Clinical hematology and hematologic examination is a strong practical curriculum, and experimental class is very important in the aspects of teaching.In order to improve the quality of teaching and train compound laboratory talents with high quality,several effective reform mea-sures were carried out based on the years of teaching practice by the department:improving the methods of experimental teaching;using modern means of teaching;emphasizing on the analysis of experimental results,formulating the rigorous experimental evaluation system and starting the second class etc.

AlM:To investigate the protective effect of astaxanthin (AST) on human retinal pigment epithelial (RPE) cells against oxidative damage induced by hydrogen peroxide (H2O2).METHODS:Human RPE cells were subcultured, cell activity was detected by MTT, rate of apoptosis was detected by flow cytometry and cell ultrastructure changes were observed under transmission electron microscope. RESULTS: MTT results showed that cell activity elevated to ( 53. 66%± 3. 25% and 70. 43%± 2. 38% after 10-8 mol/L and 10-4 mol/L AST treated. The difference had statistically significant (P<0. 05) compared with oxidative injury group (38. 76%± 3. 74%). Flow cytometry results showed that the apoptosis rate of RPE cells decreased to 30. 23%± 1. 91% and 12. 58%± 2. 12% in AST pretreated group, the difference was significant (P<0. 05) compared with oxidative injury group ( 42. 50%± 1. 94%); Electron microscopy showed that the morphology of cells gradually improved accompanied with the concentration of AST elevated.CONCLUSlON:AST may inhibit hydrogen peroxide-induced apoptosis of RPE cells, it can provide reliable evidence for pursue effective medicine to prevent and treat retina injury.

Objective To investigate the neural proliferation , differentiation and apoptosis of the developing spinal cord of the mouse and to discuss the mechanism of spinal cord ’ s development .Methods 5-Bromodeoxyuridine ( BrdU) assay was used to mark the proliferative neural stem cells , and the immunofluorescent stainings ( DCX, NeuN and Caspase8) were carried out to visualize the newborn neurons , mature cells and apoptotic cells in the spinal cord with 173 mice arrange from E18 to P90.Results BrdU positive neural stem cells appeared evenly in the spinal cord at early days . With age increasing , the neural stem cells differentiated into neuroglial cells and neurons .The newborn neurons in the subventricular zone migrated toward the intermediate zone ( putative gray matter ) and differentiated into mature neurons gradually .With neurons ’ concentrating towards the center , the gray matter formed an “H” shape .In the meantime , with neural differentiation , some apoptotic neurons appeared among the newborn neurons and mature neurons . Double immunostaining showed that most apoptotic neurons were newborn neurons , suggesting the neuroapoptosis more likely occurred in newborn neurons .The statistical data showed that the number of DCX , NeuN and Caspase-8 positive cells reduced with age increasing , suggesting neural differentiation and neuroapoptosis decreased during spinal cord ’ s development .Conclusion Neural proliferation , neural differentiation and neuroapoptosis occur in developing spinal cord . They work together to regulate the formation and development of the spinal cord .

Objective HupA is one of the potential drugs which can be used to treat Alzheimer's disease(AD).The aim of this study was to explore the pharmacokinetics and biodistribution of HupA in vivo by using 11C-HupA.Methods A total of 25 SD rats were studied.They were divided into 5 groups (5 rats in each group).All had intravenous injection of 22 MBq(in0.2 ml)11C-HupA through tail vein.Dynamic im-aging Was acquired from 5 to 90 minutes after injection.Venous blood and organ activities were collected at 5,15,30,60.and 90 minutes after injection.Percentage activity of injected dose per gram of tissue(%ID/g)was calculated to characterize the biodistribution of tracer in different brain regions: frontal,apical, temporal,occipital,cerebellum,hippocampus,striatum,thalamencephalon, and brain stem, Variance analysis using SPSS 11.5 software was performed and compared among the study groups.Results 11C-HupA was character-istic for its quick clearance from blood,with half time T1/2 of (14.61±1.77) min,and clearance rate (CL)macokinetics of 11C-HupA in rats corresponded to a one-compartment model.with an activity curve(area 11C-HupA distribution in different brain regions,being greater in cerebral cortex,hippocampus,hypothala-mus and brain stem. Conclusions Pharmacokinetic study of 11C-HupA in brain was fast.convenient and showed high specificity and sensitivity.Its ability to quantitatively evaluate brain function and its character-istic distribution in mice provided some evidence for monitoring therapy in AD patients.

Objective To investigate the onset of left ventricular remodeling (LVRM) after acute myocardium infarction (AMI) and its changes within 6 hours in dogs on echocardiography. Methods AMI was induced in 14 dogs by ligating the left anterior descending arteries. Eight myocardium infarcted models were successful and were sacrified for pathological study. The indices of LVRM: wall infarction thickness (WIT), the wall motion score index (WMSI), left ventricular end-diastolic volume (LVEDV), left ventricular end-systolic volume (LVESV) and left ventricular ejection fraction (LVEF) were evaluated before and 1 h, 2 h, 3 h, 4 h, 5 h and 6 h after operation. Results Compared with the pre-operation, WIT and LVEF were decreased (P<0.01), LVESV and WMSI were increased (P<0.01), and LVEDV was increased (P<0.05 or P<0.01) at every time point after operation. WIT had no significant difference at 1 h, 2 h, 3 h, 4 h, 5h and 6h after operation (P＞0.05). LVEDV, LVESV were higher (P<0.05) and LVEF was lower (P<0.05 or P<0.01) at 4 h, 5 h, and 6 h than at 1 h, and 2 h after operation. WMSI was higher at 3 h, 4 h, 5 h, and 6 h than at 1h (P<0.05). Conclusions In our experiment, LVRM occurred at 1 h after AMI in dogs. Thus echocardiography may evaluate early LVRM.

