Objective To explore the factors influencing pneumococcal vaccination uptake among the el-derly people in Guangzhou. Methods A survey by questionnaire was performed among 827 subjects aged 60 years or above and living in Guangzhou for five consecutive years. Chi-square test and multiple logistic regression analysis were applied to identify factors influencing pneumococcal vaccination uptake among the elderly. Results The positive factors for vaccination uptake among the elderly people included age of over 70 years (OR=1.677, 95%CI: 1.156 ~ 2.434), mental workers (OR = 1.837, 95%CI: 1.214 ~ 2.779), education background of over-three-year-course training (OR=1.769, 95%CI:1.039~3.012), and history of chronic diseases (OR=1.659, 95%CI:1.096~2.512) were positively associated with pneumococcal vaccination uptake. Monthly disposable income was not an influencing factor (OR=1.420, 95%CI: 0.895 ~ 2.251). Conclusion Strengthened publicity of pneumo-coccal vaccination among the elderly people and flexible measures tailored to the needs of different groups are rec-ommended in order to improve pneumococcal vaccination uptake among the elderly people.

Aim To construct eukaryotic expressing plasmid of hi FGF2 ( high molecular weight isoform fi-broblast growth factor-2,hi FGF2) gene and to investi-gate its effect on apoptosis after its overexpression in HEK293 cells. Methods The DNA template primer was designed and synthesized. The pDsRed1-N1 plas-mids were digested by the restriction enzymes of Nhel and Hind III. The hi FGF2 was ligated with linearized pDsRed1-N1 by T4 DNA Ligase. The recombinant plasmid was identified by endonuclease digestion and sequenced. The recombinant hi FGF2 plasmid was transient transfected into HEK293 cells by Lipofectami-neTM 2000 Reagent. The transfection efficiency was de-tected by fluorescence inversion microscope. The cell apoptosis was detected by Annexin V-FITC/PI apopto-sis detection kit with flow cytometry analysis. Results The pDsRed1-N1 eukaryotic expression vector was consistent with the design. The recombinant hi FGF2 plasmid was transfected in HEK293 cells. The trans-fection rate was more than 70%. The FITC/PI dyeing rate in hi-FGF2 over-expression HEK297 cells was a-bout ( 29. 12 ± 2. 81 )%. Conclusions pDsRed1-N1 eukaryotic expression vector is successfully constructed and transfected into HEK293 cells. Over-expression of hi FGF2 induces cell apoptosis.

OBJECTIVETo explore a simple and reliable method for intraperitoneal injection through a paravertebral approach in rabbits.

METHODSSixty New Zealand rabbits were randomized into conventional group and modified groups to receive intraperitoneal injections through conventional and paravertebral approaches, respectively. In the conventional group, the injection site was on the abdominal wall 3~4 cm lateral from the umbilicus bilaterally, while that in the modified group was located dorsally at L5/L6 level 3-4 cm lateral from the midline. Abdominal CT scan was performed in the post-injection rabbits, which were sacrificed after 24 h for abdominal dissection.

RESULTSSuccess with a single puncture was achieved in 13 out of the 20 rabbits in the conventional group, and the rest required at least two punctures, with a mean rank sum of 23.50. With the modified approach, a single attempt was successful in all the 40 rabbits, with a mean rank sum of 34.0, showing a significant difference between the two groups (P<0.01). The success rates of a single injection differed significantly between the two groups (P<0.01). CT scan and abdominal dissection showed that the injection site with the modified approach was far away from the vital organs and large vessels with less peritoneal hyperemia and exudation.

CONCLUSIONParavertebral intraperitoneal paracentesis is a convenient and reliable method for intraperitoneal injection in rabbits.

Animals ; Injections, Intraperitoneal ; methods ; Rabbits

Objective To explore a simple and reliable method for intraperitoneal injection through a paravertebral approach in rabbits. Methods Sixty New Zealand rabbits were randomized into conventional group and modified groups to receive intraperitoneal injections through conventional and paravertebral approaches, respectively. In the conventional group, the injection site was on the abdominal wall 3~4 cm lateral from the umbilicus bilaterally, while that in the modified group was located dorsally at L5/L6 level 3-4 cm lateral from the midline. Abdominal CT scan was performed in the post-injection rabbits, which were sacrificed after 24 h for abdominal dissection. Results Success with a single puncture was achieved in 13 out of the 20 rabbits in the conventional group, and the rest required at least two punctures, with a mean rank sum of 23.50. With the modified approach, a single attempt was successful in all the 40 rabbits, with a mean rank sum of 34.0, showing a significant difference between the two groups (P<0.01). The success rates of a single injection differed significantly between the two groups (P<0.01). CT scan and abdominal dissection showed that the injection site with the modified approach was far away from the vital organs and large vessels with less peritoneal hyperemia and exudation. Conclusion Paravertebral intraperitoneal paracentesis is a convenient and reliable method for intraperitoneal injection in rabbits.

Objective To explore a simple and reliable method for intraperitoneal injection through a paravertebral approach in rabbits. Methods Sixty New Zealand rabbits were randomized into conventional group and modified groups to receive intraperitoneal injections through conventional and paravertebral approaches, respectively. In the conventional group, the injection site was on the abdominal wall 3~4 cm lateral from the umbilicus bilaterally, while that in the modified group was located dorsally at L5/L6 level 3-4 cm lateral from the midline. Abdominal CT scan was performed in the post-injection rabbits, which were sacrificed after 24 h for abdominal dissection. Results Success with a single puncture was achieved in 13 out of the 20 rabbits in the conventional group, and the rest required at least two punctures, with a mean rank sum of 23.50. With the modified approach, a single attempt was successful in all the 40 rabbits, with a mean rank sum of 34.0, showing a significant difference between the two groups (P<0.01). The success rates of a single injection differed significantly between the two groups (P<0.01). CT scan and abdominal dissection showed that the injection site with the modified approach was far away from the vital organs and large vessels with less peritoneal hyperemia and exudation. Conclusion Paravertebral intraperitoneal paracentesis is a convenient and reliable method for intraperitoneal injection in rabbits.

