Effect of Ganoderma Lucidum Preparation on the Behavior,Biochemistry,and Autoimmune Parameters of Mouse Models of APP/PS1 Double Transgenic Alzheimer's Disease / 中国医学科学院学报

Objective To evaluate the efficacy of Ganoderma lucidum preparation on the behaviors,biochemistry,and autoimmunity parameters of mouse models of APP/PS-1 double transgenic Alzheimer's disease(AD).Methods A total of 44 4-month-old APP/PS-1 double transgenic AD mice were randomly divided into AD model group,Aricept group,Ganoderma lucidum middle-dose(LZ-M)group,and Ganoderma lucidum high-dose(LZ-H)group,with 11 mice in each group.In addition,10 4-month-old C57BL/6 mice were used as the control group.Water maze test was conducted to observe the behavior changes,and the protein expressions in brain tissues were detected by Western blot analysis.The autoimmune indicators were detected by indirect immunofluorescence method.Results In the navigation experiment,the time of finding the platform was gradually shortened since the 2day in the control,LZ-H,and LZ-M groups,and the time of searching the platform in the AD model group gradually increased.On the 5day,the time of finding platform was significantly shorter in control group (t=5.607,P=0.000) and LZ-H group(t=2.750,P=0.010)than AD model group.In the space exploration experiment,the number of crossing the target platform(t=2.452,P=0.025)and the residence time in the target quadrant(t=2.530,P=0.020)in AD model group mice was significantly smaller/shorter than those in control group;in addition,the number of crossing the target platform in the AD model group was significantly smaller than that in LZ-H group(t=2.317,P=0.030)and LZ-M group(t=2.443,P=0.030),while the residence time in target quadrant decreased significantly(t=2.770,P=0.020)compared with LZ-H group;the number of crossing through the target platform quadrant(t=2.493,P=0.022)and residence time in the target quadrant(t=2.683,P=0.015)in LZ-H group were significantly higher than in Aricept group.Western blot analysis showed that the expression of ApoA1 in the brain tissues of mice in LZ-H and LZ-M groups were significantly higher than those in AD model group(P<0.01,P<0.05);Aβ-40 expression in LZ-H group was significantly lower than that in AD model group(P<0.05);the expressions of Syt1,ApoE,and ABCA1 in brain tissues of mice in LZ-H group were significantly higher than those in model group(P<0.01,P<0.05).The plasma IgG level in Aricept group(t=30.945,P=0.000),LZ-M group(t=25.639,P=0.000)and LZ-H group(t=4.689,P=0.001)were significantly higher than that in the control group.Conclusion Ganoderma lucidum preparation can improve behavior disorders of AD model mice,promote the expressions of ApoA1,ApoE and Syt1,inhibit the expression of Aβ-40 protein,and improve the autoimmune function.

Effects of acute knockdown of hepatic insulin receptor on secretion of apolipoprotein B and triglyceride in mice / 中华老年医学杂志

Objective To analyze the effects of acute depletion of liver-specific insulin signaling on secretion of apolipoprotein B (apoB) and triglyceride (TG).Methods Based on Cre-LoxP principle,a promoter of hepatic tissue specific albumin gene was inserted into upstream of the cre recombinase gene.Albumin-Cre adenovirus (Ad-CRE) and GFP adenovirus (Ad-GFP) were amplified in 293A cells and purified before intravenous administration to mice.After adenovirus infection for 2 days,4 days and 6 days,blood samples from mice were collected and hepatic tissues were frozen.The secretion rates of hepatic newly synthesized apoB and very low density lipoprotein (VLDL)-TG were determined by injection of Triton WR-1339.The levels of plasma cholesterol (TC) and TG were measured.The expressions of insulin receptor and other lipoprotein metabolism related proteins in hepatic tissues were analyzed by Western blot.Results After 2 d,4 d and 6 d of the Ad-CRE injection into mice,insulin receptor expression was reduced by 30.50％ (P＜0.05),60.12％ (P＜ 0.01) and 99.54％ (P＜0.001),and VLDL-TG secretion rate was decreased by 20.43％ (P＜0.05),33.63％ (P＜0.05) and 44.21％ (P＜0.01),respectively.Expressions of sterol regulatory binding proteins 1,fatty acid synthase,and the related proteins of VLDL-formation were decreased,but there were no changes in hepatic secretion of apoB100 and hepatic lipids.The hepatic secretion of apoB48 was increased by 35.07％ (P＜0.05) 6 d after Ad-CRE injection.Conclusions Acute depletion of hepatic insulin receptor might reduce VLDL-TG secretion in manner of time-dependent,and increase the assembly and secretion of smaller apoB-containing lipoproteins in mice liver,which is probably associated with decreased lipogenesis.

Preparation of serum total cholesterol,total glycerol,free glycerol and triglycerides reference materials / 中华检验医学杂志

Objective To prepare serum cholesterol and triglycerides reference materials.Methods Blood units were collected from healthy donors and the sera separated and screened for cholesterol and triglycerides levels.Four serum pools were prepared by pooling sera grouped according to cholesterol and triglyceride levels.The materials were tested for homogeneity and stability and their values for total cholesterol,total glycerol and free glycerol and triglycerides were assigned by HPLC methods.Results The materials were tested to be homogeneous and stable for at least 4 years at-20℃.The certified values (reference value±expanded uncertainty)of the 4 materials for total cholesterol were(5.110±0.065) mmol/L,(4.761±0.062)mmol/L,(3.941±0.050)mmol/L and(3.158±0.041)mmol/L,respectively;for total glycerol(2.212±0.043)mmol/L,(1.679±0.033)mmol/L,(1.275±0.027)mmol/L and(1.067±0.023)mmol/L;for free glycerol(0.142±0.005)mmol/L,(0.149±0.004)mmol/L,(0.146±0.003)mmol/L and(0.122±0.003)mmol/L;and for triglycerides(2.069±0.043)mmol/L,(1.530±0.033)mmol/L,(1.129±0.027)mmol/L and(0.945±0.023)mmol/L.Conclusion Certified reference materials for serum total cholesterol,total glycerol,free glycerol and triglycerides have been prepared.

