Objective To explore the potential genetic variants related with late-onset Alzheimer's disease(LOAD),and to broaden the AD genetic susceptibility factor profile.Methods We targetedly sequenced four lipids metabolism related genes including apolipoprotein E(APOE),sortilin-related receptor gene (SORL 1),apolipoprotein C-I (APOC1) and clusterin (CLU),validated these candidate variants in patients with LOAD and mild cognitive impairment (MCI)and the controls with normal cognitive function,and made a bioinformatic analysis.Results The mutation in the allele of CLU rs117389184,a low-frequency single nucleotide polymorphism,would increase the risk of LOAD and MCI[OR (95 ％ CI):3.56 (1.08-4.84),P =0.026;OR (95 ％ CI):3.06 (0.98-4.03),P =0.026],suggesting CLU rs117389184 gene variant was associated with cognitive impairment.This variation could affect CLU gene expression by interfering with the binding of transcription factors.Conclusions CLU rs117389184 is a new genetic risk variant for LOAD and MCI.

BACKGROUND:Angiotensin II receptor antagonists have been found to exerct a stronger protective effect on bone than angiotensin converting enzyme inhibitors. OBJECTIVE:To investigate the effect of different concentrations of irbesartan (angiotensin II receptor antagonist) on the differentiation and mineralization of mouse preosteoblasts. METHODS:Mouse preosteoblast cel lines MC3T3-E1 in logarithmic phase were selected and cultured in the osteogenic induction medium containing 0 (control group), 0.001, 0.01, 0.1 mmol/L irbesartan, respectively. Ten days later, the cel differentiation was observed by alkaline phosphatase staining. The mineralization was observed by alizarin red staining after 21 days of culture. mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 in osteoblasts were detected by real-time PCR at 1, 4, 7, 14 and 21 days of culture. RESULTS AND CONCLUSION:The activity of alkaline phosphatase in al the irbesartan groups (0, 0.001, 0.01, 0.1) was higher than that in the control group (P<0.05), which was the most obvious in 0.01 mmol/L. The number and area of calcium nodules in each irbesartan group were significantly higher than those in the control group (P<0.05), especial y in 0.01 mmol/L. Compared with the control group, 0.01 mmol/L irbesartan significantly upregulated the mRNA expressions of osteocalcin, alkaline phosphatase and Runt-associated transcription factor 2 (P<0.05). These results suggest that 0.01 mmol/L irbesartan significantly promotes the differentiation and mineralization of osteoblasts.

To provide theoretical basis for breeding good variety of Dendrobium denneanum, agronomic and qualitative characters of 4 different lines and relationships among them were studied. The stem length, stem diameter, leaf length, leaf width, length/ width ratio and leaf area were measured. The single fresh and dry stem was weighed and drying rate was calculated. The contents of polysaccharides and total alkaloids were determined by sulfuric acid-phenol colorimetry and acid-dye colorimetry, respectively. The correlations between characters were analyzed. The results showed that differences in major agronomic characters between four lines were significant. The plant types of dq-1 and dq-2 were higher, dq-3 was medium and dq-4 was lower. The fresh weigh of stem and content of polysaccharides were the highest in dq-2, 7.81 g and 14.33%. While the highest content of total alkaloids and was 0. 486% in dq-3. There were significant correlations between agronomic characters, but these characters had low or non correlations with qualitative characters such as polysaccharides and total alkaloids. It was shown that the content of polysaccharides and total alkaloids were significantly different among 4 lines of D. denneanum, which could be selected for different uses.
Agriculture ; Dendrobium ; anatomy & histology ; chemistry

