Objective: To prepare tissue engeneered bone in the shape of human TMJ condyle. Methods: Rabbit marrow stem cells (MSCs) were in vitro cultured and induced by rhBMP2.107 Cells were seeded into each piece of natural porous coral (NC) in the shape and in the size of 4 -year-old-child mandibular condyle. After two days in vitro incubation, six cell-coral complexes were implanted subcautanrously into the back of nude mice. Two months after operation, bone formation was observed by gross inspection,X-ray examination,scanning electronic microscope observation and histological observation. Results: New bone grafts in the shape of human mandibular condyle were successfully restored two months after implantation in all the samples. X-ray examination showed large amount of X-ray blocking shadow. NC was partially absorbed. New bone formation could be observed by electronic microscope observation and hostological observation on the surface and in the pores of NC. Conclusion: It is an effective method to fabricate bone graft in specific shape by seeding osteogenesis cells into natural coral in the wanted shape.

Objective: To prepare tissue engieered bone graft loading titanium dental implant. Methods: Titanium dental implant (3 mm in diameter) was inserted into porous natural coral column((5 mm in diameter). Bone marrow derived osteoblasts were cultured and expanded in vitro. Cells were induced by recombinant human bone morphogenetic protein-2 for three days and then harvested and seeded into porous coral and onto dental implant at the density of 2 ?108/ml. Four cell-coral-implant complexs were incubated in vitro for 2 days and then implanted subcutaniously into nude mice. New bone formationre and new bone integration with dental implant were evaluated by gross inspection, X-ray examination and hitologic observation 1 and 2 months after implantation. Results: By gross observation, specimen of 1 month was red and white. X-ray examination showed that there was little radiodense shadow around the dental implant. Specimen of 2 months was red and had the gross appearance of bone. Dental implant could be observed situating in the newly formed bone graft. X-ray examination showed that coral scaffold was absorbed completely. Large amount of X -ray blocking shadow could be observed around the dental implant. Histologic examination showed that bone-like tissue formed in the pores and on the surface of natural coral and in some area new bone could be observed integrating with implant in 1 month specimen. In 2 months specimen, large amount of new bone formed around the implant and integrated well with the implant. Conclusions: Tissue engineered bone graft may integrate well with titatium dental implant.

ObjectiveTo report the first laboratory-confirmed human monocytic ehrlichiosis (HME) case in Shandong Province.MethodsThe epidemiological investigation was done by filling out questionaires.Blood sample was collected for detecting the specific 16S rRNA gene of Anaplasma phagocytophilumandEhrlichiachaffeensisbynested-polymerasechainreaction(PCR).ResultsThis case was characterized by acute onset of fever,leukopenia and thrombocytopenia.The specific 16S rRNA gene of Anaplasma phagocytophilum was negative and the specific 16S rRNA gene of Ehrlichia chaffeensis was positive by nested PCR test.The PCR product was sequenced.The homology was above 99 ％ between the acquired nucleotide acid sequence and the Ehrlichia chaffeensis sequences registered in GenBank.ConclusionThe Ehrlichia chaffeensis infection exists in Shandong Province,which warrants further research on these natural loci of HME in this area.

Objective To analyze the incidence trends and epidemiological characteristics of natural focus and vector borne infectious diseases in Zibo City between 2004 and 2015,and to provide evidence for control and prevention of the diseases.Method Surveillance data of natural focus and vector borne infectious diseases in Zibo City between 2004 and 2015 from the National Disease Reporting Information System were analyzed via descriptive epidemiological methods.Results A total of 10 natural foci and vector borne infectious diseases and 3 287 cases including 55 death cases were reported in Zibo City from 2004 to 2015.The average annual incidence was 6.24/100 000 and the case fatality rate was 1.67％ (55/3 287).The incidence rate increased in recent years after the lowest rate in 2009 (2.83/100 000),and the rate in 2015 was 8.83/100 000.The top three diseases with higher incidence were hemorrhagic fever with renal syndrome (3.44/100 000),brucellosis (1.48/100 000) and tsutsugamushi fever (0.53/100 000).The top three high-prevalence areas with higher incidence were Yiyuan County (13.22/100 000),Zichuan District (9.73/100 000) and Boshan District (6.13/100 000).The cases mainly occurred from March to May and September,October,which accounting for 27.47％ (903/3 287) and 24.64％ (810/3 287),respectively of the total.The 40-59 year-oldage group was the highest,accounting for 48.13％ (1 582/3 287)of the total.And 75.48％ (2 481/3 287) of reported cases were farmers.Conclusions The overall incidence of the natural focus and vector borne infectious diseases in Zibo City has a annual increasing trend in recent years,while the incidence of brucellosis and tsutsugamushi fever have continued to rise since 2004.These diseases have showed a significant seasonal distribution and population distribution.It is necessary to conduct targeted prevention and control strategies.

G mutation were intervened and avoided the occurrence of deafness,1 babies with 235delC homozygote was confirmed severe sensorineural hearing loss in the hearing screening.Conclusion Newborn gene screening make up the defects of missed diagnosis in simple hearing screening in finding the newborn babies with late-onset deafness or the high risk as well as the pathogenic carriers.So the hearing and gene screening were necessary in the current situation,and this screening strategy would be developed further in Henan province.

