AIM: To investigate whether a consensus oligonucleotide, which contains nuclear factor-kappa B(NF-?B) binding site, decreases TNF production stimulated by lipopolysaccharide(LPS). METHODS: The thiophosphoric acid modified oligonucleotides were transferred into rat peritoneal macrophages directly, TNF content in LPS-stimulated cell culture supernatant was determined by ELISA.RESULTS: Adding the consensus oligonucleotides with NF-?B binding site into macrophages markedly decreased TNF production following LPS stimulation. CONCLUSION: These results show that use of the consensus oligonucleotides with NF-?B binding site, which can combine with NF-?B, can block or decrease TNF production induced by LPS in macrophages.

AIM: The selective recognition of the sense peptides which are located in special regions of thyrotropin receptor (TSHR) by their corresponding antisense peptides has been investigated. Three pairs of sense and antisense peptides were named TR1 (aa37-45) and RT1 (aa45-37), TR2 (aa353-366) and RT2 (aa366-353), TR3 (aa648-655) and RT3 (aa655-648). METHODS: To prepare three affinity chromatography columns, antisense peptides were immobilized, called RT1-sepharose 4B, RT2-sepharose 4B and RT3-sepharose 4B, respectively and investigate the retardative behavior for each of native peptide TR1, TR2 or TR3 on above columns with stepwise elution. RESULTS: Each of the three immobilized antisense peptides recognized and retarded its corresponding sense peptide-TR1, TR2 or TR3 instead of those non-complementary peptides. Immobilized RT1 recognized free TSHR protein molecule as well. In additional, bovine thyrotropin was recognized by immobilized TR1. CONCLUSION: The results indicate that molecular recognition theory exsits in thyrotropin receptor system. It may be useful to isolate biological molecules and to locate epitopes of TSH on TSHR molecule. Otherwise, antisense peptide may be used for treatment of experimental autoimmunolized thyroid disease (AITD) in the rat. [

Objective To study the immune regulation of antisense peptide in rats by observing immune function of activity fragments of thyrotropin receptor (TSHR) and their corresponding antisense peptides. Methods TSHR peptides TR1, TR2, TR3 and their antisense peptides RT1, RT2, RT3, and three pairs of complementary peptides were injected into rats of different groups respectively, and the serum levels of TT_3, TT_4, TSHR antibody (TRAb), thyroid stimmulating antibody, thyroid blocking antibody and TSH antibody (TSHAb) and pathological changes in thyroid tissue were investigated. Results Serum TRAb could be induced when each of three fragments of TSHR was injected into rats; TRAb and TSHAb were induced by RT1 or RT2; epithelial hyperplasia and lymphocytic infiltration observed in thyroid tissue of rats injected with TR2 could be abated by injecting RT2 subsequently. Conclusion The results suggest that all 3 TSHR fragments are shown to be immunogenic and are capable to induce TRAb; both RT1 and RT2 show their effect on immune regulation and are idiotypic of TSHR peptides; On the other hand, the humoural and cell immunities are ameliarated by injection of antisense peptides. Therefore, it is possible that antisense peptides may be involved in immune regulation via immune network.

The cloning and expression of a?-galactosidase gene(TM_0310)from Thermotoga maritima MSB8 was studied.The gene consists of 2019 bp,and the translated protein encodes 672 amino acids and its molecular mass is approximately 78.972 kD.The homology analysis of the deduced amino acid sequences showed that the enzyme shared 95%identity with a putative?-galactosidase from Thermotoga petrophila RKU-1 and a?-galactosidase from Thermotoga sp.RQ2.The galactosidase activity was up to 2.08 U/mg after the recombinant E.coli BL21 was induced by IPTG.The crude lysate remained about 70%activity after treated at 80℃for 10 min,indicating that the recombinant enzyme is thermostable and may be used at high temperatures.