Objective To investigate the effect of Bufalin on aerobic glycolysis in human colorectal cancer cell.Methods The colorectal cancer cells were divided into Bufalin groups and control group.Bufalin groups were treated with Bufalin at various concentrations(5,10,20,40nmol · L-1) for 48 h and control group was given the same dose of complete medium.The level of intracellular ATP level and cell lacate production were determined by Kit.The C-myc and energy metabolism regulator were measured by Western blot.Results Compared with the control group(1.00),4 concentrations (5,10,20,40 nmol · L-1) Bufalin were 91.69％,78.00％,68.55％,54.03％ on the inhibition of human colon cancer cell line HCT116 in ATP level rate;the 4 concentrations Bufalin were 89.04％,77.27％,59.66％,47.52％ to inhibit HCT116 cell junction formation of lactic acid in colorectal cancer cell line rate,the difference was statistically significant (P ＜ 0.05,P ＜ 0.01).Compared with the control group (1.00),the 4 concentrations Bufalin with HCT116 cells C-myc protein colon cancer cell line expression ratio were 0.95,0.84,0.73,0.68;lactate dehydrogenase A (LDH-A) expression ratio were 0.95,0.90,0.79,0.60,the difference was statistically significant (P ＜ 0.05,P ＜ 0.01).Conclusion Bufalin can inhibit the energy metabolism of colorectal cancer cell,which may be related with the down-regulation of C-myc expression.

OBJECTIVETo explore a new approach to the management of malignant biliary obstruction using percutaneous transhepatic biliary radiofrequency and endoprothesis.

METHODSPercutaneous transhepatic biliary radiofrequency and endoprothesis were performed in 2 cases of malignant biliary obstruction, including 1 of hilar cholangiocarcinoma and 1 of pancreatic head carcinoma. The tumor was ablated with radiofrequency followed by placement of matched metal stents into the biliary duct.

RESULTSThe surgical procedures were carried out smoothly in the 2 cases. The symptoms of the patients were obviously improved after the operation with a significant decrease in the serum levels of total bilirubin, and CA-199 level decreased to the normal level in 1 case.

CONCLUSIONSThis new approach is safe for management of malignant biliary obstruction. Compared with the more conventional interventional therapy, radiofrequency can reduce the intraoperative bleeding and arrest the local tumor growth to promote the patency of the stent as well as the postoperative survival of the patients.

Adult ; Aged ; Catheter Ablation ; methods ; Female ; Humans ; Jaundice, Obstructive ; etiology ; surgery ; Male ; Prosthesis Implantation ; methods ; Stents

OBJECTIVETo investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells.

METHODTo establish a model of hypoxia four hours and reoxygenation twelve hours injury in rat cerebral microvascular endothelial cells in vitro. The injured cells were treated with geniposide (0. 128, 0.064, 0.032 micromol mL(-1), baicalin (0.028, 0.014, 0.007 micromol mL(- 1)) and berberine (0.024, 0.012, 0.006 micromol mL(-1)). The expression of p65 subunit of NF-kappaB was detected by immunocytochemical assay and techniques of image quantitative analysis. The protein expression of NF-kappaB was calculated with the mean optical density and mean area. The nuclear translocation of NF-kappaB was calculated with the percentage of positive cells and ratios of light transmittance of cytoplasm and cell nucleus.

RESULTCompared with the normal group, both the protein expression and the nuclear translocation of NF-kappaB of model group were significant increased (P <0.01). Compared with the model group, the mean optical density of all treated groups was decreased ,but these was no significant difference between them. As compared with model group, the mean area of all treated groups was significant decreased (P < 0.01). The percentage of nuclear translocation of all treated groups is not only lower than that of the model group but higher than that of the normal group (P <0.01). Compared with the model group, the ratios of light transmittance of cytoplasm and cell nucleus of all treated groups was significantly elevated (P <0.01).

CONCLUSIONThe results suggesed that geniposide, baicalin and berberine could protect hypoxia/reoxygenation injuried rat cerebral microvascular endothelial cells injury. One of the mechanism may lie in inhibiting both the protein expression and the nuclear translocation of NF-kappaB.

Animals ; Brain ; blood supply ; Cell Nucleus ; drug effects ; metabolism ; Cells, Cultured ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Endothelial Cells ; drug effects ; metabolism ; pathology ; Gene Expression Regulation ; drug effects ; Hypoxia ; complications ; Male ; Microvessels ; pathology ; NF-kappa B ; metabolism ; Oxygen ; metabolism ; Protein Transport ; drug effects ; Rats

OBJECTIVETo investigate the protective mechanism of geniposide, baicalin and berberine on hypoxia and reoxygenation injury in cultured rat cerebral microvascular endothelial cells.

