OBJECTIVE:To discuss the accuracy of detecting tacrolimus(FK506) concentration in whole blood by ELISA and its application in organ transplantation.METHODS:The ELISA was used to detect FK506 valley point concentrations in 5 liver transplanted recipients at different postoperative periods .Based on the results combined with clinical findings, the dosages were adjusted.The accuracy of the method and guidance action in FK506 clinical application were described.RESULTS: In 131 detections of 350 samples, the results of quality control samples were all in the control range .The detecting RSDs of high and low concentration quality control samples were 9.09% and 11.90%, respectively.The control range of FK506 concentrations in whole blood were similar to that reported in literature .CONCLUSION :ELISA is a sensitive and specific method for routine FK506 concentration monitoring in whole blood,which is suitable for application in hospital.

Objective To investigate blood gas analysis and lactic acid evaluation in aerobic forearm exercise and the significance of aerobic forearm exercise for the auxiliary diagnosis of mitochondrial myopathy and encephalopathy patients.Methods Forty-two patients with mitochondrial myopathy and encephalopathy patients, 40 healthy control, and 40 patients control were studied.They performed a protocol under aerobic exercise conditions, consisting of intermittent forearm exercise for 4 minutes at 40% of intented maximal voluntary contraction force.Blood samples were collected to monitor blood gas and plasma lactate before, during arid after exercise.Results During exercise venous PO_2(mm Hg, 1 mm Hg=0.133 kPa)decreased in mitochondrial myopathy and encephalopathy patients from 41.2?12.6 to 39.5?16.2, whereas PO_2 fell from 50.5?14.4 to 30.8?13.1 in healthy control and from 50.1?7.9 to 44.3?35.5 in patient control.Venous PO_2 decreased much more in healthy control group than the other 2 groups(F= 6.34,P

Objective To investigate the potential role of T follicular helper cells (Tfh) in the pathogenesis of autoimmune thyroid diseases by comparing the expression of C-X-C chemokine teceptor type 5 (CXCR5) and CD57 in Hashimoto's thyroiditis (HT) and Graves' disease (GD) thyroid tissues.Methods The expression of CXCR5 and CD57 proteins was determined by immunohistochemical analysis in 15 HT thyroid samples,18 GD samples and 10 normal thyroid samples.Results Immunohistochemical staining showed that CXCR5 and CD57 were mainly positive in cytomembrane and cytoplasm of the infiltrated lymphocytes both in HT and GD tissues,with much higher levels than that of normal thyroid tissues ( P ＜ 0.05 ).Both CXCR5 and CD57 were not significantly different between the HT and GD tissues.Conclusion CXCR5 and CD57 expressions were increased with a similar expression pattern in both of the two main autoimmune thyroid diseases( AITD),indicating that Tfh may participate inthe development and progression of AITD.

Objective To appraise and compare protein expression profiles in sera of patients without or with recurrence following liver transplantation for hepatocellular carcinoma (HCC) using SELDI-TOF-MS technique,and establish the diagnostic and predictive model. Methods A total of 76 sera (41 from disease free survival patients and 35 from recurrence individuals) were collected pretransplantation and differentially expressed proteins were identified by SELDI-TOF-MS. The intensity values for each peak were analyzed by Biomarker Wizard Software to screen serum proteome biomarkers related to the recurrence post-transplantation. By using Biomarker Patterns Software, the classification trees were generate. from randomly selected samples (30 fingerprints obtained from each group). The sensitivity and specificity of best decision tree were then chosen for blind test with 16 samples (5 from recurrence individuals and 11 from recurrence-free survival patients). Results There were significant differences only in tumor size and the presence of vascular invasion between recurrence group and recurrence-free survival group (P＜0.05). According to serum protein fingerprints, a total of 368 protein peaks were identified at the mass-to-charge ratio (M/Z) value ranging from 2000 to 300 00. There were 22 significant differential proteins between two groups. Among them, 9 proteins were up-regulated and 13 proteins were down-regulated -espectively in recurrence group. The intensity values of differential proteins were input into BPS for classification tree analysis and the best performing tree could distinguish two groups successfully. As a result of blind assessment for this model,a sensitivity of 80.0 % (4/5) and specificity of 72.7 % (8/11) were obtained. Conclusion Some of differential proteins screened by SELDI-TOF-MS technique in the serum may be correlated with the prognoses of liver transplantation patients with HCC. The decision tree may be useful for the clinical application of formulating the indication for liver transplantation, detecting extrahepatic micrometastasis and setting up the diagnostic and treatment strategies.

