Objective To observe anti-sense phosphorothioate oligonucletide (ASPSODN) targeted directly to hTERT mRNA to its inhibiting effect on aimed gene and the influence on the telomerase activity, cellular proliferation, cell apoptosis of K562 cells. Methods Human leukemia cell line K562 was transfected with anti-sense oligonucleotide ASPSODN by liposome. The proliferation activity of K562 cell line was determined by using methyl thiazolyl tetrazolium assay, and telomerase activity was detected by TRAP-PCR-ELISA. Flow cytometry was adopted to examine apoptotic rate and cell cycle. RT-PCR was used to detect the expression of target gene hTERT mRNA. Results 0.6 μmol/L ASPSODN (0.42 ±0.16) was remarkably decreased the expression of hTERT mRNA, Telomerase relative activation was decreased by 52 %. According to 0.6 (μmol/L ASPSODN caused significant the inhibition of K562 cell growth. Apoptotic rate and cell cycle was examined by 0.6 μmol/L ASPSODN with flow cytometry. The cell apoptosis rate of 0.6μmol/L ASPSODN were 10.31 %. It showed the cells treated with 0.6 uU PSASODN arrested in G_1/C_0. The ratio of cells in G_2/M and S period was reduced. But there was no characteristic apoptosis peak. Conclusion ASPSODN targeted hTERT can inhibit the expression of target gene hTERT mRNA, and decrease the telomerase activity of K562 cells. ASPSODN can inhibit strongly the proliferation of K562 cell and induce cell apoptosis by decreasing telomerase activity.

Feeding Behavior ; Humans ; Infant ; Infant Food ; Infant Nutritional Physiological Phenomena

Objective To investigate the effect of jejunal infusion of amino acids on secretion of gastrointestinal hormone in healthy dogs.Methods Six healthy adult dogs were treated with jejunal fistulas and femoral vein intubation.Twenty-four hours after the operation,solution of 8 different amino acid monomers (experimental group) or normal saline (control group) were infused into the jejunum of the dogs every 24hours.The levels of cholecystokinin (CCK),motilin,and gastrin in the peripheral plasma were measured using radioimmunoassay at the start of infusion (0 minute),and 30,60,90,and 120 minutes after infusion.Results Compared with the control group,the serum CCK level in the phenylalanine group was significantly higher 30 and 60 minutes after infusion [(1.25 ±0.19) ng/L vs.(0.66 ±0.14) ng/L,(1.23 ±0.12) ng/L vs.(0.80 ± 0.03) ng/L,both P ＜ 0.01],while that in the tryptophan group was significantly higher 30 minutes after infusion [(1.08 ±0.26) ng/L vs.(0.66 ±0.14) rig/L,P ＜0.01].The other measurement results showed no statistically significant differences.Conclusions Jejunal infusion of phenylalanine or tryptophan may stimulate the secretion of gastrointestinal hormone to some extent.Aromatic amino acids (phenylalanine and tryptophan) is more potent in triggering the release of CCK than aliphatic (leucine,isoleucine,and methionine) and charged amino acids (aspartic acid,arginine,and glutamate).The mechanism may be related to the properties of the amino acids.

Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.

Objective To investigate the effects of informed consent on emotion and immunologic function in patients with laryngopharynx cancer.Methods Sixty patients with laryngopharynx cancer were divided into group A and group B by random digits table with 30 cases each group.In group A patients were informed consent,and in group B family were informed consent.The self-rating anxiety scale (SAS) and self-rating depression scale (SDS) scores,as well as cellular immunity function were measured before informed consent and 2 weeks after surgery.Results The SAS,SDS scores and levels of cellular immunity function between two groups,differences was no statistically significant (P ＞ 0.05).The SAS,SDS scores 2 weeks after surgery in group A were significantly less than those in group B [(43.17 ± 7.63) scores vs.(47.84 ± 8.46) scores,(37.43 ± 8.69) scores vs.(42.67 ± 9.82) scores,P ＜ 0.05].The CD3,CD4,NK cells 2 weeks after surgery in group A were significantly higher than those in group B (0.5762 ±0.0941 vs.0.5160 ± 0.0821,0.3832 ± 0.0751 vs.0.3140 ± 0.0561,0.1521 ± 0.0491 vs.0.1163 ± 0.0422,P ＜ 0.05),CD8 and CD4/CD8 between two groups differences was no statistically significant (P ＞ 0.05).Conclusion The informed consent for patients with laryngopharynx cancer may relieve emotional disorders and avoid immunologic function decreasing.

