Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulins, Intravenous ; therapeutic use ; Infant ; Infant, Newborn ; Male ; Prednisone ; therapeutic use ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy

Objective To investigate the density of bladder pacemaker cells (ICC) in prostatic hyperplasia with detrusor underactivity (PDU) and the impact of stress environment on its growth. Methods 23 PDU guinea pig models and 26 pigs for control (BPH, n = 6; normal group, n = 20) were developed. After water-filled urodynamic pressure measurement of bladder for three groups , detrusor tissue was made into frozen sections. TheICC numberwere measured with immunofluorescence stainingand observed by confocal microscopy. Further ICC cells in vitro were cultured under stress environment and its counts were analyzed by colony formation assay. Results Decreased density of ICC was observed in PDU group (0.28 ± 0.52)% compared to control [(5 ± 0.36)%,P < 0.05] and the BPH group (5.4% ± 0.42)%, P < 0.05). The ratio of dimer and monomer [(6.13 ± 1.45)%] in PDU group was significantly lower than those of the control [31.55 ± 9.67)%, P < 0.05] and the BPH group [(71.52 ± 8.95)%, P < 0.05]. There is a negative correlation between the density of ICC in vitro and duration of stress. By contrast , there is a positive correlation between the density of ICC and incubation time after the stress is removed. Conclusion Distribution of decreased density of ICC is involved in the pathogenesis of PDU , which may be due to long-term stress environment. Thus eliminating stress environment may help restore ICC density in PDU.

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.

Objective To assess the efficacy and safety of microvascular decompression for treatment of trigeminal neuralgia in elderly patients. Methods The clinical data of29 elderly patients (above 65 years old) with trigeminal neuralgia treated with microvascular decompression were retrospectively reviewed to analyze the outcomes of the surgery and complications in comparison with those of younger patients below 65 years of age. Results Good therapeutic effects were achieved in both groups without obvious difference between them,the rate ofpostoperative pain relieffor the elderly group is 93.10%,96.36%for the control group.The elderly patients showed some special anatomical and systemic features in relation to aging,including the presence of brain atrophy,obvious arteriosclerosis in the culprit arteries(the elderlygroupis about 1/4,the control group is about 1/10),multiple systemic diseases(the elderly group accounted for 71.69%,41.81%for control group),and increased risk of surgical complications(one of the elderly group appears brainstem infarction,2 cases of lower extremity deep VenOUS thromboembolism disease). Conclusion Microvascular decompression provides an effective cure or pain relief for most elderly patients with trigeminal neuralgia with the application of effective perioperative treatment.

OBJECTIVETo obtain streptavidin-tagged human interleukin-15 (SA/hIL15) fusion protein and evaluate its bioactivity.

METHODSpET24a-6His-SA-hIL-15 and pET32a-hIL-15-SA-6His plasmids were constructed and expressed in BL 21(DE3) host bacteria to generate the fusion protein. The recombinant fusion protein IL-15/SA was purified using Ni-NTA affinity chromatography and refolded, and the efficiency of surface modification of the fusion protein on biotinylated cells was examined by fluorescence-activated cell sorting. CCK-8 method was used to evaluate the effect of IL-15/SA fusion protein in inducing the proliferation of human peripheral-blood lymphocyte (PBL) cells stimulated by PHA.

RESULTSThe recombinant SA-hIL-15 and hIL15-SA fusion proteins were highly expressed in BL21(DE3) at about 20% of the total bacterial proteins. The purified hIL15-SA fusion protein exhibited a bifunctionality by promoting the proliferation of PBL cells activated by PHA and high-affinity binding to biotinylated cell surface mediated by SA, with a cell surface modification efficiency exceeding 95%. SA-hIL-15 showed a 4-fold higher hIL15 bioactivity than hIL15-SA.

CONCLUSIONSA/hIL-15 bifunctional fusion protein has been successfully obtained to facilitate the future development of hIL-15-surface-modified cancer cell vaccine.

Cancer Vaccines ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Interleukin-15 ; biosynthesis ; genetics ; Lymphocyte Activation ; drug effects ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Streptavidin ; biosynthesis ; genetics

OBJECTIVEHepatic stellate cells (HSC) in hepatocellular carcinoma (HCC) transdifferentiate into extracellular matrix-producing myofibroblasts. Activated HSC can promote invasion and metastasis of HCC. To understand the differences of HSC in normal liver and HCC, we compared the gene expression patterns in HCC cell induction-activated and culture-activated rat HSC.

METHODSHSC were isolated by density centrifugation and exposed to conditioned medium from rat HCC cell line C5F. Expression of 22 012 genes in quiescent HSC, culture-activated HSC and HCC induction-activated HSC was analyzed by cDNA microarray and confirmed by real-time RT-PCR and Western blot.

