This paper combines the experience that university is in constructing center of experimental teaching.We thoroughly elaborated center of experimental teaching the guidelines of construction,develop process,and concrete conditions were introduced such as experiment resources integration,instruments management,network construction,personnel dispensation,model of managements establishment,laboratory open,and have obtained the obviously result.

Objective To investigate the etiology spectrum of hand , foot and mouth disease ( HFMD) and to analysis the molecular characteristics of three predominant human enterovirus stains in Qingdao in 2013.Methods The total enterovirus (EV) strains and strains of EV71, CVA16 and CVA6 in throat swabs of HFMD cases were detected by using multiplex real time RT-PCR.The full-length of the viral VP1 genes of the EV strains were amplified and sequenced .The sequences were phylogenetically analyzed by using the MEGA5.0 software package .Results A total of 841 patients with mild HFMD and 107 patients with serious HFMD were recruited in this study and 64 .3%of them were positive for EV .The predominant pathogens were EV71 (44.8%), CVA6 (28.2%) and CVA16 (9.5%) in 2013.CVA6 replaced CVA16 as the second most common pathogen for HFMD , accounting for 42.7% of all pathogens in children aged less than 3 years and 22.2%of all pathogens in the serious patients .The proportions of CVA6 in the etiology spectrum showed a downtrend along with the increasing age of the patients (P<0.001).Phylogenetic analy-sis of the complete VP1 gene sequences showed that all of the EV 71 strains identified in this study belonged to the subgenotype C4 (evolutionary branch C4a) and all of the CVA16 strains belonged to the subgenotype B1 (evolutionary branches B1a and B1b).There were 6 genogroups (A to F) regarding to the VP1 gene of CVA6 and all of the CVA6 strains identified in this study belonged to genogroups A and D .Among the CVA6 strains isolated in Qingdao in 2013, 83.9% belonged to genogroup A, while the rest 16.1% belonged to genogroup D.66.7%of the CVA6 strains isolated in 2012 belonged to genogroup A, while the rest 33.3%belonged to genogroup D .All of the CVA6 strains isolated from year 2008 to 2011 in Qingdao belonged to genogroup D.Conclusion EV71, CVA6 and CVA16 were the prevalent pathogens responsible for the de-velopment of HFMD in Qingdao in 2013.The proportions of CVA6 strains in the etiology spectrum showed a downtrend with the increasing age in children .C4a was the major subtype of EV71 strains circulating in Qingdao in 2013, while B1a and B1b were the major subtypes of CVA16 strains.The pattern of endemic cir-culation of CVA6 strains showed a trend of changing from genogroup D to A from year 2008 to 2013 .

Objective To analyze the etiologic spectrum of hand, foot and mouth disease (HFMD) and the molecular characteristics of coxsackievirus A10 (CVA10) strains isolated in Qingdao from year 2010 to 2012.Methods Throat swab specimens were collected from patients with HFMD to detect to-tal enteroviruses ( EVs) , EV71 and CVA16 strains by multiplex real time RT-PCR.The EV-positive speci-mens were further detected by a semi-nested RT-PCR to amplify the sequence of viral genes encoding VP1. The serotypes of EVs were identified based on the sequences of genes encoding VP1.The full-length gene se-quences encoding VP1 of CVA10 isolates were amplified and sequenced. The phylogenetic analysis was con-ducted by using MEGA5.0 software package.Results A total of 1919 outpatients with mild HFMD and 1336inpatients with serious HFMD were recruited in this study .CVA16 strains were the predominant patho-gen for outpatients in year 2010 ( prevalence rate of 53%) and 2012 ( prevalence rate of 73%) .EV71 strains were the predominant pathogen for outpatients in year 2011 ( prevalence rate of 78%) and inpatients in year 2010 ( prevalence rate of 70%) and 2011 ( prevalence rate of 86%) . Some serotypes other than CVA16 or EV71 were the predominant pathogens for inpatients in year 2012 ( prevalence rate 44%) . CVA10 strains were identified in 12 patients with HFMD in year 2010and 17 patients with HFMD in 2012. The full-length gene sequences encoding VP1 of 23 CVA10 isolates were successfully amplified and se-quenced.The phylogenetic analysis showed that the 23 CVA10 isolates all belonged to genotype C and could be further divided into six clades.The genes encoding VP1 of the 23 CVA10 strains shared 97.3% to 1000.%homologies in amino acid sequences.The CVA10 isolates were also similar to their counterparts isolated from other regions during the same period.Conclusion CVA16 and EV71 strains were the preva-lent pathogens of HFMD in Qingdao from year 2010 to 2012, co-circulated with some other serotypes of EVs, especially CVA10 strains.The CVA10 strains belonged to genotype C and shared high homologies among them and their counterparts circulated in other regions during the same period.

