Aiming at some problems existing in the cultivation of advanced studying doctors,we think that strict selection should be made to guide advanced studying doctors to have correct understanding of their specialty and systemic training to improve the training quality.

Objective To investigate the brain protection of adenosine preconditioning in infants with tetralngy of Fallot (TOF) undergoing open heart surgery. Methods 120 patients with TOF undergoing open heart surgery were randomly divided into two groups: adenosine group and control group. Total dosage of 1.5 mg/kg adenosine was intravenously dripped in the adenosine group before the cardiopulmonary bypass. Serum S-100β and neurone specific enolase (NSE) were measured before the cardipulminary bypass, 30 min and 12 hours after the aorta off clamping. Results At 30 min and 12 hour after opening the aorta, the S-100β[(0. 346±0. 162)μg/L vs. (0.717±0.260) μg/L and (0.541±0.272) μg/L vs. (1.140±0.567) μg/L,P <0.001 ]and NSE[(10.43±5.54) μg/L vs. (15.75±7.96)μg/Land (15.54±7. 98)μg/L vs. (23.67±8.00)μg/L,P<0.001]were both lower in adenosine group than that in control group. Conclusion Adenosine preconditioning can reduce the S-100β and NES in infants with TOF undergoing open heart surgery and has brain protection.

Objective The purpose of this study was to discuss the effect of visible light therapeutic equipment accompanied with electrolyzed-oxiding water (EOW) on pressure ulcer above the second phase. Methods We divided 44 patients with pressure ulcer above the second phase into the test group (23 cases, 28 spots) and the control group (21 cases, 25 spots). The test group received local decompression, debridement with EOW, exposure to visible light for 20 minutes, 10 times per day. The control group adopted local decompression, debridement with normal saline and local application of burn cream, 3 times per day. Results The effective rate of the test group was apparently higher than that of the control group (P<0.05). The healing time was also shorter than that of the control group (P<0.01). Conclusion Treatment of pressure ulcer above the second phase with visible light therapeutic equipment accompanied with electrolyzed-oxiding water proved to be an effective treatment method.

Objective To explore the pancreatic kininogenase enteric-coated tablets in the treatment of diabetic retinopathy (DR) patients and its effect on the optic disc and macula retinal hemodynamics. Methods 86 cases (140 eyes) of DR patients were randomly divided into pancreatic kininogenase group and normal group with 43 cases in each group, two groups were treated with basic therapy, pancreatic kininogenase group were combined with pancreatic kininogenase treatment. The best corrected visual acuity and the clinical effect of optic disc and macula retinal hemodynamic changes were compared between two groups. Results After treatment, the best corrected visual acuity (BCVA) in pancreatic kininogenase group was greater than the normal group (P<0.05), the retinal neovascularization and fluorescein leakage area in pancreatic kininogenase group was less than the normal group (P<0.05). The disc vascular and macular retinal blood flow volume (VOL), blood flow velocity (FLW)values in pancreatic kininogenase group were larger than those of normal group, and the clinical curative effect of pancreatic kininogenase group was better than that of normal group (P<0.05). Conclusion Pancreatic kininogenase enteric-coated tablets in the treatment of DR patients can improve the optic disc and macular retinal hemodynamic parameters, improve visual acuity and reduce the retinal neovascularization and fluorescein leakage area, so as to improve the clinical treatment effect.

A century ago the Flexner Report facilitated the process of modern medical education reform.Half a century ago,Problem-based learning initiated the student-centered medical education model.At present training medical students' competency will usher in the third reform in medical education.Over 100 years' development and in the global context of current medical education reform,China's medical education has become an important part of the world medical education.Harbin Medical University conducted an all-around reform through international cooperation and received the accreditation referenced according to global standards in medical education,which is an epitome of the medical education reform in China.

OBJECTIVE To construct a lentiviral vector of RNA interference (RNAi) of MMP-9 gene and observe its inhibitive role on the invasion of laryngeal cancer cells.METHODS The effective sequence of siRNA targeting MMP-9 gene was confirmed.Both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pLVTHM vector,which contained H1 promoter and green fluorescent protein (GFP).The resulting lentiviral vector containing MMP-9 shRNA was called LV2 shMMP-9,and it was confirmed by PCR and sequencing.After that,MMP-9 shRNA was transfected into Hep-2 cells and western blot was used to test the expression of MMP-9.At last,Boyden Chamber was used to observe the invasion of the Hep-2 cells. RESULTS PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of MMP-9 (LV2shMMP-9) producing MMP-9 shRNA was constructed successfully. The titer of concentrated virus was 8?1010TU /L. Western blot showed that the expression of MMP-9 was negative in the MMP-9 siRNA transfected Hep-2 cells. And Boyden Chamber showed the invasive capability of Hep-2 cells transfected MMP-9 siRNA were obviously decreased.CONCLUSION The lentivirus RNAi vector of MMP-9 was constructed successfully,and MMP-9 silence can inhibit invasion of laryngeal cancer in vitro.