Objective Hyperthyroidism and a diffuse goiter are the main symptoms of Graves' disease (GD) associated with autoantibodies to thyroid-stimulating hormone (TSH) receptor (TRAb). The present study was conducted to evaluate effects of autoantibodies in patients with GD (TRAb-IgG) on induction of the proliferation and c-fos mRNA expression in FRTL-5 cells (Fisher rat thyroid cell line).Methods Highly purified IgG fractions were isolated from 11 patients with GD, TRAb-IgG and 15 normal individuals (normal controls) with Protein A Sepharose CL-4B affinity column chromatograph. FRTL-5 cells, which had been grown to subconfluency and deprived of TSH for a few days. Then, these cells were used for measuring cAMP content, 3H-thymidine incorporation in cells and the expression of c-fos mRNA respectively.Results After stimulation of TRAb-IgG, the cAMP production and 3 H-thymidine incorporation in FRTL-5 cells were much higher than those from normal controls (P<0.05 respectively). Using 32P labelled v-fos probe by the Northern Blot method, the expression of c-fos mRNA could be induced by IgGs from patients with GD.Conclusions These data suggest that the stimulation of TRAb-IgG followed by cAMP production and 3H-thymidine incorporation is related to the induction of c-fos mRNA and, thus, to the growth of FRTL-5 cells.

OBJECTIVEThis study aimed to explore the impact of specimen collection and storage consumable products on trace element quantitative analysis.

METHODSDevices and consumable products of different brands used in specimen collection or storage were selected and treated separately as below:urine collection and storage tubes (Brand A, B, C and D, 2 samples for each brand) were treated with 1% of HNO(3) volume fraction for 2 - 4 h; blood taking device (Brand O, P and Q, 3 samples for each brand) were used for ultra-pure water samples collecting as simulation of blood sampling;dust sampling filters (Brand X, Y and Z, 2 samples for each brand) were cold digested by nitric acid for 12 h, followed by microwave digestion. Then cadmium, cobalt, chromium, copper, iron, manganese, molybdenum, nickel, lead, selenium, stannum, titanium, vanadium and zinc concentrations in the solutions obtained during the course of collect or storage were quantified by inductively coupled plasma mass spectrometer.

RESULTSFor the urine collection and storage consumable products, background values of elements were described as mean of parellel samples. The consentration of 14 quantified elements were relatively low for 5 ml cryogenic vials (brand B) with background values range of 0.001 - 0.350 ng/ml. The background values of copper of 50 ml centrifuge tubes (brand A), chromium of 5 ml cryogenic vials (brand C) and zinc of 1.5 ml centrifuge tubes (brand D) were relatively high, which were 1.900, 1.095 and 1.368 ng/ml, respectively. Background values of elements in blood sampling devices were described as x(-) ± s. Background values of chromium for brand O, P and Q were (0.120 ± 0.017), (0.337 ± 0.093) and (0.360 ± 0.035) ng/ml; for copper were (0.050 ± 0.001), (0.017 ± 0.012) and (0.103 ± 0.015) ng/ml; for lead were (0.057 ± 0.072), (0.183 ± 0.118) and (0.347 ± 0.006) ng/ml; for titanium were (7.883 ± 0.145), (8.863 ± 0.190) and (8.613 ± 0.274) ng/ml; zinc were (2.240 ± 0.573), (42.140 ± 22.756) and (8.850 ± 3.670) ng/ml. There were statistically differences of background values for chromium, copper, lead, titanium and zinc among the above three brands of blood sampling devices (all P values < 0.05). For air sampling filters, background values of elements were described as mean of parellel samples. Background values of chromium and nickel of sampling filters (brand X) were lowest, which were 17.000 and 15.400 ng per piece, respectively; while background values for other elements were relatively high, the quantification of cadmium, cobalt, copper, iron, manganese, molybdenum, lead, selenium, stannum, titanium, vanadium and zinc were 0.250, 0.550, 48.500, 690.000, 25.500, 0.900, 6.500, 10.550, 7.950, 10.500, 0.850, 370.000 ng per piece, respectively. Background values of chromium and nickel of sampling filters (brand Z) were highest, which were 171.000 and 29.850 ng per piece.

CONCLUSIONBackground values of trace elements varied among products of different brands, and the most noticable differences were found in chromium, manganese, nickel, lead, stannum and zinc.

Environmental Monitoring ; methods ; Quality Control ; Specimen Handling ; methods ; Trace Elements ; analysis

INTRODUCTIONFaecal incontinence (FI) is a stigmatised condition that remains a silent affliction for many populations. To date, no local study has been performed to determine its prevalence in our population. The existing literature from the West has shown highly variable rates, ranging from 0.8% to 18.0%. The aim of this study was to determine the cross-sectional prevalence of FI in Singapore and to identify at-risk groups.

METHODSA door-to-door questionnaire survey was performed between February and March 2013. We randomly selected 1,000 individuals from the electoral roll to be surveyed using the validated Comprehensive Fecal Incontinence Questionnaire.

