ABSTRACT
Objective: To explore the clinical effect of double blood supply composite tissue flap with peroneal artery perforator and sural nerve nutrient vessel carrying gastrocnemius muscle in repairing lacunar skin and soft tissue defect of ankle. Methods: From September 2015 to September 2017, 10 patients with lacunar skin and soft tissue defects of ankle were hospitalized in our unit, including 7 males and 3 females, aged 17-62 years, 8 traffic accident injuries and 2 heavy body pressure injuries. After debridement, the wound area ranged from 7 cm×4 cm to 15 cm×9 cm, and the lacunar area ranged from 3.0 cm×2.0 cm×1.0 cm to 7.0 cm×4.0 cm×2.0 cm. The defect was repaired with island composite tissue flap with peroneal artery perforator and sural nerve nutrient vessel and partial gastrocnemius muscle at stage â . The area of flaps ranged from 8 cm×5 cm to 16 cm×10 cm, and the area of gastrocnemius muscle flaps ranged from 4 cm×3 cm to 8 cm×5 cm. The donor site was repaired with ipsilateral thigh intermediate split-thickness skin graft or sutured directly. The location of vascular pedicle, survival of tissue flap, follow-up, and occurrence of complications were recorded. Results: The vascular pedicle of this group of cases was located 5-15 cm above the lateral malleolus. The tissue flaps of 10 patients survived completely after operation at stage â . Follow-up for 1-12 months showed that the area repaired with tissue flaps had good color, blood supply, and texture, without ulceration, and recovered different degrees of pain sensation and deep tactile sensation, and no short-term or long-term complications occurred. Conclusions: The double blood supply composite tissue flap with peroneal artery perforator and sural nerve nutrient vessel carrying gastrocnemius muscle has good blood supply, which can effectively fill the invalid space in repairing lacunar skin and soft tissue defect of ankle, with good effect after operation, and it is worthy of clinical application.
Subject(s)
Ankle/surgery , Nutrients/administration & dosage , Perforator Flap/blood supply , Perforator Flap/innervation , Plastic Surgery Procedures/methods , Skin Transplantation , Soft Tissue Injuries/surgery , Sural Nerve , Adolescent , Adult , Arteries , Female , Humans , Male , Middle Aged , Muscle, Skeletal , Young AdultABSTRACT
Objective: To explore the clinical effects of adjacent fasciocutaneous flaps in repairing small wounds with bone or steel plate exposure in anterior tibia. Methods: Twelve patients with small wounds of bone or steel plate exposure in anterior tibia covering area of 2 cm×2 cm to 5 cm×3 cm were admitted to our unit from January 2014 to December 2016. A circular or elliptical adjacent fasciocutaneous flap was designed on the normal skin located at the inside or outside of the wound according to the size of wound after thorough debridement. The pedicle of the flap was located at the proximal end and transferred through the subcutaneous tunnel to cover the wound. The sizes of flaps were 3 cm×3 cm to 6 cm×4 cm. Flaps were fixed with interrupted sutures and drainage rubber sheets were placed under the flaps. The drainage rubber sheets were removed within 24 to 48 hours. The donor area was repaired by medium-thickness skin graft collected from homolateral outer thigh. Results: All the flaps of 9 patients survived. Two patients had necrosis at the distal end of the flaps and were cured by changing dressing. One patient had tension blisters on the flap and was cured by removing blisters and improving microcirculation. All patients were followed up for 3 months, and the flaps were good in blood supply, appearance, and color, with hypaesthesia. Conclusions: Repair of small wounds with bone or steel plate exposure in anterior tibia by adjacent fasciocutaneous flap is simple in surgical procedure and does not damage the well-known blood vessels, and the appearance, texture, and thickness of flaps are close to the skin of anterior tibia region. It is a good choice for repairing this kind of wounds and worth promoting in clinic.
