Objective To investigate changes of serum levels of interleukin(IL)-4,-6,-8 and tumor necrosis factor-?(TNF-?),and probe their clinical significance in children with Henoch-Schonlein purpura(HSP)/ HenochSchonlein purpura nephritis(HSPN).Methods Serum levels of IL-4,-6,-8 and TNF-? of 45 children with HSP/HSPN and 43 healthy children were examined by enzyme linked immunosorbent assay(ELISA),changes and correlation between the aforementioned cytokines in children with HSP /HSPN were analyzed.Results 1.Serum levels of IL-4,-6,-8 and TNF-? of children with HSP were higher than those of control group(P0.05);The serum level of TNF-? was positively correlated to the serum levels of IL-6,-8 of patients with HSP(r=0.670 P

Objective To establish a molecular inversion probe ( MIP) method for detection of single base drug-resistance mutation in Hepatitis B virus ( HBV) gene.Methods The HBV wild type and YVDD mutant strain were isolated by Daping Hospital of the Third Military Medical University.The MIP was designed and applied to detect the HBV drug-resistance YVDD mutation in one case of wild type and one case of YVDD mutant HBV strain isolated previously.The results of MIP method were compared with that of sequencing to evaluate the detection accuracy.Results Thermal cycling single-base extension and connection reaction performed by Taq DNA Ligase and Ampligase DNA Ligase could ensure the specificity of the detection.The optimum probe concentration of MIP was 1 nmol/L.Through detection of the target gene with different DNA concentrations , the detection sensitivity of MIP was determined as 1 nmol/L.The results of MIP were consistent with that of sequencing method in detection of the clinical samples.Conclusion MIP is successfully used to detect single-base drug-resistance mutation in HBV gene.

Monitoring engraftment of donor cells after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is supposed to be important for the early diagnosis of graft failure or relapse of malignancy. Several techniques have been reported for this purpose. PCR-based assays analyzing polymorphic short tandem repeats (STR) as markers are attractive because they are sensitive and can be performed rapidly. The intent of this study was to test a novel approach for assessment of donor engraftment using denaturing high-performance liquid chromatography (DHPLC) combined with STR-PCR. The feasibility of this assay and the accuracy of semi-quantitative results were tested in-vitro by using serial DNA mixtures from unrelated individuals. The results showed that dilution experiments of the mock chimerism sample revealed a clear correlation between the percentage of donor or recipient DNA and the proportion of allele peak areas, with the limit of detection for a minor DNA percentage being 5%. Discrimination between donor and recipient was possible in all patients analyzed (n = 51) except for 5 patients whose pre-transplant samples were not available and identical twins in one case. STR results were the same as values obtained by capillary electrophoresis combined with fluorescence labeling multiply PCR. Results were also compared with data obtained with FISH analysis in a subgroup of patients receiving grafts from sex-mismatched donors or with PCR-detectable disease-specific gene products analysis. The results of the microsatellite analysis correlated well with the corresponding clinical findings. Full donor chimerism (FDC) were detected in all patients; decreasing values of donor chimerism were detected concomitantly with the appearance of relapse of disease in 3 patients. Samples from eight patients receiving HLA mismatched-haploidentical transplants from related donors together with cord blood transplants from unrelated donors were analyzed by this method. The results showed all 8 patients achieved FDC derived from related donors. It is concluded that this novel approach allows a rapid, sensitive, economical, auto-mated and non-isotopic STR-PCR testing, thus provides a reliable alternative for assessment of the status of engraftment after allo-HSCT.
Adolescent ; Adult ; Child ; Child, Preschool ; Chromatography, High Pressure Liquid ; methods ; Female ; Graft Survival ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia ; genetics ; surgery ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Transplantation, Homologous