RESULTSA total of 381 participants agreed to be enrolled in the study. The mean age of the participants was 52 (range 21-86) years, and slightly more than half of the participants were female (52.8%). Among the female participants, 73.1% had children (78.8% underwent normal vaginal delivery). The overall prevalence of FI in our study population was 4.7%. The prevalence of FI was observed to be significantly associated with increasing age (p = 0.004) and female gender (p = 0.029); women were three times more likely to suffer from FI than men.

CONCLUSIONWith the ageing population of Singapore, the results of the present study provide further impetus to continue public outreach efforts as well as develop clinical programmes that address the growing need for specialist treatments for people with FI.

Adult ; Age Distribution ; Aged ; Aged, 80 and over ; Cross-Sectional Studies ; Delivery, Obstetric ; methods ; statistics & numerical data ; Fecal Incontinence ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Risk Factors ; Sex Distribution ; Singapore ; epidemiology ; Surveys and Questionnaires ; Young Adult

Human exhaled breath has great application prospects,e.g.,monitoring pharmacokinetics,disease diagnosis,due to its advantages such as non-invasive and high-frequency sampling.Breath samples can be collected from the oral and nasal cavity.However,the oral and nasal environment affect the chemical composition of breath sample.Therefore,the investigation on the chemical composition of mouth-exhaled breath and nose-exhaled breath is crucial for selection of appropriate sampling strategy for individual studies.In this work,secondary electrospray ionization-high resolution mass spectrometry(SESI-HRMS)was applied to analysis of respiratory metabolomics in real time.A quantitative analysis approach was established for 9 kinds of volatile organic compounds(VOCs)e.g.2-butanone,2-pentanone,ethyl acetate,methyl methacrylate,toluene,styrene,mesitylene,isoprene and limonene.The limit of detection was 2.3?240.8 ng/m3.The intra-day(n=6)and inter-day(n=18)relative standard deviations were 0.6%?4.6%and 4.3%?12.2%,respectively.Nine healthy subjects were recruited to investigate the chemical composition of mouth-exhaled and nose-exhaled breath.The results showed the good performance in quantitative analysis of 9 VOCs in breath air.It was found that the number of unique component(m/z)detected in mouth-exhaled breath(167)was 2.2 times greater than that detected in nose-exhaled breath(76),which might result from the complex environment in oral cavity.The signal intensity of commun component(163)was significantly different between mouth-exhaled breath and nose-exhaled breath.Additionally,the elemental composition analysis showed that the proportion of polar compounds detected in nose-exhaled breath was higher than that in mouth-exhaled breath.This study demonstrated that there was significant differences in the chemical composition between mouth-exhaled and nose-exhaled breath,which provided a theoretical basis for selection of exhalation mode.

OBJECTIVETo explore the operative technique and clinical results of using peroneal perforator-based sural neurofasciocutaneous flaps to repair skin and soft tissue defects in lower extremities.

METHODSFrom January 2009 to March 2012, 26 patients with skin and soft tissue defects at distal region of leg and foot were hospitalized. Among them, 9 patients suffered from tendon or bone exposure at the distal region of leg after injury due to traffic accident; 4 patients suffered from skin defects in the ankle as a result of electric injury; 8 patients suffered from chronic ulcer at the distal part of leg and foot; 5 patients suffered from plantar pressure ulcer. After debridement, soft tissue defect sizes ranged from 11 cm×5 cm to 18 cm×13 cm. According to the position and size of the defects, peroneal perforator-based sural neurofasciocutaneous flaps were designed and procured to repair the skin and soft tissue defects. The size of flaps ranged from 12 cm×6 cm to 20 cm×15 cm. Flap donor sites were closed by direct suture or skin grafting.

RESULTSTwenty-five flaps survived completely. Only one flap suffered partial margin necrosis in the size of 2 cm×1 cm, which was healed after dressing change. Patients were followed up for 6 to 12 months. The appearance and sensation of flaps were satisfactory; no ulcer occurred; the movement of lower extremities was normal.

CONCLUSIONSIt is suitable to repair the skin and soft tissue defects at distal region of leg and foot with the peroneal perforator-based sural neurofasciocutaneous flap, as it possesses reliable blood supply, long and non-bulky pedicle, and sufficient available size. The operation is also easy to perform.

Adult ; Aged ; Female ; Humans ; Lower Extremity ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Soft Tissue Injuries ; surgery ; Sural Nerve ; transplantation ; Surgical Flaps ; blood supply ; innervation ; Young Adult