Objective　Standard method of testing serum fluoride was tested and verified by certified reference material CRM of free fluo ride component in cattle serum (GBW 09143～09144).The method was suited for fluor ide research to assess its practical value.Methods　ISE-standard curve,linear relationship,linear range,sensitive and precision were disired result by CRM.400 serum of fluoride in rats were measured.Results　The linear ranges of the method were 0.02～5.0 mg/L,r=0.9999,b=57.8,and test lower limit was 0.013 mg/L.The test value was equal to standard value.Conclusions　The standard method is accurate,sensitive,convenient and small sample is used;and it is easy to spread.Thus,it has been of practical value.

Anemia, Aplastic ; diagnosis ; etiology ; Benzene ; poisoning ; Expert Testimony ; Humans ; Male ; Occupational Diseases ; diagnosis

Objective To explore the acute toxicity and LD 50 of Tripterygium Hypoglaucum (Levl) Hutch(THH) solution to provide information for safe clinical application. Methods After oral administration of THH solution in mice, the mortality and the physiological and pathological changes were observed. Results The LD 50 (95% confidence limit) of THH in male and female mice was 79 g/kg(69～89 g/kg) and 100 g/kg (90～112 g/kg), respectively. No marked pathological change of the organs was found. Conclusion According to the standard of grading of acute toxicity, THH solution belongs to the moderate class. Therefore, it is safe in clinical practice and has a wide application.

Objective The aim of the study WSfl to establish the method using CDl27 as the new biomarker to identify regulatory T cells(Treg cells)and apply the CD 127 to detect the Treg cells in patients with gastric cancer.Methods The phenotypes of Treg cells were analyzed using five-color flow cytometry method Foxp3.FITC/CD127-PE/CD4-PerCP/CD25-APC/CD3-PC7.The mRNA and protein expression of Foxp3 in isolated CD+4 CD25high CD127-/low Treg cells were detected.The relationships between Foxp3 and CD127 protein expression in CD4+ T cells from aduh human peripheral blood were investigated.PBMCs,Ascltes,turnor-infihration lymphocyte and tumor-draining lymph nodes in 35 patients with gastric cancer and PBMCs in 20 normal healthy donors were evaluated for the proportion of Treg ceils,as well as the percentage ot the total CD +4 cells.Results CD4+ CD25 high CD-127 low ceils expressed the high Foxp3 in protein level(87.1%)and mRNA level.Within the CD4+CD+25 population,there was a significant correlation between Foxp3 and the CD127low phenotype(r=0.985,P<0.01).Compared with healthy olunteers,patients with gastric malignancies had a higher proportion of CD4+4 Cdhigh 25 CD-low127 cells in peripheral blood(t=2.542,P<0.05).The Dereentages of Treg cells were more abundant in ascites(t=2.357,P<0.05),TIL(t=6.174,P<0.01) and tumor-draining lymph nodes(t=5.481,P<0.01)of individuals with gastric cancer than that in their blood.There were significant differences in the prevalence of Treg ceils between the early and advanced disease stages in gastric cancer[(6.04±2.31)%in stageⅠ+Ⅱ VB(10.16±2.29)% Ⅲ+Ⅳ,t=2.473,P<0.05].Conclusions The CD127 biomarker can be used to selectively enrich human Treg cells for in vitro functional studies.The populations of CD4+ CD high25 CD127-/low Treg cells increased ith tumor stage in individuals with gastric cancer.

