Objective To detect the relationship between the genotypes of the 2642 deletion polymorphism (delta 2642) in IT15 gene and the age of onset of Huntington disease (HD). Methods Peripheral blood samples were collected from 29 patients with HD and 38 gender- and age-matched controls. All patients with HD were diagnosed by gene diagnosis. The CAG trinucleotide repeats of the 29 patients with HD outnumbered 40. Polymerase chain reaction-restriction fragment length polymorphism and polyacrylamide gel electrophoresis technique were used to detect the genotypes of delta 2642. Results No B/B genotype was detected in both 2 groups. The genotype frequencies of A/A and A/B in the IT15 delta 2642 polymorphism was 65.5% and 34. 5% in HD patients,92. 1% and 7. 9% in the controls respectively (x2 = 7. 435, P =0. 006). The frequency of the B allele was 17.2% in the HD group and 3. 9% in the control group (P = 0. 010, OR = 5.07, 95% CI 1.47-15.12). Analysis showed no significant difference between A/A genotype patients and A/B genotype patients for CAG trinucleotide repeats(P =0. 188). HD patients with A/B genotype (37.33±6. 46) had an earlier onset than the patients with A/A genotype (47.10± 10. 86, t = 2. 491, P = 0. 019). Conclusions These data demonstrated that variations in IT15 delta 2642 polymorphisms may be a genetic factor that influences the variability in HD age of onset. HD patients with delta 2642 A/B genotype have an earlier onset than the patients with A/A genotype.

Objective To investigate the regulation of electroacupuncture(EA)on gastric mucosal blood flow(GMBF) and its relation to the contents of plasma and gastric mucosal calcitonin gene-related peptide(CGRP) in dogs.Methods Twenty dogs were randomly divided into four groups:blank control group,no meridian point group,Shangjuxu group and Zusanli point group.By using laser doppler flowmeter(LDF)and radioimmunoassay method,GMBF,contents of CGRP were simultaneously measured,with or without EA stimulation at different acupoints in expermental dogs.Results In the Zusanli group,after EA stimulation, an increase was observed in plasma CGRP content(P

BACKGROUND:Study on antisense drug is still one of hotspots in the current field of biomedicine. Due to high-efficiency and specificity, antisense drug used for gene therapy has been paid more attention by many scholars.OBJECTIVE: To observe the effect of liposome-mediated keratin 14 (K14) antisense oligonucleotide on K14 gene and protein expressions as well as in vitro proliferation activities in human keratinocytes (KC).DESIGN: Single sample observation.SETTING: Department of Dermatology, Xijing Hospital, Fourth Military Medical University of Chinese PLA.MATERIALS: Human KC, K14 oligonucleotide gene fragments (modified with phosphrothioate, and above sequence was synthesized by Shanghai Shenggong Bioengineering Company). Reverse transcriptase and TaqDNA polymerase were purchased from Invitrogen Company, K14 monoclonal antibody was purchased from Antibody Company, and SABC kit was purchased from Boster Company. EPICS-PRO-FILE Ⅱ flow cytometer was purchased from Coulter Company (USA).METHODS: Human epidermal KCs were primarily cultured, and their 3rd to 10th generations were used for the experiment.Artificially synthesized sense and antisense as well as mismatched K14 oligonucleotide gene fragments were introduced into KCs by means of liposome. Blank control group were set. The effects of antisense oligonucleotide on the cell cycle,K14 gene and protein expressions of KCs were detected by flow cytometer, reverse transcription polymerase chain reaction and SABC methods.MAIN OUTCOME MEASURES: Effect of oligonucleotide transfecting human KCs on the proliferation of KCs and K14 expression.RESULTS: [1]The electrophoresis of reverse transcription polymerase chain reaction products: Specific K14 gene band appeared in each group, and K14 gene expression in the antisense group was significantly lower in the sense group,missense group and blank control group. K14/β-actin value was similar among sense group, missense group and blank control group (P ＞ 0.05), But K14/β-actin value was significantly lower in the antisense group than in the above-mentioned 3 groups (F =47.554, P ＜ 0.01). ②K14 protein expression detected by immunohistochemical method:K14 was expressed in all the cultured KCs at different levels, and was obviously reduced after antisense oligonucleotide being added. 20 μmol/L antisense oligonucleotide could markedly inhibit K14 expression; K14 expression did not change in the control group. ③ DNA level change detected by flow cytometer: After being treated by K14 antisense oligonucleotide for 48 hours, human epidermal KCs were significantly increased at G1 stage (74.6%), and were markedly decreased at S stage (19.4%). Such changes were not found in the antisense group, missense group and blank control group.CONCLUSION: Antisense oligonucleotide can specifically inhibit K14 synthesis, and thereby, inhibit the proliferation of human KCs.

Otolaryngology ; Periodicals as Topic

0.05). After EA at Tsusanli, a significant increase was observed in ir SP and ir VIP content in the pituitary gland and peripheral blood, CD4 +, RBC C 3b RR, RBC ICR in the peripheral blood of the normal rats (P0.05). SP and VIP contents in the pituitary gland and the peripheral blood increased (P

Objective To knock down angiopoietin-1 expression in human gastric cancer cell line BGC-823 and to observe its effect of reversing tumor invasion. Methods siRNA sequence fragments was designed to target angiopoietin-1 and transferred into human gastric cancer cell line BGC-823. RT-PCR was used to assess the transcription level of angio-poietin-1 mRNA, then western blot and immunofluorescence were used to examine the expression level of three invasion-as-sociated proteins include integrinβ1, CD44V6 and Ang-1. Cell adhesion ability was evaluated by cell adhesion assay and cell invation was determined by matrigel and transwell plastic dual-chamber culture system. Results Ang-1 mRNA was knocked down by siRNA showed by RT-PCR. The expression of integrinβ1, CD44V6 and Ang-1 were significantly lower than control group(P<0.05), so did the cellular adhesion and invasion abilities(P<0.05). Conclusion Knocking down angiopoietin-1 by siRNA can reverse invasion of human gastric cancer cell line BGC-823 and may provide new ideas and reference for gene therapy of gastric cancer in the future.