Subject(s)
Plastic Surgery Procedures/methods , Skin Transplantation , Soft Tissue Injuries/surgery , Surgical Flaps , Tibia , Humans , Steel , Treatment Outcome , Wound HealingABSTRACT
Objective: To summarize the measures and experience in diagnosis and treatment of extremely severe burn patients with severe inhalation injury in dust explosion accident. Methods: The medical records of 13 patients with extremely severe burn complicated with severe inhalation injury in August 2nd Kunshan factory aluminum dust explosion accident who were treated at the First Affiliated Hospital of Soochow University (hereinafter referred to as our hospital) on August 2nd, 2014, were retrospectively analyzed. All the patients were transferred to our hospital 3-8 hours after injury under the condition of inhalation of pure oxygen. Twelve patients underwent tracheotomy within 5 hours after admission, while 1 patient underwent tracheotomy before admission. All the patients were in ventilator-assisted respiration, with synchronized intermittent mandatory ventilation combined with positive end expiratory pressure. All the patients underwent thorax or limbs escharotomy on the second day after admission, so as to reduce the restrictive ventilatory dysfunction caused by the contraction of thorax eschar and the terminal circulation disorder caused by the contraction of limbs eschar. All the patients underwent electronic bronchoscopy within 48 hours after admission, airway secretion were cleared and airway lavage were carried out under electronic bronchoscope according to the patients' condition, and the sputum, lavage solution, pathological tissue were collected for microbiological culture. All the patients underwent chest X-ray examination on the second day after admission and reexamination as required. Patients were all treated with a combination of broad-spectrum antibiotics early after admission to control lung and systemic infection. One patient was treated with extracorporeal membrane oxygenation for acute respiratory distress syndrome 1 week after admission. Results: One patient suffered from cardiopulmonary arrest during tracheotomy, which recovered autonomous respiration and cardiac impulse after cardiopulmonary resuscitation. Three patients showed decreased pulse oxygen saturation (SpO(2)) within 48 hours after injury, and the SpO(2) returned to normal after sputum aspiration, scab removal and lavage under electronic bronchoscope. During the course of disease, bacteria were cultured from wound exudate of 7 patients, bacteremia occurred in 10 patients, and sputum microbiological culture results of 13 patients were positive. Eight of the 13 patients in this group survived, and 5 died. One patient died 19 days after injury, and 4 patients died 33-46 days after injury. The main cause of death was multiple organ dysfunction syndrome induced by severe septic shock eventually. Conclusions: For this batch of patients with extremely severe burn complicated with severe inhalation injury caused by dust explosion accident, the treatment and cure measures including early definite diagnosis and timely tracheotomy, the application of effective ventilation, the effective treatment of respiratory system complications, and rational use of antibiotics for the control of lung infection obtained quite good curative effect.
Subject(s)
Aluminum/toxicity , Burns, Inhalation/surgery , Burns/therapy , Explosions , Tracheotomy/methods , Accidents, Occupational , Airway Obstruction/etiology , Airway Obstruction/surgery , Bacteremia/epidemiology , Blast Injuries , Burns/complications , Burns, Inhalation/complications , Burns, Inhalation/mortality , China/epidemiology , Dust , Extracorporeal Membrane Oxygenation , Humans , Lung/physiopathology , Multiple Organ Failure/epidemiology , Positive-Pressure Respiration , Respiration, Artificial , Respiratory Distress Syndrome , Retrospective Studies , Sepsis/epidemiologyABSTRACT
Objective: To investigate the role of "Hong's single-stitch duct to mucosa pancreaticojejunostomy(HSDMP)" in laparoscopic pancreaticoduodenectomy (LPD). Methods: The clinical data including perioperative and short-term outcomes of 51 cases of LPD with HSDMP which performed in Zhejiang Provincial People's Hospital(33 cases) and Frist Clinical Hospital of Jilin University(18 cases) between April and October 2016 were reviewed retrospectively. There were 31 male patients and 20 female patients. The mean age was(59±11)years. Body mass index (BMI) was 18 to 28 kg/m(2) and the average BMI was (23.2±4.4)kg/m(2). Preoperative diagnosis: 18 cases with pancreatic mass, 26 cases with peri-ampullary tumor, 3 cases with intra-ductal papillary