Objective To reveal the trend of change incidence of hepatitis A(HAV)in Shandong province and explore dynamic analysis methods.Method Dynamic analysis was made on data of hepatitis A in Shandong province from 1990 to 2006.Results The reported incidence of hepatitis A patients dropped from 53.97/100,000 to 1.14/100,000 in Shandong province from 1990 to 2006,which showed a marked downward trend;the groups under the age of 10 dropped from 55.60% to 5.12%,the average age of incidence moved from 14.40 years old to 40.19 years old.Conclusion The incidence of hepatitis A has been changed from prevalent to sporadic in Shandong province,and there was a further downward trend;the age of the high-risk population has been shifted to older age group;the sex ratio of incidence between males and females in the population above 10 years of age had an expansion trend,the age distribution curve differences between males and females was also expanded.

Objective To observe the influence of Fibronectin-Thrombopoietin(FN-TPO) gene modified human bone marrow mesenchymal stem cells(MSCs) on the engraftment of cord blood hematopoietic stem cells.Methods FN-TPO gene modified human bone marrow MSCs combined with cord blood mononuclear cells(CB-MNC) were transplanted to sublethal dose treated severe combined immunodeficiency disease(SCID) mice.After transplantation,these mice were observed for 4 weeks.Peripheral blood cell counts were performed at different time point to assay the hematopoietic system status of the mice.Four weeks after the transplantation,human-sourced cell integration was assayed by flow cytometry(FCM) and polymerase chain reaction(PCR).Results One week after the cell transplantation,every main index of the peripheral blood cell counts in the gene modified group was higher than that in the control groups(P

Objective To study the relationship between macular pigment optical density (MPOD) and serum concentration of lutein and zeaxanthin in an adult population.Methods Twenty patients with mild cataract and 39 healthy subjects were enrolled in this study,including 15 males and 44 females.The average age was 43.75 years.Fifty-three subjects were non-smokers and 6 male subjects were smokers.Two subjects preferred meat diet,22 preferred meat-less diet,and 35 have balanced diet.MPOD was measured using heterochromatic flicker photometry at 0.25,0.5,1.0 and 1.75 degrees,and serum concentration of lutein and zeaxanthin was measured using high-performance liquid chromatography.The relationship between MPOD and serum concentration of lutein and zeaxanthin was analyzed.The differences of serum lutein and zeaxanthin between different gender,smokers and non-smokers and subjects with different dietary pattern were also analyzed.Results MPOD at 0.25,0.5,1.0 and 1.75 degrees were 0.59,0.48,0.34 and 0.18,and the average concentration of lutein and zeaxanthin were (0.45± 0.16) μmol/L and (0.11 ± 0.04) μmol/L respectively.Serum concentration of lutein and zeaxanthin in males were slightly higher than that in females,but it was not statistically significant (t=1.13,0.86;P=0.27,0.40).The differences of serum lutien and zeaxanthin between smokers and non-smokers (t=0.15,-0.11;P=0.87,0.91),among subjects of 3 dietary patterns groups were not statistically significant (Flutein=3.87,4.05,0.18;P=0.83,0.81,0.99.Fxeaxanbin=0.99,1.51,0.52;P=0.85,0.68,0.72).There was no correlation between MPOD and serum concentration of lutein (r=-0.06,-0.02,-0.07,0.03;P＞0.05) and zeaxanthin(r=0.02,0.12,0.09,0.11;P＞0.05).Conclusion MPOD was not statistically significantly correlated with serum concentration of lutein and zeaxanthin in the studied population.