OBJECTIVE:To investigate the effect of the extracts of Polygonum lapathifolium on the activity of ?-glucosidase.METHODS:Soxhlet extraction was applied to extract overground part of P.lapathifolium,and the inhibition effect of extracts on ?-glucosidase was determined with 96-microplate-based method,and the inhibitory kinetic characteristics of methanol extract was investigated using Lineweaver-Burk method.RESULTS:The extracts from P.lapathifolium showed good inhibitory activity.The methanol extract had the highest inhibitory activity (IC50=2.13 ?g?mL-1)followed by ethyl acetate extract (IC50=3.826 ?g?mL-1),petroleum ether extract (IC50=230.86 ?g ? mL-1).And they were all higher than that of acarbose (IC50=1 103.01 ?g?mL-1) as positive control.Inhibitory kinetics test indicated that methanol extract was noncompetitive inhibitor.CONCLUSION:The extracts of P.lapathifolium have good inhibition effect on the activity of ?-glucosidase with a good prospect of development and utilization.

Objective To investigate the changes of bone density,bone microstructure,bone BMP-2 expression in rats at early period of diabetes.Methods Thirty SPF 4-month-old male Wistar rats were randomly divided into 3 groups(n=10 in each group):A(fed with common forage as normal control group),B(fed with high-lipid forage and intraperitoneally injected with STZ to establish type 2 diabetes with insulin resistance),C(fed with common forage and intraperitoneally injected with STZ to establish type 2 diabetes with insulin secretion defaulting).The bone histomorphology,bone histomorphometry,bone BMP-2 expression were investigated 12 weeks later.Results As compared with group A,bone microstructure was of no obvious changes and bone BMP-2 expression was higher in group B(P

Objective To evaluate the biological toxicity of heavy metals by using Caenorhabditis elegans. Methods The C. elegans at L4 stage were exposed to CdCl2 CrCl3 As2O3 PbCl2 HgCl2 with low concentrations and M9 buffer the control group for 72 h respectively and the effects of heavy metals with different concentrations on the survival time and reproduction of C. elegans were evaluated. Results After exposure to 2.5 10μmol/L HgCl2 and PbCl2 10μmol/L CdCl2 and 50μmol/L CrCl3 for 72 h respectively the life spans and survival curves of the C. elegans were different from those in the control group the differences were statistically significant all P<0.05 . After exposure to CdCl2 CrCl3 As2O3 PbCl2 and HgCl2 with the con?centrations of 2.5 50 100μmol/L for 72 h respectively the generational time and brood size of C. elegans were all different from those in the control group all P<0.01 . Among the 5 heavy metals at low concentrations the reproduction toxicity of Hg was bigger than Pb Cd Cr and the toxicity of As was the weakest. Conclusion Heavy metal exposure can affect the life span and reproductive toxicity of C. elegans.

AIM: To study the effect of hydrogen sulfide on wound healing of skin ulcer in diabetic rats . METHODS:Male SD rats were randomly divided into 3 groups, including non-diabetic control (NDC) group, untreated diabetic control ( UDC) group, and treated diabetic administration ( TDA) group.Diabetic rats were induced by intraper-itoneal injection of streptozotocin (STZ).After 1 week, wound healing model was prepared by making a round incision ( 2.0 cm in diameter ) on the dorsal skin in full thickness .The rats from TDA group received 2%sodium bisulfide ointment on the skin ulcer wound , and the animals from UDC and NDC groups received control cream .After 21 d of treatment with sodium bisulfide , blood samples were collected for biochemical analysis , including prothrombin time ( PT) , thrombin time (TT), and fibrinogen (FIB) in plasma, as well as the activity of superoxide dismutase (SOD) and the content of malondi-aldehyde ( MDA) in the serum.White blood cells ( WBC) and lymphocytes were also counted .Granulation tissues from the wound were processed for histological examination and Western blot analysis was used to detect heme oxygenase -1 ( HO-1) and tumor necrosis factor α(TNF-α) expression.RESULTS:Compared with UDC group, sodium bisulfide treatment accelerated wound healing of skin ulcer (P<0.01), and increased the activity of SOD in serum (P<0.01) in the diabet-ic rats.The declined number of WBC and lymphocytes , prolonged PT and TT , and decreased FIB levels in rats treated with sodium bisulfied were also confirmed .Pathological section showed that there were inflammatory cell infiltration , and irregu-lar and loose fibril alignment in the granulation tissue of rats from the UDC group , but there were regular fibril alignment and increased angiogenesis in the granulation tissue of rats from the TDA group (P<0.05).Furthermore, sodium bisulfide treatment raised HO-1 protein expression , and decreased TNF-αprotein expression in the diabetic rats .CONCLUSION:Hydrogen sulfide accelerates the wound healing of skin ulcer in the rats with diabetes .The beneficial effect of H 2 S may be associated with formation of granulation , anti-inflammation , and antioxidation .