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Objective To evaluate the effects of urotensin Ⅱ on cell proliferation of and α-smooth muscle actin (α-SMA) expression in normal human dermal fibroblasts (NFs),and to explore their regulatory mechanisms.Methods NFs were isolated from foreskin tissues and subjected to primary culture in vitro.Reverse transcription PCR and Western blot analysis were performed to measure the mRNA and protein expression of urotensin Ⅱ and its receptor,respectively.Cell counting kit-8 (CCK-8) assay was conducted to estimate the proliferation of NFs,which were treated with urotensin Ⅱ at different concentrations of 0,10-10,10-9,10-8,10-7 and 10-6 mol/L for 0,6,12,24 and 48 hours separately,and then the optimal concentration and duration of urotensin Ⅱ exposure were selected to be 10-8 mol/L and 24 hours respectively.Some cultured NFs were divided into 5 groups:control group receiving no treatment,U Ⅱ group treated with 10-8 mol/L urotensin Ⅱ,U Ⅱ + nicardipine group treated with 10-8 mol/L urotensin Ⅱ and the calcium channel blocker nicardipine at the concentration of 10-5 mol/L,U Ⅱ + PD98059 group treated with 10-8 mol/L urotensin Ⅱ and the mitogen activated protein kinase (MAPK)inhibitor PD98059 at the concen-tration of 10-5 mol/L,and U Ⅱ + cyclosporine group treated with 10-8 mol/L urotensin Ⅱ and the calcium-dependent protein kinase (CaM PK) inhibitor cyclosporine at the concentration of 10-5 mol/L.After 24-hour treatment,CCK-8 assay was conducted to evaluate the proliferation of NFs in the above groups,real-time fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of α-SMA respectively.Results Urotensin Ⅱ receptor was expressed in NFs,but urotensin Ⅱ was not.The proliferative activity of NFs significantly differed among the control group,U Ⅱ group,U Ⅱ + nicardipine group,U Ⅱ + PD98059 group and U Ⅱ + cyclosporine group (the mean absorbance value at 405 nm:1.036 ± 0.046,1.405 ± 0.158,1.121 ± 0.109,1.192 ± 0.089 and 1.141 ± 0.056,respectively;F =9.587,P < 0.01),and the U Ⅱ group showed significantly higher proliferative activity of NFs compared with the control group,U Ⅱ + nicardipine group,U Ⅱ + PD98059 group and U Ⅱ + cyclosporine group (q =8.263,6.355,4.774 and 5.912,respectively,all P < 0.05).There were significant differences in the mRNA and protein expression of α-SMA among the 5 groups (F =6.351,7.045,both P < 0.01),and the mRNA and protein expression of α-SMA was significantly higher in the U Ⅱ group than in the other 4 groups (all P < 0.05).Conclusion Urotensin Ⅱ may induce the proliferation of and α-SMA expression in NFs through calcium channels,MAPK and CaM PK pathways.
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In this research,Microtox technology was used to evaluate the comprehensive toxicity of Shen-Fu injection,which was an exclusive product of the pharmaceutical company.Firstly,vibrio fischeri was used as test strain.The best test system and methodology were identified.Under the best test conditions,vibrio fischeri was firstly used in the luminescent bacteria comprehensive toxicity of the exclusive product Shen-Fu injection.The results showed that in the 2 mL reaction system,the best recovery liquid volume was 0.9 mL/ freeze-dried vial,50 μL bacterial suspensions/ sample,the detection time was 10 min after adding bacterial suspensions,the pH was 5-10,the luminous intensity was 800 000-12 000 000 for 10 rin.The RSD of replication experiment and intermediate precision test was ? and ?,respectively,which fitted to the requirements.There was no significant difference on EC50 values among 9 batches of tested Shen-Fu injection (41.07%,RSD < 5%).It was concluded that there was significant concentration-effect relationship between Shen-Fu injection and the toxicity of vibrio fischeri.There was no significant difference on EC50 values among different batches.It indicated that there was small quality volatility among different batches of Shen-Fu injection.The control on product manufacturing was strong.
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Microtox technology is one of quick toxicity test methods for determining the poisonous or toxic substances in the environment by luminous bacteria as a indicator organism.This paper reviewed the appearance and development of microtox technology.Besides,we expounded the methods and stepwise technique map and addressed key questions of microtox technology for the evaluation of the comprehensive toxicity of Chinese medicinal materials.In conclusion,microtox technology is a promising method from a vantage point of the toxicity detection of Chinese medicinal materials.