METHODA model of four hours hypoxia and twelve hours reoxygenation injury in rat cerebral microvascular endothelial cells in vitro was established. The injured cells were treated with geniposide (0.128, 0.064, 0.032 mmol x L(-1)), baicalin (0.028, 0.014, 0.007 mmol L(-1)) and berberine (0.024, 0.012, 0.006 mmol L(-1)), respectively. The immunocytochemical method and techniques of image quantitative analysis were used to detect the mean optical density and mean area in order to match the protein expression of VCAM-1. The method of RT-PCR was adopted to observe and match the mRNA expression of VCAM-1.

RESULTAs compared with the normal group, the mean optical density, the mean area and the mRNA expression of VCAM-1 of model group were significant increased (P < 0.01, P < 0.01, P < 0.01). As compared with the model group, both the mean optical density and the mean area of all treated groups were decreased, and there was significant difference between them (P < 0.01, P < 0.01). As compared with normal group, the mean optical density of baicalin (0.007 mmol x L(-1)) and berberine (0.012, 0.006 mmol x L(-1)) were significant decreased (P < 0.05, P < 0.01, P < 0.01), but there was no significant difference between the other groups and the normal group. As compared with normal group, the mean area of baicalin (0.0014 mmol x L(-1)) was significant decreased (P < 0.05), but there was significant difference between the other groups and the normal group. The mRNA expression of all treated groups was not only lower than that of the model group but also higher than that of the normal group (P < 0.05, P < 0.05).

CONCLUSIONThe results suggest that geniposide, baicalin and berberine, which are effective compositions of huanglian jiedu decoting, can protect hypoxia-reoxygenation injuried rat cerebral microvascular endothelial cells. One of the protected mechanisms is that they can inhibit the expression of VCAM-1.

Animals ; Berberine ; pharmacology ; Cell Hypoxia ; drug effects ; Cells, Cultured ; Cerebrum ; drug effects ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Endothelium, Vascular ; drug effects ; metabolism ; Flavonoids ; pharmacology ; Gene Expression ; drug effects ; Humans ; Hypoxia ; drug therapy ; genetics ; metabolism ; Iridoids ; pharmacology ; Male ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; drug therapy ; genetics ; metabolism ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

BACKGROUND:Insulin analogues have been extensively applied in the treatment of diabetes mellitus.Insulin glargine has a higher affinity for insulin like growth factor 1 receptor compared with human insulin.Further research is needed to ensure whether insulin and its analogues exert same effects on fracture healing in type 2 diabetes mellitus.OBJECTIVE:To observe the osteocalcin expression and callus formation in the healing of fracture in type 2 diabetic rats induced by human insulin and insulin glargine,to observe the difference between two treatment methods,and to explore the related mechanisms.METHODS:Sixty-four Sprague-Dawley rats were randomly assigned into human insulin group (group A),insulin glargine group (group B),diabetes mellitus group (group C) and control group (group D).Rats in the groups A,B and C were fed with high-sugar and high-fat diet for 4 weeks followed by intraperitoneal injection of small-dosage streptozotocin twice,to establish the rat models of type 2 diabetes mellitus.After the right tibia of each rat was broken,insulin glargine and Novolin 30R were used in the groups A and B,respectively.Fracture healing was observed on X-ray,callus formation and number of osteoblasts were observed by microscope,and serum level of osteocalcin was measured by ELISA method at 1,2,4,and 6 weeks after modeling.RESULTS AND CONCLUSION:X-ray results revealed better fracture healing in the groups A,B and D than the group C.Osteoblast proliferation in callus was significantly better in the groups A,B and D than in the group C.Serum level of osteocalcin in each group was on the rise,which was significantly higher in the groups A,B and D than the group C (P ＜ 0.05),but had no significant difference among groups A,B and D (P ＞ 0.05).In summary,insulin glargine can increase the serum level of osteocalcin,accelerate the callus formation,and improve the healing of fracture in type 2 diabetic rats.Furthermore,there is no significant difference in the therapeutic efficacy between insulin glargine and human insulin.

