This work was directed at obtaining a better gene carrier to improve the effects of gene delivery. Neutral liposomes made from cholesterol, lecithin and DOPE by reverse evaporation technique were used for encapsulating DNA-HAP complex which was made from DNA and optimized HAP. The sizes of complexes and the efficiency of encapsulation were detected. The efficiency of transfection into Hela cells was shown by observation of X-gal staining and measurement of transfection efficience. The average size of complexes was 643nm, the average encapsulating efficiency of DNA in microspheres reached 11.67%. These Lipid-Hydroxyapatite-DNA complex (LHD) could be transfected into mammalian cells. The Lipid-Hydroxyapatite-DNA complex prepared by reverse evaporation technique could be applied availably in DNA delivery system, and it gave another thinking to increase the gene transfection of non- viral genetic vector.
DNA ; administration & dosage ; Durapatite ; administration & dosage ; Genetic Therapy ; Lipids ; administration & dosage ; Transfection ; methods

OBJECTIVETo monitor the 3-dimensional (3D) morphological changes of C6/36 cells during dengue virus (DENV) infection using a live-cell imaging technique based on digital holographic microscopy and provide clues for better understanding the mechanisms of DENV infection.

METHODSC6/36 cells were seeded in 6-well plates to determine the optimal imaging density under a holographic cell imager, and the morphological changes of the cells were recorded in response to a culture temperature change from 28 degrees celsius; to 37 degrees celsius; C6/36 cells were infected with 4 DENV strains with different serotypes at 28 degrees celsius; and incubated at 37 degrees celsius; for 24 h, and the 3D holograms and relevant morphological parameters were recorded at different time points using HoloMonitor M4 holographic cell imaging and analysis system.

RESULTSThe holograms of C6/36 cells inoculated at the optimal density for imaging (4×10per well) showed unified 3D morphologies of the single cells with minimal dispersions in the cell area, thickness and volume (P<0.05), which did not undergo obvious changes when the cells were incubated at 37 degrees celsius; for 24 h (P>0.05). The cell area and volume of the cells infected with the 4 DENV strains all increased and the cell thickness was reduced during incubation. Among the 4 strains, DENV-1 and DENV-2 caused reduced cell thickness while DENV-3 and DENV-4 increased the cell thickness, and the pattern and degree of such changes differ among the 4 strains.

CONCLUSIONSDigital holographic microscopy allows monitoring of the complex morphological changes of cells during DENV infection. The 4 DENV strains with different serotypes causes characteristic cell damages during infection.

Objective To identify the presence of candidate pathogenicity island 89K DNA sequence of Streptococcus suis serotype 2 (SS2) strains isolated from patient in Zhejiang province. Methods Genes and DNA fragments were amplified by PCR, using specific primers, and three amplified fragments of the89K sequence were directly sequenced. The results were analyzed using software related to bioinformaties and epidemiology. Results 8 strains of SS2 all contained 89K sequence, cps2J and mrp virulent genes, and species-specific 16S rDNA. 3 amplified fragments of 89K candidate pathogenicity island of SS2 ZJ0501 were above 99% similar to SS2 strain identified from outbreaks in Jiangsu in 1998, and the gene fragment of coding DNA recombinant protein in the 89K sequence was highly homological with that of S. dysgalactiae and S. agalactiae. Conclusion In recent years SS2 strains isolaed from patients with clinical symptoms in Zhejiang province had been detected to have contained candidate pathogenicy 89K DNA fragment.

OBJECTIVE: To establish a simple, fast and economical technique for multiplex-typing SNPs and to explore its potential forensic application. METHODS: Five Y-SNP loci (IMS-JST164520, IMS-JST021354, IMS-JST003305, M119 and M134) were selected and the allele specific primers of each locus were designed with the universal reporter primers tailed at their 5' end. Alleles of these loci were amplified first by allele specific primers, then amplified by universal reporter primers tagged by fluorescent dye. RESULTS: A fluorescent-multiplex PCR system of the five Y-SNP loci was established. The typing results showed that two different colors of product peaks denoted two different alleles of a SNP locus, and the fragment sizes of alleles among different SNP loci were different. The haplotype diversity of these five loci was estimated to be 0.8655 in Wuhan Han population. CONCLUSION The multiplex-typing SNPs based on the universal reporter primers is a simple, fast, and economical technique, and may have good application value in forensic medicine.
Alleles ; Asian People/genetics* ; China/ethnology* ; Chromosomes, Human, Y/genetics* ; DNA Primers ; Female ; Forensic Genetics ; Gene Frequency ; Genetic Markers ; Genetics, Population ; Haplotypes ; Humans ; Male ; Nucleic Acid Amplification Techniques/methods* ; Polymorphism, Single Nucleotide/genetics*

