Objective:To explore the expression of TNF a and IL-10 in different layers of laryngeal tissues after laryngeal transplantation and its relationship with acute rejection.Methods:Laryngeal heterotopic transplantation was performed in Wistar and SD rats(Wistar→SD rats).The SD rats were divided into 4 groups:GroupⅠ:Sham control(receive no transplantation): GroupⅡ(receive transplantation,without cyelosporine A treatment);GroupⅢ(receive transplantation.with 5 mg?kg~(-1)?d cyelosporine A treatment):and GroupⅣ(receive transplantation.with 10 mg?kg~(-1)?d~(-1)cyelosporine A treatment).The transplanted larynx was harvested on 3,7 and 11 days after transplantation for pathological examination.The expression of TNF-?and IL-10 in different layers of grafts was detected immunohistochemically.Results:Pathological observation showed mild,moderate and severe acute rejection in GroupⅡandⅢon 3.7 and 11 days after transplantation,respectively:there was no obvious rejection in GroupⅣ.Immunohistochemical staining showed expression of TNF-?and IL-10 in GroupⅡ.Ⅲ.andⅣ,not in GroupⅠ.The ratios of the positive areas of TNF-?and IL-10 in the mucosal and submucosal layers were significantly higher than those in the muscle and cartilage layers of laryngeal tissues(P

Objective To optimize the enamel electron paramagnetic resonance (EPR) spectral processing by using the EPR spectral simulation method to improve the accuracy of enamel EPR dosimetry and reduce artificial error.Methods The multi-component superimposed EPR powder spectral simulation software was developed to simulate EPR spectrum models of the background signal(BS) and the radiation- induced signal (RS) of irradiated enamel respectively.RS was extracted from the multi-component superimposed spectrum of irradiated enamel and its amplitude was calculated.The dose-response curve was then established for calculating the doses of a group of enamel samples.The result of estimated dose was compared with that calculated by traditional method.Results BS was simulated as a powder spectrum of gaussian line shape with the following spectrum parameters:g =2.00 35 and Hpp =0.65 - 1.1 mT,RS signal was also simulated as a powder spectrum but with axi-symmetric spectrum characteristics.The spectrum parameters of RS were:g(1) =2.0018,g (11) =1.996 5,Hpp =0.335 - 0.4 mT.The amplitude of RS had a linear response to radiation dose with the regression equation as y =240.74x + 76 724 ( R2 =0.9947 ).The expectation of relative error of dose estimation was 0.13.Conclusions EPR simulation method has improved somehow the accuracy and reliability of enamel EPR dose estimation.

Eustachian Tube ; surgery ; Foreign Bodies ; surgery ; Humans ; Male ; Middle Aged

In this study, OVA-induced asthma mice was taken as the model, and orally administered with different concentration of ethanol extracts of crude and processed Stemona tuberosa, in order to determine the cytokine level released from Th1 and Th2 in splenocytes. RT-PCR was carried out to determine the genetic expression of T-bet/GATA-3 in lung, and compare the differentiation between ethanol extracts of crude and processed S. tuberosa in therapeutic effect on asthma in mice. According to the results, compared with the crude samples, processed samples significantly increased the levels of inflammatory factor INF-gamma (P < 0.05) and decreased IL-5 (P < 0.05) in splenocytes. According to the RT-PCR results, the administration of processed samples could increase the ratio of T-bet/GATA-3 (P < 0.05). The experiment showed that ethanol extracts of both crude and processed S. tuberosa could treat asthma by regulating Th1/Th2 ratio, but processed samples showed more notable effect. This indicated that crude and processed S. tuberosa had significant pharmacological difference. Therefore, it was more rational to apply processed S. tuberosa in clinical treatment of asthma and chronic cough, which layed a foundation for further revealing the processing mechanism of S. tuberosa.
Administration, Oral ; Animals ; Asthma ; drug therapy ; immunology ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; GATA3 Transcription Factor ; metabolism ; Gene Expression Regulation ; drug effects ; Mice ; Mice, Inbred BALB C ; Stemonaceae ; chemistry ; T-Box Domain Proteins ; metabolism ; Th1 Cells ; drug effects ; secretion ; Th2 Cells ; drug effects ; secretion

Aged ; Female ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; surgery ; Humans ; Immunohistochemistry ; Spleen ; chemistry ; pathology ; surgery ; Splenectomy ; Splenic Neoplasms ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

The purpose of this study is to investigate the transdermal delivery characteristics of Gentiana macrophylla complex components system through different parts of the skin under micro-needles conditions. Two-chamber diffusion cells were used, different parts of isolated skin and micro-needle pretreated isolated mouse skin were applied separately, high performance liquid chromatography (HPLC) similarity evaluation methods were used to evaluate transdermal delivery characteristics of Gentiana macrophylla complex components system on receiving pool and the permeation rate and penetration amount of Gentiopicroside at different parts of mouse skin. In the 24 h, the similarity between receiving fluid which was on passive transdermal delivery and micro-needle transdermal delivery conditions and original fluid were ranged from 83.0% to 98.9%; By the micro-needle pretreatment with different parts of the mouse skin, the time that Gentiana macrophylla complex components system though abdominal skin to the receiving fluid which reached 90% similarity compared with that of original fluid was 4 h, which was 18 h at back skin and 12 h at neck skin separately. Micro-needles can be used as the ideal ingredients for traditional Chinese medicine complex transdermal delivery; transdermal absorption time delay could be greatly reduced and its bioavailability was improved. The permeation rate and similarity to original liquid of Chinese medicine complex components increased significantly in the abdominal skin relative to the neck and back skin under micro-needle conditions.
Administration, Cutaneous ; Animals ; Biological Availability ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacokinetics ; Gentiana ; chemistry ; Iridoid Glucosides ; pharmacokinetics ; Mice ; Needles ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Skin Absorption

OBJECTIVETo investigate the difference of serum high molecular weight alkaline phosphatase (HMAP) levels between biliary atresia (BA) and neonatal hepatitis (NH), and to develop a new differential method and early diagnostic indicators for cholestatic jaundice in neonates.

