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 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.

Objective To investigate the effects of electroacupuncture at Sanyinjiao (SP6) and Yinlingquan (SP9) on muscle repair in the gastrocnemius muscle damaged rats and its possible mechanism. Methods 30 male Sprague-Dawley rats were randomly divided into blank group (n=10), model group (n=10) and electroacupuncture group (n=10). The latter 2 groups were modeled with gastrocnemius muscle crush injury. The electroacupuncture group received electroacupuncture at Sanyinjiao and Yinlingquan for 14 days. Their gastrocnemius was observed with HE staining, and the expressions of basic fibroblast growth factor (bFGF) and insulin-like growth factor-1 (IGF-1) in gastroc-nemius tissues and the serum were determined with immunochemical assay and enzyme-linked immunosorbent assay (ELISA). Results Compared with the model group, there was less damage in the electroacupuncture group, with more expression of bFGF and IGF-1 in tis-sues (P<0.01) and serum (P<0.05). Conclusion Electroacupuncture can promote muscle repair after gastrocnemius muscle damage in rats, which may be related to the increase of the bFGF and IGF-1 expression in gastrocnemius tissue and serum.

Objective To research the endometrial receptivity by established the endometriosis rat model in intimal window period.Methods The autologous endometrial tissues of 22 sexually mature SD rats were transplanted into the homonymy abdomi-nal wall and offside uterine mesometrium.After four weeks,we randomly dissected two rats and observed the growth of etopic en-dometrium.The EMs rats were mated with male rats in the same cage and vaginal plug was observed in the next day.The histomor-phology of eutopic and ectopic endometrial was observed in the fifth day of pregnancy.Results Ectopic endometrium was grown in the abdominal wall and mesometrium,which formed a small bulge cystic mass in 20 rats.All foci was showed adhesion to the sur-rounding tissues in a different extent.In the light microscope,the morphology of ectopic endometrial was as the same as nomal en-dometrium.The establishment of the endometriosis rat mode is successful.Vaginal plug was positive the next day.Nonoperative side uterine was full of fluid in the fifth day of pregnancy.Eutopic and ectopic endometrium were in the secretory phase..Conclusion It is a successful method to establish a rat model of endometriosis by autotransplantion of endometrium.The endometrium of EMs model rats is in embryo implantation window period,and it provides a convenient model for studying endometrial receptivity.

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.