Objective To analyze the levels and speciation of arsenic metabolites in urine of rats treated with sodium arsenite and sodium arsenate in order to investigate the different aspects of metabolism between sodium arsenite and sodium arsenate,thus to understand further the basic data about relationship between it's metabolism and mechanism of toxicity. Methods Seventy Wistar rats,weighting 80-120 g,were divided into 7 groups of 10 each,such as normal control group,high,middle and low sodium arsenite group and high,middle and low sodium arsenate group. After the animals were fed for one month,the urine was collected by metabolic cage in 12 hours. Applying the high efficiency liquid chromatography and hydride genesis atomic fluorescence spectroscopy (HPLC-HGAFS),the levels and speciation of arsenic metabolites were determined in urine of rats. Meanwhile,the recovery rate of dimethyl arsinic acid(DMA) would be determined to estimate the degree of accuracy of results. Results The levels of iAs~(3+),iAs~(5+) and DMA in middle sodium arsenite group[(121.66±1.26),(10.26±2.68),(200.91±0.56) μg/L]were higher than the high sodium arsenite group[(113.20±0.75),(5.16±1.32),(147.70±μ0.77)μg/L,all P < 0.05]and low sodium arsenite group[(79.35±2.12),(5.13±2.25),(56.35±1.23)μg/L,all P < 0.05]. The levels of iAs~(3+) and DMA in middle sodium arsenate group[(315.81±1.69),(245.12±1.18)μg/L]were higher than the high sodium arsenate group[(85.03±0.56),(110.34±1.04)μg/L,all P< 0.05]and low sodium arsenate group[(22.97±2.67),(15.75±2.15)μg/L,all P < 0.05]. Compared with sodium arsenate group,the levels of iAs~(3+) and DMA in high and low sodium arsenite group were higher(all P < 0.05) ; and the levels of iAs~(3+) and DMA in middle sodium arsenite group were lower(all P < 0.05). Meanwhile,the average urinary recovery rate of DMA of rats in different sodium arsenite group were 94.80%-102.70%,and the average urinary recovery rate of DMA of rats in different sodium arsenate group were 95.33%-108.40%. Conclusion The speciation and levels of arsenic are influenced by the external exposure dose,and some distinction appeared in the metabolism and metabolic path between sodium arsenite and sodium arsenate in urine in vivo.

Lycii Cortex, a popular herb medicine in traditional Chinese medicine, is used to treat different inflammation-related diseases. The aim of our work is to find the key constituents inhibiting NF-kappaB, a key regulator of inflammation. In the investigations of cell-based in vitro assays of extracts, we found that both ethyl acetate extract and methanol extract of Lycii Cortex inhibited the TNF-alpha-induced activation of NF-kappaB. Through bioassay-guided fractionation, we identified 4 phenolic amides including trans-N-(p-coumaroyl) tyramine (1), trans-N-feruloyltyramine (2), trans-N-caffeoyltyramine (3), and dihydro-N-caffeoyltyramine (4). Four phenolic amides showed differently inhibitory activities on TNF-alpha-induced NF-kappaB activation. Trans-N-caffeoyltyramine (3) was identified as the key component with an IC50 of 18.41 micromol x L(-1). It was suggested that the hydroxyl group at C-3 in trans-N-caffeoyltyramine might be a key binding site and its C-7,8-double bond might play an important role on NF-kappaB inhibitory activities as the link of the conjugation of pi electrons leading to a partial planar conformation. It might be inferred that the biological activity of compound 3 is attributed to the structure of Michael reaction acceptor containing alpha, beta-unsaturated ketones and benzene along with hydroxyl group in o-diphenol.
Biological Assay ; Cell Line ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Humans ; Inflammation Mediators ; antagonists & inhibitors ; immunology ; Lycium ; chemistry ; Molecular Structure ; NF-kappa B ; antagonists & inhibitors ; immunology