To study feasibility of HLA-DRB1 matching by using denatured high performance liquid chromatography (DHPLC), 20 pairs of DNA samples from donors and recipients of hematopoietic cell transplantation (HCT) for DRB1 matching and 2 pairs of samples from donors and recipient of HCT for DRB1 mismatching were studied by DHPLC and PCR-SSP. After being amplified and annealed slowly to produce heteroduplex, PCR products for exon 2 of DRB1 were detected by DHPLC to find matched or mismatched peaks in chromatogram. The results showed that DHPLC and PCR-SSP were consistant with matched or mismatched HLA-DRB1 typing. The results of DHPLC and PCR-SSP for matching were compared by using kappa test (kappa = 0.776, P = 0.00), which suggested DHPLC for HLA-DRB1 matching was in agreement with PCR-SSP. In conclusion, DHPLC for HLA-DRB1 matching is economic and convenient, moreover, will not be affected by unknown genes in HLA-DRB1 locus.
Base Sequence ; Blood Donors ; Chromatography, High Pressure Liquid ; methods ; Feasibility Studies ; HLA-DR Antigens ; genetics ; immunology ; HLA-DRB1 Chains ; Hematopoietic Stem Cell Transplantation ; Histocompatibility Testing ; methods ; Humans ; Molecular Sequence Data ; Polymorphism, Genetic ; genetics

OBJECTIVETo investigate the effects of ammonium perchlorate (AP) on thyroid functions and mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes of rats.

METHODSThirty SD male rats were randomly divided into six groups: control group, iodine-deficient group, low dose AP group (130 mg/kg), moderate dose AP group (260 mg/kg), high dose AP group (520 mg/kg) and high iodine-combined group. After the rats were exposed orally for 90 days, serum free-thyroxine (FT(4)), free-triiodothyronine (FT(3)) and thyroid stimulating hormone (TSH) were measured using radioimmunoassays. mRNA expression levels of thyroglobulin (Tg) and thyroperoxidase (TPO) genes were detected by real-time quantitative PCR.

RESULTSSerum FT(4) levels in moderate dose AP group and high dose AP group were [(9.540 ± 1.327) fmol/ml] and [(6.509 ± 1.949) fmol/ml] respectively, which were significantly lower than that [(13.505 ± 1.276) fmol /ml] in control group (P < 0.05 or P < 0.01). Serum TSH level in high dose AP group was [(1.227 ± 0.295) mIU/L], which was significantly higher than that [(0.545 ± 0.282) mIU/L] in control group (P < 0.05). The mRNA expression levels of thyroglobulin (Tg) gene in all groups exposed to AP were significantly lower than that in control group (P < 0.01). The mRNA expression level of thyroperoxidase (TPO) gene in high dose AP group was significantly higher than that in control group (P < 0.05).

CONCLUSIONAP can reduce the serum FT(3) and FT(4) levels of rats, increase the serum TSH level of rats and decrease obviously the mRNA expression levels of Tg and TPO genes. In addition, high iodine can reduce the toxic effects of AP on thyroid gland of rats to some extent.

Animals ; Iodide Peroxidase ; genetics ; metabolism ; Iodine ; administration & dosage ; Male ; Perchlorates ; toxicity ; Quaternary Ammonium Compounds ; toxicity ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Thyroglobulin ; genetics ; metabolism ; Thyroid Gland ; drug effects ; metabolism ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood

A device to produce low temperature plasma ( LTP) was designed and constructed to serve as the ion source of a high resolution mass spectrometry, and was applied to qualitatively analyze the steroid samples. In comparison with conventional electrospray ionization mass spectrometry, low temperature plasma mass spectrometry ( LTP-MS) had some advantages such as simple sample pretreatment and less interference. Mass spectrometry and tandem mass spectrometry were used to characterize the steroid samples in this research, and it was found that the structural stability of each steroid sample was presented in its mass spectrum, while in the tandem mass spectra there were more fragments of H2 O lost. And then the fragmentation process of typical steroid samples in collision induced dissociation ( CID ) was discussed based on theoretical calculation. In addition, by comparing tandem mass spectrometry and the fragmentation process, a pair of isomers of testosterone and dehydroepiandrosterone could be distinguished successfully.

