ABSTRACT
Aim To screen the potential inhibitors of post-transcriptional activity of pro-inflammatory media-tor TNF-α from the lipophilic constituents in Chinese Medicine Salvia miltiorrhiza Bunge ( Danshen) , we es-tablished dual luciferase reporter gene system pGL3-TNF-α3′UTR ( 3′untranslated region ) co-transfected with Renilla control gene. Methods Complementary DNA ( cDNA) template was obtained from human um-bilical vein endothelial cells ( HUVECs ) . The full length DNA of TNF-α 3′-UTR was amplified through PCR, and then connected the luciferase reporter vector pGL3-control after enzyme digestion. pGL3-TNF-α 3′UTR constructs were co-transfected with pSVRenilla into the mononuclear macrophages RAW264. 7 cells. The relative activity of reporter genes was measured by dual luciferase reporter ( DLR ) assay system after the stimulus of lipopolysaccharide ( LPS ) in presence or absence of tanshinones compounds. Results The pGL3-TNF-α3′UTR luciferase reporter gene was suc-cessfully constructed. The cloning DNA fragment and sequence were both consistent with the GENBANK da-tabase. LPS significantly induced the relative reporter activityof RAW264 . 7 cells transfected with pGL3-TNF-α 3′UTR. Among four tanshinones compounds, we found only cryptotanshinone could significantly de-crease LPS-induced relative reporter activity. Conclu-sion The pGL3-TNF-α 3′UTR construct combined with DLR assay system was successfully established, which can be used to discover the agents such as cryp-totanshinone that regulate the post-transcription of TNF-α in treatment of inflammatory and malignant dis-eases.
ABSTRACT
Objective To explore the diagnostic value of ultrasonography for the residual gallbladder after cholecystectomy.Methods Application of the ultrasound diagnostic apparatus in cholecystectomy patients was to be screened to find the residual gallbladder.Results Ultrasound resuts showed 18 cases of residual gallbladder,which combined stones in 12 cases(66.7％ ),polyps in 1 case(5.6％ ) ;Atrophic cholecystitis,acute cholecystitis,postoperative residual gallbladder probability were high,accounting for 83.0％.Conclusion Ultrasound for the preoperative prevention and postoperative diagnosis of the residual gallbladder has better value,and the cholecystectomy patients should be routinely review the ultrasound.
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Objective To investigate re-innervation in the neovaginal mucosa of patients underwent sigmoid colon vaginoplasty in treatment of Mayer-Rokitansky-Kistner-Hauser Syndrome (MRKHS).Methods Biopsies in the upper third of the posterior neovagina were taken in 20 patients treated by sigmoid colon vaginoplasty at 1, 2 and 3 years after surgery, respectively. Protein gene product 9. 5 ( PGP 9. 5 ),vasoactive intestinal peptide (VIP) and neuropeptide Y (NPY) were detected by immunohistochemical method and compared with those in intact sigmoid colon mucosa. Results ( 1 ) Density of nerve fiber:abundant distribution of PGP 9. 5 nerve fibers were observed in the mucosal muscle layer, submucosa, and smooth muscle layer of the neovagina. The nerve fibers of VIP and NPY immunoreactivity were mainly distributed around blood vessels and in the smooth muscles. In the neovagina, the density of nerve fibers of PGP 9. 5 of 17 ± 6 were much more than VIP of 2. 9 ± 1.0 and NPY of 2. 5 ± 0. 8 significantly ( P ＜ 0. 05 ).( 2 ) Expression of PGP 9. 5 in neovagina: at 1 year after surgery, PGP 9. 5 positive expression of 14 ± 4 was significantly lower in the neovagina than 28 ± 7 in the intact sigmoid colon( P ＜ 0. 05 ). However, after 2 to 3 years, its expression displayed an upgrade tendency in the neovagina and was significantly higher at the 3 year postoperatively than that at the 1 years postoperatively ( 22 ± 7 vs. 14 ± 4, P ＜ 0. 05 ). The changes were much more obvious in submucosa. (3) The expression of VIP and NPY in neovagina: at 1 year after surgery, VIP and NPY positive nerve fibers were also decreased in the neovagina when compared with those in the intact sigmoid colon ( 2. 3 ± 0. 7 vs. 5.3 ± 1.4, P ＜ 0. 05; 2.5 ± 1. 1 vs. 5.5 ± 1.1, P ＜ 0. 05 ) . At 2 to 3 years after surgery, the positive VIP fiber showed initially decreased and subsequently increased tendency. The density of VIP of 3.7 ± 0. 7 in the neovagina at 3 years postoperatively was higher than 2. 3 ±0. 7 at 1 years postoperatively (P ＜ 0. 05 ). No significant up-regulation was observed in NPY-positive expression in the neovagina within 3 years after operation. Conclusions Distribution of sensory PGP 9. 5,VIP and NPY immunoreactive nerve fibers was similar to the pattern observed within the intact sigmoid colon wall. The number of nerve fibers in the neovagina decreased after surgery and then increased subsequently within 3 years after surgery.
ABSTRACT
quamous metaplasia occurred, the expression of occludin and ZO-1 in cells were similar to those in control group, which might play a role in the defense function of neovaginal mucosa.