mucinous neoplasms, 2 cases with duodenal carcinoma, 2 cases with serous cystadenoma. Results: Fifty-one patients accepted LPD using HSDMP. One patient underwent LPD combined with resection of superior mesentery vein. The mean operation time was (307±69)minutes, the mean diameter of pancreatic duct for reconstruction was (3.1±1.1)mm.The mean operation time for HSDMP was (34±5) minutes, the estimated blood loss was (170±127)ml. Twelve cases(23.5%) had pancreatic fistula according to International Study Group definition, including 9 cases(17.6%) of grade A and 3 cases (5.9%) of grade B. Five cases(9.8%) had delayed gastric empty, 5 cases(9.8%) had bile leakage and 2 cases(3.9%) had pulmonary infection postoperative.All these complications were treated by non-surgical strategies. One patient(2.0%) suffered from postoperative intra-abdominal bleeding and recovered after reoperation. Pathologic results showed pancreatic ductal adenocarcinomas in 20 cases(39.2%), non-pancreatic original peri-ampullary tumors in 23 cases(45.1%), intra-ductal papillary mucinous neoplasms in 3 cases(5.9%), duodenal carcinoma in 2 cases(3.9%), serous cystadenoma in 2 cases(3.9%) and neuroendocrine tumors in one case(2.0%). Conclusions: HSDMP could not only reduce the incidence of clinical pancreatic fistula, but also save operation time. It is a feasible and safe method for pancreaticojejunostomy.
Subject(s)
Pancreatic Neoplasms/surgery , Pancreaticoduodenectomy , Pancreaticojejunostomy , Aged , Anastomosis, Surgical , Carcinoma, Pancreatic Ductal , Duodenal Neoplasms , Female , Humans , Laparoscopy , Male , Middle Aged , Operative Time , Pancreatectomy , Pancreatic Ducts , Pancreatic Fistula , Postoperative Complications , Retrospective Studies , Treatment Outcome , Pancreatic NeoplasmsABSTRACT
OBJECTIVE: To investigate the effect of the application of pulse contour cardiac output (PiCCO) monitoring technology on delayed resuscitation of patients with extensive burn in a mass casualty. METHODS: The clinical data of 41 patients injured in Kunshan dash explosion hospitalized in the First Affiliated Hospital of Soochow University, the 100th Hospital of the People's Liberation Army, and Suzhou Municipal Hospital were retrospectively analyzed. The patients were divided into traditional monitoring group (T, n=22) and PiCCO monitoring group (P, n=19) according to the monitoring technic during delayed resuscitation. The input volumes of electrolyte, colloids, and water of patients in the two groups within 2 hours after admission, the first, second, and third 8 hours post injury (HPI), and the first 24 HPI were recorded. The fluid infusion coefficients of patients in the two groups within 2 hours after admission, the first, second, and third 8 HPI, and the first, second, third, and fourth 24 HPI were calculated. The urine volume, mean arterial pressure (MAP), and central venous pressure (CVP) of patients in the two groups at post injury hour (PIH) 8, 16, 24, 48, 72, and 96 were recorded. The blood lactate, base excess, hematocrit (HCT), and platelet count of patients in the two groups at PIH 24, 48, 72, and 96 were recorded. Complications and death of patients in the two groups were recorded. Data were processed with analysis of variance for repeated measurement, Chi-square test, t test, and Wilcoxon test. The deviations between figure 2 and the fluid infusion coefficients of the first or second 24 HPI, and the deviations between figure 1 and the fluid infusion coefficients of the second, third or fourth 24 HPI were calculated, and the three groups deviations were analyzed by Pearson correlation analysis. RESULTS: (1) The input volumes of electrolyte of patients in group P were significantly more than those in group T within the first 8 and 24 HPI (with Z values respectively -3.506 and -2.654, P<0.05 or P<0.01), and the input volumes of electrolyte of patients in the two groups were similar within the other time periods (with Z values from -1.871 to -0.680, P values above 0.05). The input volumes of colloid of patients in group P were significantly less than those in group T within the second, third 8 HPI, and the first 24 HPI (with Z values from -4.720 to -2.643, P<0.05 or P<0.01), and the input volumes of colloid of patients in the two groups were similar within the other time periods (with Z values respectively -2.376 and -2.303, P values above 0.05). The input volumes of water of patients in the two groups were similar within each time period (with Z values from -1.959 to -0.241, P values above 0.05). (2) The fluid infusion coefficients of patients in group T within 2 hours after admission, the first, second, and third 8 HPI, and