AIM:To explore the effects of atorvastatin (Atorv) on atherosclerosis in streptozotocin (STZ)-in-duced diabetic apolipoprotein E knockout ( ApoE-/-) mice with fat-rich diet and the possible mechanism .METHODS:C57 mice served as control.ApoE-/-mice (n=34) fed with high-fat diet were randomly divided into ApoE-/-group, STZ-ApoE-/-group and STZ-ApoE-/-+Atorv group.Intraperitoneal injection of streptozotocin was performed to create di-abetic animal model .Blood glucose was determined by glucose oxidase method .Blood lipid levels were detected by enzymic method or selective homogeneous method .The plaque area in the thoracic aorta was measured by HE staining .The protein level of nicotinamide-adenine dinucleotide phosphate ( NADPH) oxidase subunit gp91phox in the thoracic aorta was deter-mined by Western blotting .The levels of reactive oxygen species ( ROS) in blood and thoracic aorta homogenates were de-tected by Fenton reaction and Griess reagent .Human umbilical vein endothelial cells ( HUVECs ) were isolated from healthy umbilical cords by collagenase I and cultured .ROS production was detected by flow cytometry .NADPH oxidase ac-tivity was measured using lucigenin assay .Effects of retinoid X receptor α( RXRα) on inhibition of oxidative stress by ator-vastatin were evaluated by RNA interference and plasmid transfection .RESULTS: (1) Compared with C57 group, the plaque areas of the thoracic aorta in ApoE-/-group were increased .No difference of the fasting glucose between the 2 groups was observed.The levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in ApoE-/-group than those in C57 group.(2) Compared with ApoE-/-group, the plaque areas of the thoracic aorta in STZ-ApoE-/-group were further enlarged [(314.13 ±35.72) μm2 vs (215.88 ±34.19) μm2, P<0.05].The levels of blood glucose, TG, TC and LDL-C, thoracic aorta gp91phox protein and ROS in blood and thoracic aorta homogenates were higher in STZ-ApoE-/-group than those in ApoE-/-group (P<0.05).(3) Compared with STZ-ApoE-/-group, the plaque areas of the thoracic aorta in STZ-ApoE-/-+Atorv group were reduced [(217.47 ±24.56) μm2 vs (314.13 ±35.72) μm2, P<0.05].The levels of blood glucose , LDL-C, TC, HDL-C and TG showed no significant difference between the 2 groups.Thoracic aorta gp91phox protein level and ROS production in blood and thoracic aorta homogenates were lower in STZ -ApoE-/-+Atorv group than those in STZ-ApoE-/-group (P<0.05).(4) High glucose-induced increases in NADPH oxidase activity and gp91phox expression were significantly inhibited by atorvastatin (10-6 mol/L) in HUVECs.The inhibitory effects of atorvasta-tin on high glucose-induced ROS production and NADPH oxidase activation were largely impaired when the cells were trans -fected with RXRαsiRNA.However , the effect of atorvastatin was significantly strengthened when RXRαwas over-expressed in the HUVECs transfected with RXRαplasmid.CONCLUSION: Atorvastatin inhibits atherogenesis by depressing high glucose-induced oxidative stress in diabetic ApoE-/-mice with fat-rich diet.The anti-oxidative stress effect of atorvastatin is mediated by RXRα.

Objective:To investigate the whole genome characteristics of coxsackievirus A4 (CVA4) circulating in Qingdao city.Methods:Four CVA4 isolates circulating in Qingdao city during 2013 to 2015 were selected. Whole genome sequences of these strains were amplified by one-step reverse transcription-polymerase chain reaction (RT-PCR). Sequence alignment and phylogenetic analysis were performed using MEGA7.0 software package. Genetic recombination analysis was performed using similarity plots 3.5.1 software package.Results:Phylogenetic analysis showed that based on the sequences of the whole genome and P1, P2 and P3 regions, HS312/QD/CHN/2013 and HS605/QD/CHN/2014 strains together with the early domestic isolates belonged to the same clade, while FY218/QD/CHN/2015 strain and CV-A4/P1033/2013/China strain collected in Wenzhou in 2013 formed another clade in each phylogenetic tree. HS144/QD/CHN/2014 strain belonged to the same clade as HS312/QD/CHN/2014, HS605/QD/CHN/2014 and the early domestic CVA4 isolates in the phylogenetic tree based on the P1 region, but formed a separate clade in the phylogenetic trees based on the whole genome, P2 region and P3 region. Genetic recombination analysis revealed that there was genetic recombination between HS144/QD/CHN/2014 strain and the CVA2 strain of CV-A2/P373/2013/China isolated in mainland China in 2013 in the region of 2C-3D (5 081-7 301); FY218/QD/CHN/2015 and CV-A4/P1033/2013/China strains were highly homologous and recombination signal sequences were detected in the region of 2A-2B (3 821-4 161) between the two strains and the CVA2 strain of CV-A2/P373/2013/China.Conclusions:The CVA4 isolates circulating in Qingdao city presented obvious genetic diversity at the genome-wide level.