OBJECTIVETo detect the most prevalent mutations, R778L and P992L of ATP8B gene, in Chinese Wilson disease(WD) patients by high resolution melting (HRM) analysis after polymerase chain reaction (PCR).

METHODSGenomic DNA was extracted from peripheral blood samples obtained from 30 cases of WD by the standard phenol/chloroform method. DNA fragments encompassing ATP7B exons 8 and 13 were produced by PCR amplification. The amplicons containing the R778L or P992L mutations were then generated by nested PCR. The nested PCR products were subjected to HRM analysis using the HR-1 instrument. Mutations detected in HRM analysis were verified by restriction analysis using restriction enzyme (MspI or AluI or AfaI) or DNA sequencing.

RESULTSHRM analysis of the fragments encompassing ATP7B exon 8 showed four curve patterns. Subsequent restriction analysis and DNA sequencing proved that the four different curves represent four different genotypes: the wild type, the R778L/R778L homozygote, the R778L heterozygote, and the R778L/752.33delG compound heterozygote. Three HRM curve patterns were observed for the fragments encompassing ATP7B exon 13, representing the wild type, the P992L heterozygote, and the P992L/S975Y compound heterozygote. In our studied samples, allele frequencies of the R778L, P992L and S975Y mutations were 25%, 15% and 1.67%, respectively.

CONCLUSIONHRM analysis is a simple, accurate and sensitive approach for rapid detection of the ATP7B mutations and could be used as an optimized method for genetic testing in WD.

Adenosine Triphosphatases ; genetics ; Base Sequence ; Cation Transport Proteins ; genetics ; Copper-transporting ATPases ; DNA ; genetics ; metabolism ; DNA Mutational Analysis ; methods ; DNA Restriction Enzymes ; metabolism ; Exons ; genetics ; Freezing ; Gene Frequency ; Genotype ; Hepatolenticular Degeneration ; genetics ; Humans ; Nucleic Acid Denaturation ; Polymerase Chain Reaction ; Time Factors

OBJECTIVETo examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease.

METHODSRat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain.

RESULTSThe model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models.

CONCLUSIONCerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

Alzheimer Disease ; pathology ; Amyloid beta-Peptides ; chemistry ; Animals ; Brain ; pathology ; Cerebral Amyloid Angiopathy ; pathology ; Disease Models, Animal ; Rats ; Staining and Labeling

Objective To examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease. Methods Rat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain. Results The model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models. Conclusion Cerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

Objective To examine cerebral pathologies in cerebral amyloid angiopathy in a rat model of Alzheimer's disease. Methods Rat models of Alzheimer's disease was established by stereotactic Aβ1-42 fiber injection in the bilateral hippocampus. The cognitive function of the rats was evaluated with water maze test. HE staining, Congo red staining and double-labeling indirect immunofluorescence were used to examine the dynamic distribution of Aβ fiber deposit in the brain. Results The model rats showed significant differences from the control rats in the escape latency and the times of crossing platform in waster maze test. HE staining revealed a decreased number and degeneration of the granular cells with increased glial cells in the model rats. Congo Red staining showed that the Aβ fiber was deposited gradually in the small vessels in the brain parenchyma to cause thickening, stenosis or occlusion of the small vessels. Immunofluorescence staining detected Aβ fiber migration from the parenchyma to the walls of the small arteries in the rat models. Conclusion Cerebral amyloid angiopathy is a major pathological feature in Alzheimer's disease.

OBJECTIVETo identify the causative mutation in a Chinese family affected with dentinogenesis imperfecta shields type II (DGI-II).

METHODSWith informed consent obtained from all participants, peripheral blood or chorionic villi samples were collected from the family members. Genomic DNA was extracted using a standard SDS-proteinase K-phenol/chloroform method. The whole coding region and exon/intron boundaries of the DSPP gene were amplified with polymerase chain reaction (PCR) and subjected to Sanger sequencing. To confirm the pathogenicity of the identified mutation, an Alu I recognition sequence was introduced into the mutant allele using mismatch primers by semi-nested PCR. Restriction fragment length polymorphism (RFLP) analysis was then carried out for all family members and 60 unrelated healthy controls. Meanwhile, mini-DSPP constructs were conducted to confirm the effect of the mutation in vitro.

RESULTSA splicing site mutation, c.52-1G>A, which was located upstream of exon 3, was found in all three patients and the fetus of the proband. Restriction analysis confirmed that all unaffected individuals and the 60 healthy controls did not carry the same mutation. The expression of minigene showed that the exon 3 of the DSPP gene was skipped during the transcription.

CONCLUSIONA novel pathogenic splicing-mutation c.52-1G>A has been detected in a Chinese family affected with DGI-II, which enabled prenatal diagnosis for the fetus of the proband.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child, Preschool ; Dentinogenesis Imperfecta ; genetics ; Exons ; Extracellular Matrix Proteins ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Phosphoproteins ; genetics ; Point Mutation ; RNA Splicing ; Sialoglycoproteins ; genetics