the first, second, third, and fourth 24 HPI were respectively (0.59±0.18), (0.70±0.23), (0.94±0.24), (0.74±0.14), (2.38±0.44), (1.70±0.56), (1.35±0.67), and (0.92±0.46) mL·kg(-1)·%TBSA(-1,) and the values in group P were respectively (0.59±0.29), (0.82±0.37), (0.86±0.38), (0.59±0.24), (2.27±0.85), (2.13±0.68), (1.59±3.78), and (1.46±0.56) mL·kg(-1)·%TBSA(-1). The fluid infusion coefficients of patients in the two groups were similar within 2 hours after admission, the first, second 8 HPI, and the first, third 24 HPI (with t values from -1.262 to 0.871, P values above 0.05). The fluid infusion coefficient of patients in group P was significantly lower than that in group T within the third 8 HPI (t=2.456, P<0.05), and the fluid infusion coefficient of patients in group P were significantly higher than that in group T within the second and fourth 24 HPI (with t values respectively -2.234 and -3.370, P<0.05 or P<0.01). There was obviously negative correlation between the deviations of figure 2 and the fluid infusion coefficient of the first 24 HPI and that of the second 24 HPI (r=-0.438, P<0.01). There was no obvious correlation between the deviations of figure 1 and the fluid infusion coefficient of the second 24 HPI and that of the third 24 HPI (r=0.091, P>0.05). There was obviously positive correlation between the deviations of figure 1 and the fluid infusion coefficient of the second 24 HPI and that of the fourth 24 HPI (r=0.695, P<0.01). (3) The urine volumes and MAP of patients in the two groups were similar at each time point (with Z values from -1.884 to 0, P values above 0.05). The CVP of patients in group P were significantly higher than that in group T at PIH 16, 24, 48, and 72 (with Z values from -4.341 to -2.213, P<0.05 or P<0.01), and the CVP of patients in the two groups were similar at the other time points (with Z values respectively -0.132 and -1.208, P values above 0.05). The blood lactate of patients in group P was significantly higher than that in group T at PIH 72 (Z= -2.958, P<0.01) , and the blood lactate of patients in the two groups were similar at the other time points (with Z values from -1.742 to -0.433, P values above 0.05). The base excess of patients in group P were significantly lower than that in group T at PIH 24, 48, 72, and 96 (with Z values from -4.970 to -4.734, P values below 0.01). The HCT of patients in the two groups were similar at PIH 24, 48, 72, and 96 (with Z values from -2.239 to -0.196, P values above 0.05). There were significant differences in the platelet count of patients in the two groups at PIH 24, 72, and 96 (with Z values from -4.578 to -2.512, P<0.05 or P<0.01). (4) There were 15 cases in group T accompanied by complications, and 7 cases died, while 13 cases in group P accompanied by complications, and 9 cases died. The occurrence of complications and death of patients in the two groups were similar (with χ(2) values respectively <0.001 and 1.306, P values above 0.05). CONCLUSIONS: On the basis of traditional burn shock monitoring index, the effect of fluid resuscitation in patients with severe burn monitored by PiCCO technology is not so good and still needs further clinical research.
Subject(s)
Burns/therapy , Cardiac Output , Mass Casualty Incidents , Monitoring, Physiologic/methods , Resuscitation , Fluid Therapy , Hematocrit , Humans , Retrospective Studies , Shock/therapyABSTRACT
The aim of this study was to investigate the pharmacokinetic interaction between ritonavir (RTV) and an anti-HIV agent 3-cyanomethyl-4-methyl-DCK (CMDCK). CMDCK was administered orally (8 mg/kg) and intravenously (2 mg/kg) to rats in the absence or presence of RTV (1 or 2.5 mg/kg). By comparing the pharmacokinetic parameters between the control and the RTV treated groups, it was found that co-administration with RTV could significantly increase the plasma exposure of CMDCK, through improving the hepatic and intestinal availabilities. The AUCinf of CMDCK was increased by 2.4 or 8.7 times for intravenous or oral route, respectively. The oral bioavailability of CMDCK was increased from 15% of the control group to 45% of the RTV concomitant group (2.5 mg/kg). In the in vitro studies with liver and intestinal microsomes, the K i values of RTV on the CMDCK metabolism were determined and found to be 0.22 and 0.48 µM for human, 0.33 and 1.60 µM for rat, respectively. Caco-2 cells study showed that CMDCK is not a P-glycoprotein (P-gp) substrate and its transepithelial transport is mainly through passive diffusion. The in vitro and in vivo results indicate that RTV could improve the bioavailability of CMDCK by inhibiting CYP3A mediated metabolism in both liver and intestine.
