Objective To study the expression of RUNX3 mRNA in primary liver cancer (PHC) tissue and its surrounding normal tissue,and its clinical significance.MethodsReverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expression of RUNX3 mRNA in tumor and peritumor tissues in 51 patients with PHC.The relationship between RUNX3 mRNA expression and some clinical pathological parameters was analyzed.ResultsThe relative expression values of RUNX3 mRNA in the tumor tissue and the surrounding normal tissue were 0.4509±0.0963 and 0.9147± 0.0222,respectively.The difference was significant (t=33.6087,P＜0.001).The RUNX3 mRNA expression in tumor tissue correlated with some clinical pathological parameters including low tumor differentiation,positive cancer embolus and intrahepatic invasion and metastasis.The RUNX3 mRNA expression was not correlated with other clinicopathological parameters including gender,cancer diameter,cancer location,hemorrhage and necrosis of cancer,and histotype.ConclusionRUNX3 may be a new tumor suppressor gene for PHC.

Objective To monitor the resistance of Streptococcus pneumoniae following the 2004 in Qingdao area, and to provide a reasonable basis for clinical application of antimicrobial agents. Methods Collecting respiratory tract, blood, cerebrospinal fluid and other specimens from out-patient and in-patients of some hospital in Qingdao from January 2005 to December 2008. According to the recommendation of NCCLS, antibiotic resistance analysis of 11 kinds of antibiotic to the isolated 231 Streptococcus pneumoniae by micro-agar dilution method, and analysis resistance trends and age differences. Results The results showed that the rate of Streptococcus pneumoniae not sensitive to penicillin is 23. 38% (PRSP: 9.52% , PISP: 13. 85% ) , resistant to cefotaxime is 9. 96% (23/231), resistant to amoxicillin is 12. 55% , resistant to erythromycin is 90. 48% (209/231). PRSP rate of patients younger than 14 years of age 27. 91% (12/43), significantly higher than that of the PRSP rate of adults 5. 38% (10/186). Conclusion The rate of resistant to penicillin Streptococcus pneumoniae increased significantly from 2004, and an increasing trend year by year, the resistance of Streptococcus pneumoniae is also a rising trend year by year. For patients infected low penicillin-resistant Streptococcus pneumoniae in this region, cefotaxime, amoxicillin are preferred drugs.

Brucellosis is an infectious-allergic anthropozoonosis caused by invasion of Brucella.It belongs to category B infectious disease in China.The epidemic situation of brucellosis is raging again in China just like the rest of world.Especially after 2000,brucellosis became one of the fastest growing infectious diseases.By 2012,all Chinese administrative units at provincial levels except Taiwan had reported the cases of brucellosis.In this paper,we analyzed the epidemic features,prevention and control of brucellosis.We elucidated deep social reasons and the root problems,which impede the prevention and control of brucellosis.We have also discovered the important problems,which should be focused on and solved in the process of prevention and control of brucellosis.

Objective To evaluate the safety and efficacy of radiofrequency catheter ablation(RFCA)in chil-dren with tachycardia arrhythmias. Methods One hundred and twenty - three children under 3 years old with drug -refractory tachycardia underwent an interventional electrophysiology procedure and RFCA in Medical Center of Tsinghua University from April 1994 to December 2014,including 98 male and 25 female,and their average age was(22. 9 ± 9. 6)months(2 - 35 months)and body weight was(13. 64 ± 2. 80)kg(4. 66 - 19. 40 kg). The mechanism,RFCA methods,the success rate,recurrence rate and complication rate of the different types of tachycardia were investigated. Results Among these children,interventional electrophysiological study confirmed 94 cases(76. 4% ,94 / 123 cases) with atrioventricular reentrant tachycardia(AVRT),7 cases(5. 7% ,7 / 123 cases)with atrioventricular nodal reentrant tachycardia(AVNRT),3 cases(2. 4% ,3 / 123 cases)with focal atrial tachycardia(FAT),8 cases(6. 5% ,8 / 123 ca-ses)with atrial flutter(AF),5 cases(4. 1% ,5 / 123 cases)with idiopathic left ventricular tachycardia(ILVT),6 ca-ses(4. 8% ,6 / 123 cases)arrhythmia was not induced. RFCA was performed in 109 cases(88. 6% ,109 / 123 cases). The success rate of RFCA was 94. 5%(103 / 109 cases)and the recurrence rate was 6. 8%(7 / 103 cases). In 88 cases of AVRT,the success rate was 96. 6%(85 / 88 cases),and the recurrence rate was 8. 2%(7 / 85 cases). In 5 cases of AVNRT,the success rate was 100. 0%(5 / 5 cases). In 3 cases of FAT,the success rate was 33. 3%(1 / 3 cases). In 8 cases of AF,the success rate was 87. 5%(7 / 8 cases). In 5 cases of ILVT,the success rate was 100. 0%(5 / 5 cases), and there was no recurrence. The complication of RFCA was puncture vascular occlusion in 2 cases(1. 6% ,2 / 123 ca-ses),without other serious complications and death cases. In this group of children,the fluoroscopy exposure time was (11. 2 ± 5. 8)min(5. 7 - 18. 2 min)and dose area product(DAP)exposure was(954. 5 ± 117. 4)mGy × cm2(707 -2 201 mGy × cm2 ). Compared with the elder children(379 cases over 3 years old)of the exposure fluoroscopy time (10. 8 ± 6. 4)min(3. 9 - 20. 5 min)and DAP exposure(927. 5 ± 193. 4)mGy × cm2(439 - 3 201 mGy × cm2 ),there was no statistically significant difference(t = 0. 616,1. 463,all P ﹥ 0. 05). Conclusions AVRT was the most common type of arrhythmia in infants and a higher incidence of multiple accessory pathways was found in infants. Application of radiofrequency ablation to children with tachycardia arrhythmias can be relatively safe and effective to the infants with drug - resistant tachycardia.

