Objective To establish an ELISA method for detection of the concentration of TAP in human urine samples.Methods TAP-BSA was used as coating antigen. The TAP was a competitor to TAP-BSA. They reacted to the limited amount of monoclonal antibody against TAP.Results The optimal concentration of the coating antigen was 250ng/ml.The dilutions of monolclonal antibody against TAP and sheep anti-mouse IgG were 25?g/ml and 1∶4 000 respectively. The optimal range was from 0.69 to 1 000ng/ml.The assay provided a sensitivity of 0.69ng/ml.The coefficients of variation of intra-assay and inter-assay were 9.10% and 10.33% respectively. The average recovery rate of TAP was 97.70%.Conclusion The indirect competitive enzyme-linked immunosorbent assay is established.

Objective To compare the body fluid count results detected by Sysmex XE-5000 automatic blood cell analyzer and manual method .Methods A total of 300 cases of body fluid specimens (including cerebrospinal fluid and fluid of serous cavity ) were analyzed .RBC ,WBC counting and classification were respectively detected by XE-5000 and manual method of improvement Neubauer counting plate .Results The fresh specimens without contain a large number of cell clusters ,which RBC counts(RBC-BF)(100-10 000)× 106/L ,and WBC counts(WBC-BF) (9-50)× 106/L ,showed there were a linear relationship between the in-strument method(r=0 .998 5 ,0 .986 3) .In the range ,there was no significant difference between XE-5000 and manual method(t=9 .96 ,P>0 .05) .Also in this range the results of instrument correlated with those of manual method(r= 0 .989 3 ,0 .971 7 , 0 .924 9) .For those specimens which contain a large number of cell clusters ,RBC-BF and WBC-BF were a badly linear relationship between the instrument method(r=0 .564 8 ,0 .456 1) .Conclusion Body fluid specimens which are fresh and do not contain a large number of cell clusters ,in the range of RBC-BF (100 -10 000) × 106/L ,WBC-BF (9 -50) × 106/L ,Sysmex XE-5000 automatic blood cell analyzer could ensure the results have good accuracy .

Animals ; Atherosclerosis ; Humans ; Serine Proteinase Inhibitors ; Serpins

Acupuncture Points ; Acupuncture Therapy ; Adolescent ; Adult ; Ankle Injuries ; therapy ; Female ; Humans ; Male ; Sprains and Strains ; therapy ; Treatment Outcome ; Young Adult

Humans ; Interferons ; administration & dosage ; therapeutic use ; Lymphoma, Mantle-Cell ; drug therapy ; Male ; Middle Aged ; Thalidomide ; administration & dosage ; therapeutic use

Objective To observe the intervention effect of electro-acupuncture(EA) on mammalian target of rapamycin(mTOR) expression in hypoxic-ischemic encephalopathy(HIE) newborn rats. Methods Seven-day-old SD rats were randomly assigned into 3 groups, namely sham-surgery group, HIE model group, and EA group. All of the groups were subdivided into 4 subgroups in terms of 4 time periods of 1, 3, 7 and 21 day(s). Hematoxylin-eosin staining was used for the observation of histological changes in the affected cerebral region. Expression levels of cerebral mTOR were detected with Western blotting method. Results The results of HE staining showed that EA group at experimental day 21 had clear organizational structure, ordered arrangement of neurons, relieved cellular swelling, well cell integrity with clear nucleolus and proliferative glial cells. Compared with the sham-surgery group, mTOR level of the model group was dramatically increased on the 1st, 3rd, 7th and 21st day(P<0.05), and the mTOR expression level of EA group was significantly higher than that in the model group at the above time points (P < 0.05). Conclusion EA can promote mTOR expression in brain tissue and can protect the brain of newborn rats from HIE.

Objective To study the clinical features of follicular occlusion triad, and whether it is a hereditary disease. Methods Based on the clinical examination of a case who developed squamous cell carcinoma secondary to follicular occlusion triad, the pedigree of the patient was surveyed and analyzed. Results There were a total of thirteen patients in this pedigree, the age of onset was about 20 years old. The clinical features and laboratory examination of the proband was consistent with follicular occlusion triad. Conclusions Hereditary factor is important in the pathogenesis in follicular occlusion triad,and the disease maybe an autosomal dominant inherited disease.

