Animals ; Cardiomegaly ; Constriction ; Heart ; Mice

Objective:To explore a rapid and cost-effective method for identification of Candida gla-brata through the comparison of two different methods , using molecular methods of sequencing as gold standard.Methods:From our clinic, 200 strains of suspected Candida glabrata were collected during the last 3 years and selected after incubation in CHROMagar Candida medium for 48 h.By comparing the results of the CHROMagar Candida medium, the identification of the rapid trehalose test for different kinds of strains were analyzed under incubation in the tubes for 3 h, 6 h, and 24 h at 37 ℃and 42 ℃, respectively .All the strains were identified to species level by methods of sequencing .The optimal time and temperature of the trehalose test for the identification of Candida glabrata were assessed .Two different methods, CHROMagar Candida medium and the rapid trehalose test , in identification of Candi-da glabrata were compared.Results:In all the 200 strains, Candida glabrata ferment trehalose with 3 h incubation under 42 ℃ were the optimal time and temperature for fermenting trehalose .The accuracy , sensitivity, and specificity of the rapid trehalose test were 99.00% (198/200), 98.66% (147/149) and 100.00%(51/51).The accuracy rate of CHROMagar Candida medium was 79.50%(159/200), the sensitivity and specificity were only 89.93%(134/149) and 49.02%(25/51), however, compared with the domestic current popular methods , the rapid trehalose test had better time efficiency ratio .Con-clusion:The evaluation results suggest that the rapid trehalose test has advantages in terms of operational convenience and low cost , and the results can be obtained in 3 h.Therefore, it has application value in clinical laboratory .

Objective To investigate the prevalence of low T3 syndrome in patients with acute myocardial infarction(AMI) and explore the effect of low T3 syndrome on outcome of AMI.MethodsThree hundred and thirty-eight patients with AMI admitted to cardiac care unit(CCU) underwent examinations of thyroid function and cardial ultrasound,and were further categorized according to thyroid hormone profile.The records of noninvasive bi-level positive airway pressure(BiPAP)ventilation utilization,length of hospital stay,mortality during hospitalization were evaluated,and the related factors were analysed.ResultsOne hundred and thirty-nine of the 338 patients(41.12%) with AMI complicated with low T3 syndrome.Free triiodothyronine(FT3) was the independent influential factor for length of hospital stay.Low FT3 was significantly correlated with noninvasive BiPAP ventilation utilization and mortality during hospitalization.The average time of follow-up was(21.4?8.1) months.It was revealed by multivariate Cox regression analysis that FT3 was the chief predictor for cumulative death(risk ratio,4.25;95% confidential interval,2.30-7.87),followed by age and left ventricular ejection fraction.ConclusionThe recognition of AMI complicated with low T3 syndrome plays an important role in predicting the disease severity and outcome.

Objective To build predictive model of secondary mild cognitive impairment (MCI) in patients with painful diabetic neuropathy (PDN), and analyze its apply. Methods The patients with PDN were consecutively selected from March 2013 to March 2016. The relevant clinical data were recorded, and the patients were followed up for 1 year. According to the results of follow-up, secondary MCI risk indicators were predicted, and the time window of adverse outcomes event was validated. Results A total of 82 PDN patients completed the study, and secondary MCI occurred in 16 cases. Sixty-six cases had not secondary MCI. The Cox regression model multivariate analysis results showed that the independent influencing factors of secondary MCI was course of PDN, brief pain inventory (BPI) score and neuron-specific enolase (NSE) in patients with PDN (HR = 1.238, 1.336 and 1.450; P<0.05). The secondary time window of the MCI in PDN patients with the course of PDN ≥3.367 years, BPI score≥4.704 scores and NSE ≥ 7.420 μg/L was shorter, in whom BPI score and NSE had a higher evaluation ability. Conclusions The courses of PDN, BPI score and NSE are independent influencing factors of secondary MCI in PDN patients, and the BPI score≥4.704 scores and NSE≥7.420μg/L have a higher evaluation ability.

