In this study, a new ileal orthotopic bladder (Urumqi Bladder) modified by our center based on the ileal "W" type orthotopic bladder and Studer bladder was used on 8 patients with invasive bladder cancer. All of patients were male and aged between 54 and 66 years. The history of disease ranged from 1 month to 3 years, including 5 patients with initial onset, 3 patients with ≥2 TURBT history. 6 patients had multiple tumors, tumor size from 0.5 cm to 2.5cm. There were 2 patients with single tumor. Preoperative PET-CT examination showed no distant metastasis and pelvic lymph node enlargement, no urinary tract hydronephrosis, and cystoscopy showed no suspected tumor in the urethra. Preoperative pathological results: high-grade invasive urothelial carcinoma was found in 6 cases and muscular invasive urothelial carcinoma in 2 cases. In 8 patients, 50cm ileum was taken from 15cm away from ileocecum after radical cystectomy, which was crimped clockwise inward from the right end into a nearly circular shape, with 10cm left at the left end. The remaining 40cm ileum was formed into 3 sections of about 13cm each, which were decanted to form a storage capsule. The last 10cm intestinal tube was crossed from the front of sigmoid colon. The end of intestine was anastomosed with the left ureter. The right ureter was anastomosed with the top of the right intestine pouch, and the urethra was anastomosed with the pouch to complete the diversion of urine flow. During 3-12 months of postoperative follow-up, 4 patients had short-term mild urinary incontinence. All had complete urinary control at 12 months. 1 patient still had mild left ureter reflux 12 months after surgery, and the other 7 patients had no ureter reflux. In this group of 8 patients, postoperative excretory cystography showed satisfactory effect of bladder voiding, residual, and bladder capacity. Follow-up review of chest CT, urinary CT and abdominal ultrasound showed no hydronephrosis, and no tumor recurrence or distant metastasis.

OBJECTIVE: To explore the pathogenesis of two fetuses from one family affected with Joubert syndrome (JS). METHODS: Whole exome sequencing was employed to screen potential mutations in both fetuses. Suspected mutations were verified by Sanger sequencing. Impact of intronic mutations on DNA transcription was validated by cDNA analysis. RESULTS: Two novel TCTN1 mutations, c.342-8A>G and c.1494+1G>A, were identified in exons 2 and 12, respectively.cDNA analysis confirmed the pathogenic nature of both mutations with interference of normal splicing resulting in production of truncated proteins. CONCLUSION The genetic etiology of the family affected with JS has been identified.Above findings have enriched the mutation spectrum of TCTN1gene and facilitated understanding of the genotype-phenotype correlation of JS.
Abnormalities, Multiple ; diagnosis ; genetics ; Cerebellum ; abnormalities ; Eye Abnormalities ; diagnosis ; genetics ; Humans ; Kidney Diseases, Cystic ; diagnosis ; genetics ; Membrane Proteins ; genetics ; Mutation ; Pedigree ; Retina ; abnormalities ; Whole Exome Sequencing

Objective To investigate the effect of sleeve gastrectomy (SG) on the distal ileum apical sodium-dependent bile acid transporter (ASBT) and ileum bile acid binding protein (IBABP)in obese type 2 diabetic rats.Methods Thirty 8-week-old male Sprague-Dawley rats were divided into groups O,F and N according to the random number table method.Among them,groups O and F were given high-sugar and high-fat diets,and group N were fed with normal diets before and after surgery.After 1,2 and 4 weeks,the body weight,blood glucose and plasma total bile acid levels were measured.The experimental animals were sacrificed 4 weeksafter operation and the terminal ileum tissue was harvested.Two types of ASBT and IABAP were detected by Western blot and immunohistochemistry.Bile acid transporters were tested.Results Blood glucose in group O was significantly lower than that in group F at 4 weeks postoperatively,(5.8±1.77) and (19.55±2.16) mmol/L,respectively (P＜0.05).Body weight in group O was significantly lower than that in group F at 4 weeks postoperatively,(182±10.66) g and (362±23.54) g,respectively (P＜0.05).Plasma total bile acid levels in group O were significantly higher than those in group F and N at 4 weeks after operation (52±18.48),(6.39±1.18),(5.89±1.21)μmol/L,respectively(P＜0.05).At the end of the 4th postoperative period,the expression of ASBT and IBABP in the distal ileum of group O increased,but there was no significant change in group F and group N.Conclusions The effect of SG on the hypoglycemic and weight-loss of obese T2DM rats is definite.The level of bile acid in plasma increases significantly after SG,and the expression of bile acid transporter in the terminal ileum of rats is one of the mechanisms.