Objective To explore the effects of sodium arsenie on Metallothionein(MT) isoforms genes expression,and to study the relevance between the MT expression and cell survival percentage. Methods Healthy persons blood was extracted aseptically and the lymphocytes were separated. The lymphocytes were treated by 0 (control) ,2,5,10,15,30,60 μmol/L sodium arsenites, respectively. The cell survival percentage was determined by methyl thiazolyl tetrazolium(MTT) reduction assay at 24,48,72 h intervals, while the expression of MT-1 and MT-2 were examined by RT-PCR in 72 h. Results The cell survival percentage in 2,5μmoL/L groups were (115.50± 11.80)% and (130.49±8.28)%,which were all higher than those in the control group [(100.00±0.00)%,all P < 0.05]at any intervals. In 10,15,30,60 μmol/L groups,the cell survival percentage[(78.12±9.33)%,(71.62± 10.82)%,(52.06±3.05)%,(40.98±5.41)%]increased along with the decrease of concentration,which showed a significant difference between these groups and the control group and 2,5μmol/L groups(all P < 0.05). There was no significant difference on MT-1 expression after exposed to different concentration sodium arsenites (0,2,5,10,15,30,60 μmol/L) for72 h(0.925±0.123,1.082±0.504,1.103±0.170,0.927±0.056,0.730±0.307,0.604± 0.173,0.540±0.075,all P > 0.05). The expression of MT-2 in 2,5μmol/L groups(1.503±0.212,1.557±0.377) was up regulated compared with that in control group(0.702±0.112) and MT-2 also expressed more than that in other groups(all P < 0.05). However,the expression of MT-2 declined when the concentraion was 10,15,30,60 μmol/L(0.814±0.139,0.679±0.201,0.607±0.229,0.533±0.102). There was no significant difference among the groups (all P > 0.05). The expression level oE MT-1,MT-2 was positively correlated with the cell survival pereentage(r = 0.955,0.909,all P < 0.05). Conclusions The sodium arsenite at concentration of 10 μmoL/L might inhibit the expression of MT of lymphocytes and low concentration sodium arsenite (2,5 μmol/L) might stimulate the lymphocytes to regulate the expression of MT-2 to higher levels,which can increase the cell survival percentage and exert the function of protecting cells.

OBJECTIVETo compare the regional load deflection rate (LDR) of multiloop edgewise arch wire (MEAW) of three dimensions with coupled use of two dimension brackets in the individual interbracket span, to understand the mechanical properties of MEAW.

METHODSThe MEAW arch wires of stainless steel of three dimensions, 0.41 mm x 0.56 mm, 0.43 mm x 0.64 mm and 0.46 mm x 0.64 mm, were bent into single L-loop. The study was performed with the coupled use of 0.41 mm x 0.56 mm wires with 0.46 mm x 0.64 mm bracket (A bracket) and 0.43 mm x 0.64 mm, 0.46 mm x 0.64 mm wires with 0.56 mm x 0.71 mm bracket (B bracket). The LDR of each L-loop at the individual interbracket span when loading and unloading was measured. The data were analysed by SPSS 11.0.

RESULTSCompared the regional LDR for the couple of 0.41 mm x 0.56 mm L-loop with A bracket with those for the couple of 0.43 mm x 0.64 mm L-loop with B bracket, the former showed lower value than the latter at regions between the upper central and lateral incisor, the lower central and lateral incisor, and between the lower lateral incisor and canine (P < 0.05). For the rest regions, the two couples exhibited similar value to the regional LDR (P > 0.05). The regional LDR for the couple of 0.41 mm x 0.56 mm L-loop with A bracket were lower at all regions than those for the couple of 0.46 mm x 0.64 mm L-loop with B bracket (P < 0.05) except that at the region between the lower first and second molars which showed similar value between the two couples.

CONCLUSIONThe coupled use of B bracket with 0.43 mm x 0.64 mm MEAW arch wire and A bracket with 0.41 mm x 0.56 mm MEAW arch wire exhibited similar mechanical properties.

Humans ; Molar ; Orthodontic Brackets ; Orthodontic Wires ; Stainless Steel

OBJECTIVETo set up random amplified polymorphic DNAs (RAPD) method in genotyping Neisseria gonorrhoeae on DNA level, and to explore its use to trace the source of infection.

METHODSFour different pretreatments were used to extract the Neisseria gonorrhoeae genomic DNA with its advantages and disadvantages compared. Arbitrary sequence was then used to amplify the genomic DNA of Neisseria gonorrhoeae and RAPD fingerprint maps was applied to distinct the Neisseria gonorrhoeae strains. Finally, RAPD fingerprint of Neisseria gonorrhoeae strain between patient and his/her sexual partner was compared.

RESULTSCetyltrimethylammonium bromide method was classical in extracting genomic DNA, and could get integrated genomic DNA and good fingerprint maps, since main segments were common to all the Neisseria gonorrhoeae but some were different among strains so that the fingerprint of different Neisseria gonorrhoeae were distinctive. However, fingerprint maps of Neisseria gonorrhoeae collected from sex partners were quite similar.

CONCLUSIONBased on genomic levels, effective fingerprint maps could be identified and to classify the Neisseria gonorrhoeae into different genotypes. RAPD fingerprint maps could be used to trace the source of infection.

DNA Fingerprinting ; DNA, Bacterial ; analysis ; Genotype ; Humans ; Neisseria gonorrhoeae ; classification ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVEUsing molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.

METHODSA random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.

RESULTSSelected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.

CONCLUSIONRAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.