Objective To investingate the effect of low-density lipoprotein (LDL) on epithelial -mesenchymal transition and extracellular matrix (ECM) accumulation in human peritoneal mesothelial cells (HPMCs).Methods (1)HPMCs were randomly divided into control group,LDL group (100 mg/L) and LDL (100 mg/L) + lactoferrin (100 mg/L,LDL receptor blocking agent) group.After co-cultured for 24 h,the expression of LDL receptor in HPMCs was examined by immunofluorescence staining,and the LDL uptake by HPMCs was observed with oil red O staining.(2)HPMCs were cultured with different concentrations of LDL (0,25,50,100 mg/L).After co-cultured for 24 h,the change of cell morphology was observed by inverted phase contrast microscope,and the expression of α-smooth muscle actin (α-SMA) was examined by immunofluorescence.(3) HPMCs were randomly divided into control group (5.6 mmol/L glucose),mannitol group (M,2.18％ mannitol),low glucose group (LG,30 mmol/L),high glucose group (HG,120 mmol/L) and HG + LDL group (120 mmol/L glucose + 100 mg/L LDL).Cocultured for 48 h,the mRNA expression of α-SMA,E-cadherin and type 1 plasminogen activator inhibitor (PAI-1) was detected by real-time quantitative PCR,the protein expression of α-SMA was detected by Western blotting,the content of type I collagen (Col I) and PAI-1 in supernatant was detected by ELISA.Results (1) After co-cultured with LDL for 24 h,the expressin of LDL receptor was found on the cell membrane of HPMCs.Oil red staining showed that LDL could be uptaken into the cells and abolished by LDL receptor blocker.(2) HPMCs tended to be loosely intercellular connected to each ofher,and prsesnted significant formation of fibroblast-like spindle morphology.The cytoplasm immunofluorescence intensity of α-SMA gradually increased with the increase of LDL concentration.Compared to the control group,the expressions of α-SMA mRNA and protein were significantly increased,and the expression of E-cadherin mRNA was decreased in HG + LDL group(all P ＜ 0.05).But the expressions of the parameters above-mentioned were not significant different between HG group and HG + LDL group or between HG group and control group.(3) Compared with HG group or control group,the concentrations of Col Ⅰ [(19.27±0.17) μg/L vs (14.09±0.30) μg/L or (14.81±0.91) μg/L,all P ＜ 0.05] and PAI-1 [(498.24±76.91) ng/L vs (342.19±30.43) ng/L or (220.39±33.82) ng/L,all P ＜ 0.05] in supernatant of HPMCs were significantly up-regulated in HG + LDL group,meanwhile the expression of PAI-1 mRNA was significantly higer than that in control group (P =0.022).Conclusions HPMCs uptake LDL into cells via LDL receptors.LDL can induce HPMCs transdifferentiation in the condition of high glucose,increase the secretion of Col Ⅰ,inhibit the degradation of ECM through up-regulating the expression of PAI-1,and lead to ECM accumulation.