Subject(s)
Anti-HIV Agents/pharmacokinetics , Bridged Bicyclo Compounds, Heterocyclic/pharmacokinetics , Coumarins/pharmacokinetics , Reverse Transcriptase Inhibitors/pharmacokinetics , Ritonavir/pharmacology , Animals , Biological Availability , Caco-2 Cells , Drug Interactions , Humans , Intestinal Mucosa/metabolism , Liver/metabolism , Male , Rats , Rats, Sprague-DawleyABSTRACT
The fluorescence spectral characteristics and the interaction of several water-soluble metal complexes of Schiff-base with DNA are described. Among the complexes tested, Mn-Schiff-base bound to DNA showed a marked decrease in the fluorescence intensity with a blue shift of the excitation and emission peaks. Some hypochromism in the UV absorption spectra was also observed. KI quenching and competitive binding to DNA between Mn-Schiff-base and ethidium bromide (EB) were studied in connection with other experimental observations to show that the interactive model between Mn-Schiff-base and DNA is an intercalative one. The pH and salt effect on the fluorescence properties was also investigated. The linear relationship between F/F0 and the concentration of calf thymus DNA covers 3.0 x 10(-6)-2 x 10(-4) mol L-1, which can be utilized for determining traces of calf thymus DNA with a detection limit of 8.0 x 10(-7) mol L-1 in base pairs.
Subject(s)
DNA/chemistry , Metals/chemistry , Schiff Bases/chemistry , Fluorescent Dyes , Hydrogen-Ion Concentration , Indicators and Reagents , Solubility , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet , WaterABSTRACT
A renewable amperometric immunosensor based on the sol-gel technique has been constructed by dispersing graphite, complement 3 (C3) antiserum, and sol-gel at low temperature. The prepared immunosensor is rigid, porous, and has a renewable external surface. A competitive binding assay has been used to determine C3 in human serum with the aid of C3 labeled with horseradish peroxidase. The enzyme-labeled antigen can readily diffuse toward the encapsulated antibody, which retains its binding properties. The experimental conditions for the assay with the biocomposite, including the loading of C3 antiserum in the biocomposite, the amount of labeled C3 in incubation solution, incubation time, and temperature, have been optimized. Using C3 labeled with horseradish peroxidase, and o-AP as the substrate, amperometric detection at -150 mV (relative to the SCE) results in a linear detection range of 1.17-35.1 microg mL(-1), with a detection limit of 0.56 microg mL(-1). Serum samples have been assayed and the results demonstrate the feasibility of the proposed immunosensor for clinical analysis. The surface of the immunosensor can be renewed simply by polishing to obtain a fresh immunocomposite ready to use in a new competitive assay.
Subject(s)
Biosensing Techniques , Complement C3/analysis , Immunoassay/methods , Electrochemistry , HumansABSTRACT
A renewable amperometric immunosensor has been proposed for the determination of Schistosoma japonium antibody (SjAb) in rabbit serum. A paraffin-graphite-Schistosoma japonium antigen (SjAg) biocomposite, which needs no additional curing, was directly used to construct the immunosensors. The analytical sample containing the desired SjAb was mixed with SjAb labeled with horseradish peroxidase (HRP) to form the incubation solution for the competitive binding assay. Amperometry was used to determine the amount of HRP fixed on the sensor surface, which was related to the content of desired SjAb. Assay conditions were optimized, including the selection of substrate, the loading of SjAg in the biocomposite, the amount of labeled SjAb in the incubation solution, the incubation time, and the temperature. Using o-aminophenol (o-AP) as a substrate, amperometric detection at -200 mV (vs SCE) resulted in a pseudolinear detection range of about 0.36 to 14 microg/mL, with a detection limit of 0.36 microg/mL. Rabbit serum samples with varying infection degrees were analyzed, and the results demonstrated that the concentration that is detectable in this system meets the demands of clinical analyses. A new surface on the immunosensor for use in another competitive assay can be obtained by removing the original one and polishing the surface.
