Objective To investigate the relationship of apolipoprotein E (ApoE) allelic frequency and serum lipid levels in patients with Alzheimer′s disease (AD). Methods DNA microarray was used to detect the ApoE genotypes of AD patients (n = 200) and age-matched non-demented elderly control subjects (n = 159). Serum lipid levels was measured by Immunoturbidimetric assay at the same time. We analyzed the ApoE genotype distribution and the relationship of apolipoprotein E ( ApoE ) allelic frequency and serum lipid levels . Results The ApoE ε4 allelic frequencies (25.5%) in AD group is higher than that of the control group (7.9%) (P < 0.05). The ε2 allele was associated with a higher ApoE concentration, whereas with a mediate concentration in ε3 and the lowest concentration ( P < 0 . 05 ) in ε4 . Serum concentrations of ApoE showed no significant difference between AD patients and the healthy population who were with the same genotype (P > 0.05). Conclusion The ApoE levels are negatively related to ApoE ε4 allele frequency and have no significant differences with the same genotype in AD and the control group,which suggests that lower serum ApoE levels in AD patients is caused by higher ApoE ε4 allelic frequency in AD than in healthy population.

Objective To investigate the application value of apolipoprotein E (ApoE) genotyping by DNA microarray technology and the relationship between ApoE allelic frequency and serum ApoE levels in both healthy individuals and patients with Alzheimer ′s disease (AD).Methods This research is case-control study.DNA microarray was used to detect the ApoE genotypes of AD patients (n =280) and age-matched non-demented elderly control subjects ( n =230) .The cases and controls were collected in Guangzhou Huiai Hospital during July 2014 to September 2015.The accuracy of genotype results was verified by DNA sequencing.Serum ApoE levels were measured by immunoturbidimetric assay .The ApoE genotype distribution and the relationship between ApoE allelic frequency and serum ApoE levels were analyzed.The “t” test was used to compare the ApoE levels of AD patients and controls , variance analysis was used to analyze ApoE levels in the persons with different genotype .Results DNA microarray technology genotyping results were completely consistent with the results of DNA sequencing .In AD group, the ApoE genotype distribution were 2.9%(8 /280) for ε2ε3, 1.8% (5/280) for ε2ε4, 46.8% (131/280)for ε3ε3,45.4%(127 /280) for ε3ε4 and 3.1%(9 /280) for ε4ε4.While in the control group, the ApoE genotype distribution were 0.9%(2 /230) for ε2ε2, 12.6% (29/230)for ε2ε3, 1.3%(3 /230) forε2ε4, 70.0% (161 /230) for ε3ε3 and 15.2% (35 /230) for ε3ε4.The average serum concentrations of ApoE were (33.29 ±10.87)mg/L in AD patients and (41.28 ±10.95)mg/L in the controls.Among all participants, the average serum levels of ApoE were (50.86 ±6.21) mg/L for ε2 carriers, (38.78 ± 12.07)mg/L for ε3 carriers and (30.47 ±7.68)mg/L for ε4 carriers.In AD group,ApoE level of ε2, ε3,ε4 carriers is (50.31 ±9.08)mg/L, (38.30 ±7.60) mg/L and (32.86 ±5.93)mg/L respectively.In the control group, the ApoE level of ε2, ε3, ε4 carriers is (51.00 ±5.53)mg/L, (41.01 ±10.09)mg/L and (32.86 ±5.93)mg/L respectively.The ApoE levels of persons with different ApoE alleles are ε2 >ε3 >ε4. The difference is significant (F =89.6, P <0.05).However, the ApoE levels in persons with the same ApoE genotype between healthy individuals and AD patients have no significant difference ( t =0.981, 2.878 and 1.732 respectively, P >0.05) .Conclusions DNA microarray technology possesses high efficiency and favorable accuracy.The ε2 allele is associated with a higher ApoE concentration , ε3 allele with a mediate concentration and ε4 allele with a lowest concentration.Serum concentrations of ApoE showed no significant difference between AD patients and the healthy groups who have the same genotype .The primary cause of the low serum ApoE levels in AD patients is that the ApoE ε4 allelic frequencies of them are higher than that of the healthy persons.