METHODSTotally 31 patients with cholestatic jaundice seen between Aug. 2000 and Feb. 2002, including 15 cases with BA, 16 cases with NH, 30 healthy infants and 30 infants with non-cholestatic jaundice were enrolled in this study. Serum samples were obtained from each subject by using venipuncture. The samples were stored at -80 degrees C and analyzed within 6 months. A murine hybridoma producing monoclonal antibody to human high molecular weight alkaline phosphatase (MoAb HMAP-1) was prepared by using partially purified HMAP from human serum as the immunogen. The antibody did not cross-react with other alkaline phosphatase (ALP) isozymes. A monoclonal antibody immunocatalytic assay for HMAP in serum was developed by using MoAb HMAP-1 bound to nitrocellulose membrane discs. The serum total ALP (TALP) and gamma-GT were determined in the meantime, the hepatobiliary ultrasonography and scintigraphy were performed too. The data were analyzed with t test, chi-square test and percentage. Comparisons were made between BA and NH with their sensitivity and specificity in different methods.

RESULTSSerum HMAP was detected in 14 of 15 patients of BA, in 2 of 16 NH patients, while in none of the healthy control group. The positive ratios of serum HMAP in BA and NH were 93.3% and 12.5%, respectively (P < 0.005). The sensitivity and specificity of serum HMAP in BA and NH were 93.3% and 87.5%, respectively. The sensitivity and specificity of TALP, gamma-GT and hepatobiliary scintigraphy were 80.0%, 73.3%, 86.7% and 62.5%, 68.8%, 62.5%, respectively, which were clearly lower than those of serum HMAP.

CONCLUSIONSThe determination of serum HMAP was more sensitive and specific than the other methods tested. Therefore the method can be used as a useful indicator for cholestatic jaundice in neonates, although it needs further study.

Alkaline Phosphatase ; blood ; Diagnosis, Differential ; Female ; Humans ; Immunoenzyme Techniques ; methods ; Infant ; Infant, Newborn ; Jaundice, Obstructive ; diagnosis ; Male ; Sensitivity and Specificity ; gamma-Glutamyltransferase ; blood

This study was purposed to investigate the inhibitory effect of bactericidal permeability-increasing protein (BPI) on lipopolysaccharide (LPS)-mediated activation of platelets. Venous blood samples were obtained from 10 healthy volunteers and were prepared into platelet-rich plasma (PRP, 1 × 10(8)/ml). Experiments were divided into four groups: normal platelet group (untreated group); LPS group, BPI group and BPI+LPS group. PRP were stimulated by LPS (10 µg/ml) in the presence and absence of BPI (100 µg/ml) or BPI alone. Then platelets were harvested and determined for Toll-like receptor-4 (TLR-4) with flow cytometry (FCM), the supernatant was used for detection of cytokines including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) with enzyme-linked immunosorbent assay (ELISA). The results showed that as compared with normal platelet group, TLR-4 expression on platelets was significantly increased under LPS stimulation (P < 0.001); the levels of TNF-α and IL-6 in the supernatant were also remarkably elevated (P < 0.001). However, either TLR-4 expression or the cytokine levels significantly decreased in the presence of BPI when platelets underwent LPS-challenge (P < 0.05), but still were higher than that in normal platelet group. Stimulating the platelets with BPI alone could not enhance the TLR-4 expression and cytokine levels. It is concluded that BPI has the ability to inhibit the LPS-induced platelet activation.
Antimicrobial Cationic Peptides ; pharmacology ; Blood Proteins ; pharmacology ; Humans ; Inflammation ; Lipopolysaccharides ; adverse effects ; Platelet Activation ; drug effects ; Platelet-Rich Plasma ; metabolism ; Toll-Like Receptor 4 ; metabolism

Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal disorder of hematopoiesis due to the inactivation of PIG-A gene. However, the presence of mutant PIG-A gene in a group of hematopoietic cells is not enough for the development of PNH, immunologic injury and apoptotic effects are considered to play an important role in clonal expansion. Knowledge of the molecular mechanisms leading to PNH has substantially increased in the past decades, which remarkably advances the diagnostic modalities and treatment approaches of patients with PNH. Though great progress has been made because of targeted therapy method, the challenges are still ahead. In this review the advances of studies on mechanism, laboratorial diagnosis and therapeutic protocols of PNH are summarized.
Complement System Proteins ; Hemoglobinuria, Paroxysmal ; diagnosis ; genetics ; pathology ; therapy ; Humans ; Membrane Proteins ; genetics ; T-Lymphocytes, Regulatory

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.