Objective To optimize the serum bactericidal assay (SBA) , detect and analyze the bactericidal antibody level against Neisseria meningitidis serogroup C strains after divalent polysaccharide (A plus C) vaccine immunization. Methods Two Neisseria meningitidis serogroup C strains, vaccine candidate strain (C11) and epidemic strain (053442), were selected as targets. The national Neisseria meningitidis standardized serum was used as reference serum. Pel-Freez infant rabbit complements was available. The optimized SBA method was used to detect bactericidal antibody against strain C11 and 053442 for 122 pairs of sera before and after immunization with a divalent polysaccharide (A and C) vaccine. Results The strain C11 and 053442 both could be used as targeted strain for SBA. The optimized concentration of targeted strain was achieved when a whole-cell suspension of 0.35 A at 600 nm was diluted 4×104 times. Before immunization, SBA geometric mean titers(GMT) of 122 sera against strain C11 and 053442 were 1: 1.75 and 1:2.63 respectively, and the protective rates were 9.8% and 17.2% respectively. After immunization, the GMTs and the protective rates of 122 sera both rose significantly (P<0.01), the GMTs against strain C11 and 053442 were 1:483.73 and 1:412.57 respectively. The protective rates against strain C11 and 053442 were 100% and 95.9% respectively. Conclusion Immunization with a divalent polysaccharide (A and C) vaccine could elevate remarkably the population SBA titer against Neisseria meningitidis serogroup C strains of different subtypes, but the surveillance of vaccine effect against different targeted strains remains necessary.

To investigate the effect of a new reactive oxygen metabolites (ROM) scavenger as immune adjuvant in NK cell-mediated killing effect on K562 cell, IL-2 and PHA were used to activate monocyte to produce ROM, and different concentrations of tiopronin as ROM scavenger was used in the cultivated systems with different ratio of monocytes plus NK cells and K562 cells, while histamine dihydrochloride (DHT) with different concentrations was used as positive control. The reuslts indicated that after IL-2 and PHA were supplemented in the cultivated systems mixing with NK cells and K562 cells as the E/T ratio was 10/1, the ROM production increased from 33.17 +/- 25.02 U/ml to 223.59 +/- 59.41 U/ml (P < 0.05) while K562 cell inhibition rate (KIR) increased from 65.56% to 85.89% (P < 0.05). When the monocytes as the E/MO ratios of 10/2, 10/5 and 10/10 were supplemented respectively, ROM production increased correspondingly (ROM production was 389.79 +/- 43.83 U/ml, 456.74 +/- 42.77 U/ml, 601.42 +/- 21.92 U/ml, respectively), and KIR was on the other round (KIR was 82.36%, 81.36%, 48.09% respectively). Tiopronin, DHT were used in the K562 + NK + MO + IL-2/PHA cultivated systems as the E/MO ratio was 10/2, the ROM production also decreased from 389.79 +/- 43.83 U/ml to -1.20 +/- 60.70 U/ml, 50.21 +/- 22.4 U/ml (P < 0.05), respectively, however KIR increased from 82.53% to 96.09% and 94.64% either (P < 0.05). Higher concentrations of tiopronin and DHT were used, ROM production decreased accordingly. There showed a reverse correlation between ROM production and KIR (r = -0.518). When E/MO ratio was 10/5 or 10/10, tiopronin at any testing concentration and DHT at the higher testing concentration could reduce the ROM production (P < 0.05), but did not improve KIR significantly (P > 0.05). Tiopronin was as good as DHT in ameliorating KIR (P > 0.05) and better than DHT in scavenging ROM (P < 0.05). It is concluded that (1) Monocytes are the major resources of ROM, and the ROM derived from monocytes can disable NK cells in killing neoplasm cells (K562 cells); (2) A new ROM scavenger, tiopronin, can scavenge ROM effectively, and reverse the ROM induced inhibition of NK cell-mediated killing of K562 cell in a certain extent. And tiopronin is better than DHT in scavenging ROM, and as good as DHT in up-regulating KIR. The new ROM scavenger tiopronin with less side effect may take the place of DHT as adjuvant during the adoptive immuno-therapy in leukemia.
Coculture Techniques ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; immunology ; Free Radical Scavengers ; pharmacology ; Humans ; K562 Cells ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; Reactive Oxygen Species ; antagonists & inhibitors ; metabolism ; Tiopronin ; pharmacology