Objective To evaluate the methylation status of RASSF2A in epithelial ovarian cancer (EOC) tissues and plasma, and to explore the correlations in two kinds of samples with clinicopathological characteristics. Methods The frequency of methylation of the RASSF2A gene in tissues and corresponding plasma samples from 60 EOC patients, 20 patients with benign ovarian tumors and 10 patients with normal ovarian tissues were detected respectively. The methylation-specific PCR was used to determine the methylation status. Results The frequency of aberrant methylation of RASSF2A was 58.3% (35/60) in EOC tissues and 43.3%(26/60) in corresponding plasma samples respectively, however, such hypermethylation was not detected in the benign ovarian tumors and normal ovarian samples, and there was significant difference between them (P<0.05). Besides, RASSF2A methylation levels in serum of EOC patients correlated reasonably well with methylation status in tumor samples (r=0.739, P=0.000). No significant differences were observed between the promoter hypermethylated status of the RASSF2A and clinicopathological characteristics (P>0.05). Conclusions The promoter methylation of RASSF2A gene is an early and frequent epigenetic event in EOC. Detection of promoter hypermethylation in plasma specimens can help the early diagnosis of EOC.

BACKGROUND: Embryonic stem cell (ESC) is a kind of highly undifferentiated totipotent cell. It can proliferate and maintain its totipotency in the system cultured in vitro. It is one of most promising stem cells in thetreatment of central nerve injury.OBJECTIVE: To observe the survival and migration of induced transplanted ESC in mice with spinal injury and hypoxic-ischemic encephalopathy.DESIGN: A completely randomized grouping design, controlled animal experiment.SETTING: Laboratory of Developmental Biology Research Center of Shanghai Second Medical University.MATERIALS: Sixty C57/BL6J mice, of clean grade and either gender, aged 6 to 8 weeks (n =30) and 7 days (n =30)were provided by the Shanghai Experimental Animal Center, Chinese Academy of Sciences [Permission No, SCXK (hu)2003-0003]. This animal experiment was approved by Animal Ethics Committee. Mouse ESC strain S8, labeled LacZ marker gene (Provided by Shanghai Developmental Biology Research Center). X-gal dyeing reagent (Sigma Company).METHODS: This experiment was carried out in the laboratory of Shanghai Developmental Biology Research Center (Shanghai Key Laboratory) from October 2002 to December 2003. ① Experimental grouping of spinal injury: Sixteen C57/BL6J successful mice models, aged 6-8 weeks, were randomized into 2 groups: experimental group (n =8), in which, following right spinal semi-sectioning, derivated cell suspension for inducing the in vitro differentiation of ESC was injected at 1 cm away from injury through vertebral canal, and control group (n =8), in which, following right spinal semi-sectioning, phosphate buffer solution (PBS) was injected at the peripheral region of injury. ② Hypoxic-ischemic encephalopathy experimental grouping: Sixteen successful C57/BL6J mice models, aged 7 days, were randomized into 2 groups: experimental group (n =8), following ligation of right common carotid artery, mice were placed in the closed container containing 0.08 volume fraction of oxygen and 0.92 volume fraction of Nitrogen gas, and taken out 1.5 hours later; 3 μL ESCs were injected into the right cerebral ventricle at about 1 week, and control group (n =8), in which, the same amount of PBS was injected into the right cerebral ventricle. ③ At 12 weeks after transplantation, the survival and migration of induced ESCs labeled by Lac-Z in the spinal cord and brain were observed by zymologic method.MATN OUTCOME MEASURES: Survival and migration of ESCs in the central nervous system.RESULTS: ①After being induced in vitro and transplanted to spinal injured region, ESCs differentiated into neural precursor cells. Neural precursor cells could survive in the injured region and migrate to 5 mm away from injured region.Immunohistochemistry proved that the neural precursor cells of transplanted ESCs could differentiate into neurons.Morphologically, it was proved that neural precursor cells-derived from ESCs could well integrate peripheral tissue. ② The induced ESCs were injected into the lateral cerebral ventricle of mice. Derived ESCs widely distributed in the injured hippocampal region, cerebral cortex ventricle choroid plexus, vascular endothelium and other regions, and integrated peripheral tissue, which were similar to adjacent cells in morphology, suggesting that induced ESCs also could survive for long time and far migrate.CONCLUSION:The induced ESC can survive and migrate in the host injured brain and spinal cord, and the migration of ESCs is more obvious in the brain than in the spinal cord.