Estrogen receptor alpha (ERalpha) has been a primary target of treatment as well as a prognostic indicator for breast cancer. The level of human X-box binding protein 1 (XBP-1) mRNA was related with that of ERalpha in breast tumors and was over-expressed in some breast tumors. These previous studies suggested that XBP-1 may interact with ERalpha. XBP-1 has two isoforms, XBP-1S and XBP-1U, as the result of unique splicing. GST pull-down assay showed that both XBP-1S and XBP-1U bound to ERalpha in vitro. The binding of XBP-1S to ERalpha was stronger than that of XBP-1U to ERalpha. Co-immunoprecipitation revealed that the binding was in a ligand-independent manner. XBP-1S and XBP-1U interacted with the region of ERalpha that contains a DNA-binding domain. The ERalpha-interacting regions on XBP-1S and XBP-1U have been mapped to two regions, the N-terminal basic region leucine zipper domain (bzip) and the C-terminal activation domain. These findings suggest that XBP-1S and XBP-1U may participate in ERalpha signaling pathway through the mediation of ERalpha.
Breast Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; metabolism ; Estrogen Receptor alpha ; genetics ; metabolism ; Female ; Humans ; Protein Interaction Domains and Motifs ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Regulatory Factor X Transcription Factors ; Signal Transduction ; Transcription Factors ; genetics ; metabolism ; X-Box Binding Protein 1

OBJECTIVETo evaluate clinical results of implant supported telescopic overdenture.

METHODS21 patients with edentulous jaws underwent telescopic overdenture restoration. A total of 28 prostheses were fabricated. Of them, 13 were in the upper jaws, 15 in the lower jaws. Among 139 placed implants, 74 were Camcog, 28 IMZ, and 37 Frialit-2. The secondary crown was fabricated by electroforming technique and wax-lost cast method. Clinical examination and radiographs were conducted. Changes in the marginal bone level around the implants were evaluated with radiograph. The mean follow-up time was 26.5 months (range 12 - 39 months).

RESULTSThis type of restoration could provide sufficient stability and maintain peri-implant hygiene easily. Peri-implantitis and prosthetic complications were not observed. Marginal bone around implants was stable. No implant was lost during the loading time.

CONCLUSIONSThe preliminary clinical results of this research showed that implant supported telescopic overdentures were predictable for edentulous patients.

Aged ; Crowns ; Dental Prosthesis, Implant-Supported ; methods ; Denture, Overlay ; Female ; Follow-Up Studies ; Humans ; Jaw, Edentulous ; surgery ; Male ; Middle Aged

AIMTo investigate the effects of fungal elicitors on inophyllums production in suspension cultured cell of Calophyllum inophyllum Linn.

METHODSThe pathogen of leaf spot disease of C. inophyllum L. was isolated and prepared as fungal elicitor. The fungal elicitor was added to the medium with different concentrations and culture period. Their effects on biomass and inophyllums content of the suspension of cultured cells were studied.

RESULTSThe optimum effects of S-I fungal elicitor concentrations on inophyllums content was 60 mg GE x L(-1). Adding the fungi elicitor into the cell suspension culture system at stationary phase (being cultured for 18 days) resulted in a highest inophyllum content of 59.174 mg x L(-1) at the 3rd day with 27% higher than control. Fungal elicitor treatment promoted the inophyllums accumulation in medium.

CONCLUSIONAdding the Stagonospora curtisii (Berk.) Sacc. to the medium was effective approaches to enhance inophyllums yield in the suspension of C. inophyllum L culture cell.

Calophyllum ; cytology ; metabolism ; Cell Division ; Cells, Cultured ; Coumarins ; metabolism ; Culture Media ; Mitosporic Fungi ; physiology

OBJECTIVETo observe the expression and effect of human calcitonin gene-related peptide (hCGRP) gene mediated by recombinant adeno-associated virus (rAAV) in primary cultured corporal cavernosum smooth muscle cells of the rat and explore the possibility of using CGRP gene for gene therapy in erectile dysfunction.

METHODSThe primary cultured corporal cavernosum smooth muscle cells of the rat were randomly divided into 4 groups and infected with recombinant virus VssHGCMV-hCGRP, VssHGCMV, VssC-MV-GFP and the untreated, respectively. CGRP-like immunoreactivity was measured by protein dot blot assay in the 24 h-culture medium, and intracellular cAMP and cGMP levels in the cultured cells were also determined using radioimmunoassay to ascertain bioactivity of transduced CGRP.