Molecular biological methods were used to construct the eukaryotic expression vectors of precore and core gene named EBO PreC/C. Then T1862, A1896, A1899 and A1896+A1899 variants were constructed by site mutagenesis in vitro. By PCR RFLP and sequencing ,T1862,A1896,A1899 and A1896+A1899 variants were obtained. Wild type and variants were transfected to HepG2 cells, and HBeAg was tested to observe the difference of HBeAg expression between wild type and variants. After stable expression in HepG2 cells, HBeAg was detected to be positive in cells transfected wild type and A1899 variants, and negative in cells transfected with T1862, A1896,A1896+A1899 variants. The construction of these variants will play an important pole in studying the relation of PreC/C mutations and HBV expression and replication of HBV genome.

Objective To study HBeAg change in patients infected hepatitis B virus(HBV) with pre-C signal enzyme cleavage site mutation. Methods Mutation in pre-C signal enzyme cleavage site was detected by PCR-RFLP. The PreC/C gene with mutation was amplified by PCR and was cloned to EB viral eukarotic expression vector. Then transfect the vector with wild type or mutant PreC/C gene to HepG2 cell. SEAP reporter system was used to monitor the efficiency of transfection. HBeAg and its precursor in the supernatant and HepG2 cell were detected by ELISA and Western blot. Results HBeAg was positive in the supernatant of wild type and negative control in T1862 vaniant by ELISA. In HepG2 cell transfected with wild type, three proteins were detected by Western blot, they were HBeAg(17 000) and two HBeAg precursor(22 000 and 25 000). And in HepG2 cell transfected T1862 vaniant, only two HBeAg precursor was detected. The precursor in cells transfected withT1862 vaniant were significantly stronger than cells transfected with wild type. Conclusion Mutation in pre-C signal enzyme cleavage site may affect the decoration of HBeAg, which may cause great of HBeAg precursor locating in cells and lead to HBeAg negative in serum of patients infected with HBV.

Objective To investigate the value of percutaneous transhepatic cholangioscopy (PTCS) for diagnosis and treatment of biliary cast after liver transplantation. Methods Data of 11 patients with biliary cast after liver transplantation, who underwent PTCS from April 2008 to November 2010, were retrospectively analyzed. Results In 11 patients , one had biliary cast in common bile duct, 3 in right intra-hepatic bile duct, 4 in left intra-hepatic bile duct, and 3 distributed in intra- and extra-hepatic bile ducts. A total of 68 times of PTCS were performed in 11 patients, achieving significant decrease in levels of serum transaminase and bilirubin in 10. Occasional fever occurred in 1 patient after closure of drainage tube,which was managed by replacement with a thinner one. There were no severe complications such as biliary fistula or uncontrollable bleeding. Partial rupture of fistula occurred in 1 case. All patients were followed up for 10-30 months and were all in good condition except one patient died from other disease during the followup. Conclusion PTCS is a safe, effective and applicable method to treat the biliary cast after liver transplantation.

OBJECTIVE: To assess the expression of NGF, BDNF, NT-3 mRNA in the peripheral blood of patients with allergic rhinitis (AR). Meanwhile, to analysis whether the expression of NGF, BDNF, NT-3 mRNA correlate with the severity of rhinitis. METHOD: This study is a group controlled trial, which takes the healthy adults as control group. The total RNA have been extracted from the peripheral blood of AR patients. The expression of NGF, BDNF and NT-3 mRNA have been tested by real-time quantitative RT-PCR. RESULT: Comparing with the healthy adults, the expression of NGF, BDNF and NT-3 mRNA as 2(-deltadeltaCt) are 2.436 8, 4.4588 and 1.781 8 respectively. The increasing expression of NT-3 correlated positively with the scores of visual analog scale. CONCLUSION The expression of NGF, BDNF and NT-3 mRNA are as high as 2.4368, 4.4588 and 1.7818 times to healthy adults. We propose NGF, BDNF and NT-3 may contribute to the pathogenesis of AR. NT-3 could reflect the severity of rhinitis as a molecular biological index.
Adolescent ; Adult ; Brain-Derived Neurotrophic Factor ; blood ; genetics ; Child ; Female ; Humans ; Male ; Nerve Growth Factor ; blood ; genetics ; Neurotrophin 3 ; blood ; genetics ; RNA, Messenger ; genetics ; Rhinitis, Allergic ; blood ; Young Adult