Objective To explore the relationship between lumbar disc degeneration (LDD) of lumbar spinal stenosis(LSS)and the dural sac cross-sectional area(DSCA)by MRI measurement. Methods 91 patients with central degenerative LSS were randomly selected and 91 age-and sex-matched people without LSS were select-ed as a control group. LDD was classified into five grades by MRI detection according to the method proposed by Pfirrmann and DSCA were measured. Results LDD was not associated with age in LSS. The proportion of severe degenerated disc in lower lumbar levels were higher than that of L2/3 in the two groups;DSCA in severe degenerat-ed disc group was significantly smaller than that in light degenerated group only in L2/3 and L3/4 in LSS. There were no statistical differences in every lumbar level in the control group. Conclusions LDD in L4/5 and L5/S1 of LSS is more severe than that of the normal people. DSCA and LDD are positively correlated in L2/3 and L3/4,but not in L4/5 and L5/S1 for LSS.

Objective To explore the protective mechanism of HSP70 protein in traumatic brain injury (TBI)‐related acute gastric mucosal lesions in mice .Methods Forty adult male Balb/c mice were randomly divided into sham (A) ,TBI (B) ,TBI+ geranylgeranylacetone (GGA) (C) ,and TBI+saline (D) groups .TBI was induced via the Feeney impact model .GGA (800 mg/kg) was administered via oral tube beginning before the model was built in group C .The expressions of HSP70 protein in brain and gastric mucosa were determined by immunohistochemistry , and the apoptotic index was detected by TUNEL method .Results The injury area in mouse brain and gastric mucosa was greater in group B than in groups A and C (P<0 .05) .After model induction ,the content of HSP70 protein in group B was markedly higher in the brain and gastric mucosa ,which was notably higher than in group A (P<0 .05) .Obviously apoptotic cells were observed in groups B and D ,which were significantly higher than in groups A and C .GGA pretreatment enhanced the up‐regulated expression of HSP70 and decreased the apoptotic index distinctly ;HSP70 expression was higher in group C than in groups B and D ,but the apoptotic index was lower (P<0 .05) .Conclusion GGA can induce HSP70 protein expression in mouse brain and gastric mucosa .HSP70 is involved in the process of apoptosis inhibition .GGA can be used in the prevention and therapy of TBI‐related acute gastic mucosal lesions .

Objective To evaluate the clinical effects of thoracic close drainage with thin drainage tube assisted to thick drainage tube after video-assisted thoracic surgery(VATS)lobectomy. Methods We ret-rospectively reviewed 89 patients received VATS lobectomy in Chinese PLA General Hospital from January 2014 to September 2014. The patients with non-small cell lung cancer were divided into two groups:treatment group (50 patients)and control group(39 patients). Treatment group took thin tube assisted to thick tube of thoracic close drainage and control group took general thoracic closed drainage tube. We studied the operation time,the bleeding of operation,the number of lymph node dissection,time of first activity out of bed,the hospitalization time of post-operation,post-operative complications,the days of post-operative drainage,drainage volume,the effect of drainage,the VAS evaluation score of post-operative pain in the two groups. Results Compared with control group,there was no statistical significance in the differences of the time of operation[(2. 58 ± 0. 57)h vs(2. 57 ± 0. 50)h;t = 0. 127,P = 0. 681],bleeding of operation[(108. 00 ± 52. 84)ml vs(114. 10 ± 107. 18)ml;t = 0. 352,P = 0. 334],the number of lymph node dissection[(14. 20 ± 5. 95)vs(11. 21 ± 4. 71);t = 2. 576,P = 0. 068)],the staying time of drainage[(5. 66 ± 2. 53)d vs(5. 82 ± 2. 02)d;t =0. 324,P = 0. 219],the postoperative drainage volume[(1 141. 76 ± 819. 26)ml vs(1 022. 95 ± 464. 84) ml;t = 0. 889,P = 0. 367]and the occurrences of the post-operative complications(8. 00% vs 10. 25% ;χ2 =1. 750,P = 0. 726). There was statistical significance in the differences of the post-operative time of off-bed [(11. 28 ± 8. 78)h vs(13. 97 ± 7. 83)h;t = 4. 027,P = 0. 045],the time from surgery to discharge [(8. 36 ± 2. 63)d vs(9. 56 ± 2. 89)d;t = 2. 952,P = 0. 043]and the drainage effect(costophrenic angle sharp:72. 0% vs 46. 2% ;χ2 = 5. 329,P = 0. 017). In the two groups,there were statistical significance differences in scores of VAS for the 24 to 72 hours resting and coughing of post-operation:24 h[(2. 78 ± 1. 13)vs(3. 74 ± 1. 68);t = 3. 226,P < 0. 001)],48 h[(1. 98 ± 0. 59)vs(3. 33 ± 1. 72);t = 5. 189,P <0. 001)],72 h[(1. 94 ± 0. 55)vs(3. 15 ± 1. 60);t = 5. 010,P < 0. 001)],coughing[(3. 64 ± 1. 23)vs (5. 33 ± 1. 95);t = 5. 005,P < 0. 001)]. Conclusion The thin drainage tube assisted to thick drainage tube for thoracic close drainage make the drainage more effective,release the pain,shorten the hopital stay;moreo-ver,it is simple and safe for operation and easy to popularize with high modified value.