Subject(s)
Antibodies, Helminth/blood , Biosensing Techniques/methods , Schistosoma japonicum/immunology , Animals , Antigens, Helminth/immunology , Biosensing Techniques/instrumentation , RabbitsABSTRACT
An acryloyl group was attached to fluorescein through the phenol hydroxyl group to make it photopolymerizable with 2-hydroxypropyl methacrylate and covalently immobilized on an optode glass surface modified with gamma-(methacryloxy)propyltrimethoxysilane. The optode system with a plastic-clad fused silica bifurcated fiber optic bundle shows satisfactory analytical performance characteristics for determining picric acid in terms of selectivity and reproducibility with a linear range from 8.0 x 10(-7) to 4.0 x 10(-3) mol L-1. The quenching mechanism was investigated. The optode membrane can be applied to the indirect determination of cinchonine drug and the results obtained were satisfactory.
Subject(s)
Picrates/analysis , Fiber Optic Technology , Hydrogen-Ion Concentration , Indicators and Reagents/analysis , Optical Fibers , Spectrometry, FluorescenceABSTRACT
For preparing a berberine-sensitive electrode, 5,10,15,20-tetrakis[2-(2,3,4,6-tetraacetyl-beta-D-glucopyranosyl)-1-O- phenyl]porphyrin (T(o-glu)PPH2) was synthesized from the reaction of pyrrole with ortho-acetylglycosylated benzaldehyde by Lindsay's method. The electrode based on T(o-glu)PPH2 with an optimized membrane composition exhibits Nernstian response to berberine in the concentration range 2.4 x 10(-7)-5.0 x 10(-3) mol L-1, with a pH range from 3.9 to 10.2, and a fast response time of 30 s. The electrode shows fair selectivity towards berberine with respect to common co-existing species. T(o-glu)PPH2 shows better potentiometric response characteristics comparing to chloro[5,10,15,20-tetrakis[2-(2,3,4,6-tetraacetyl-beta-D- glucopyranosyl)-1-O-phenyl]-porphinato]-manganese (MnT(o-glu)PPCl) and better selectivity towards berberine than tetraphenylporphyrin (TPPH2). The effect of the composition of the electrode membrane has been studied and the experimental conditions optimized. The contents of berberine in pharmaceutical tablets were determined by direct potentiometry and the results agreed with values obtained by the pharmacopoeia method.
Subject(s)
Berberine/analysis , Electrophoresis/methods , Porphyrins/chemical synthesis , Electrochemistry , Glycosylation , Hydrogen-Ion Concentration , Ion-Selective Electrodes , Membranes/chemistry , Pharmaceutical Preparations/chemistry , Tablets/chemistryABSTRACT
Lanthanide porphyrin complexes synthesized by a solid state method were used to prepare a novel ethacrynic acid (EA) sensor. The sensor, based on pentane-2,4-dionato(meso-tetraphenylporphinato)terbium [TbTPP(acac)] with an optimized membrane composition, exhibits a Nernstian response to EA- ion in the concentration range 7.4 x 10(-6)-1.0 x 10(-1) mol l-1 with a pH range from 3.2 to 6.8 and a fast response time of 30 s. The electrode shows improved selectivity towards EA- ion with respect to common co-existing ions compared with the previously reported EA sensor. As electroactive materials, lanthanide porphyrin complexes show better potentiometric response characteristics than copper porphyrin complexes. The effect of solvent mediators and lipophilic ion additives was studied and the experimental conditions were optimized. The electrode was applied to the determination of EA in human urine samples with satisfactory results.