RESULTS1672 genes were differentially expressed in culture-activated HSC, including proinflammatory factors, cell adhesion molecules, cell surface receptors, signaling transduction molecules and immune factors. 711 genes were differentially expressed in HCC induction-activated HSC. Some of them were identical to those in culture-activated HSC. HCC Induction-activated HSC showed specific gene expression patterns, including Raf1, Rac2, Adam17, Wnt6, MMP-9 and TNF, suggesting that HCC cells can specifically induce HSC activation.

CONCLUSIONThe gene expression patterns in HCC induction-activated HSC are different from those in culture-activated HSC. HCC induction-activated HSC may play a major role in the invasion and metastasis of HCC. In vivo activation should be considered as the standard for the study of HSC biology. HCC induction-activated HSC should be considered as the standard for HSC biology studies.

Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Hepatic Stellate Cells ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Oligonucleotide Array Sequence Analysis ; Rats ; Rats, Inbred F344

The essence of endogenous turbidity in Chinese medicine (CM) is different from cream, fat, phlegm, retention, damp, toxicity, and stasis. Along with the development of modern scientific technologies and biology, researches on the essence of endogenous turbidity should keep pace with the time. Its material bases should be defined and new connotation endowed at the microscopic level. The essence of turbidity lies in abnormal functions of zang-fu organs. Sugar, fat, protein, and other nutrient substances cannot be properly decomposed, but into semi-finished products or intermediate metabolites. They are inactive and cannot participate in normal material syntheses and decomposition. They cannot be transformed to energy metabolism, but also cannot be synthesized as executive functioning of active proteins. If they cannot be degraded by autophagy-lysosome or ubiquitin-prosome into glucose, fatty acids, amino acids, and other basic nutrients to be used again, they will accumulate inside the human body and become endogenous turbidity. Therefore, endogenous turbidity is different from final metabolites such as urea, carbon dioxide, etc., which can transform vital qi. How to improve the function of zang-fu organs, enhance its degradation by autophagy-lysosome or ubiquitin-prosome is of great significance in normal operating of zang-fu organs and preventing the emergence and progress of related diseases.
Autophagy ; Humans ; Medicine, Chinese Traditional ; Proteasome Endopeptidase Complex

The present study investigated whether a co-application of tramadol (TRA) and dihydroetorphine (DHE) would exert a synergy in analgesic effect and delay acute tolerance development. Intraperitoneal injection of TRA (in mg) and subcutaneous injection of DHE (in ng) were delivered in fixed proportions (1:6.25, 1:12.5, 1:25, 1:50, 1:100, and 1:200). The effect of analgesia was accessed by tail-flick test and analyzed with isobolographic analysis. For test of acute tolerance, six successive injections of either TRA (20 mg/kg) alone, DHE (1 000 ng/kg) alone, or a combination of TRA (20 mg/kg) and DHE (250 ng/kg) were administered. We found that (1) except for 1 mg : 6.25 ng and 1 mg : 50 ng, combinations, all the other ratios produced a significant synergy in their analgesic effect; (2) the effect of analgesia induced by repeated TRA plus DHE injections lasted significantly longer, indicating a slower onset of acute tolerance. These results indicate that TRA and DHE injections in certain dose ratios can induce synergistic analgesia, which is resistant against the development of acute tolerance.
Analgesics, Opioid ; administration & dosage ; pharmacology ; Animals ; Drug Synergism ; Drug Tolerance ; physiology ; Etorphine ; administration & dosage ; analogs & derivatives ; pharmacology ; Female ; Male ; Pain ; drug therapy ; physiopathology ; Rats ; Rats, Wistar ; Tramadol ; administration & dosage ; pharmacology

Different factors including hybridization solution components, hybridization temperature, and the concentration and proportion of the labelled primer, which affected the sensitivity and specificity of single mutation identification, were exploited. Asymmetric PCR increased the hybridization sensitivity, and the asymmetric multi-PCR did not affect the specificity, while the sensitivity was improved a little. Among 30 lung cancer samples detected with the oligonucleotide microarray, 12 was found P53 gene mutations and 5 had K-ras gene mutations. The P53 gene mutations identified by the oligonucleotide microarray was proved 80% same as the sequencing results. The obvious statistical relations of K-ras and P53 gene mutations with tumor type, tumor stage and smoking were not obtained because of less samples and mutation sites.
Genes, ras ; genetics ; Humans ; Lung Neoplasms ; genetics ; pathology ; Mutation ; Oligonucleotide Array Sequence Analysis ; methods ; Oligonucleotides ; genetics ; Sensitivity and Specificity ; Tumor Suppressor Protein p53 ; genetics