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This study aimed at exploring the application of microtox technology to the evaluation of comprehensive toxicity of Sheng Mai injections.Characteristic parameters of vibrio fischeri (CS234) were measured by methodology inspection under different conditions to optimize the reaction condition with reliable technology.It was the first experiment to accomplish the comprehensive toxicity of Sheng Mai injections from various manufacturers using vibrio fischeri under the target condition.It was found that parameters of the optimum condition contained 0.9 mL recovery liquid volume in each freeze-dried vial and 50 μ L bacteria suspension of each sample,being included in 2 mL solution in aggregate under 10 mins' detection time after adding bacteria suspension with the favorable pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million within 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15%.Under this optimum detection condition,it was found that the EC50 values were 22.10%,34.10% and 46.04%,respectively,presenting significant statistical differences (P<0.05).These results demonstrated that the growth of biological detection standards of Sheng Mai injection was highlighted a long way to go.Besides,the microtox-based fast test for the evaluation of the comprehensive toxicity of Sheng Mai injection showed a prospective application to controlling the quality of different products from manufacturers.
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In this research,we chiefly explored a new fast test method--microtox technology to evaluate the comprehensive toxicity of Shen Mai injection.We investigated characteristic parameters of vibrio fischeri (CS234) under different conditions in the pursuit of the optimum test parameters of the reliable method;and then locked the best parameters for Shen Mai injection assay with the products from different manufacturers by microtox fast test.As a result,the optimum reaction condition included 0.9 mL recovery liquid in each freeze-dried vial and 50 μL bacteria suspension of each sample within 2 mL solution in aggregate with 10-mins detection after adding bacteria suspension which pH value ranging from 5 to 10 and luminous intensity from 0.8 to 1.2 million in 10 mins.The relative standard deviation values of replication experiment and precision test were all below 15%.Under this optimum detection condition,it was found that the EC50 values were 35.60%,92.34% and 146.57%,differing among the samples from three representative drug manufacturers,respectively (P < 0.01).In conclusion,the concentration-effect relationship of Shen Mai injection existed in the toxicity assessment of vibrio fischeri with various ECs0 values from the three manufacturers.These results suggested that biological detection standards of Shen Mai injection presented great growth potential.Microtox-based fast test in the evaluation of comprehensive toxicity of ShenMai injection showed favorable application prospects over controlling the quality of different sources of products.
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This study aimed at exploring a new method--microtox technology to evaluate the comprehensive toxicity of safflower injections from different drug manufacturers.In the experiment,we investigated the characteristic parameters of vibrio fischeri (CS234) under different conditions,and then locked the optimum parameters for safflower injection assay:recovery liquid volume 0.9 mL in freeze-dried vial,50 μL bacteria suspension in each sample,and the detection time of 10 mins after adding the bacteria suspension.Under this optimum detection condition,we found that the EC50 values were 3.36%,5.58% and 4.33%,being significantly different between 3 representative drug manufacturers (P < 0.05).In conclusion,it was implied that the biological detection standards of safflower injections need improving,while the microtox-based fast test of the comprehensive toxicity of safflower injection showed a favorable and prospective application to controlling the product qualities of different manufacturers.
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Objective To evaluate the effect of a vasoactive substance urotensin Ⅱ on the expression of type Ⅰ collagen and migration of human skin fibroblasts,and to explore the underlying mechanisms of signal transduction.Methods Fibroblasts were isolated from human foreskin tissues and subjected to primary culture.After a series of subculture,fibroblasts were classified into several groups to be treated with different concentrations (10-10 to 10-6 mol/L) of urotensin lⅡ for 24 hours,urotensin Ⅱ of 10-s mol/L for different durations (0,4,12,24 hours),or pretreated with PD98059 (a mitogen-activated protein kinase kinase inhibitor),nicardipine (a calcium channel blocker) and ciclosporin (a calcineurin inhibitor) of 10-5 mol/L respectively for 30 minutes followed by treatment with urotensin Ⅱ of 10-8 mol/L for 24 hours.The cells receiving no treatment served as the control.Subsequently,enzyme-linked immunosorbent assay was performed to determine the level of urotensin Ⅱ in the supernatant of fibroblasts,and Transwell assay to estimate the migration activity of fibroblasts.Statistical analysis was carried out by t test and analysis of variance.Results Urotensin Ⅱ promoted the expression of type Ⅰ collagen in a time-and concentrationdependent manner.The level of type Ⅰ collagen was increased by 21.2% (P > 0.05),52.2% (P < 0.05),84.4% (P <0.05),83.6% (P < 0.05) and 77.1% (P < 0.05) in the supernatant of fibroblasts treated with 10-10,10-9,10-8,10-7 and 10-6 mol/L of urotensin Ⅱ for 24 hours respectively,by 23.2% (P > 0.05),69.5% (P < 0.05) and 84.1% (P <0.05) in the supernatant of fibroblasts treated with urotensin Ⅱ of 10-8 mol/L for 4,12 and 24 hours respectively,compared with the untreated control fibroblasts.The migration activity was markedly enhanced in fibroblasts treated with urotensin Ⅱ of 10-8 mol/L for 24 hours compared with the control fibroblasts (P < 0.05).PD98059,nicardipine and cyclosporin A inhibited the secretion of type Ⅰ collagen by 18.2%,15.9% and 19.7% respectively,and suppressed the migration of fibroblasts by 38.3% (P < 0.05),20.7% (P < 0.05) and 81.4% (P < 0.05) respectively in the groups receiving pretreatment compared with those treated with urotensin Ⅱ alone.Conclusions Urotensin Ⅱ can promote the secretion of type Ⅰ collagen by and migration of fibroblasts,which may be realized through the Ca2+,calmodulin kinase,and mitogen-activated protein kinase pathways.