Purpose To analyze the MMP-9,MMP-13,HIF-1α expression in lung adenocarcinoma tissue and to explore the relationship with clinical pathologic features,EGFR mutation and prognosis of the patients.Methods The expression of MMP-9,MMP-13,HIF-1α in 629 cases of lung adenocarcinoma were detected by using immunohistochemical of SP method.50 cases of normal tissue adjacent to carcinoma and 50 cases of pneumonia pseudotumor hyperplasia tissues were selected as controls.629 patients with lung adenocarcinoma were detected by real-time fluorescence quantitative PCR and all of them were followed-up.Results The MMP-9,MMP-13,HIF-1α expression in lung adenocarcinoma tissues were higher than controls (P ＜ 0.001).The MMP-9 expression was correlated with lymph node metastasis and the size of the tumor (P ＜ 0.05).The MMP-13 expression was correlated with smoking history,TNM stage,lymph node metastasis and the size of the tumor (P ＜0.05).The HIF-1α expression were correlated with smoking and lymph node metastasis.Statistically significant differences were found in all these above groups (P ＜ 0.05).But the expression of all has nothing to do with the EGFR mutation (P ＞0.05).The Kaplan-Meier survival analysis results showed that MMP-9,MMP-13,HIF-1α positive expression groups of 12,24 and 36 months cumulative survival were significantly lower than the negative expression groups.COX multiple factor analysis results showed that EGFR mutation,the expression of MMP-9,MMP-13,HIF-1α,tumor size,TNM stage were independent risk factors for the development of lung adenocarcinoma patients living conditions.Statistically significant differences were found in these groups (P ＜0.05).Conclusion MMP-9,MMP-13,HIF-1α are overexpressed in lung adenocarcinoma tissues,and its expression are associated with lymph node metastasis,but has nothing to do with EGFR mutations.Patients with positive expression of MMP-9,MMP-13,HIF-1α and with no EGFR mutation have lower survival rates,and they are independent risk factors for the development of lung adenocarcinoma patients living conditions.

Objective To evaluate the characteristics of and correlation between the inclination of occlusal plane (IOP) and masticatory path in young adults with skeletal malocclusion.Methods Thirtythree individuals were eligible for this study based on the inclusion criteria.Individuals were grouped by ANB angles.There were ten,six and seventeen cases categorized as group Ⅰ ( normal occlusion with skeletal Class Ⅰ ),Group Ⅱ ( skeletal Class Ⅱ ) and Group Ⅲ ( skeletal Class Ⅲ ),respectively.The following parameters of sagittal anatomical landmarks were studied by lateral cephalometric analysis:ANB,MP-FH,OP-CP (IOP),and OP-FH.Mandibular movement of the incisal point during unilateral-sided gum-chewing was recorded by the BioEGN mandibular kinesiography analyzer and both maximal distances and direction (OP-opening/closing angles ) in sagittal plane were analyzed.Correlation between IOP and maximal distances,direction of masticatory path were analyzed,respectively.Statistical analysis was performed with PASW 17.0 software.Results IOP varied among three groups:group Ⅰ ( - 1.78 ± 5.16) °,group Ⅱ( 0.13 ± 2.31 ) °,group Ⅲ ( - 5.28 ± 4.31 ) ° ( P ＜ 0.05 ).Maximal distances of masticatory path in Group Ⅰ,Ⅱ and Ⅲ were as follow:Vertical length:(20.30 ± 8.92),( 17.60 ± 3.29),and ( 18.25 ± 4.09),Anterior-Posterior width:(2.99 ±7.90),(9.60 ±5.80) and (4.67 ±4.64) mm,Lateral width:(8.79 ±2.22),(7.11 ± 2.03 ) and ( 8.47 ± 1.70 ) mm,respectively.The variation among groups was not ofstatistical significance (P ＞ 0.05 ).OP-opening/closing angles of masticatory path in each level were found increasing from group Ⅱ,group Ⅲ to group Ⅰ.There was significant difference between Group Ⅱ and Group Ⅰ in each level of OP-opening angle.OP-opening/closing angles were not correlated with IOP in group Ⅰ and group Ⅱ ( P ＞ 0.05 ).IOP and OP-opening/closing angles were shown to be positively correlated in group Ⅲ with statistical significance (P ＜ 0.05 ).Conclusions The discrepancy of craniofacial structure has an impact on IOP.Skeletal malocclusion and IOP have no significant influence on the maximal distances of masticatory paths.The direction of masticatory path is influenced by IOP in skeletal malocclusion.

OBJECTIVETo evaluate the epidemiological and serological efficacy after 10 years of vaccination against hemorrhagic fever with renal syndrome (HFRS) vaccines in Zhejiang province.