Subject(s)
Ethacrynic Acid/urine , Electrochemistry , Electrodes , HumansABSTRACT
Dendritic cells (DC) are special subsets of antigen presenting cells characterized by their potent capacity to activate immunologically naive T cells. By subtracting the mRNAs expressed by the mouse epidermus-derived DC line XS52 with the mRNAs expressed by the J774 macrophage line, we identified five novel genes that were expressed selectively by this DC line. One of these genes encoded a type II membrane-integrated polypeptide of 244 amino acids containing a putative carbohydrate recognition domain motif at the COOH-terminal end. This molecule, termed "dectin-1," was expressed abundantly at both mRNA and protein levels by the XS52 DC line, but not by non-DC lines (including the J774 macrophage line). Dectin-1 mRNA was detected predominantly in spleen and thymus (by Northern blotting) and in skin-resident DC, i.e. Langerhans cells (by reverse transcription-polymerase chain reaction). Affinity-purified antibody against dectin-1 identified a 43-kDa glycoprotein in membrane fractions isolated from the XS52 DC line and from the dectin-1 cDNA-transfected COS-1 cells. His-tagged recombinant proteins containing the extracellular domains of dectin-1 showed marked and specific binding to the surface of T cells and promoted their proliferation in the presence of anti-CD3 monoclonal antibody at suboptimal concentrations. These in vitro results suggest that dectin-1 on DC may bind to as yet undefined ligand(s) on T cells, thereby delivering T cell co-stimulatory signals. Not only do these results document the efficacy of subtractive cDNA cloning for the identification of unique genes expressed by DC, they also provide a framework for studying the physiological function of dectin-1.
Subject(s)
Dendritic Cells/metabolism , Membrane Proteins/chemistry , Membrane Proteins/genetics , Nerve Tissue Proteins/chemistry , Nerve Tissue Proteins/genetics , Amino Acid Sequence , Animals , Blotting, Northern , COS Cells , Cell Culture Techniques/methods , Cell Line , Cell Membrane/metabolism , Cloning, Molecular , DNA, Complementary/metabolism , Epithelial Cells/metabolism , Female , Flow Cytometry , Immunoblotting , Lectins/metabolism , Lectins, C-Type , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Phenotype , Rabbits , Recombinant Fusion Proteins/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Sequence Homology, Amino Acid , T-Lymphocytes/metabolism , Tissue DistributionABSTRACT
Using a subtractive cDNA cloning strategy, we isolated previously five novel genes that were expressed abundantly by the murine dendritic cell (DC) line XS52, but not by the J774 macrophage line. One of these genes encoded a unique, DC-associated C-type lectin, termed "dectin-1." Here we report the characterization of a second novel gene that was also expressed in a DC-specific manner. Clone 1B12 encoded a type II membrane-integrated polypeptide of 209 amino acids containing a single carbohydrate recognition domain motif in the COOH terminus. The expression pattern of this molecule, termed "dectin-2," was almost indistinguishable from that for dectin-1; that is, both were expressed abundantly at mRNA and protein levels by the XS52 DC line, but not by non-DC lines, and both were detected in spleen and thymus, as well as in skin resident DC (i.e. Langerhans cells). Interestingly, reverse transcriptase-polymerase chain reaction and immunoblotting revealed multiple bands of dectin-2 transcripts and proteins suggesting molecular heterogeneity. In fact, we isolated additional cDNA clones encoding two distinct, truncated dectin-2 isoforms. Genomic analyses indicated that a full-length dectin-2 (alpha isoform) is encoded by 6 exons, whereas truncated isoforms (beta and gamma) are produced by alternative splicing. We propose that dectin-2 and its isoforms, together with dectin-1, represent a unique subfamily of DC-associated C-type lectins.
