Age-related macular degeneration ( AMD ) is a kind of age-related blinding degenerative fundus lesions, totally about 30 million patients suffering from AMD all over the world, with about 500 000 people blind for it yearly. As the development of economy and the aging of the population intensified, incidence of AMD indicates a trend of rising year by year, being the third major cause of blindness in our country. At present, the pathogenesis of AMD is not fully clear, as reported it may be related to oxidative stress, inflammatory immune response, VEGF and genetic manipulation. Clinical treatments mainly include photodynamic therapy, drug therapy, radiation therapy, laser photocoagulaory operation, the pupil warm treatments, Chinese medicine and intravitreous injection VEGF antagonists such as Ranibizumab, Conbercept and so on. ln this issue, we mainly expound on the progress in the epidemiological studies of AMD, especially elaborate the progress made on genetic manipulation in recent years.

Ahigh-resolutionmethodforhumanleukocyteantigen-B(HLA-B)genotypingwasestablished based on the optimized polymerase chain reaction, cloning and sequencing technology. The exon2 and exon3 of HLA-B gene were amplified with primers based on the HLA-B gene sequence. These produced heterozygous alleles were effectively cloned into plasmid DNA based on the principle of plasmid incompatibility, and were followed by bacterial culture. Then Sanger sequencing was carried out and after analyzing the result by software ClustalX2 and IMTG/HLA database comparison, the HLA-B genotype of the samples was achieved. Seven clinical samples were detected, and the results were consistent with those of PCR-SBT genotyping method. The method was cost-effective, high-resolution and it did not require technical software. The use of universal primers simplified the cumbersome design and optimization process of specific primers.

Objective To study a simple and manual contrast board in Double Contrast Examination,so as to decrease labor intensity and the X-ray influence to the work staff.Methods The integrated design consisted of two parts:filled gas and oppressive container.The gas and Barium was injected into the large intestine by manual control and its volume may be adjusted casually according to the requirement.Results It is proved in clinical application that the design is reasonable,the structure is simple and the operation is convenient.Conclusion The device makes the relationship between medical staff and patients more harmonious.It worth popularizing.

Objective To forward the concept of solution space of pharmacokinetics for studying radiophannaceutical distributions in animal models. Methods On the basis of special solutions of differential equations of pharmacokinetics, the solution space was established using the characteristics of linearly independent particular solutions and used to express the pharmacokinetics of pharmaceuticals in vivo. 0. 2 ml (7.4 MBq) 2β-carbomethoxy-3β- (4-corophenyl)-8-(2-18F-fluoroethyl) nortropane (18F-FECNT) was injected through tail vein into normal mice. The mice were sacrificed by decapitation at 5, 15, 30, 60, 120 and 180 min post-injection. Brain tissues were removed and weighed, and radioactivity was counted with the γ-counter. The solution space theory was used to study pharmacokinetics of 18F-FECNT in brain tissues of mice. Results The result showed that all solutions of pharmacokinetics models, based on differential equations, were included in the solution space. The solution of any organ or tissue could be linearly expressed by bases of the solution space. When the dimension number of the solution space was no more than 3, the solution could be directly expressed with coordinate picture. By this rule in our theory, the quantity of 18F-FECNT in brain tissues of mice changed with time, which was accorded with the experiment. The coordinates of striatum, frontal cortex, temporal cortex, occipital cortex, parietal cortex, hippocampus and cerebellum in the solution space were ( 10.13, 1.49), (4.27, 0. 84), (4.48, 0.81 ), (2.89, 0.98), (3.65, 0. 83),(3.55, 0. 98) and (2.03, 1.25 ), respectively. Conclusion The theory of solution space could be used to study pharmacokinetics of 18 F-FECNT in mice brain.

OBJECTIVETo investigate whether the ERK/FoxO3a signal axis could induce the inhibitory effect of vitexin 1 (VB-1) in HepG2 cell proliferation.

METHODThe MTT method was adopted to observe the effect of different concentrations of VB-1 on human hepatoma carcinoma cell line HepG2 and immortalized human embryo liver cell line L-02. The cell growth was assessed by the clone formation assay. The protein phosphorylation levels of ERK1/2 and FoxO3a were measured by the western blot.

RESULTVB-1 inhibited the viability of HepG2 cell line in a concentration-dependent manner, with a weak effect on L-02 cell line. VB-1 could effectively inhibit the anchorage-dependent growth of HepG2 cells, and reduce the expression levels of pERK1/2 and pFoxO3a in a concentration-dependent manner. MEK1/2 inhibitor PD98059 could enhance VB-1' s effect in inhibiting HepG2 cell proliferation and ERK1/2, FoxO3a phosphorylation.

CONCLUSIONVB-1 inhibits the proliferative activity of hepatoma carcinoma cell line HepG2 by blocking the ERK/FoxO3a signal axis.