Objective To explore the risk factors for childhood death from pneumococcal meningitis.Methods The data of 32 hospitalized children were retrospectively analyzed,who were diagnosed as pneumococcal meningitis and enrolled in the Affiliated Children′s Hospital of Soochow University from November 201 0 to December 201 5.The subjects were divided into the death group and survival group according to their prognosis.The clinical characteristics and laboratory data were compared between 2 groups.Results Between the death group and survival group,there were significant statistically differences in shock within 24 hours after admission(63.6% vs 1 4.3%,P =0.01 3),as well as endotracheal tube intubation(1 00.0% vs 23.8%,P <0.001 ),the levels of cerebrospinal fluid(CSF)IgG[(491 .27 ± 203.53)mg/L vs (267.24 ±1 88.07)mg/L,P =0.006],IgM[(1 1 5.72 ±79.1 9)mg/L vs (32.80 ±28.52)mg/L, P =0.006],IgA[59.52(1 5.51 ,75.69)mg/L vs 1 8.77(9.33,27.54)mg/L,P =0.023],CSF leukocyte[330.00 (1 50.00,380.00)×1 06 /L vs 870.00 (403.00,6 1 60.00)×1 06 /L,P =0.009 ],CSF protein [(4 047.00 ± 1 942.1 6)mg/L vs (2 470.62 ±1 259.94)mg/L,P =0.009],CSF adenosine deaminase (ADA)[35.20(1 8.90, 87.20)U /L vs 8.80(3.05,23.78)U /L,P =0.001 ],serum sodium[(1 30.21 ±2.85)mmol/L vs (1 32.83 ±3.69) mmol/L,P =0.049],serum lactic acid (LA)[4.40 (2.60,5.70)mmol/L vs 2.40 (1 .75,4.50)mmol/L,P =0.01 3],serum C -reactive protein (CRP)[(95.87 ±65.40)mg/L vs (1 65.61 ±83.05)mg/L,P =0.022],serum lactate dehydrogenase (LDH)[81 3.40(465.20,2 31 0.70)U /L vs 359.20(257.85,405.90)U /L,P =0.001 ], platelet[(1 63.82 ±1 64.86)×1 09 /L vs (295.71 ±1 30.29)×1 09 /L,P =0.01 9]and positive rate of blood culture (90.9% vs 47.6%,P =0.023)between the death group and survival group.Conclusions The risk factors associated with mortality in pediatric SPM include shock within 24 hours after admission,endotracheal intubation,hyponatremia, thrombocytopenia,as well as high serum LA level,high serum LDH level,lower serum CRP level or cultures of blood and CSF double positive.

OBJECTIVETo establish an optimized method for extracting saponins from Radix et Rhizoma Notoginseng (Sanqi).

METHODHPLC coupled with charged aerosol detector (CAD) was used for saponins determination. Peak areas of the main saponins notoginsenoside R1, ginsenosides Rg1, Re, Rb1, Rg2, Rh1 and Rd in Sanqi were monitored to evaluate the extract effectiveness. One-factor analysis was conducted to investigate the factors of particle size, solvent, extraction method and soaking time. Orthogonal experiment design L9 (3(4)) with different level of concentration of solvents, extraction time, extraction times and volume of the solvent, was implemented to determine the optimized extract condition.

RESULTThe optimum extract technique was that the dried and well pulverized (passed through a 80 mesh (180 +/-7.6) microm sample 1.0 g was extracted in 20 mL methanol by reflux (1.5 h) after soaked at room temperature for 0.5 h.

CONCLUSIONThe optimized technique is simple and high extraction efficiency. It provide basic data for further study on Sanqi.

