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 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.

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.

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 explore the response of rat bone marrow mesenchymal stem cells (MSCs and calvarial osteoblasts to mechanical strain and the consequent changes of cytoskeleton F-actin.

METHODSBone marrow MSCs and calvarial osteoblasts were isolated from SD rats and cultured in vitro. Mechanical stretch was performed on passage 3 cells at 2 000 microepsilon for 0, 2, 6 and 12 hours using four-point bending system. The response of cells and the distribution of F-actin were observed using fluorescent staining under laser scanning confocal microscope and the morphological parameters were quantified using image analysis software Laserpix.

RESULTSUnder mechanical stretch, the fluorescent staining decreased obviously at both MSCs and osteoblasts, and F-actin filaments were rearranged and became tenuous, thinner, and abnormally distributed. The outline of nucleus became unclear and apoptotic changes were observed at some of both cells. Cellular size decreased more significantly in MSCs than in osteoblasts. Quantity analysis showed that total area of cells, total fluorescent density and green fluorescent density (F-actin) were all significantly decreased in MSCs (P < 0.05 or P < 0.01), and total fluorescent density, green fluorescent density and red fluorescent density (nuclei) did also in osteoblasts (P < 0.05 or P < 0.01).

CONCLUSIONMechanical stretch caused extensive response on both MSCs and osteoblasts which led to the rearrangement of F-actin filament and apoptosis in some of these cells. MSCs were more sensitive to mechanical strain than osteoblasts.

Actin Cytoskeleton ; metabolism ; Actins ; metabolism ; Animals ; Bone Marrow Cells ; Cells, Cultured ; Cytoskeleton ; Mesenchymal Stromal Cells ; Microtubules ; Osteoblasts ; Rats ; Stress, Mechanical

OBJECTIVETo study the relationship of protein intake and energy supply with the physical growth in very/extremely low birth weight infant at their early life.

METHODRetrospective survey was performed in Neonatal Intensive Care Unit (NICU) in Peking University First Hospital. Inclusion criteria were preterm infant, birth weight < 1500 g, hospitalization for longer than 2 weeks, discharge with body weight greater than 1800 g. The infants were divided into two groups according to gestational age (GA). GA < 32 weeks and ≥ 32 weeks. Physical growth and its relation with the protein intake and energy supply were analyzed. The predictive value of serum blood urea nitrogen (BUN) on protein intake was studied.

RESULTNinety-three very/extremely low birth weight infants were involved, 69 in GA < 32 weeks group and 24 in GA ≥ 32 weeks group.Compared with GA ≥ 32 group, GA < 32 weeks preterm infants had more weight loss, (9.2 ± 4.4)% vs. (5.0 ± 3.1)%, P = 0.000; slower birth weight recovery (10.6 ± 3.8) d vs. (7.1 ± 2.6) d, P = 0.000; poorer weight gain at 1, 4, 5 weeks of life, (-4.5 ± 9.3) g/ (kg·d) vs. (3.4 ± 6.9) g/ (kg·d), P = 0.000 , (13.5 ± 7.3) g/ (kg·d) vs. (19.2 ± 4.9) g/ (kg·d), P = 0.001, (14.6 ± 5.6) g/ (kg·d) vs. (18.2 ± 4.5) g/ (kg·d), P = 0.031; less energy supply at 1 to 5 weeks (P value was 0.000,0.000,0.025,0.001,0.008 respectively) and less protein intake at 1, 4, 5 weeks of life (P value was 0.009,0.006,0.032). Extrauterine growth retardation (EUGR) was still predominant in our subjects, 47.8% in GA < 32 weeks group, and 95.8% in GA ≥ 32 weeks group, P = 0.000. The incidence increased greater in GA < 32 weeks infants, 43.5% vs. 20.8%, P = 0.000.The duration of weight loss and mechanical ventilation correlated negatively with weight gain rate, respectively β = -0.591, P = 0.000 and β = -0.281, P = 0.005; the average energy supply and time taken to reach full enteral feeding were factors improving weight gain, respectively β = 0.202, P = 0.021 and β = 0.354, P = 0.000. After birth, serum BUN declined gradually. Positive relation showed between average protein intake at 3(rd) week and BUN level at the end of 3 weeks, r = 0.420, P = 0.000. Serum BUN 1.44, 1.49 mmol/L at the end of 3(rd) and 4(th) week were cut-off predictors for protein intake less than 3 g/(kg·d) at related period, sensitivity and specificity were 65.3%, 83.3% and 60%, 80% respectively.

CONCLUSIONNo enough protein intake and energy supply, poor weight gain are critical problems in the management of very/extremely low birth weight infants. Prevention from NEC, appropriate parenteral/enteral nutrition transforming will benefit their physical growth. Low serum BUN after 3 weeks of life is a valuable predictor of low protein intake.

Blood Urea Nitrogen ; Dietary Proteins ; administration & dosage ; Energy Intake ; Enteral Nutrition ; Humans ; Infant ; Infant Nutritional Physiological Phenomena ; Infant, Low Birth Weight ; growth & development ; Infant, Newborn ; Infant, Premature ; Infant, Premature, Diseases ; epidemiology ; etiology ; Infant, Very Low Birth Weight ; growth & development ; Intensive Care Units, Neonatal ; Nutritional Status ; Parenteral Nutrition ; Retrospective Studies ; Weight Gain

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

OBJECTIVETo study the effects of glial cell derived neurotrophic factor (GDNF) on regeneration of facial nerve defects by autogenous facial vein conduit.

METHODSThirty-six rabbits were used in this study and 10 mm-length facial nerve defects were made on both sides of all animals. The nerve gaps were bridged using autogenous posterior facial vein graft of the same side. The animals received injection of either saline (group A, n=16) or GDNF (group B, n=16) into the veins. Nerve function was evaluated by evoking nerve action potential immediately after operation and 4, 8 and 16 weeks after operation. Regenerated nerve samples were harvested at 4, 8, and 16 weeks after operation and processed for histology and transmitting electron microscopic examination (TEM).

RESULTSAction potential did not exist immediately after operation but it was evoked at 4, 8, and 16 weeks in both groups. At 4 and 8 weeks after operation, the amplitude and width of action potential were significantly higher in group B than group A (P < 0.01), except wave width at 4 weeks, which showed no significant differences, while the latency period was significantly shorter in group B than that in group A (P < 0.01). At 16 weeks, action potential was similar between two groups, except wave amptitude, which was higher in group B than group A (P < 0.01). Morphologic and TEM examinations showed more matured myelinated nerve fibers and active Schwann's cells in group B when compared group A during the whole regeneration process.

CONCLUSIONGDNF can promote nerve regeneraat early stage during reconstruction of facial nerve defects by autogenous facial vein conduit and combination of GDNF and autogenous vein graft provides a valuable method for clinical reconstruction of facial nerve defects.

Animals ; Facial Nerve ; Nerve Growth Factors ; Nerve Regeneration ; Neuroglia ; Rabbits ; Regeneration