OBJECTIVETo investigate the predominance ticks and the infectious status of severe fever with thrombocytopenia (SFTSV) in Penglai and Laizhou counties, Shandong province.

METHODSTwo towns with high incidence rate were selected in Penglai and Laizhou, respectively, then three villages were selected in each towns. Parasitic ticks were collected from the host skin by hand manually and free ticks manually with white cloth from the grassland, monthly, during April to December in 2011. Samples were classified by original, varieties, developmental stages, then extracted RNA, using Realtime RT-PCR to test severe fever thrombocytopenia syndrome virus, S fragments were amplified with nested PCR, then isolated virus. By neighbor joining method in the phylogenetic tree, the minimum infection rate (MIR) was used to represent the infection status of ticks in novel bunyavirus.

RESULTSA total of 3 145 ticks were collected totally from 5 categories, there were 3 048(96.92%) of Haemaphysalis longicornis, 73(2.32%) of Rhinpicephalus sanguineus, 10(0.32%) of microplus Boophilus, 9(0.29%) of Haemaphysalis campanulata, 5(0.16%) of Dermacentor sinicus, respectively. The positive rate of nucleic acid of 2 044 samples was 6.16% (126/2 044), minimum infection rate (MIR) was 4.01%, there were 122(96.83%) of Haemaphysalis longicornis, 3(2.38%) of Rhinpicephalus sanguineus, and 1(0.79%) of microplus Boophilus, MIR was 4.00%, 4.11%, and 10.00%, respectively. There were no nucleic acid positive samples in Haemaphysalis campanulata and Dermacentor sinicus. The 11 S segments were amplified in 126 positive samples, the homology of S fragment was 95.6%-99.9% with 11 strains isolated from the identified SFTS cases in local area, 3 strains isolated from animals, and 11 strains isolated from other areas. There was no significant difference among original, varieties and developmental stages.

CONCLUSIONHaemaphysalis longicornis was the predominant species in Penglai and Laizhou counties, it could be propagation medium with Rhipicephalus sanguineus and microplus Boophilus, S sequence in ticks was higher homology with virus isolated from local SFTS cases.

Objective: Compare the detection result of blood samples of severe fever with thrombocytopenia syndrome (SFTS) patients using different detection techniques, and observe the dynamic characteristics of the virus specific RNA, IgM antibody and IgG antibody, to provide theoretical basis for selection of diagnostic methods of disease. Methods: Acute phase serum of suspected SFTS cases and convalescent serum samples of lab-confirmed cases were collected. Real-time fluorescence quantitative PCR and enzyme-linked immunosorbent assay (ELISA) were used to detect the virus specific RNA, IgM antibody and IgG antibody. The detection results of different methods, the relationship between positive results and the acquisition time, and the dynamic characteristics of viral nucleic acid and antibodies were analyzed. Results: A total of 87 serum samples of the suspected SFTS patients were collected, the positive rate of virus specific RNA, IgM antibody and IgG antibody were 53.41%, 31.03% and 3.41%, respectively. Among 55 confirmed cases of SFTS, the consistent rate of virus specific RNA and IgM antibody detection methods was 36.36%, and the difference between the two methods was significant (χ²=6.82, P=0.009), kappa=-0.257. The sampling intervals of RNA positive samples were all within 12 days, of which the positive detection rate was highest after 7-9 days, and the difference was statistically significant (χ²=10.35, P=0.016). In 34 SFTS convalescent serum samples, all the nucleic acid tests were negative, the positive rate of IgM antibody was 41.18%, which was not significantly different from the acute phase serum samples (P=1.00). The positive rate of IgG antibody was 94.12%, which was significantly higher than that of acute IgG antibody (0%). The dynamic characteristics of IgM and IgG antibody showed that IgM antibody could be detected on the second day after onset, the latest detection time was 74 days after onset, and the highest absorbance value and antibody detection rate occurred in 30-60 days. The earliest detection time of IgG antibody was 12 days after onset, and the last detection time was 100 days.The detection rate of IgG antibody and absorbance value increased rapidly after 30 days, and maintained in a high level. The detection rate of IgG antibody was 100% in 30-60 days. Conclusions Blood samples taken from SFTS suspected patients within two weeks of onset may be prioritized for detection of viral nucleic acids using Real-time fluorescence PCR or for detection of IgM antibodies by ELISA. Although IgM antibody can be detected 2 days after the onset, the peak appeared much later, so the negative result can’t rule out the diagnosis. IgG antibody has a high seroconversion rate in convalescent samples, and can be used as an auxiliary tool for disease diagnosis.

