project in Shanxi province in order to provide scientific basis for speeding up the prevention and control of endemic fluorosis.Methods According to "The National Technical Scheme for Endemic Disease Control" from 2005 to 2009, the investigation points were selected in the counties that implemented the measures of water improvement and defluoridation,the status of drinking water defluoridation Project was investigated, and the water fluoride levels were determined by fluoride selective ion electrode. Results The primary status was surveyed in 1658 water improvement and defluoridation projects in 51 counties. The resource of drinking water for water improvement and defluoridation projects was mostly ground water[accounting for 93.12% (1544/1658)]. Among 1658 water improvement and defluoridation projects 1405 projects worked well(accounting for 84.74%) and 190 projects intermittently worked (accounting for 11.46%). Sixty three projects abandoned (accounting for 3.80%), in Datong basin the abandoned projects accounted for 36.36% (12/33). Water fluoride content of 1595 water improvement and defluoridation projects had been determined, among them water fluoride content of 874 projects were above 1.0 mg/L (accounting for 54.80%). The situations of exceeded national standard in the five basins was different(H = 33.22,P ＜ 0.01). The rate of over national standard of fluoride levels in drinking water was 88.37%(38/43) in Datong basin. Therefore, in Datong basin water improvement should be strengthened. Conclusions In Shanxi province the water improvement and defluoridation projects are basically running normally. However, the qualified rate is lower for the water improvement and defluoridation projects. The water improvement status varies dramatically among areas.The situation is still grim in Shanxi province. Water improvement and defluoridation needs to be strengthened to improve the effect of prevention and control of the disease.