Apigenin ; pharmacology ; Carcinoma, Hepatocellular ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; genetics ; metabolism ; Forkhead Box Protein O3 ; Forkhead Transcription Factors ; genetics ; metabolism ; Growth Inhibitors ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Signal Transduction ; drug effects

The aim of this study is to establish stable transfected cell lines which could produce SPAG4L protein labeled with Myc and His tags in vitro. The open reading frame (ORF) of human SPAG4L was amplified by PCR and the fragments were cloned into eukaryotic expression vector pcDNA3.1/myc-His(-)A. The recombined plasmids of pcDNA3.1/myc-His(-)A/SPAG4L were verified by sequencing and digestion with enzymes. Then, the recombined plasmids were introduced into HeLa cells and screened by G418. Western blotting was performed to detect the expression of SPAG4L and its tags in stable transfected cell lines. SPAG4L and its tags were expressed in the stable cell lines transfected with pcDNA3.1/myc-His(-)A/SPAG4L, but not in the control group. Further study showed that SPAG4L colocalized with the endoplasmic reticulum(ER) marker PDI by immunofluorescence. The stable transfected cell lines established in this study will provide a powerful tool for further studies such as co-immunoprecipitation and pull-down.
Carrier Proteins ; biosynthesis ; genetics ; Cloning, Molecular ; Female ; Genetic Vectors ; genetics ; HeLa Cells ; Histidine ; genetics ; Humans ; Male ; Proto-Oncogene Proteins c-myc ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Transfection

Objective To investigate the feasibility of the low tube voltage setting and personalized contrast agent application in 64-row multi-slice spiral CT pulmonary angiography.Methods Ninety patients with high risk of pulmonary artery embolism were sequentially enrolled in the study and divided into 3 groups employing completely randomized design: (1) Regular group included 30 patients using 120 kV and fixed dose of 70 ml contrast agent, (2)Another 30 patients were in 120 kV group, using 120 kV and the contrast amount was determined according to the patient weight (1.0 ml/kg), (3) The remaining 30 patients were included in 100 kV group, using 100 kV and the contrast amount was also determined according to the patient weight(1.0 ml/kg).Administration of contrast agent was completed within 20 seconds for all the patients, followed by 20 ml of saline.The objective and subjective indexes for assessing CT image quality, CT dose index volume (CTDIvol) and effective received dose (ERD) were compared between 120 kV group and 100 kV group; then the contrast media volume, injection rate, objective CT image indexes and subjective indexes for image quality was compared between the 100 kV group and regular group.The variance analysis and post hoc test were employed for the statistical analysis.Results Compared with 120 kV group(3.4± 0.7), the image quality of 100 kV group(5.2±1.8)had higher noise(52.9%), but subjective index for the image quality demonstrated no differences(q=0.272 ,P=0.063)in mediastinum window while CTDIvol and ERD decreased for 34.9%[(9.5±0.0) vs (14.6±0.0) mGy]and 36.8%[(3.8±0.6) vs (2.4± 0.4) mSv].The mean CT values on pulmonary artery of 100 kV group[(269.2±54.7) HU]were 13.4% (31.8/237.4) higher than the 120 kV group[(237.4±62.9) HU], but there was no statistical differences eornpared to normal group(q=0.172,P=0.260).Conclusion Using low kV setting (100 kV) to reduce radiation dose is proved to be effective and feasible in 64-MSCT pulmonary angiography.Personalized contrast agent injection has clinical application value for specific patient group.

Pyrosequencing is one of the important genetic polymorphism detection methods currently, but the complicated pretreatment procedure limits its application in clinical test. To simplify the whole process of pyrosequencing, on the basis of the linear_after_the_exponential_polymerase chain reaction ( LATE_PCR) , we improved the primer design method of LATE_PCR, increased the length and the concentration of the excess primer, applied direct amplification technology with whole blood, and established a whole blood_imLATE_PCR method based on common rTaq polymerase and “HpH Buffer” ( High pH buffer ) . The amplification system was optimized, and the influences of blood anticoagulant and the amount of whole blood template were investigated. The single stranded template for the pyrosequencing was obtained by PCR amplification using a single tube in one_step process, and the alcohol dehydrogenase gene polymorphisms of 24 clinical blood samples were then detected successfully. The results could be used to guide clinical individualized medication. The genotypes of ADH1B locus of 24 samples were 6 cases of AA homozygote, 14 cases of AG heterozygote, and 4 cases of GG homozygote. The genotypes of ADH1C were 20 cases of GG homozygote, 4 cases of AG heterozygote, and no cases of AA homozygote.

Objective To evaluate the relationship between serum matrix metalloproteinase-9(MMP-9), white blood cell(WBC)count and type 2 diabetes mellitus with macroangiopathy and to investigate the mechanism of the protective effects of rusiglitazone(RSG)on blood vessel.Methods Serum MMP-9 was determined by ELISA in 30 normal controls and 80 type 2 diabetic patients(including 40 cases with macroangiopathy and 40 without maeroangiopathy).WBC count and other clinical parameters were also determined.32 type 2 diabetic patients received RSG(4rag qd)for 12 weeks.All parameters were determined after 4 weeks(17 patients)and after 12 weeks(all patients)to observe the changes in MMP-9,WBC and other parameters.Results In the diabetic patients,serum MMP-9 and WBC were markedly higher as compared with normal controls;and MMP-9 and WBC in patients with macroangiopathy[579(440-949)?g/L,(7.51?1.47)?10~9/L]were higher than those [324(275-423)?g/L,(6.22?0.79)?10~9/L]in the cases without macroangiopathy(P