Funding of research projects within the category of Endemiology (H2401) funded by National Natural Science Foundation of China (NSFC) from 2010-2019 was summarized in this paper. Granted projects were categorized and analyzed based on the funding type, support institute and disease classification. The characteristics and trends of granted projects supported by NSFC were analyzed and summarized in order to provide reference for researchers to apply for projects in the future.

Objective:To investigate the effects of ovarian cancer ascites-derived exosomes on the stem cell properties and invasion ability of ovarian cancer stem-like cell (OCS-LC).Methods:(1) A2780 cells were induced into OCS-LC in serum-free medium, and authenticating their stem-like properties by sphere-forming test, differentiation test and CD 133 marker detection. (2) Exosomes from ascites and A2780 cell were extracted by ultracentrifugation, then authenticating them. The morphology of exosomes was observed by the transmission electron microscope, exosome particle size was measured by nanoparticle tracking analysis (NTA). The expressions of heat shock protein 70 (HSP-70), CD 63 and CD 9 were detected by western blot. (3) The exosomes from ovarian cancer ascites and tumor cell supernate were co-cultured with OCS-LC. The groups were divided into control group, ascites-derived exosomes (ADE) group (ADE+OCS-LC group), and cells-derived exosomes (CDE) group (CDE+OCS-LC group). The sphere-forming ability was evaluated by sphere-forming cycle, maximum sphere diameter and sphere-forming rate of each group; the expression of CD 133 was detected by immunofluorescence staining under microscope; quantitative real-time (qRT)-PCR was used to detected the expression levels of octamer-4 (Oct-4), Nanog mRNA of the signature genes in the stem cells of each group; the metastasis ability of each group was measured by transwell assay. Results:(1) Identification of OCS-LC: sphere-forming experiment showed that the suspension of OCSC single cells was grown in serum-free medium in secondary sphere-forming. Differentiation function experiment showed that OCS-LC were differentiated into adherent A2780 cells by changing the growth mode in serum containing medium. Flow cytometry showed that the proportion of CD 133 positive ( CD133+) cells in OCS-LC group was (18.9±0.9)%, significantly higher than that of control group (0.6±0.5)% ( t=38.570, P<0.01). (2) Under transmission electron microscope, clear lipid bilayer structure was observed in ADE and CDE, and one side presented a concave hemispheric or cup like structure. NTA showed that the diameter of exosomes mainly ranged from 30 to 100 nm, with an average diameter of 67.2 nm. Western blot analysis showed that the specific protein molecules HSP-70, CD 63 and CD 9 were positive. (3) Three groups′ OCS-LC could continue to grow into spheres, and the group of ADE+OCS-LC showed two growth modes, most of the cells continued to grow into spheres, while a small part of cells grew in adherent differentiation. The sphere-forming rate of OCS-LC in the control group, ADE+OCS-LC group, and CDE+OCS-LC group were (1.05±0.20)%, (4.15±0.10)%, and (10.45±0.25)%, the sphere-forming cycle of OCS-LC in the three groups were (15.3±1.5), (10.3±0.6), and (6.7±0.6) days, and the maximum diameters of OCS-LC were (100.3±3.2), (145.2±5.1) and (170.0±2.1) μm, respectively. And the differences were statistically significant (all P<0.05). After co-culture of exosomes with OCS-LC, the sphere-forming ability of cells in the group of CDE+OCS-LC was prior to ADE+OCS-LC group (all P<0.05). Immunofluorescence staining showed that the number of CD 133 green fluorescent chromophore cells in OCS-LC groups [(46.2±2.1)%, (58.4±2.2)%] was significantly higher than that in the control group [(26.6±1.5)%] after the addition of exosomes in co-culture, the positive rate of CD 133 was higher than that in the control group( F=187.588, P<0.05). The qRT-PCR results showed that the expression levels of Oct-4 mRNA in ADE+OCS-LC and CDE+OCS-LC groups were 3.46±0.24, 4.03±0.31, compared with that in control group (1.04±0.12), the differences were statistically significant ( F=134.932, P<0.05). The mRNA expression levels of Nanog were 1.57±0.32, 2.66±0.15, which were significantly higher than that in the control group (1.00±0.07), and the differences were statistically significant ( F=49.329, P<0.05). And the expression of both in CDE+OCS-LC group increased more significantly than ADE+OCS-LC group (all P<0.05). The number of invasive cells in the three groups of OCS-LC were: control group 30±5, ADE+OCS-LC group 102±4, CDE+OCS-LC group 210±7, and there were statistically significant differences among three groups ( F=820.800, P<0.05). Compared with the control group, the number of invaded cells in the co-culture group were significantly increased ( P<0.05), and the CDE+OCS-LC group had the higher cell invasion ability then the ADE+OCS-LC group ( t=23.202, P<0.05). Conclusions:Exosomes derived from ovarian cancer ascites could enhance and maintain the stemness of OSC-LC, and promote the invasion of tumor cells. Moreover, CDE is superior to ADE.