Head and Neck Neoplasms ; diagnosis ; therapy ; Humans ; Neck ; Paraganglioma ; diagnosis ; therapy

OBJECTIVETo construct recombinant plasmid pEGFP-BMP7 and determine its expression in rat bone marrow mesenchymal stem cells (MSCs) in vitro.

METHODScDNA of target gene was obtained from neonatal rat kidney by RT-PCR. After sequencing the target gene, the cDNA was subcloned into a eukaryote plasmid pEGFP-N1 by directed cloning and then digested with two restrictive endonucleases to verify the correctiveness of the recombinant plasmid pEGFP-BMP7. Rat bone marrow MSCs were transiently transfected with the pEGFP-BMP7 and transfection efficiency of the Green Fluorescent Protein (GFP) was determined. RT-PCR and immunocytochemical analysis were also performed to detect the expression of BMP7 in rat MSCs.

RESULTS1 311 bp cDNA fragment was obtained by RT-PCR and sequence analysis showed it matched perfectly with that of rat BMP7 gene except a single nucleotide change at 756 bp from T to A. Digestion of the recombinant plasmid showed two 1.3 kb and 4.7 kb fragments and their size were same as those of BMP7 and pEGFP. This indicated that BMP7 cDNA was successfully subcloned into pEGFP. Transient transfection showed an efficiency of 33% at day 2 in rat MSCs. After transfection, transcription of BMP7 was detected in MSCs and expression of BMP7 protein was also verified.

CONCLUSIONRecombinant eukaryote plasmid pEGFP-BMP7 was successfully constructed and expressed in rat bone marrow MSCs. This procedure may provide a unique method for stimulation of callus formation in distraction osteogenesis and reconstruction of craniofacial bone defects.

Animals ; Bone Marrow Cells ; Bone Morphogenetic Protein 7 ; Genetic Vectors ; Green Fluorescent Proteins ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Plasmids ; Rats ; Transfection

OBJECTIVETo express and purify of the BC097361 recombinant protein, and to prepare the BC097361 specific rabbit polyclonal antibody.

METHODSBC097361 cDNA was ligated into the prokaryotic expressive vector pET-32a (+), and the resulting plasmid was transformed into E.coli BL21 (DE3). The protein expression was induced with IPTG and the protein was analyzed with SDS-PAGE and western blotting. The expressed product was purified using Ni+ affinity column chromatography.Then the purified pET-32a (+) -BC097361 fusion protein was used to immunize New Zealand rabbits to gain polyclonal antibody. The specificity and potency of polyclonal antibody were evaluated by Western blot and ELISA.

RESULTSThe BC097361 fusion protein was highly expressed.The protein was mainly in the inclusion body. ELISA indicated the titer of polyclonal antibody more than 1:320000. The high specificity was confirmed with Western blot.

CONCLUSIONSThe recombinant BC097361 fusion protein and the BC097361 specific polyclonal antibody will be valuable tools for the investigation on the biological function of BC097361.

Angiotensin II ; genetics ; Animals ; Antibodies ; immunology ; isolation & purification ; metabolism ; Antibody Specificity ; Blotting, Western ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Liver Cirrhosis ; genetics ; Male ; Plasmids ; genetics ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; immunology ; Reverse Transcriptase Polymerase Chain Reaction