BACKGROUND:Brain-derived neurotrophic factor (BDNF) has widespread effects on dopaminergic neurons, cholinergic neurons and other neurons, which can promote more differentiation of stem cells into neuron-like cells. Hypoxia inducible factor-1α(HIF1α) can improve tissue and cellviability in the ischemic environment and maintain the local microenvironment of hemangioblasts, which has a more far-ranging physiological role than genes. OBJECTIVE:By constructing double gene recombinant adenovirus vector with BDNF and three points mutant HIF1αto complete the transfection of adenovirus vector into rats mesenchymal stem cells and then study the promoting nerve regeneration and angiogenesis effect of these two genes to spinal cord injury in vitro in constant oxygen conditions. METHODS:(1)We finished the site-directed mutagenesis of 402, 564 and 803 amino acids in human HIF1αgene CDS region by PCR method and we finished recombination of mutation posterior HIF1αgene and BDNF into adenovirus pAdEasy-1 system. Packaging viral and titration determination were also finished. (2)Four kinds of virus fluids and blank group were selected for subsequent experiments. We observed transfection efficiency after transfection of virus into bone marrow mesenchymal stem cells and detected the expressions of BDNF and HIF1αmRNA and protein in transfection cells. (3) The protein expression of vascular endothelial growth factor, downstream formation vascular gene of HIF1α, in cells in al groups was detected by western blot assay. RESULTS AND CONCLUSION:(1)The site-directed mutagenesis of 402, 564 and 803 amino acids to alanine in coding sequence region was successful. The construction of four kinds of adenoviral recombinants was successful and identification of packaging was completed. (2)The level of BDNF gene mRNA and protein expression in experimental and positive control 1 groups was significantly higher than that in the other groups (P<0.05). The level of HIF1αmRNA and protein expression in experimental and positive control 2 groups was significantly higher than that of the other groups (P<0.05). The level of vascular endothelial growth factor protein was also increased in the experimental and positive control 2 groups, which was significantly different from other groups (P<0.05). These findings indicate that the BDNF and HIF1αproteins are largely and efficiently expressed in constant oxygen conditions after single vector double gene adenovirus system transfection into mesenchymal stem cells and high-efficiency expression of vascular endothelial growth factor protein is promoted so that this is a new direction for treatment of spinal cord injury by gene therapy combined with celltransplantation.

BACKGROUND:Whether transplanted bone marrow mesenchymal stem cells under hypoxic conditions can survive is crucial for the successful celltransplantation. Therefore, studies on the growth of bone marrow mesenchymal stem cells under hypoxic conditions in vitro can provide experimental evidence for in vivo celltransplantation. OBJECTIVE:To observe the expression of vascular endothelial growth factor in bone marrow mesenchymal stem cells under hypoxia. METHODS:Rat bone marrow mesenchymal stem cells were obtained and cultured, and observed under light microscopy. Passage 3 cells were cultured under normoxia (21%O2) and hypoxia (3%O2 hours. Then cellcounting kit-8 assay and flow cytometry were employed to detect cellproliferation in the two groups. Western blot assay was adopted to detect the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the two groups. RESULTS AND CONCLUSION:(1)Rat bone marrow mesenchymal stem cells were obtained and cultured successful y, which were fusiform cells and had uniform shape under the light microscope. (2)The results of cellcounting kit-8 assay showed that the number of cells in the hypoxic group was higher than that in the normoxic group at each time point, and cellviability increased significantly at hours 36 and 48 (P<0.05). (3)The results of flow cytometry demonstrated that the proportion of cells in S phase and cellproliferation index in the hypoxic group were significantly increased, compared with the normoxic group (P<0.05). (4)Western blot results showed ), respectively, for 72 that there was a smal amount of the expression of hypoxia-inducible factor-1αand vascular endothelial growth factor in the normoxic group, but the expression of these two proteins in the hypoxic group was increased in a time-dependent manner (P<0.05). These findings suggest that hypoxia can induce proliferation of rat bone marrow mesenchymal stem cells cultured in vitro, and also raise hypoxia-inducible factor-1αand vascular endothelial growth factor expression in a time-dependent manner.

Adolescent ; Fetus ; parasitology ; pathology ; Humans ; Male ; Teratoma ; parasitology ; pathology