Objective To learn the prevalence of infection of human and animals severe fever with thrombocytopenia syndrome bunyavirus(SFTSV)in Yantai,Shandong province,and to analyze the pathogenic features of SFTSV as well as its relationship between human and animal hosts. Methods From April to November in 2011,3 576 serum samples were collected from domesticated animals,including sheep,cattle,pigs,dogs,chickens,in Laizhou and Penglai areas where fever with thrombocytopenia syndrome frequently occured among local residents. Total SFTSV antibodies and virus-specific nucleic acids of the serum were tested by ELISA and Real time RT-PCR,respectively. SFTSV infection on each animal was observed in different months. 2 590 human serum samples were also collected in Laizhou and Penglai areas,with IgG antibodies tested by ELISA. Virus was isolated with Vero cells from the serum which SFTSV viral nucleic acids were positive. S fragments were amplified by RT-PCR and sequenced,with homology analysis conducted on these sequences. Results The overall positive rate of serum samples from animals on the total SFTSV antibodies was 40.24%(1 439/3 576)while the positive rate for specific nucleic acids was 4.56%(163/3 576). The positive rates for SFTSV antibodies were 62.78%,52.97%,45.56%,28.73%,1.45%and the positive rates for specific nucleic acids were 5.72%,4.63%,3.02%,5.25%and 3.73%,in sheep,cattle,chickens,dogs, pigs,respectively. The antigens/antibodies for SFTSV in animals changed seasonally. The overall positive rate for SFTSV IgG antibody from 2 590 human samples was 5.41%. Thirteen virus strains were isolated from these serum samples(10 strains from human and 3 strains from animals). The nucleotide homology of 13S fragments’sequences ranged from 95.23%to 100.00%and the nucleotide homology with the isolates from other provinces were between 94.72%and 99.13%. The homology was considered to be high. Conclusion High prevalence of SFTSV infections occured both in human and domestic animals in Yantai city. The nucleotide sequences of SFTSV were highly homologous among human and domestic animals. The findings suggested that domesticated animals might serve as SFTSV proliferation and the hosts for transmission thus should be attached great importance.

Objective To investigate the association between ten single nucleotide polymorphisms (SNPs) in the peroxisome proliferator-activated receptors and pulse pressure (PP) as well as the relationships between gene-gene interaction between PPARα/δ/γ genes and PP.Methods A total of 820 subjects,with 550 females and 270 males,were recruited from a cohort study of “Prevention of Metabolic Syndrome and Multi-metabolic Disorders in Jiangsu Province of China Study (PMMJS)”.Ten SNPs of PPARα/δ/γ genes were selected.GMDR software (version 1.0.1) was used to evaluate the gene-gene interactions among PPARs SNPs associated with PP.Results The mean levels of PP in people with mutant genotype of rs1805192 in PPARγ genes (PA+AA) showed a significant increase by 1.341 mmHg (95％CI:0.431-2.252 mmHg) when compared to the persons with wild genotype (PP).In the subgroup of subjects with more than 30 mmHg levels of PP,a six-locus model comprised rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγshowed a highest level of prediction accuracy (0.577) and displayed a better cross-validation consistency (10/10).In the subgroup of subjects with less than 40 mmHg levels of PP,a two-locus model was statistically associated with PP with 0.628 of prediction accuracy and 10/10 of cross-validation consistency.Conclusion PPARγrs1805192 was associated with the occurrence of PP.Gene-gene interactions among rs135539 of PPARα,rs2016520 of PPARδ,rs10865710,rs1805192,rs709158 and rs3856806 of PPARγ were all significantly related to PP.

Objective:To learn about the epidemiological and spatial clustering characteristics of brucellosis in Shandong Province, and to provide a reference for scientific prevention and control of brucellosis.Methods:The epidemic data of human brucellosis in Shandong Province from January 2015 to December 2020 were collected from the Infectious Disease Reporting Information Management System of China Disease Control and Prevention Information System, and the data were analyzed by descriptive epidemiology and spatial clustering analysis.Results:A total of 18 811 cases of human brucellosis were reported in Shandong Province from 2015 to 2020, and the average annual incidence rate was 3.16/100 000. Human brucellosis occurred in every month of the year, and the peak incidence was from March to August, accounting for 66.31% (12 474/18 811). The top 5 counties (districts) with average annual incidence rates were Lijin County (32.39/100 000), Kenli District (11.02/100 000), Wudi County (10.35/100 000), Zhanhua District (9.59/100 000) and Shanghe County (8.80/100 000). There were 13 436 males and 5 375 females, with a male-female sex ratio of 2.50 ∶ 1.00; the age was mainly concentrated in 30-69 years old, accounting for 83.23% (15 656/18 811); farmer was the main occupation, accounting for 85.82% (16 144/18 811). The results of global spatial autocorrelation analysis showed that the annual incidence rates of brucellosis in Shandong Province showed a spatial clustering distribution from 2015 to 2020; and the local spatial autocorrelation analysis showed that the high incidence of human brucellosis was mainly concentrated in the north of Shandong Province.Conclusions:The incidence of brucellosis in Shandong Province is mainly concentrated in spring and summer, most of them are farmers, and the high incidence areas have spatial clustering. Key prevention and control measures should be taken for high incidence seasons, high-risk population and northern high clustering areas to reduce the incidence of brucellosis.