RESULTSThe exogenous gene was transferred into primary corporal cavernosum smooth muscle cells by VssHGCMV-hCGRP infection and efficiently expressed. Compared with the control group, intracellular cAMP level in the cell infected by VssHGCMV-hCGRP was significantly increased (48.7 +/- 1.1 nmol/L vs 7.8 +/- 1.4 nmol/L, P < 0.01), whereas cGMP level remained unchanged in two groups, and CGRP-like immunoreactivity was also detected in the culture medium infected by VssHGCMV- hCGRP.

CONCLUSIONThe system of secretory expressing bioactive peptide rAAV mediated gene transfer may be used to express efficiently exogenous gene in corporal cavernosum smooth muscle cells and affect cAMP level in the corporal cavernosum smooth muscle cells of the rat.

Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Cells, Cultured ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Dependovirus ; genetics ; Male ; Muscle, Smooth ; cytology ; metabolism ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Recombination, Genetic ; Transfection

OBJECTIVETo determine whether recombinant adeno-associated virus-mediated overexpression of hCGRP in the corpus cavernosum can affect the continuous production of hCGRP in the penile tissue and enhance erectile responses in STZ-induced diabetic rats.

METHODSDiabetes mellitus was induced by a single intraperitoneal injection of 60 mg/kg streptozotocin in male SD rats. VssHGCMV-hCGRP, VssCMV-GFP and rAAV solution were injected into the corporal cavernosum of STZ-induced diabetic rats, respectively. The corporal tissue was obtained from groups of 8 rats on day 5 post-injection, and the expressions of CGRP and GFP in cavernosal tissue were detected using immunohistochemistry and laser scanning confocal microscopy, respectively. Cavemosal tissue cAMP and cGMP levels were measured using radioimmunoassay. On day 5 post-injection, intracavernous pressure induced by electrostimulation of penile dorsal nerves was measured and recorded with a biological signal processing system in each group rat.

RESULTSrAAV transduction efficiency of GFP reporter gene was measured by laser scanning confocal microscopy and was observed in the penile tissue, especially in the corporal cavernosum and the vessel 5 days after transfection with VssCMV-GFP. Immunohistochemistry showed that the CGRP increased in the corporal cavernosum. In addition, both cAMP and cGMP levels in the corpora cavernosa transfected with VssHGCMV-hCGRP were significantly increased, compared with controls [(48.4 +/- 6.5) nmol/L and (21.2 +/- 13.6) nmol/L vs (16.7 +/- 2.5) nmol/L and (0.42 +/- 0.12) nmol/L, respectively]. More importantly, 5 days after administration of VssHGCMV-hCGRP,a significant increase was observed in the erectile response to penile dorsal nerve stimulation in the diabetic rat [(60.5 +/- 4.5) mm Hg vs (22. 3 +/- 1.3) mm Hg].

CONCLUSIONSThis results demonstrate that rAAV-mediated transfer of the CGRP gene can increase production of endogenous CGRP, cAMP and cGMP in corpora cavernosa of STZ-induced diabetic rats. Moreover, overexpression of CGRP enhances ICP and the erectile response to penile dorsal nerve stimulation in the diabetic rat.

Adenoviridae ; genetics ; Animals ; Calcitonin Gene-Related Peptide ; biosynthesis ; genetics ; Diabetes Mellitus, Experimental ; physiopathology ; Green Fluorescent Proteins ; biosynthesis ; Humans ; Male ; Penile Erection ; physiology ; Penis ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo devise a multiplex PCR system of three X-chromosome specific short tandem repeat (X-STR) loci and study the genetic polymorphism.

METHODSDXS6799, DXS6804 and DXS6854 were amplified simultaneously using a multiplex system and were typed by polyacrylamide gel electrophoresis and silver staining.

RESULTSA total of 262 male and 255 female individuals from Guangdong Han population were tested; each locus showed 7 alleles. 73 haplotypes were detected in the male individuals. The haplotype diversity reached 0.9674.

CONCLUSIONThe 3 X-STR multiplex system is relatively abundant in polymorphic information for forensic identification and paternity testing.

Chromosome Mapping ; Chromosomes, Human, X ; Female ; Gene Amplification ; Haplotypes ; Humans ; Male ; Paternity ; Polymorphism, Genetic ; Tandem Repeat Sequences