Objective To investigate the populations of CD+4CD25high Foxp3+ regulatory T cells(Treg cells)and mRNA expression of Foxp3 in peripheral and the marginal region of tumor from patients with gastric cancer,and to evaluate the prevalence of Treg cells from patients with gastric cancer in relation to the TNM stages.Methods PBMC,Ascites,tumor-infiltration lymphocyte and tumor-draining lymph nodes in 47 patients with gastric cancer(TNM I11,Ⅱ18,Ⅲ10,Ⅳ8)and PBMC in 20 normal healthy donors were evaluated for the proportion of Treg cells,as a percentage of the total CD4+ cells,by flow cytometric analysis.Levels of mRNA for Foxp3 were measured witll a real-time quantitative PCR Results The percentage of CD4+CD25high Foxp3+ Treg cells in PBMC for cases of gastric cancer were significantly higher than those for healthv donors(P<0.05).The percentage of Treg cells were more abundant in ascites(P< O.05),1rIL(P<0.01)and tumor-draining lymph nodes(P<0.01)of individuals with gastric cancer than in their blood.And the Foxp3 mean fluorescent intensity (MFI) of Treg cells in TIL and tumor draining lymph nodes with gastric caners were significantly higher than PBMC and ascites(P<0.05).The proportion 0f Treg cell in patients of gastric cancers with stage I,II,Ⅲ,Ⅳ were(5.72±1.95)%,(6.45± 2.23)%,(9.58±3.13)%,(11.70±2.30)%,respectively.There were significant correlation between Drevalenee of Treg cells and disease stages in gastric cancer(r=0.784,P<0.01).Foxp3 mRNA levels were higher in PBMC from the group with gastric cancers than in the group with healthy donors.Foxp3 mRNA levels were correlated with TNM stages(r=0.623,P<0.05).Conclusions Our results suggest that the populations of CD4+CDhigh25 Foxp3+Treg cells and Foxp3 mRNA levels in patients with gastric cancer are signifieantlv higher in comparison to those in control donors.In addition,the expression of Fox：p3 mRNA increased with tumor stage.The increased prevalence of Treg cells may be one of the explanations for impaired cell-mediated immunity in cancer bearing hosts.

OBJECTIVETo study interaction between a novel centrosomal protein TACP1 and mitotic kinase Nek2A.

METHODSNek2A305-446 protein was expressed and purified in E.coli and TACP1 protein was expressed in transfected 293T cells. Pull-down assay was used to examine the interaction between Nek2A305-446 and TACP1. TACP1 and Nek2A complex was tested by co-immunoprecipitation assay with polyclonal anti-TACP1 antibody. The localization of those two proteins in Hela cells was verified by immunofluorescence.

RESULTSTACP1 was pulled down by Nek2A305-446 protein but not by GST control. Nek2A was co-precipitated with TACP1 protein by polyclonal anti-TACP1 antibody but not by pre-immunization serum. The Immunofluorescence test showed that these two proteins formed a complex at centrosome during mitosis.

CONCLUSIONCentrosomal protein TACP1 is a novel interacting protein with Nek2A, both of which are localized in centrosome during mitosis.

Cell Line ; Centrosome ; metabolism ; Escherichia coli ; genetics ; Fluorescent Antibody Technique ; HeLa Cells ; Humans ; Immunoprecipitation ; Mitosis ; NIMA-Related Kinases ; Protein Binding ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Telomere-Binding Proteins ; genetics ; metabolism ; Transfection

RNA-Sequencing (RNA-Seq) is a newly-developed method in transcriptome research, it can afford more accurate transcription information and be more quickly by using Next-generation Sequencing (NGS) technology. RNA-Seq has been widely used in various biological fields. Genuine traditional Chinese medicines (TCM), with good quality and therapeutic effect, were always praised highly and used by famous physicians. The geo-herbalism formation of TCM is based on the product of the gene expression at specific space and time. So it has been a research hotspot to analyze the mechanism of biosynthesis through RNA-Seq in the study on the secondary metabolism of medicinal plant. This article mainly illustrates the RNA-Seq and its advantages, it also discusses the potential application in genuine TCM, and it can provide useful information for other researchers.
Drugs, Chinese Herbal ; Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Medicine, Chinese Traditional ; Plants, Medicinal ; genetics ; RNA ; Sequence Analysis, RNA ; Transcriptome

OBJECTIVERadial corrective osteotomy is an established but challenging treatment for distal radial malunion. There is an ongoing discussion about whether an opening or closing-wedge osteotomy between should employed. The purpose of the present study was to retrospectively compare the clinical and radio graphic results between conventional opening-wedge osteotomy and closing-wedge technique.

METHODSFrom January 2004 and December 2012,42 patients with extra-articular distal radial malunion were managed with corrective osteotomy and were followed for a minimum of one year. Twenty-two patients (5 males and 17 females, ranging in age from 25 to 75 years old) were managed with radial opening-wedge osteotomy and implanting of interpositional bone graft or bone-graft substitute, and twenty patients (4 males and 16 females, ranging in age from 19 to 79 years) were managed with simultaneous radial closing-wedge and ulnar shortening osteotomy without bone graft. The selection of the surgical procedure was determined by the surgeon. Each patient was evaluated on the basis of objective radio graphic measurements, and functional outcomes were determined on the basis of clinical examinations, including range of wrist motion, grip strength, pain-rating score, Mayo wrist score, and Disabilities of the Arm, Shoulder and Hand (DASH) score.