Objective To investigate the effect of continuous positive airway pressure(CPAP)during one lung ventilation on pulmonary function in patients undergoing video-assisted thoracoscopic repair of atrial septal defect.Methods Twenty ASA Ⅱ patients of both sexes,aged 16-30 yr,weighing 41-64 kg,scheduled for video-assisted thoracoscopic repair of atrial septal defect,were randomly divided into 2 groups(n = 10 each): control group and CPAP group.One lung ventilation(VT 8 ml/kg,RR 12-16 bpm,I:E 1:2,PETCO2 35-40 mm Hg)was perform in both groups.CPAP(6 cm H2O)was perform during one lung ventilation in group CPAP.The oxygenation index,pulmonary compliance and airway pressure were recorded during operation.The cardiovascular events,hyoxemia and extubation time were recorded.Results The oxygenation index and pulmonary compliance were significantly higher,extubation time was shorter,and the incidence of hyoxemia was lower in CPAP group than in control group(P ＜ 0.01).The airway pressure was in the normal range in both groups.No cardiovascular events was found in both groups.Conclusion CPAP(6cm H2O)during one lung ventilation can improve the pulmonary function in patients undergoing video-assisted thoracoscopic repair of atrial septal defect.

ObjectiveTo establish an analytical method with high sensitivity and wide range in biotin-avidin and time-resolved fluoroimmunoassay system for the quantitative detection of antinuclear antibody (ANA)-Ig (GAM).MethodsANA in standard preparation or sample was combined with solid nuclear antigen,biotinylation antibody and the europium(Eu3+)-labelled avidin to form the compounds of solid nuclear antigen-antibody-biotinylation antibody-SA-Eu3+.The fluorescence enhancement fluid was added to dis-sociate the Eu3+,the content of ANA was directly proportional to fluorescence intensity,the BAS-TRFLA was established for the quantitative detection of ANA-Ig (GAM).The sensitivity,specificity,reliability and range of detection were evaluated.Semm of 50 blood donor,105 patients with systemic lupus erythematosus (SLE),109 patients with rheumatoid arthritis(RA),25 patients with PBC,29 patients with Sj(o)gren's syndrome (SS),23 patients with scleroderma,25 patients with MCTD were included in this study.The positive rate was calculated.ResultsThe inner-group difference between high,medium and low densities mixture serum was 3.13％,3.74％ and 5.76％ and the inter-group precision rate was 5.31％,6.25％ and 7.46％ in BAS-TRFLA.The sensitivity of TRFIA was better than that of the ELISA method.The low detection limit was 2.24 U/ml.The mean recovery rate was 100.74％.The results measured by the TRFIA and ELISA methods were closely correlated(R2=0.978).The positive rate of blood donor,and patient with SLE,RA,PBC,SS,scleroderma and MCTD were 0,100％,23％,96％,38％,91％,92％ respectively.When compared with ELISA,the detection range of TRFIA was wider,and stability was better.ConclusionBAS-TRFIA is a stable method for detection of ANA with high sensitive and wide range of detection.It is important for the early diagnosis of autoimmune disease and monitoring the treatment efficacy of autoimmune disease.And this method may be widely used in clinical laboratories in the future.

