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.

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 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.

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 compare the feasibility and safety of pancreaticogastrostomy versus pancreaticojejunostomy after pancreaticoduodenectomy. Methods A retrospective study was performed on 37 patients who underwent pancreaticoduodenectomy for duodenal carcinoma or pancreatic head tumors at the First Hospital of Sun Yat-sen University from April 2006 to December 2010.Pancreatic anastomosis was carried out either using pancreaticogastrostomy (n＝ 19) or pancreaticojejunostomy (n=18).The operative time,intraoperative bHood Hoss,postoperative pancreatic Heak,mortaHity and Hength of hospitaH stay were compared between the two groups. ResuHts The mean operative time,intraoperative bHood Hoss,incidence of pancreatic fistuHa,mortaHity rate and mean Hength of postoperative hospitaH stay were (372.1 ±79.5) min vs (351.0±69.2) min; (693.5± 412.8) mH vs (645.1±488)ml; 10.5％ (2/19) vs 11.1％ (2/18); 5.3％ (1/19) vs 5.6％ (1/18); and (17.5± 8.9)d vs (16.1± 7.6)d,respectively.The differences between the two groups were not statistically significant.Conclusion Pancreaticogastrostomy appears to be a feasible and safe alternative to pancreaticojejunostomy for the pancreatic remnant after pancreaticoduodenectomy.

OBJECTIVE: To detect the expression of NGF, BDNF, NT-3 mRNA in the peripheral blood of patients with allergic rhinitis (AR). And to analyze the correlation between NGF, BDNF, NT-3 mRNA expression and the epidsode of rhinitis through Th-2 Hypothesis. METHOD: This study was a group controlled trial. The expression of NGF, BDNF and NT-3 mRNA were tested by real-time quantitative RT-PCR and the concentrations of IL-4, IL-6, IL-10 and INF-alpha were tested by ELISA. RESULT: The expression of NGF, BDNF and NT-3 mRNA in AR patients were 2.44, 4.46 and 1.78 times the amount of those in the healthy adults, respectively. The increased expression of NT-3 correlated positively with the scores of visual analog scale of AR. The concentrations of IL-4, IL-6 and IL-10, which were 2198 +/- 472 pg/mL, 9407 +/- 703 pg/mL and 3917 +/- 323 pg/mL respectively, were higher than those in the healthy adults. The concentration of INF-alpha was 2198 +/- 472 pg/mL and less than the healthy adults. The increased expressions of NGF, NT-3 were positively related to the increase of IL-4, IL-6 and IL-10. CONCLUSION The expressions of NGF, BDNF and NT-3 mRNA in AR patients are higher than those in the healthy adults. NGF, BDNF and NT-3 may contribute to the pathogenesis of AR. Moreover, NGF and NT-3 may induce the episode of rhinitis through Th-2 Hypothesis.
Adolescent ; Adult ; Brain-Derived Neurotrophic Factor ; blood ; Case-Control Studies ; Child ; Female ; Humans ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Nerve Growth Factor ; blood ; Nerve Growth Factors ; blood ; genetics ; Neurotrophin 3 ; blood ; RNA, Messenger ; genetics ; Rhinitis, Allergic ; blood ; immunology ; Th1-Th2 Balance ; Young Adult

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 effect of autophagy inhibitor 3-methyladenine (3-MA) on phenotype transformation and proliferation of rat pulmonary arterial smooth muscle cells (PASMCs).Methods Cultured PASMCs were treated with different concentrations of 3-MA (low-dose group,2.0 mmol/L;middle-dose group,4.0 mmol/L;high-dose group,8.0 mmol/L;control group,0 mmol/L).The protein expression of LC3 Ⅱ,OPN and Vimentin was detected by Western blotting.Cell proliferation was detected by MTT assay.Results The autophagy of PASMCs was decreased with the increase of the concentration of 3-MA.Compared with the control group,significantly down-regulated protein expression of LC3 Ⅱ,OPN and Vimentin was observed in 3-MA-treated cells,with significantly lower proliferation activity.Conclusion The autophagy inhibitor 3-MA significantly down-regulated the expression of LC3 Ⅱ,OPN and Vimentin in PASMCs,with inhibiting the proliferation of PASMCs.