Current studies have demonstrated that SLC38A1 proteins play a causal role in neoplastic cell transformation.The twofold aim of this study was to provide insight into whether a variance in the expression of SLC38A1 exists between human colorectal cancer and healthy human tissues and to determine how silencing or overexpressing the SLC38A1 gene could affect the proliferation,viability and migration of colorectal cancer cells.Immunohistochemical staining was used to analyze the expression of SLC38A1 in colorectal cancer tissues and adjacent normal mucosa in 77 patients who underwent surgical resection.The expression of SLC38A1 in colorectal cancer tissues and cell lines was detected using RT-PCR and Western blotting.Two colorectal cancer cell lines SW480 and HCT116 were used to examine whether silencing SLC38A1 with siRNA and overexpressing SLC38A1 with shRNA could affect cell viability and migration.As a result,the SLC38A1 protein was very low or undetectable in the normal colon mucosa.In contrast,strong staining of SLC38A1 protein was found in the cytoplasm in 79.2％ colorectal cancer samples.More pronounced SLC38A1 expression in colorectal cancer tissues was significantly associated with tumor node metastasis (TNM) stage.Inhibition of SLC38A1 reduced tumour growth and suppressed proliferation and migration of SW480 cells.In contrast,overexpression of SLC38A1 had the opposite effects on HCT116 cells.S LC38A1 is overexpressed in colorectal cancer,which suggests that it is associated with tumour progression.These results encourage the exploration of SLC38A1 as a target for intervention in colorectal cancer.

BACKGROUNDAtypical meningioma is one of the rare subtypes of meningioma, which is lacking of optimal consensus on treatment strategies. This study aimed to investigate the radical treatment strategies to improve the long-term outcome of recurrent atypical meningiomas.

METHODSThe prognostic factors including the age and gender of patients; the location, histology, recurrence pattern and mitotic cell rate of the tumors; and the resection extents, surgical strategies and adjuvant therapies of 15 cases of recurrent atypical meningiomas were analyzed retrospectively.

RESULTSThe age and gender of patients were not associated with tumor recurrence. However, high recurrence rates and poor prognosis for atypical meningiomas were associated with the high mitotic cell rate, failure to achieve Simpson grade I-II resection, and without the dura and bone flap replacement intraoperatively. Post-operative radiotherapy improved the outcomes of tumors in patients after the second surgery.

CONCLUSIONRadical treatment strategies such as dura and bone flap replacements and radiotherapy should be considered in patients diagnosed with atypical meningiomas.

Adult ; Aged ; Female ; Humans ; Male ; Meningeal Neoplasms ; radiotherapy ; surgery ; Meningioma ; radiotherapy ; surgery ; Middle Aged ; Neoplasm Recurrence, Local ; radiotherapy ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effect of lentiviral vector mediated herpes simplex virus-thymidine kinase/ganciclovir (HSV-TK/GCV) on graft- versus-host disease (GVHD) after allogeneic bone marrow transplantation (allo- BMT) in mice.

METHODSDonor splenic lymphocytes from C57BL/6 which were infected by lentiviral vectors carrying HSV-TK were transplanted into 60Co gamma ray irradiated recipient mice with donor bone marrow cells. GCV 25 mg x kg(-1) x d(-1) was administered in 3 groups on day 0, +7, +12 respectively after transplant for 7 days by intraperitoneal injection. Survival time, severity of GVHD, incidence of GVHD, T lymphocytes immune reconstruction and of allogeneic chimerism ratio were detected after allo-BMT.

RESULTSThe average survival times for GCV 0 day, +7 day and +12 day group were (30. 10 +/- 5.21) d, (36.40 +/- 5.28) d and (28.20 +/- 4.82) d respectively, being significantly longer than that in the control group [(15.10 +/- 0.43) d] (P < 0.05). The 50 d-survival rate for TK/GCV + 7 day group was 60%. While for 0 day and +12 day group was 40% and 30% respectively. The incidence of grade III approximately IV GVHD in the control group was 100%, and the dead mice in experimental groups showed pathological changes of II approximately III GVHD. Long-term alive recipient mice only developed grade I approximately II GVHD after allo-BMT. The number of CD4+ lymphocytes in experimental groups was higher than that in control group (P <0.05), but CD8+ lymphocytes was lower on day +5, +10, +15 day (P <0.05). Allogeneic chimerism rate of recipient mice on +30 d was 100%.

CONCLUSIONSHSV-TK/GCV induced by the lentiviral vectors has a definite effect in prevention of GVHD after allo-BMT. GCV administrated from 7 days post-transplantation showed the best effects.

Animals ; Bone Marrow Transplantation ; immunology ; Ganciclovir ; pharmacology ; Genetic Vectors ; Graft vs Host Disease ; prevention & control ; Lentivirus ; genetics ; Lymphocyte Transfusion ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Simplexvirus ; enzymology ; Thymidine Kinase ; genetics ; Transfection ; Transplantation, Homologous