Objective:In this study,a prokaryotic expression of the 3-ketosteroid-Delta (1)-dehydrogenase (KSDD) which came from Arthrobacter simplex was built.Moreover,in order to investigate the catalytic mechanism of KSDD and improve its stability,the structure of KSDD was predicted by computer and the critical sites were confirmed by site-directed mutations.Methods:The recombinant plasmid was constructed by eukaryotic expression vector pET-22b and the recombinant strain was constructed and expressed in Escherichia coli BL21 (DE3).High-performance liquid chromatography was used to determine the transformation rate of 4-AD to ADD.The KSDD structure and key sites were predicted by SWISS-MODEL.Site-directed mutations for the amino acid residues of key sites were constructed and activities of the mutations were detected.Results:The recombinant strain E.coli pET-22-ksdd was successfully constructed.It was induced to express the dehydrogenase by IPTG and the conversion rate of 4-AD to ADD was 27％ at 21 ℃.The structure of 3-ketosteroid-Delta (1)-dehydrogenase and the four key sites was analyzed by SWISS-MODEL.Four mutants,Y120R,Y320L,Y488F and G492Y were constructed.Mutants Y120R and Y488F were inactivated,so they were proved to be the key active sites of KSDD.The conversion rate of mutant Y320L was consistent with that of wild type,but the stability at 37 ℃ was improved.The conversion rate of mutant G492Y was 1.2 times of the wild type and the stability has been improved at 37 ℃.Conclusions:At present,there are few studies about the structure and catalytic mechanism of dehydrogenase.The active sites of the enzyme were verified by this study,which laid the foundation for the further study of the properties of the enzyme KSDD.

Objective To observe the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on patients with type A thoracolumbar fractures treated by pedicle screw fixation via dorsal-approach and vertebral body bone graft.Methods A retrospective case-control study was conducted to analyze the clinical data of 64 patients with type A thoracolumbar fractures treated from June 2012 to July 2015.The patients were divided into control group (32 cases) and research group (32 cases) according to the random number table.There were 22 males and 10 females aged (48.2 ± 11.2)years old in control group.The injury was located at T11 in 2 cases,T12 in 4,L1 in 9,L2 in 7,L3 in 7 and L4 in 3 in control group.There were 24 males and 8 females aged (50.7 ± 11.4) years old in research group.The injury was located at T11in 2 cases,T12in 4,L1in 8,L2 in 8,L3 in 7 and L4 in 3.The control group was treated with posterior pedicle screw reduction,internal fixation and allograft cancellous bone grafting through pedicle of vertebral arch.The research group was treated with rhBMP-2 (mixed with cancellous bone implants) on the basis of the control group.The time of operation,the amount of bleeding during operation,the volume of postoperative bleeding,the amount of postoperative drainage,the recovery of the injured vertebra,the Lane-Sandhu score,the time of fracture healing,and the postoperative complications were compared.Results There was no difference between two groups on operation time,amount of bleeding during operation or amount of postoperative drainage (P ＞ 0.05).No difference existed in two groups about Cobb angle and injured vertebral leading edge height at one week after operation (P ＞ 0.05).At the time of last follow-up (12 months),in research group,Cobb angle was lower while injured vertebral leading edge height was much higher than that of control group (P ＜ 0.05),both of which were better than before (P ＜ 0.05).At 12 months after surgery,the lost Cobb angle of [(2.0 ± 0.7) °] and lost height of injured vertebral leading edge [(3.2 ± 1.0) ％] were smaller than those in control group [(5.6 ± 1.7) ° and (6.8 ± 2.4) ％ respectively] (P ＜ 0.05).The Lane-Sandhu score and healing time of fracture in research group were (8.9 ± 0.8) points and (6.9 ± 0.9) months,which were better than that (6.8 ±0.8)points and (8.4 ± 1.6)months in control group respectively (P＜0.05).The complication rate of research group was lower than that of control group,with no significant difference (P ＞ 0.05).Conclusion Compared with simple vertebral bone graft,posterior thoracolumbar vertebrae pedicle screw fixation with vertebral bone graft and hBMP-2 treatment for type A thoracolumbar fractures can restore vertebral stability,shorten the time of fracture healing and reduce the incidence of complications.

Clinic demonstration is the important part of medical education.There are more and more medical students with enlarged university enrolment in recently year.However,the patients for demonstration are relatively fewer.In this condition,we make the best of the available resources to demonstrate clinic pediatrics by teaching at bedside;displaying video products,using human analogue and case record and role playing,thus ensuring the effect of clinic pediatric teaching.