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<p><b>OBJECTIVE</b>To assess the value of short tandem repeat (STR) analysis with fluorescence labeling polymerase chain reaction (PCR) in evaluating the status of engraftment and predicting the occurrence of graft-versus-host disease (GVHD) following orthotopic liver transplantation.</p><p><b>METHODS</b>The method of STR analysis with fluorescence labeling PCR was established. DNA was extracted by monoclonal magnetic beads from the peripheral blood nucleated cells of 23 recipients and labeled with 4 fluorescence dyes before and at 7 days after orthotopic liver transplantation.</p><p><b>RESULTS</b>In the 23 patients studied, 5 patients showed mixed chimeras after orthotopic liver transplantation. Two of the 5 patients with mixed chimeras developed GVHD and died. The other 18 patients did not show mixed chimeras, and only one of them developed GVHD. The incidence of GVHD was significantly different between the patients with and without mixed chimeras (40% vs 5.6%, P<0.05).</p><p><b>CONCLUSION</b>PCR-STR analysis after orthotopic liver transplantation can be indicative of engraftment and help to predict the occurrence of GVHD following orthotopic liver transplantation.</p>
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Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedad Injerto contra Huésped , Diagnóstico , Trasplante de Hígado , Repeticiones de Microsatélite , Reacción en Cadena de la Polimerasa , MétodosRESUMEN
Objective To discuss endoscopic surgical treatment and its effect of intranasal contact point headaches.Methods Twe(n)ty-five patients with intranasal contact point headaches were treated by endoscopic rhinoplasty of nasal cavity,including middle turbinoplasty,functional resection of ostiomeatal complex,endoscopic submucous septoplasty.Achieved organic combination of the above surgery methods according to the different results of the CT scan and endoscope,the surgical procedure waft designed individually.Results All patients were followed up for 9 to 12 months.Recovery was 21 cases(84.00%),efficacy was 3 cases(12.00%),inefficacy was 1 case(4.00%),the rate of fully recovered without serious complication was 96.00%(24/25).Conclusions Intranagal contact point headaches is a sort of the nasal headaches as a result of multi abnormality of nasal cavity structure.Endoscopic rhinoplagty is an effective treatment by means of reconstructing the balance of bilateral nasal cavity and improving its function.
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Functional MRI data is analyzed by using temporal independent component analysis confined to the potential activation extent which is firstly detected by using correlation analysis. Based on the prior information about the activation paradigm, the functional signal component is identified automatically by ordering the resultant independent components with canonical correlation analysis. Finally, the functional MRI data is reanalyzed adaptively by using correlation analysis with the recognized functional signal component as the reference function. The experimental results of analyzing actual functional MRI data show the validity and the reliability of the presented method.