BACKGROUND:Platelet rich plasma contains various growth factors, such as platelet-derived growth factor, metastatic growth factor, insulin-like growth factor, epidermal growth factor as wel as vascular endothelial growth factor. Therefore, it can directly or indirectly promote cel differentiation and proliferation in different stages of bone regeneration.
OBJECTIVE:To study the effects of coral bone with platelet rich plasma in the repair of mandibular defects.
METHODS: Totaly 24 New Zealand white rabbits were randomly divided into three groups (n=8 per group) including test group, control group and blank control group. Coral bone with autologous platelet rich plasma, coral bone or nothing was implanted, respectively, after establishing unilateral mandibular defect models. The defects were evaluated by imaging observation and bone his-tomorphometric analysis at 2, 4, 8, 12 weeks after surgery.
RESULTS AND CONCLUSION:At 12 weeks after surgery, by imaging observation, density of the defect increased in the blank control group, which was lower than that of the normal bone; the bone density in the test group was higher than that in the control group, both of which were similar with the normal bone. Besides, the materials were closely combined with the new tissues. By bone his-tomorphometric analysis, area of the new bone in the test group was significantly larger than that in the control and blank control group (P< 0.05). In conclusion, coral bone with platelet rich plasma has good biocompatibility and bone conductivity, which can induce bone regeneration to promote defect repair.

BACKGROUND:MATLAB has capabilities of large numerical calculation, mathematical drawing and simple finite element analysis. It can establish models rapidly and can be able to identify the grayscale with BMP and JPG
format, and it can directly transform the identified data into ANSYS finite element software-readable format, thus avoiding personal error produced by the repositioning and secondary treatment in the Autocad software.
OBJECTIVE:To find a simple, convenient and accurate method to construct the model of lumber fusion and to analyze the biomechanics of lumbar motion segment after lumbar fusion.
METHODS:Lamel ar CT and Matlab (Matrix Laboratory) scientific computing software combining Ansys finite
element software was used to construct the models of lumber fusion. Then the models were loaded to analyze the biomechanical change of the fusion model.
RESULTS AND CONCLUSION:The established models were loaded with axial, bending and stretching loads, and the biomechanical analysis showed that interbody fusion had the best stability among al the fusion models. Combined with joint fusion, the axial displacements of interbody, rear side and rear fusion models were
decreased by 5%, 1%and 4%than that of simple interbody fusion, posterolateral fusion and posterior fusion models. Under the stretch-buckling load conditions, the rotation angles were reduced by 23%, 11%and 45%. Stress concentration to the fusion parts showed fusion block could increase the load displacement. The
technology of lamel ar CT, Matlab software and Ansys finite element software can accelerate the construction of lumber fusion model and make the model more accurate. Facet joint fusion combined with interbody fusion,
posterolateral fusion and posterior fusion can get better lumbar stability, and this increased stability is more significant in the rear fusion. Stress distribution of posterior fusion is more reasonable.

OBJECTIVE To study and evaluate the value of the antibiotic-neutralized culture bottle applied by manual operations.METHODS A total of 711 blood samples were collected from clinical high fever patients in Qingdao Central Hospital from 2002 to 2005.To compare the bacteria detected rates,cultural time and bacteria types in 399 samples cultured in manual blood-culture bottles with those in 312 samples cultured in antibiotic-neutralized culture bottles applied by manual operations.RESULTS The bacteria detected rates were 10.28% in manual blood-culture bottles and 20.83% in antibiotic-neutralized culture bottles applied by manual operations.There was significant difference between both of them(P72h were no statistical difference between manual blood-culture bottles and antibiotic-neutralized culture bottles.Twenty species of bacteria were detected in manual blood-culture bottles and 29 species of bacteria in antibiotic-neutralized culture bottles applied by manual operations.CONCLUSIONS The bacteria detected rate and species of bacteria were more obviously raised in antibiotic-neutralized culture bottles applied by manual operations than in manual blood-culture bottles.