To investigate the characteristics of the allele distribution of HLA-B*15 gene family in Chinese Han population and to study its influence on the selection of clinical transplantation donor, population of a 815 Han in north China from Shaanxi sub-registry of Chinese National Marrow Donor Project was randomly selected and out of them 206 HLA-B*15 positive samples according to the previous known low-resolution typing results were acquired. HLA-B*15 gene polymorphisms of above-mentioned samples and other 17 individuals were analyzed for the first time by polymerase chain reaction sequence-based typing (PCR-SBT) at high-resolution level. The structure differentiation of all HLA-B*15 alleles were analyzed by HLA three-dimensional structure modeling and software Swiss-PdbViewer. The results showed that the distribution of HLA-A, -B, -DRB1 gene of randomly selected 815 samples accorded with Hardy-Weinberg equilibrium and the gene frequency of HLA-B*15 was 0.1379. There were a total of 16 kinds of alleles of HLA-B*15 gene family to be obtained, which belonged to 7 kinds of serologic specificities. HLA-B*1501, B*1511, B*1502 and B*1518 were the major alleles with a frequency of 0.0485, 0.0215, 0.0178 and 0.0160 respectively, and the constituent ratio of their accumulated frequencies was 75.11%. The each frequency of the other 12 kinds of B*15 alleles was lower than 0.0100. Among the homozygote of 10 samples at low/medial-resolution level, there were only 4 samples to be pur sang homozygote of HLA-B*15xx, --at high-resolution level, and all the homozygote were constituted by respective dominating alleles. HLA three-dimensional structure modeling demonstrated that within the same specificity, gentle structure differentiation not only existed, such as B*1501, 1505, 1507, 1525, 1527, 1532 (each RMSD Asian Continental Ancestry Group ; genetics ; China ; ethnology ; HLA-B Antigens ; genetics ; HLA-B15 Antigen ; Humans ; Polymerase Chain Reaction ; methods ; Polymorphism, Genetic ; Tissue Donors

OBJECTIVETo evaluate autogenous vein grafts and inside-out vein grafts as conduits for the defects repair in the rabbit facial nerves.

METHODSThe 10 mm segments of buccal division of facial nerve were transected for 48 rabbits in this study. Then the gaps were immediately repaired by autogenous vein grafts or inside-out vein grafts in different groups. All the animals underwent the whisker movement test and electrophysiologic test during the following 16 weeks at different time points postoperatively. Subsequently, the histological examination was performed to observe the facial nerve regeneration morphologically.

RESULTSAt 8 weeks after operation, the facial nerve regeneration has significant difference between the experimental group and the control group in electrophysiologic test and histological observation. However, at the end of this study, 16 weeks after operation, there was no significant difference between inside-out vein grafts and standard vein grafts in enhancing peripheral nerve regeneration.

CONCLUSIONThis study suggest that both kinds of vein grafts play positive roles in facial nerve regeneration after being repaired immediately, but the autogenous inside-out vein grafts might accelerate and facilitate axonal regeneration as compared with control.

Animals ; Axons ; physiology ; Facial Nerve ; physiology ; surgery ; Facial Nerve Injuries ; surgery ; Male ; Nerve Regeneration ; physiology ; Rabbits ; Transplantation, Autologous ; methods ; Veins ; transplantation

OBJECTIVETo study the response of rat bone marrow mesenchymal stem cells (MSCs) to a single period of mechanical strain and expression patterns of transforming growth factor-beta (TGF-beta) and insulin-like growth factor-II (IGF-II) after mechanical stretch.

METHODSBone marrow MSCs were isolated from SD rats and cultured in vitro. A four-point bending apparatus were used to perform a single period of mechanical strain (2000 microepsilon, 40 min) on MSCs. Cellular proliferation and alkaline phosphatase (ALP) activity of MSCs were examined and gene expression patterns of TGF-beta and IGF-II were detected by SYBR green quantitative real-time RT-PCR.

RESULTSCell proliferation, ALP activity and expression of TGF-beta and IGF-II were all significantly up-regulated in stretched MSCs when compared with their controls. The mRNA levels of TGF-beta and IGF-II got top increase immediately after mechanical loading and increased about 51.44 and 8.92 folds, respectively, when compared with control cells. Expression of TGF-beta and IGF-II decreased with time and returned to control level at 12 h after mechanical stimulus, despite of a small increase at 6 h.

CONCLUSIONThe mechanical stretch can promote MSCs proliferation, up-regulate its ALP activity and induce a time-dependent expression increase of TGF-beta and IGF-II which in turn result in osteogenic differentiation of MSCs. Mechanical stimulus is a key stimulator for osteogenic differentiation of MSCs and vital for bone formation in distraction osteogenesis.

Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Proliferation ; Insulin-Like Growth Factor II ; Mesenchymal Stromal Cells ; Osteogenesis ; Osteogenesis, Distraction ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Somatomedins ; Transforming Growth Factor beta