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Humanos , Algoritmos , Artefactos , Mapeo Encefálico , Métodos , Interpretación Estadística de Datos , Imagen por Resonancia Magnética , Métodos , Análisis de Componente Principal , Métodos , Procesamiento de Señales Asistido por Computador , Factores de TiempoRESUMEN
Objective To establish a method for recognizing nasal bleeding sites and hemorrhagic foci of hidden epistaxis.Methods The bleeding sites and hemorrhagic foci,as well as the used surgical techniques and the curative effects were studied retrospectively in 122 patients with spistaxis from Jan.2005 to Oct.2007,in whom the bleeding sites and hemorrhagic foci were not found by routine nasoscope examination.Under nasal endoscopic monitoring,electric heat cauterization(EHC),microwave coagulation(MC) and micro-traumatic nasal packing(MTNP) were applied respectively to treat the hidden epistaxis.Results The hemorrhagic foci were found in the following sites:Olfactory cleft 48 cases(39.3%),superior wall of inferior nasal meatus 20 cases(16.4%),posteroinferior wall of middle nasal meatus 6 cases(4.9%) and so on.Epistaxis was well controlled in 113 of 122 cases(92.6%),in whom the hemorrhagic foci were found by endoscope,by laser soldering,MC and EHC.Packing with mini gel foam was used in 9 cases,for whom the hemorrhagic foci were not found.No complications occurred during a 1-2 months of follow up after treatment.Of the 122 cases,106 cases(86.9%) stopped bleeding by treatment once and 16 cases(13.1%) stopped by treatment twice.Conclusion The lateral or posterior area of middle and inferior nasal meatus,and olfactory cleft area are the frequent sites of hidden epistaxis.Examination with endoscopy,combined with the findings on the middle and inferior turbinate and the features of blood flow in different sites,will be important on recognizing the hidden epistaxis and locating the hemorrhagic foci.
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Objective To provide new diagnose informations for clinician by using image fusion. Methods 30 cases with brain lesion (men 18,women were studied 12). Twenty cases were examined by CT or MR in 1_2 weeks, the others were checked by MR, after being diagnosed with CT. These image data were input into computer the center and main axis of image were located by Legendre moment then the CT and MR images were registered by translation, scaling and rotation. Results Of 30 image fusions, supplementary informations were provided to each other in 28 cases,prediction of disease progress in 19 cases. There were 4 cases confirmed at surgery. But in 2 cases no obvious advantage was obtained by image fusion.Conclusion CT and MR image fusion provides useful information for definitive diagnosis, planning of surgery and radiotherapy. To fuse the images of CT and MR, the Legendre moment approach is direct, simple and fast.
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Objective: To investigate the combined inhibitive effect of TNP-470 and rhES on the growth of lung adenocarcinoma LA795 in T739 mice. Methods: The purified rhES was acquired by using methanol to induce the recombinant pichia pastoris. GS115 and heparin affinity chromatography. The T739 mice inoculated with LA795 cells were randomized into three groups, 10 mice per group, one group was injected with PBS for 14 days, the other two groups were respectively treated with rhES and TNP-470+rhES. To observe the tumor growth in different groups, and the tumor volume was measured with caliper. The microvessel density(MVD) of tumors were measured by using immunohistochemistry. Results: The purified rhES was acquired. In compared with PBS group, the tumor growth of other two groups was inhibited significantly. And the tumor volume of TNP-470+rhES group are smaller than the rhES group (P
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DICOM is a standard for data format and transmission of digital medical image. DICOM Data Set is a binary data stream using DICOM encoding rule. DICOM Nesting Data Set is a kind of complex Data Set with a tree structure, and is widely used in DICOM services and encoding of DICOM files for its special structure. In this article, the functions and encoding rule of Data Set and Nesting Data Set in DICOM format are presented, and the way of parsing and organizing of them is put forward. The realization method and practical application are also discussed.
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The standardization of data transfer plays an important role in precise radiotherapy,not only boosting the speed but also ensuring the treatment quality.In this article,a method for beam data generation and transfer is introduced based on DICOM RT Standard.Firstly,the realization procedure of this module is presented.Secondly,the realization means of two different IMRT methods including SMLC and DMLC are explained respectively.Finally,the system's validity is proved with instances.
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OBJECTIVE:To standardize the writing of outpatient prescriptions and set up monitoring mechanism on rational use of antibiotics and drug costs.METHODS:A total of 400 prescriptions collected from 8 departments(50 from each department)of our hospital in the fourth quarter of 2005(before the examination)and from first to fourth quarter in 2006(after the examination)were examined statistically according to Inspection Items and Writing Requirements of Clinical Prescriptions,Guiding Principle of Rational Application of Antibiotics in Hospital,Principles of Classified Management of Antibiotics which were formulated by our hospital.RESULTS:Before prescription examination,the qualified rate of prescriptions was only 57.5%,but rose to an average of 85.6% after the examination.CONCLUSION:To include the standardized writing of prescriptions and reasonable application of antibiotics in the assessment of hospital comprehensive goals was proved to be an effective management method.