RESULTSThe mean duration of follow-up was 36 months (ranged, 12 to 101 months) for the opening-wedge cohort and 28 months (ranged, 12 to 87 months) for the closing-wedge cohort. The two techniques were comparable in terms of complications. Post-operative volar tilt and ulnar variance were improved significantly in each cohort. The ulnar variance was more frequently restored to within defined criteria (22.5 to 0.5 mm) in the closing-wedge cohort than that in the opening-wedge cohort. The post-operative mean extension-flexion are of the wrist and Mayo wrist score were significantly better in the closing-wedge cohort. Differences in the pronation-supination arc, grip strength, pain-rating score, and DASH scores between these two cohorts were not significant.

CONCLUSIONThe closing wedge osteotomy technique is an effective reconstructive procedure for the treatment of extra-articular distal radial malunion. It is significantly better than the opening-wedge osteotomy technique in terms of the restoration of ulnar variance, the extension-flexion arc of wrist motion, and the Mayo wrist score.

Adult ; Aged ; Bone Nails ; Case-Control Studies ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Osteotomy ; Radius Fractures ; surgery ; Range of Motion, Articular ; Retrospective Studies ; Wrist Joint ; surgery ; Young Adult

Objective:To report the preliminary results of intensity modulated radiation therapy(IMRT)for 20 nasopharyngeal carcinoma patients.Methods:A total of 20 patients with nasopharyngeal carcinoma received IMRT in our hospital between January 2007 and April 2008.Five patients were of stage Ⅱ,13 patients were of stage Ⅲ,and 2 patients were of stage Ⅳ.The prescribed dose 69.96 Gy was delivered to the gross tumor volume(PTV)and positive neck nodes(PTVnd);59.36 Gy to the clinical target volume(PTV1),covering the upper neck and area around the nasopharynx;and 50.96 Gy to the low neck and supreclavicular area (PTV2).The dose to 50% of the parotid was≤35 Gy.The maximum dose to the lens.pituitary gland,temporo-mandibular joint,mandible,and temporal lobe was 9,54,60,70,and 60 Gy.The maximum dose to the brainstem,spinal cord,optic nerve and optic chiasma(PRV)was 54,40,54,and 54 Gy,respectively.All of the patients received 1 or 2 circles of chemothrapy before IMRT.Results:The median follow-up time was 14 months.The one-year overall survival was 94.1%.One patient died of osseous metastasis and respiratory failure and 3 patients developed distant metastasis.Acute toxicity was mostly Grade Ⅰ to Grade Ⅱ.Seventeen patients had grade Ⅰ xerostomia.Three patients suffered from grade Ⅰ acute oral mucosa reaction and 12 patients had grade Ⅱ acute oral mucosa reaction.Analysis of the dose-volume histograms (DVHs) showed that the mean dose delivered to the PTV,PTVnd,PTV1 and PTV2 was 73.4,74.1,67.8,and 54.1 Gy,respectively.The median dose to 50% of the right and left parotid glands was 43.9 Gy and 41.9 Gy,respcetively.The average value of maximum dose to the left and right lens was 8.06 and 8.12 Gy,respectively.The average value of maximum dose to the brainstem,spinal cord,left and right optic nerve and optic chiasma PRV was 60.6,46.6,50.0,55.0,and 56.0 Gy,respectively.Conclusion:IMRT can achieve satisfactory dose distdbution to nasopharyngeal carcinoma and surrounding tissues in NPC patients,protect normal tissues during the treatment and improve local control rate.

This work reports the properties of dextran-spermine polycation (DSP) as a gene vector and its gene transfection efficiency in vitro. Oxidized dextran was reacted by reductive amination with spermine to obtain DSP, the resulted polycation was then incubated with plasmid pEGFP to form polyplexes by electrostatic interactions. DSP formed stable polyplexes when the weight ratio (DSP/DNA) varied from 4 : 1 to 20 : 1. The particle size and zeta potential of polyplexes were in the range of 162.6 - 187.9 nm and increased from + 8.45 mV to + 39.6 mV, respectively. DSP could effectively protect condensed DNA from DNase I degradation, and it showed strong buffering capacity in a certain pH range. The highest yields of transfection efficiency were found to be as high as Lipofectamine 2000 when the polyplexes were transfected to SMMC-7721 and BHK-21 cells at the weight ratio of 8 : 1. This research indicates that dextran-spermine polycation is a highly active gene vector in vitro.
Animals ; Cell Survival ; Cricetinae ; DNA ; administration & dosage ; Dextrans ; administration & dosage ; Genetic Vectors ; Spermine ; administration & dosage ; Transfection ; methods