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

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.

Otolaryngology ; Periodicals as Topic

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.

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 study the iodine nutritional status and the thyroid function of pregnant women during different periods,and provide scientific basis for iodine supplementation.Methods Totally 728 pregnant women who visited the obstetric outpatient of Nanjing Drum Tower Hospital for routine prenatal care from December 2014 to August 2016 were recruited in this study,and at the same time 182 non-pregnant women were recruited as control group.The thyroid stimulating hormone (TSH) and free thyroxine (FT4) were measured by Roche 601.The urinary iodine level was measured by SR-I-100 kit.Results The median urinary iodine of 728 pregnant women was 168.24 g/L,and the median urinary iodine of those women in the T1,T2 and T3 period were 186.31,162.65 and 148.76 g/L,respectively.The TSH at T1 period was lower than T2 and T3 period (t=3.429,3.135,P＜0.05).The FT4 at T1 period was higher than T2 and T3 period (t=5.251,5.965,P＜ 0.05).The prevalence rate of thyroid disease in normal urinary iodine group was lower than that in low urinary iodine group and high urinary iodine group (x2 =4.139,4.131,P＜0.05).Conclusion There was no iodine deficiency among those pregnant women groups,but only 34.75 ％ individuals reached the appropriate iodine nutritional level,and the ratio of iodine insufficient increased with the extension of pregnancy.The whole prevalence rate of thyroid disease in abnormal urinary iodine pregnant women was obviously higher than that in normal.It is necessary to improve the pregnant women's knowledge of iodine nutrition,moreover it is suggested that urinary iodine monitoring and thyroid function should be conducted in pregnant women.

To explore the mechanism of electroacupunture (EA) of Zusanli (ST36)in protecting gastric mucosa of dogs.Twenty mongrel dogs were randomly allocated to four groups: blank control group (Group A), non acupoint group (Group B), Shangjuxu (ST37) group (Group C) and Zusanli (ST36) group (Group D). Dynamic gastric mucosal blood flow (GMBF) was monitored by laser Doppler flowmeter and calcitonin gene related peptide (CGRP) contents in plasma and gastric mucosa were measured simultaneously by radioimmunoassay method. After sixty minutes of EA, GMBF and CGRP contents in plasma and gastric mucosa were increased in Group D (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 probe into the gene therapy of psoriasis using antisense oligonucleotides to attenuate the expression of K14 gene and protein in keratinocytes and evaluate the inhibitory effects of liposome conjugated antisense oligonucleotides on the proliferation of keratinocytes. Methods The antisense, sense and mismatched oligonucleotides for K14 gene were synthesized and conjugated with lipofectin respectively. Finally they were subsequently transfected into cultured keratinocytes in vitro. The expression of K14 gene was tested by reverse transcription polymerase chain reaction (RT-PCR). The expression of K14 protein was measured by immunohistochemistry. The variation of cell growth cycle was detected by flow cytometry. Results The expression of K14 gene and protein was markedly decreased in keratinocytes treated with K14 antisense oligonucleotides. The cell growth cycle was inhibited effectively by antisense oligonucleotides with lipofection, but not by sense and mismatched oligonucleotides. Conclusions Antisense oligonucleotides conjugated with lipofectin might be a hopeful method to inhibit the proliferation of keratinocytes by inhibiting the expression of K14 mRNA and protein.

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 .