Subject(s)
Alternative Splicing , Dendrites/chemistry , Lectins/genetics , Amino Acid Sequence , Animals , Base Sequence , Cell Line , Cloning, Molecular , Lectins, C-Type , Mice , Mice, Inbred BALB C , Molecular Sequence Data , RNA, Messenger/metabolismSubject(s)
Cytogenetic Analysis , Genome, Human , Human Genome Project , Physical Chromosome Mapping/methods , Chromosomes, Bacterial/genetics , Chromosomes, Human, Pair 22/genetics , Chromosomes, Human, Pair 7/genetics , Cloning, Molecular , Contig Mapping , Humans , In Situ Hybridization, Fluorescence , Sequence Tagged SitesABSTRACT
Several mu-oxo-bismetalloporphyrin complexes were synthesized for preparing fluoroborate sensors. The electrode based on mu-oxo-bis[tetra(p-chlorophenyl)porphinatomanganese(III)] with an optimized membrane composition shows a potentiometric response towards fluoroborate ion over the concentration range from 4.3 x 10(-7) to 1.0 x 10(-1) mol l-1 with a Nernstian slope, a wide working pH range from 5.5 to 12.0 and a fast response time of 30 s. The electrode shows anti-Hofmeister selectivity towards BF4- with respect to common co-existing ions, which is an improvement over methods reported so far. As electroactive materials, several mu-oxo-bismetalloporphyrins with different substituents on the benzene rings were compared for potentiometric response characteristics. The interaction between BF4- and bismetalloporphyrin was investigated using UV/VIS spectrophotometry. The electrode was applied to the determination of fluoroborate in electroplating solutions.
Subject(s)
Borates/analysis , Ion-Selective Electrodes , Metalloporphyrins , Spectrophotometry, UltravioletABSTRACT
A fluorescence probe of 3-methoxybenzanthrone (MBA) exhibits significant solvatochromic characteristics correlated with the polarity of solvents. The interaction of the solvatochromic fluorescence probe with calf thymus DNA (ct-DNA) has been investigated. In the presence of ct-DNA the fluorescence of MBA is strongly quenched with a blue-shift of emission peak and a hypochromism in absorption spectra. The absorption spectra, fluorescence quenching and fluorescence polarization experiments show that the MBA molecule as an intercalator is inserted into the base-stacking domain of the ct-DNA double helix, and the interaction of the nucleobases with the MBA molecule causes quenching of fluorescence and hypochromism in the absorption spectra. The intrinsic binding constant and the binding site number were determined to be 1.70 x 10(5) mol l-1 in base pairs and six, respectively. The I0/I versus [ct-DNA] plot shows linear relationship in the range covering 4.3 x 10(-7)-1.02 x 10(-4) mol l-1 in base pairs which can be used for ct-DNA determination. The limit of detection was found to be 4.3 x 10(-7) mol l-1 in base pairs (0.5 microgram ml-1).
Subject(s)
Benz(a)Anthracenes/chemistry , Benz(a)Anthracenes/metabolism , DNA/chemistry , DNA/metabolism , Fluorescent Dyes/chemistry , Fluorescent Dyes/metabolism , Absorption , Acetone/chemistry , Animals , Anions , Base Pairing , Cattle , Fluorescence Polarization , Hydrogen-Ion Concentration , Salts , Solvents , Spectrometry, Fluorescence , Spectrum Analysis/methods , Temperature , Thymus Gland/physiologyABSTRACT
The response characteristics of some iodide-selective solvent polymeric membrane electrodes based on with N,N'-bis(salicylaldehyde-n-octyl) diimine cobalt(II) (Co(II)SAODI) which is a more lipophilic substitute for a previously reported iodide-carrier are described. The electrode doped with Co(II)SAODI into a plasticized poly(vinyl chloride) membrane exhibits an anti-Hofmeister selectivity pattern with high selectivity toward iodide, long lifetime and small interference from H(+). Quartz crystal microgravimetric measurements and ac impedance experiments show that the excellent selectivity for iodide is related to the unique interaction between the carriers and iodide and steric effect associated with the structure of the Schiff base ligands.
ABSTRACT
A comparison has been made for the quenching effect of three classes of organic solvents on the porous silicon photoluminescence, including hydrocarbons, solvents containing oxygen and nitrogen. Among them organoamines possess the strongest quenching effect. The experiments show that the solvent molecules interact synergistically with porous silicon and the main pathways include the surface hydrophobic interaction, dipole-dipole interaction and hole trapping, the later being the dominant pathway in quenching of porous silicon photoluminescence.