Objective:To investigate the correlation between inositol level and glycolipid metabolism in gravidas with gestational diabetes mellitus (GDM).Methods:A cross-sectional study was conducted on 80 GDM cases undergoing routine examination at Fujian Provincial Maternity and Children's Hospital from November 2018 to August 2019, who were selected as GDM groups. Another 50 women with uncomplicated pregnancies during the same period were selected as the control group. Blood and urine inositol level and serum glycolipid profiles were compared between the two groups, and their association was analyzed. Independent or paired-sample t test, Mann-Whitney test, Chi-square (or Fisher's exact) test, and Pearson correlation test were performed for statistical analysis. Results:The serum inositol concentration and high-density lipoprotein (HDL) cholesterol in the GDM group were significantly lower [322.1 ng/ml (279.1-364.1 ng/ml) vs 403.8 ng/ml (391.8-425.3 ng/ml), Z=-7.879; 1.8 mmol/L (1.5-2.0 mmol/L) vs 2.0 mmol/L (1.7-2.2 mmol/L), Z=-2.419; both P<0.05], while the concentration of urine inositol, lipoprotein-a (lipo-a), 0 h-, 1 h-, 2 h-oral glucose tolerance test (OGTT) glucose, fasting insulin, and glycosylated hemoglobin (HbA1c) were significantly higher when comparing to the control group [192.2 ng/ml (171.0-219.9 ng/ml) vs 143.8 ng/ml (121.1-158.6 ng/ml), Z=-6.834; 253.2 mg/L (65.0-349.0 mg/L) vs 148.5 mg/L(46.5-159.3 mg/L), Z=-0.187; 5.0 mmol/L (5.1-5.6 mmol/L) vs 4.4 mmol/L (4.2-4.6 mmol/L), Z=-5.547; 10.0 mmol/L (9.1-11.3 mmol/L) vs 7.8 mmol/L (7.0-8.4 mmol/L), Z=-6.987; 8.6 mmol/L(7.6-9.4 mmol/L) and 6.6 mmol/L (5.7-7.1 mmol/L), Z=-7.100; 18.2 mU/L(10.6-25.9 mU/L) vs 11.0 mU/L (6.3-12.7 mU/L), Z=-4.537; 5.4%(4.5%-5.5%) vs 5.1%(4.9%-5.4%), Z=-3.468; all P<0.05]. (2) Serum inositol concentration was negatively correlated with fasting insulin and 0 h-, 1 h-, 2 h- OGTT glucose level ( r=-0.386, -0.416, -0.350 and -0.407, respectively); urinary inositol concentration was positively correlated with 0 h-, 1 h-, 2 h-OGTT glucose levels ( r=0.402, 0.389 and 0.429, respectively) (all P<0.05). Conclusions:Serum inositol concentration was decreased, and urinary inositol excretion was increased in women with GDM. Measurement of changes in inositol levels during the second trimester may be helpful to assess the metabolic status of pregnant women.