Adolescent ; Adult ; Anti-Bacterial Agents ; pharmacology ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Female ; Genotype ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; Random Amplified Polymorphic DNA Technique

Adolescent ; Adult ; Aged ; Ankle Injuries ; surgery ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Middle Aged ; Soft Tissue Injuries ; surgery ; Surgical Flaps

Objective　To evaluate the therapeutic effect of rapid angulation rotation traction for lumbar intervertebral disc protrusion. Methods　A total of 88 patients (66 male and 22 female;age: 18-65), with a history of 2 days to 10 years were analyzed. Among them 20 cases were central protrusion 68 cases were lateral protrusion. Traction range: 60-65 mm, angle: 20°-25°, rotation degree: 20°-25°, traction time: 3 s-1 min. The process of treatment was computer designed. The traction was then 1 to 3 times, with a interval of 4-7 days between two treatments. Results　The effective rate was 96% for the case with history <1 month 96% and 85% for cases with history >1 month (P<0.01). The effective rate was 62% for central protrusion, and 87% for lateral protrusion respectively. χ2 test showed the difference was not significant. Conclusion　The rapid angulation rotation is an effective treatment for lumbar inter-vertebral disc protrusion and this non operative method should be used as early as possible.

OBJECTIVESTo identify and clone human genes transactivated by HCV F protein by constructing a cDNA subtractive library using the suppression subtractive hybridization technique.

METHODSSuppression subtractive hybridization (SSH) and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV F protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1 (-)-F or with pcDNA3.1(-) empty vector as a control, and SSH method was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small sized cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 or adaptor 2. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5 alpha. The cDNA was sequenced and analyzed in GenBank with blast search after PCR.

RESULTSThe subtractive library of genes transactivated by HCV F protein was constructed successfully. The amplified library contains 71 positive clones. Colony PCR shows that 56 clones contain 200-1000 bp inserts. Sequence analysis was performed on 28 clones randomly, and the full length sequences were obtained with using the bioinformatics method. Altogether 19 coding sequences were obtained, consisting of 17 known and 2 unknown.

CONCLUSIONSThe obtained sequences may be target genes transactivated by HCV F protein, and some gene coding proteins are those involved in cell cycle regulation, metabolism, and cell apoptosis.

Cloning, Molecular ; Hepacivirus ; genetics ; Humans ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; biosynthesis ; genetics ; Transcriptional Activation ; Viral Core Proteins ; biosynthesis ; genetics

BACKGROUNDImmunological sensitization remains a major problem following renal transplantation. There is no consensus for the management of sensitized renal allograft recipients. The patients become tethered to dialysis while waiting for compatible donors. This study was designed to evaluate the efficacy and safety of preoperative single-bolus high-dose antithymocyte globulin (ATG) as induction therapy in sensitized renal transplant recipients.

METHODSA total of 56 patients were divided into two groups according to the level of panel reactive antibody (PRA): non-sensitized group (PRA < 10%, n = 30) and sensitized group (PRA > or = 10%, n = 26). The characteristics of the recipients and donors were comparable between the two groups. Mycophenolate mofetil (MMF, 1 g) or ATG (iv. 9 mg/kg) were given preoperatively in the two groups as induction therapy. After the transplantation, the patients were treated with standard triple therapy regimen consisting of tacrolimus (FK-506) or cyclosporine A, MMF, and prednisolone. Acute rejection (AR) and infection episodes were recorded and renal function was monitored during a 12-month follow-up. Chi(2) test and t test were used to analyze the data.

RESULTSDuring the follow-up, 6 patients (20.0%) suffered AR episodes in the non-sensitized group and 4 (15.4%) in the sensitized group (P = 0.737); 8 patients (26.7%) experienced 11 infection episodes (average, 1.4 episodes per infected patient) in the non-sensitized group, and 6 (23.1%) experienced 10 infection episodes (average, 1.7 episodes per infected patient) in the sensitized group (P = 0.757, 0.890). The safety of the drugs, which was assessed by the occurrence of side effects, was comparable between the two groups. The hospital stay was 13 - 25 days (mean, 16.7 +/- 3.3) in the non-sensitized group and 14 - 29 days (mean, 16.2 +/- 3.1) in the sensitized group, respectively (P = 0.563). No delayed graft function (DGF) was observed in all the patients. Both the 12-month actuarial patient and graft survival rates were 100% in the two groups.

CONCLUSIONPreoperative single-bolus high-dose ATG is an effective and safe induction therapy yielding acceptable acute rejection rate in sensitized renal transplant recipients.

Adult ; Antilymphocyte Serum ; adverse effects ; therapeutic use ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Humans ; Immunosuppressive Agents ; Kidney Transplantation ; Male ; Middle Aged