Relationship between plasma protein expression profiles and states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis / 中西医结合学报

To investigate the relationship between the plasma biomarker proteins and the states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis.

Analysis of low-density lipoprotein receptor gene mutations in a Chinese patient with clinically homozygous familial hypercholesterolemia / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To screen the point mutation of the low-density lipoprotein receptor (LDL-R) gene in Chinese familial hypercholesterolemia (FH) patients, characterize the relationship between the genotype and the phenotype and discuss the molecular pathological mechanism of FH.</p><p><b>METHODS</b>A patient with clinical phenotype of homozygous FH and her parents were investigated for mutations in the promoter and all eighteen exons of the LDL-R gene. Screening was carried out using Touch-down PCR and direct DNA sequencing; multiple alignment analysis by DNASIS 2.5 was used to find base alteration, and the LDL-R gene mutation database was searched to identify the alteration. In addition, the apolipoprotein B gene (apo B) was screened for known mutations (R3500Q) that cause familial defective apo B100 (FDB) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>Two new heterozygous mutations in exons 4 and 9 of the LDL-R gene were identified in the proband (C122Y and T383I) as well as her parents. Both of the mutations have not been published in the LDL-R gene mutation database. No mutation of apo B100 (R3500Q) was observed.</p><p><b>CONCLUSION</b>Two new mutations (C112Y and T383I) were found in the LDL-R gene, which may result in FH and may be particularly pathogenetic genotypes in Chinese people.</p>

Cloning and characterization of cholesteryl ester transfer transfer protein isolated from the tree shrew / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To obtain the nucleotide sequence and deduced amino acid sequence of cholesteryl ester transfer protein (CETP) cDNA from the tree shrew (Tupaia glis).</p><p><b>METHODS</b>The cDNA sequence of the tree shrew CETP was obtained by utilizing the technique of switching mechanism at 5' end of RNA transcript (SMART) and rapid amplification of cDNA end (RACE) from the first strand of the cDNA. The amino acid sequence of CETP was deduced from the cDNA sequence and its primary and secondary structures were predicted.</p><p><b>RESULTS</b>The sequence of CETP cDNA from tree shrew (GenBank accession number AF334033) covers 1636 bp, including 178 bp at the 3' end of the untranslated region and a 1458 bp fragment in a coding region, which provides the complete sequence of mature tree shrew CETP, although not the initiator methionine. The first 24 bp encodes a partial signal peptide. The mature protein consists of 477 amino acids and is longer than the human version by one amino acid (Gly318). Comparing this amino acid sequence with those of other animals' CETPs, the identity between tree shrew and human and rabbit CETP is 88% and 82%, respectively. The protein is extremely hydrophobic as it contains many hydrophobic residues, especially at the C-terminal, consistent with its function in the transfer of neutral lipids. The amino acid residues concerning with binding and transferring neutral lipids are highly conserved. There is a deletion of an N-linked glycosylation site at Asn342 in the tree shrew CETP protein that may participate in the removal of peripheral cholesterol and cholesteryl ester by increasing its activity of transferring cholesteryl ester.</p><p><b>CONCLUSION</b>The possible glycosylation in the tree shrew CETP may be involved in the molecular mechanism of its insusceptibility to atherosclerosis.</p>

Establishment and comparative genomic hybridization analysis of human esophageal carcinomas cell line EC9706 / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a long-standing cell line of esophageal squamous cell carcinoma (ESCC) in pursuit of a model for in vitro study of carcinogenesis.</p><p><b>METHODS</b>Small tissue blocks taken from resected specimens of esophageal cancer were cultured, and cell line EC9706 was established. The biologic properties of EC9706 were characterized. Comparative genomic hybridization(CGH) was performed on the cell line.</p><p><b>RESULTS</b>The growth curve of EC9706 was detected. The cell generation time was 26 hours. The plate colony forming efficiency is 91.9%, with the capacity of forming clones in soft agar. EC9706 cells show high tumorigenecity as indicated by the rapid regeneration of moderate-poor-differentiated squamous cell carcinomas after injection into nude mice. CGH analysis indicated copy number gains of 1p1, 1q2-4, 2p1, 2q1, 5p, 7p14, 7q21, 11q1, 15q2, 20q and losses of 2p2, 2q2, 3p, 4, 9p, 14, 18, Xq. High-level gain of 5p was observed.</p><p><b>CONCLUSION</b>Established cell line EC9706 can serve as a useful tool for studying the carcinogenesis of ESCC.</p>

Effects of 5-HT and electrolytes on the airway remodeling of bronchial asthma in guinea pigs / 中国病理生理杂志

AIM: To explore the effects of 5-HT and electrolytes on the airway remodeling in guinea pigs with bronchial asthma.METHODS: 70 guinea pigs were divided into 7 groups: control group,model group,continued model group,5-HT group,anti-5-HT group,high Mg~(2+) group,low Mg~(2+) group.Remodeling model was established with ovalbumin.RESULTS: ① In model group,5-HT of serum and thickness of airway walls were significantly increased compared with control group(P