Objective To evaluate the possibility of deep vein thrombosis defluvium of lower limb using ultrasonography to provide important reference for the implantation and retrieve of retrievable inferior vena cave (IVC ) filters.Methods Sixty-four patients who were diagnosed as lower limb deep vein thrombosis using ultrasonography were enrolled.Fourteen patients with critical values were allocated to high risk group,50 patients without critical value to low risk group.All the patients underwent inferior vena cava filter operation.Of all these patients,ultrosonography were performed once again before IVC filters were retrieved.Seventeen patients with broken thrombosis were allocated to unstable group,47 patients with stable thrombosis to stable group.Relationships between conditions of thrombosis and ultrasonography results were analysed.The conditions of thrombosis were recorded.The rates of thrombosis defluvium were compared.Results In high risk group,10 patients (71 .43%)had thrombosis.In low risk group,8 patients (16%)had thrombosis.There were significant differences in the rates of thrombosis defluvium beteen the high risk group and low risk group(P <0.05).In unstable group,12 patients (70.59%)had thrombosis.In stable group,6 patients (12.77%) had thrombosis,there were significant differences in the rates of thrombosis defluvium between the unstable group and stable group (P < 0.05 ).Conclusions Ultrasonography can be used to evaluate the possibility of lower extremity deep vein thrombosis defluvium which will guide the pratice of the retrievable inferior vena cava filter.

ABSTRACT:Objective To analyze the correlation between plasma nitric oxide (NO)level and atherosclerotic lesion in high cholesterol-fed (HCD ) rabbits.Methods Twenty male Japanese white rabbits were divided randomly into two groups,which were fed with normal diet (control group,n =10)or HCD (experimental group, n =10 )for 1 6 weeks.At the end of the experiment,plasma lipid and NO levels were measured.The gross atherosclerotic lesions in each group were detected by Sudan IV staining while intimal lesion area was measured by hematoxylin/eosin (HE)and elastica van gieson (EVG).Moreover,the macrophages (MΦ)and smooth muscle cells (SMC)were detected by immunohistochemical staining.The correlation analysis was made to reveal the relationship between atherosclerotic lesions and plasma NO level.Results Compared with those in control group, the total cholesterol (TC),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C)and NO levels all increased significantly in experimental group.Atherosclerotic lesions appeared on the vascular wall in the latter group.The area of atherosclerotic lesions and MΦ in the plaque had a positive association with plasma NO level. Conclusion There is a relationship between plasma NO level and the size of HCD-induced atherosclerotic lesions in rabbits.Meanwhile,the MΦ positive area in the atherosclerotic plaque is also associated with plasma NO level in cholesterol-fed rabbits,suggesting that plasma NO level may be associated with the occurrence and progress of early atherosclerosis.

Objective To investigate the effect of allicin on the development of atherosclerosis in apoE-/-mice and explore its underlying mechanism from the perspective of protein S-nitrosylation.Methods Thirty male apoE-/- mice were randomly divided into 3 groups:control group (saline,ig),low-dose group (allicin,9 mg/kg·d, ig)and high-dose group (allicin,18 mg/kg·d,ig).They were fed with high cholesterol diet for 12 weeks.The levels of plasma lipids,oxidized-LDL (ox-LDL),malondialdehyde,tumor necrosis factor-alpha and nitric oxide (NO)were measured.The atherosclerotic lesions in aortic root were evaluated after hematoxylin and eosin staining and elastica van Gieson and immunohistochemical staining,respectively.Furthermore,in vitro experiments were performed using human umbilical vein endothelial cells (HUVECs).The HUVECs were treated with allicin (10μmol/L or 20 μmol/L)for 24 hours in the presence of ox-LDL (50 μg/mL).The level of NO in supernatant was measured by a nitrate/nitrite assay. The protein S-nitrosylation of the HUVECs was detected through immunofluorescence.Results The histological analysis revealed that allicin treatment not only significantly decreased the areas of the atherosclerotic lesion (all P <0.05)but also suppressed the macrophage accumulation and smooth muscle cell proliferation in the lesion.There was no significant difference in the levels of plasma lipids between control and treated groups.However,allicin exerted obvious anti-oxidative and anti-inflammatory effects. Interestingly,the allicin treatment led to marked increase of the plasma NO level (P <0.05)and aortic protein S-nitrosylation.The experiments in vitro further proved that the allicin up-regulated the levels of NO and protein S-nitrosylation in HUVECs treated with ox-LDL (P < 0.01 ).Conclusion Allicin can inhibit the development of atherosclerosis.The mechanism is associated with the up-regulation of protein S-nitrosylation in endothelial cells, which plays an important role in anti-oxidization and anti-inflammation.