Adult ; Bone Marrow Transplantation ; Fractures, Ununited ; therapy ; Humans ; Male ; Transplantation, Autologous

To systematically evaluate the efficacy and safety of tripterygium glycosides (TG) combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV. The computer retrievals were made in Cochrane Libarary, PubMed, Embase, SCI, Sinnomed, CNKI, Chinainfo and VIP, and hand retrievals were conducted for meeting and academic papers (updated to December 30, 2014), in order to collect randomized controlled trials and quasi-randomized control trials for TG combined with ACEI/ARB preparation in treating diabetic nephropathy stage IV and set the literature inclusion and elimination standards. Eligible literatures were included and evaluated according to standards in Cochrane Handbook. RevMan 5.3 and Stata 12.0 were used for a Meta-analysis. A total of 13 randomized controlled trials and quasi-randomized control trials involving 1119 patients with diabetic nephropathy were included. The Meta analysis result showed that compared with the control group, the combination group showed better effects in reducing the 24-hour urinary protein [MD = -0.84, 95% CI (-1.02, -0.66)], raising albumin [SMD = 0.98, 95% CI (0.81, 1.16)], the total efficiency [OR = 4.23, 95% CI (2.77, 6.46)] and the significant efficiency [OR = 5.35, 95% CI (2.70, 10.60)], with no statistical difference in Serum Creatinine between Both groups [MD = -0.82, 95% CI (-4.30, 2.66), P = 0.64]. However, the risk of adverse reactions increased by 7% [RD = 0.07, 95% CI (0.03, 0.12)]. The Egger's test showed no publication bias. Tripterygium Glycosides combined with ACEI/ARB in treating diabetic nephropathy stage IV is supper than the single administration of ACEI/ARB, with a good prospect in clinical application. Nevertheless, due to the small-size and low-quality samples in this study, more high-quality and large sample-size randomized controlled trials shall be conducted to verify the findings.
Angiotensin Receptor Antagonists ; administration & dosage ; Angiotensin-Converting Enzyme Inhibitors ; administration & dosage ; Diabetic Nephropathies ; drug therapy ; Drug Therapy, Combination ; Glycosides ; administration & dosage ; Humans ; Tripterygium ; chemistry

There is growing interest in biodiesel and this results in the accumulation of glycerol. The exploitation and application of glycerol has attracted more and more attention. In the current study, glycerol was biotransformed to produce 3-hydroxypropionaldehyde by genetic engineering bacteria. It is known that 3-hydroxypopionaldehyde has been widely used as an important intermediate for chemicals, effective antimicrobial agent, and fix agent for tissues. A pair of primers was designed on the basis of the sequence of both NH2-terminus and the amino acid sequence of glycerol dehydratase reported by NCBI, and a fragment about 1.6 kb was obtained by PCR amplification using the total genome DNA of Lactobacillus reuteri as template, then the fragment was cloned to the pMD18-T vector and sequenced. Two specific primers were designed according to the obtained sequence, and a fragment with length of 1674 bp was amplified using PCR with these two specific primers. Consequently, the resulting products were digested with EcoR I and Hind III and ligated using T4 DNA ligase to the pET28b vector digested with the same enzymes. The recombinant plasmid, named pET28b-dhaB, was transformed into E. coli BL21. The positive clones were induced with IPTG and the expression products were further analyzed by SDS-PAGE, indicating that protein with a molecule weight of around 65 kD was obtained. Furthermore, the glycerol dehydratase activity was evaluated and compared with the wild type strain as well.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glyceraldehyde ; analogs & derivatives ; chemistry ; metabolism ; Hydro-Lyases ; biosynthesis ; genetics ; Lactobacillus reuteri ; enzymology ; genetics ; Propane ; chemistry ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Objective To clone ,construction ,express and purify Tp47 of Treponema pallidum (Tp) ,and assess the immunoreac‐tivity by Western‐Blot .Methods Tp47 was amplified by polymerase chain reaction ,and then cloned to the vector pGEX‐6P‐1 .The correct sequence of the recombinant plasmids pGEX‐6P1‐Tp47 was transformed into Escherichia coli BL21 (DE3) and induced .The expression product was analyzed by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western‐Blot .The expression protein was purified .Serum of different clinical stages of syphilis was used as the antibody to detect the immunoreactivity of the protein by Western‐Blot .Results A fusion protein with molecular weight about 71 × 103 was attained .Western‐Blot proved that the recombinant protein can react with Tp IgG positive sera .And the specificities and sensitivities of the diagnostic reagent detected by sera were 100% .Conclusion The recombinant protein Tp47 was expressed and purified with good antigen activity ,which could provide the basis of theory and practice for the development of early diagnostic kit applying to detect Tp infection .

Objective To study the effect of single chain variable fragment (ScFv) antibody to EC3-4 fragment of desmoglein (Dsg) 3 in a mouse model of pemphigus vulgaris.Methods The ScFv an- tibody to EC3-4 fragment of Dsg-3 was injected subcutaneously into neonatal BALB/c mice at different time points;the mice were then evaluated clinically,histopathologically and by direct immunoflorescence exami- nation for the development of lesions.Results When injected alone,the ScFv antibody did not induce the appearance of key clinical features of pemphigus vulgaris.The antibody also did not prevent the develop- ment of pemphigus vulgaris features induced by sera of patients with pemphigus vulgaris,regardless of the time point of injection of ScFv antibody.Conclusion The ScFv antibody to EC3-4 fragment of Dsg-3 lacks pathogenicity in neonatal BALB/c mice,and also could not inhibit the development of lesions induced by sera from patients with pemphigus vulgaris.