BACKGROUND:To in vitro isolate neural stem cel s with high purity and uniform biological properties and to establish a complete set of neural stem cel culture system is the basis for neural stem cel research.
OBJECTIVE:To establish an isolation and culture system for neural stem cel s from newborn mouse hippocampus, olfactory bulb and cortex and to analyze the biological properties of cel s.
METHODS:Neural stem cel s were isolated from the hippocampus, olfactory bulb and cortex tissue of newborn Kunming mice by mechanical separation and trypsin digestion. Serum-free culture technology, mechanical pipetting and trypsin digestion were used for subculture of neural stem cel s. 10%fetal bovine serum was used to induce differentiation of neural stem cel s. Neural stem cel s and their differentiated products were identified by
immunofluorescent staining of Nestin, CD133,β-TubulinIII, glial fibril ary acidic protein.
RESULTS AND CONCLUSION:The neural stem cel obtained from newborn mouse hippocampus, olfactory bulb and cortex had the capacity of self-renewal and differentiation which were positive for Nestin and CD133. After induction with fetal bovine serum, neural stem cel could differentiation toβ-tubulinIII or glial fibril ary acidic protein positive cel s that were neurons and astrocytes. This experiment has successful y established the neural stem cel isolation, culture, identification and induction system, providing experimental basis for subsequent studies of neural stem cel s.

Salidroside, the 8-O-beta-D-glucoside of tyrosol, is a novel adaptogenic drug extracted from the medicinal plant Rhodiola sachalinensis A. Bor. Due to the scarcity of R. sachalinensis and its low yield of salidroside, there is great interest in enhancing the production of salidroside by biotechnological process. Glucosylation of tyrosol is thought to be the final step in salidroside biosynthesis. In our related works, three UGT clones were isolated from the roots and the cultured cells. Our intention was to combine the catalytic specificity of these UGTs in vitro in order to change the level of salidroside in vivo by over-expression of the above UGTs. However, as the aglycone substrate of salidroside, the biosynthetic pathway of tyrosol and its regulation are less well understood. The results of related studies revealed that there are two different possibilities for the tyrosol biosynthetic pathway. One possibility is that tyrosol is produced from a p-coumaric acid precursor, which is derived mainly from phenylalanine. The second possibility is that the precursor of tyrosol might be tyramine, which is synthesized from tyrosine. Our previous work demonstrated that over-expression of the endogenous phenylalanine ammonia-lyase gene (PALrs1) and accumulation of p-coumaric acid did not facilitate tyrosol biosynthesis. In contrast, the data presented in our recent work provide in vitro and in vivo evidence that the tyrosine decarboxylase (RsTyrDC) is most likely to have an important function in the initial reaction of the salidroside biosynthesis pathway in R. Sachalinensis.
Genetic Engineering ; Glucosides ; biosynthesis ; Glycosylation ; Phenols ; Phenylethyl Alcohol ; analogs & derivatives ; chemistry ; metabolism ; Rhodiola ; metabolism ; Tyrosine ; metabolism ; Tyrosine Decarboxylase ; metabolism

The effect of AMP-activated protein kinase (AMPK) on KiSS-1 mRNA levels was detected by realtime PCR in the hypothalamic GT1-7 neurons. The promoter activity of KiSS-1 gene was detected by DualLuciferase Reporter Assay System.The effects of AMPK on the protein expression and subcellular distribution of SP1 were determined by Western blot.The results showed that AMPK reduced the mRNA expression and promoter activity of KiSS-1 gene while SP1 increased the promoter activity of KiSS-1 gene. Besides,AMPK alse decreased the translocation of SP1.These results suggest that AMPK may inhibit the expression of KiSS-1 gene by decreasing the translocation of SP1 from cytoplasm to nucleus in the hypothalamus GT1-7 neurons.

To develop a safe and efficient recombinant subunit vaccine to foot-and-mouth disease virus(FMDV)type Asia 1 in sheep,a tandem repeated multiple-epitope gene consisting of residues 137-160 and 197-211 of the VP1 gene of FMDV was designed and artificially synthesized.The biologically functional molecule,the ovine IgG heavy constant region(oIgG)as a protein carrier was introduced for design of the multiple-epitope recombinant vaccine and recombinant expression plasmids pET-30a-RE and pET-30a-RE-oIgG were successfully constructed.The recombinant proteins,RE and RE-oIgG,were expressed as a formation of inclusion bodies in E.coli.The immune potential of this vaccine regime in guinea pigs and sheep was evaluated.The results showed that IgG could significantly enhance the immune potential of antigenic epitopes.The recombinant protein RE-oIgG could not only elicit the high levels of neutralizing antibodies and lymphocytes proliferation responses in the vaccinated guinea pigs,but confer complete protection in guinea pigs against virus challenge.Although the recombinant protein RE could not confer protection in the vaccinated animals,it could delay the appearance of the clinical signs and reduce the severity of disease.Inspiringly,the titers of anti-FMDV neutralizing antibodies elicited in sheep vaccinated with RE-oIgG was significantly higher than that for the RE vaccination.Therefore,we speculated that this vaccine formulation may be a promising strategy for designing a novel vaccine against FMDV in the future.