METHODSOne county was randomly chosen as the research unit with all the healthy people between 16 and 60 years old were equally divided into study and control groups. The study group was vaccinated. Immunofluorescent antibody assay was used to test specific IgG antibody and Mcro-CPE method was used to test the titer of neutralizing antibody.

RESULTSTwo weeks after the full-course immunization, the seroconversion rate became 100% (67/67, with 95% CI as 96.3%-100%) by immunofluorescent antibody test (IgG) and 44.4% (8/18 with 95% CI as 22.0%-69.0%) by neutralization test with GMT titers as 72.1 and 4.6 respectively. Booster immunization was provided one year later. Time span as two weeks prior to, one year, one and half years, two years, three years and five years after booster immunization, the rates of seroconversion on immunofluorescent antibody using IFAT method, were 28.6%, 83.3%, 75.0%, 53.1%, 22.6%, 10.0% and 55.0% respectively, and rates of seroconversion of neutralizing antibody by Mcro-CPE method were 14.8%, 55.6%, 35.0%, 31.3%, 26.0%, 10.0% and 50.0% respectively. Nine years after the reinforcement, the rates of seroconversion of immunofluorescent antibody by IFAT method was only 7.1%. The vaccinated group had no patient seen but the control group appeared 34 patients including 3 deaths. According to the ten-year observation, the vaccine seemed effective with the protection rate in population reached 100%.

CONCLUSIONHFRS vaccine was effective on epidemiological, social and economical efficacy.

Adolescent ; Adult ; Animals ; Female ; Hemorrhagic Fever with Renal Syndrome ; epidemiology ; immunology ; prevention & control ; Humans ; Immunization, Secondary ; methods ; Male ; Middle Aged ; Rats ; Vaccination ; methods ; Viral Vaccines ; administration & dosage ; therapeutic use ; Young Adult

Adolescent ; Adult ; Aged ; Female ; Follow-Up Studies ; Humans ; Lung ; physiopathology ; Male ; Middle Aged ; Oxygen ; blood ; Radiography, Thoracic ; Severe Acute Respiratory Syndrome ; diagnostic imaging ; physiopathology ; Tomography, X-Ray Computed

OBJECTIVETo investigate the clinical usefulness of noninvasive diagnostic methods in evaluating liver fibrosis in hepatitis B virus (HBV) patients.

METHODS102 patients with chronic hepatitis B (CHB) were enrolled from Beijing Friendship Hospital Affiliated to Capital University of Medical Sciences. Noninvasive diagnostic methods including ultrasonography, CT, serum markers of liver function and fibrosis, and HBV DNA were performed and compared with histological fibrotic changes in order to establish a noninvasive method for detecting the degree of liver fibrosis.

RESULTSThe total score of liver surface, edge, parenchyma echogenicity, intrahepatic vessels, and the size of spleen had a coefficient of 0.822 with fibrotic stage. By receiver operating curve (ROC) analysis, the sensitivity to distinguish cirrhosis from CHB was 86.1% and the specificity was 95.5% if the total ultrasonic score was more than 10. The CT imaging diagnosed liver cirrhosis with a specificity of 100% and a sensitivity of 48.5%. The change of CT values in cirrhotic patients was lower than that in controls and no cirrhotic patients (F=5.805, P<0.01), when the voltage was increased from 100 KV to 140 KV. Except normal controls and S1 group, S2 and S3 group, the level of HA and collagen IV between the other groups were statistically different. The cut-off value of HA to diagnose cirrhosis was 108 (microg/L) with a sensitivity of 72.2% and a specificity of 80.3%. The cut-off value of collagen IV to diagnose cirrhosis was 188 (microg/L) with a sensitivity of 72.2% and a specificity of 78.8%. When ultrasonography was combined with serum markers, the sensitivity was 72.2% and the specificity was 80.3%.

CONCLUSIONBoth ultrasonography and serum markers are useful to diagnose cirrhosis. The combination of the two examinations is more valuable than any one alone. The characteristic CT imaging has high specificity but low sensitivity in diagnosing early cirrhosis. HA and collagen IV are correlated more closely with the stage of fibrosis, and can reflect the severity of fibrosis.

Adolescent ; Adult ; Aged ; Biomarkers ; blood ; Collagen Type IV ; blood ; Female ; Hepatitis B, Chronic ; diagnostic imaging ; pathology ; Humans ; Hyaluronic Acid ; blood ; Liver Cirrhosis ; diagnostic imaging ; pathology ; Male ; Middle Aged ; Sensitivity and Specificity ; Ultrasonography

Child ; Consensus ; Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional ; Respiratory System