Objective To establish a novel multiplex amplification system which comprises 24 Y-STR loci.Methods Total 24 Y-STR gene loci, concluding DYS531,DYS630,DYS622,DYS552,DYS510, DYS449, DYS459a/b, DYS446, DYS443, DYS635, DYS587, DYS527a/b, DYS460, Y-GATA-A10, DYS520, DYS557,DYS522,DYS481,DYS570,DYS385a/b,DYS444, were chosen for establishing the fluorescence multiplex amplification system. The specificity, identity, sensitivity, balance of the amplification, anti-in-terference and accuracy of the system were detected and the gene diversity was investigated in the popu-lation of Guangdong.Results No band was found in nonhuman and female samples that were tested by the established multiplex amplification system. The same genotyping results were obtained from different tissues of the same person. Complete profiles could be obtained from more than 0.1 ng of the standard sample 9948. The loss of alleles was found when the common inhibitors such as hemoglobin and calci-um ion were added 120-200μmol/L and 1.5-2.0 mmol/L respectively to the system which with a strong anti-interference to the indigo, humic acid and EDTA. The typing of 24 Y-STR system could give the reliable results when 146 unrelated male individuals were detected and compared with the Yfiler system parallelly. The haplotype diversity(HD)of the population in Guangdong reached 0.99972 that was better than the result retained from Yfiler system, which the HD was 0.99858.Conclusion The fluorescence amplification system with 24 Y-STR loci established in present study has a wildly application prospect and can be used for cases inspection, paternity tests and Y-STR database construction.

Objective To investigate the effects of low density lipoprotein receptor (LDLr) pathway on podocyte injury in diabetic nephropathy (DN) under inflammatory stress.Methods Male db/db mice and db/m mice were randomly divided into four groups (8 mice in each group):db/m group (control),casein injected db/m group (db/m + casein),db/db group (db/db),and casein injected db/db group (db/db + casein).An inflamed model of DN was established according to our previous study.24-hour urinary protein was measured every week.The plasma lipid profile was detected by clinical biochemistry assay.Podocyte changes were evaluated by electron microscope and immunofluorescent staining.Lipid accumulation in the kidney was evaluated by oil red O staining and intracellular cholesterol quantitative assay.The protein expression of Wilm's tumor-1 (WT-1),nephrin,α-smooth muscle actin (t-SMA),and molecules correlated with LDLr pathway were examined by immunohistochemical staining or Western blotting.The colocalized protein expression of LDLr with WT-1 was examined by immunofluorescent staining and laser confocal microscopy.Results There were no differences in plasma levels of LDL and HDL among four groups.Compared with db/db group,the db/db+ casein group showed markedly increased 24-hour urinary protein,more significant podocyte foot process effacement and podocyte damage,increased lipid droplet accumulation in kidneys,increased protein expressions of LDLr,SCAP and SREBP-2 in kidneys (all P ＜ 0.05).Interestingly,increased LDLr protein expression in kidneys of db/db mice was negatively correlated with decreased nephrin protein expression (r =-0.855,P ＜ 0.01) and positively correlated with increased α-SMA protein expression (r=0.768,P ＜ 0.01).Conclusions The disruption of LDLr pathway induced by inflammation contributes to podocyte injuries in diabetic nephropathy.

Objective To investigate the effects of different doses Alpha-Linolenic acid ( ALA ) on the expressions of fatty acid synthesis-related genes in HepG2 cells.Methods HepG2 cells were divided into two groups, normal control group (NC) and high fat group (HF) in which cells were firstly cultured for 36h in the medium contained 0.5mmol/L stearic acid.Real-time quantitative PCR was taken to detect mRNA expression of genes SREBP1C, FAS and ACC which are related to fatty synthesis.While there are significant differences in fatty synthesis, 10%, 20%, 50%, 70% and 100%ALA took the place of stearic acid, 36h later, real-time quantitative PCR and Western blotting were taken to detect mRNA and protein expression of genes related to fatty synthesis.Results SREBP1C mRNA expression of ALA substitution groups were significantly lower than the high-fat group ( P <0.001) .The FAS of 0.5 mmol/L ALA group and 0.35 mmol/L ALA group were significantly lower than the high-fat group (P <0.001).ACC genes mRNA level was not significantly different from high-fat group.SREBP1C and FAS protein expression were significantly lower than the high-fat group, but ACC showed no significant difference.Conclusions Saturated fatty acids promote hepatocyte fatty acid synthesis, ALA abates fatty acid synthesis by inhibiting FAS and SREBP1C gene expression.

Objective To build an antibody microarray based on direct labeling strategy for microalbnrninuria measurement, and evaluate it's technical potentiality for clinical application. Methods Urine samples of diabetic patients were collected. Antibody microarrays were constructed by preparation of array support, array fabrication, then protein assay and data analysis were performed. Procedure conditions for each step especially the labeling of samples were optimized. The set-ups were evaluated in terms of sensitivity, specificity and reproducibility. Urinary albumin excretion in the samples was detected by fabricated protein array, which was compared to that detected with immunoturbidimetry. Results The signal intensity was best when protein quality ratio of pure albumin or urine sample against NHS-biotin was 2:1. A calibration curve with a correlation coefficient of 0.9995 was established. The lower limit of detection was 0.0617 mg/L. Interehip and intrachip variation studies conducted on patient urine demonstrated CVs as 6.78%-9.22% and 3.35%-7.59%, respectively. Compared with the immunoturbidimetry, the antibody microarray was able to detect the extremely lower grade albumin in urine samples. The correlation coefficient of the results obtained by the two methods was 0.9199 (P <0.01). Conclusion An antibody microarray based on direct labeling strategy for microalbuminuria measurement is successfully established, which is comparable to immunoturbidimctry in its accuracy and will have great potential for clinical use with its high throughput, sensitivity, specifity and reproducibility.