Objective To investigate the molecular characteristics of coxsackievirus A12 ( CV-A12) and to understand the clinical manifestations of severe hand, foot and mouth disease (HFMD) caused by CV-A12 in Qingdao. Methods Throat swabs of HFMD, herpangina and influenza-like cases from 2011 to 2016 were detected for enteroviruses ( EVs) in Qingdao. Human rhabdomyosarcoma ( RD) and human la-ryngeal carcinoma (Hep-2) cells were used for virus proliferation and CV-A12 strains were identified through a semi-nest RT-PCR. The full-length of VP1 gene of CV-A12 strains was sequenced and phylogenetically an-alyzed using MEGA7. 0 software package. Clinical data of severe HFMD cases positive for CV-A12 were col-lected and analyzed. Results CV-A12-positive HFMD, herpangina and influenza-like cases accounted for 0. 3％(18/6798), 1. 2％(2/169) and 0. 1％(1/676) in Qingdao, respectively. Most of the HFMD caused by CV-A12 in children were mild before 2013 (84. 6％, 11/13), while hospitalized severe cases with neurological symptoms (100％, 5/5) became more common after 2013. Phylogenetic analysis of the VP1 region revealed that CV-A12 strains worldwide could be divided into two genotypes, A and B. All of the CV-A12 strains successfully sequenced in Qingdao from 2011 to 2016 belonged to genotype B, and 88. 9％(16/18) of them belonged to subgenotype B2. All hospitalized severe cases of CV-A12-caused HFMD after 2013 were associated with strains in branch B2b of subgenotype B2. Conclusion CV-A12 was one of the pathogens causing HFMD, herpangina and influenza-like illness in children in Qingdao. Strains of genotype B2 were the predominant CV-A12 strains circulating in Qingdao in recent years. CV-A12-caused HFMD might complicated by nervous system damage.

Objective: To illuminate the gene characteristics and clinical characterization of Coxsackievirus B5 (CV-B5) strains isolated from patients with sevre hand, foot and mouth disease (HFMD) in Qingdao city. Methods: A total of 1 844 patients of HFMD were consecutively admitted to Qingdao Women and Children's Hospital from 2013 to 2014. Information of the study population described above was collected retrospectively. The samples were collected from at least 1 site (throat swab, cerebrospinal fluid), which viral nucleic acid extracted and the entire VP1 gene sequences of CV-B5 isolates were amplified and sequenced, then the homology and phylogeny analysis were conducted by MEGA7.0. The prototype Faulkner strain and other VP1 amino acid sequences were derived from the GenBank database. Results: A total of 8 CV-B5 positive cases were obtained, including 4 males and 4 females; 6 severe hospitalized cases and 2 outpatients. The age of 6 hospitalized patients ranged from 3 to 48 months, with a median of 26 months. For the six inpatients, fever, convulsions vomiting, diarrhea and rash were the main clinical manifestation, and all combined with viral encephalitis. Compared with the prototype strain Faulkner, in the VP1 region,the nucleotide and the amino acid homologies was 77.3%-78.8% and 95.5%-97.0% respectively. Five out of the six severe cases with substitution of serine (S) to asparagine (N) at amino acid site 95 in the VP1 region. The sequences of 8 CV-B5 strains were classified into genogroup D. Conclusion Hand, foot and mouth disease associated with CV-B5 virus infection can result in nervous system involvement and the main complication was viral encephalitis. The CV-B5 strains associated with severe hand, foot and mouth disease had high nucleotide homology and present a certain regional aggregation.

Objective:To investigate the overall clinical performance of nursing students and analyze its influencing factors to provide evidence for the improvement.Methods:A questionnaire survey was carried out among 288 nursing students from Qilu Hospital of Shandong University in March to May 2020. The survey included a general data questionnaire, Holistic Clinical Assessment Tool, Clinical Learning Environment, Supervision and Nurse Teacher evaluation scale, Medical Student Safety Attitudes and Professionalism Survey of nursing students. The factors associated with overall clinical performance of nursing students were analyzed by t test, analysis of variance, Pearson correlation analysis and multiple linear regression. Results:The total score of Holistic Clinical Assessment Tool was 114.95 ± 22.91. The total score of Clinical Learning Environment, Supervision and Nurse Teacher evaluation scale was 156.70 ± 26.98. The total score of Medical Student Safety Attitudes and Professionalism Survey was 123.65 ± 31.00. The influencing factors included social practice experience and clinical learning environment, which accounted for 28.0% of the total variation of overall clinical performance of nursing students.Conclusions:Educators and managers of nursing students' schools and internship hospitals should actively explore the educational reform practice to promote the improvement of clinical ability, so as to promote the improvement of the overall clinical ability of nursing students.