Objective To explore a new surgical treatment for severe inverted nipple and to reduce the possibility of recurrence. Methods Two opposite long six-side polygon de-epithelialized areas were designed within the range of areola on both sides of the nipple. The four sides in lateral portion were dissected and two wedge-shaped dermal and glandular tissue flaps were created, completely releasing the contractive fiber causing nipple retraction while reserving breast ducts. The flaps were reversed 180 degrees through the tunnel bilateral sides of breast ducts to support the nipple. It was tracted for 3-4 weeks by series 7 needle penetrating the nipple basis. Results All the 14 patients were followed up for 3-18 months, and they were satisfied with the appearance and the aesthesia of the nipple and no recurrence was observed, breast-feed function reserved in 2 cases after childbirth. Conclusions This operation is easy to perform with satisfying aesthetic outcome and breast-feed function remaining.It is an ideal method for correcting serious inverted nipple.

Objective To analyse the sonographie characteristics of thyroid nodules after injection and investigate the contrast enhanced ultrasonography. Methods Analysed the sonographic characteristics in 32 cases and observed the dynamic enhancement pattern of nodules by the time-intensity curve(TIC). The evaluated parameters were arrival time (AT), time to peak(TTP), peak intensity(PI). Results Thyroid cancer (12 cases) showed early wash in an early wash out in comparison with the adjacent thyroid parenchyma, and the enhancement intensity [AT(7.49 ± 4.45), TIP( 17.41 ± 2.84), PI( 18.56 ± 5.24) s] were lower than nodular goitre (20 cases) [ AT ( 13.22 ± 3.30), TIP (20.92 ± 2.59), PI ( 19.15 ± 5.47 ) s ] (P < 0.05 ). Conclusion The contrast enhanced ultrasonography may provide a useful method for the diagnosis of thyroid cancer.

Objective To select an efficient method to inhibit telomerase activity, antisenseoligodeoxynucleotide and plasmid-vector mediated RNAi against hTERT were used to inhibit telomerase activity. The inhibiting effects of the two methods were compared. Methods Against hTERT mRNA, siRNA and oligodeoxynucleotide were designed and transfected into K562 cells by liposome. Effective and specific siRNA strands were selected and then plasmid was constructed and transfected into K562 cells; followed by analysis of the results. Results hTERT mRNA were detected after the three chemo-synthesized strands were transfected. It was found that si-hTERT-1 and si-hTERT-2 were effective, but si-hTERT-3 had no effect. The inhibiting effect of hTERT mRNA lasted only 48 h and disappeared at 72 h. Two siRNA strands were sieved and plasmids were constructed and transfected into K562 cells. In the P-1 group, hTERT mRNA was 0.39±0.13 at 48 h, 0.57±0.32 at 72 h. In the P-2 group, hTERTmRNA was 0.55±0.20 at 48 h, 0.88±0.23 at 72 h.In the P-1 group, the relative telomerase activity was 0.42±0.07 at 48 h, 0.31±0.08 at 72 h. In the P-2 group was 0.49±0.27 at 48 h, 0.39±0.03 at 72 h. The best concentration of siRNA was 100 μmol/L. The best concentration of ASODN was 0.6 μ mol/L. hTERTmRNA was 0.42±0.16 at 24 h, 0.71±0.18 at 48 h. Relative telomerase activity was 0.52±0.002 at 24 h, 0.482±0.018 at 48 h. Conclusion Both ASODN and RNAi targeting hTERT can inhibit the expression of hTERT mRNA, and then inhibit telomerase activity. The inhibiting effect is closely relative to the targeting site. The inhibiting effect of RNAi is better than that of ASODN. RNAi has better efficiency and lasts for a longer time. Plasmid mediated RNAi has better inhibiting effect than the chemo-synthesized siRNA.