Nature is abundant in protein scaffolds.By selecting suitable protein scaffold,display and screening methods,the rational and constrained random peptide library(RPL)can be constructed.Compared with the non-constrained RPL,it offered more opportunities for obtaining novel protein structures and more higher affinity ligands against the target molecules.At present,the protein scaffold constrained RPLs have been shown great potential in applications such as target selection,basic research,clinical diagnosis,medical therapy and so on.It is systematically introduced the structure bases,classification and construction of constrained RPL based on scaffolds,as well the recent great advances of application in selection against target molecules with S-S constrained scaffolds,antibodies,Zinc finger protein,Z domain,FN3 domain as important examples.

Adult ; Chronic Disease ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Occupational Diseases ; Radiation Injuries

Objective To summarize our experience in prevention and treatment of stricture after esopageal burns in the past thirty years.Methods There were 168 cases in this series.Of them,158 cases underwent surgical management in this study.Modified intraluminal stenting was used in 34 cases, colon interposition without resection of strictured esophagus in 77 cases,gastric transposion with resection of the stricture in 27,repair of cervical stricture with platysma myocutaneous flap in 22,and miscellane- ous operation in 12.Eleven cases experienced operation twice or more at our department.Results Twenty-nine cases recovered after treatment with intraluminal stenting,and 5 re-experienced stricture after stent removal.One of the 5 cases with failed stent responded to bougienage,and the remaining 4 cases re- quired esophageal reconstruction later.Of the 77 colon interpositions,5 cases died postoperatively,and complications of cervical anastomotic fistula occurred in 14 cases,anastomotic stenosis in 4,and abdomi- nal incision dehiscence in 2 cases.In the 27 cases with gastric transpositions,postoperative complications of anastomotic stricture occurred in 2 cases and empyema in 1 patient.There was a cervical leak in 3 ca- ses of the 22 cases treated with the repair of cervical esopageal or anastomotic stricture with a platysma myocutaneous flap.In the 12 cases treated with miscellaneous operation,one died of intestinal obstruc- tion.All the survivors had regular diet after discharge.Conclusions Intraluminal stenting can prevent the formation of caustic esophageal stricture.The location of the cicatricial esophagus dictates whether to perform concomitant esophagectomy during esophageal reconstruction.Platysma myocutaneous flap repair is an excellent method for the treatment of severe cervical esophageal or anastomotic stricture.

ObjectiveTo investigate the etiology and management of male urethral stricture at 8 medical centers in China during the period from 2004 to 2009 years,and to investigate whether there were any changes in etiology and management of urethral stricture with time change.MethodsThe database on 3455 male patients with urethral stricture who underwent treatment at 8 medical centers in China between January 2004 and December 2009 were prospectively collected.The databases were analyzed for possible cause of stricture and treatment techniques for urethral stricture,and for the changes in etiology and management with time change.ResultsThere were 3455 operations for urethral stricture during the study period.The main causes of urethral strictures were traumas in 1833 patients (53.05％),among which pelvic fractures were in 1327 (38.41％) and perineal trauma in 506 (14.65％).The second cause was iatrogenic causes in 1181 patients (34.18％ ),among which transurethral operations or examinations were in 602 (17.42％),hypospadias surgery in 291 (8.42％) and urethral catheterization in 164 (4.75％ ).Less common causes were urethritis in 201 patients (5.82％),lichen sclerosus in 149 (4.31％),undefined in 91 (2.63％).The treatments of urethral strictures were endourological surgery including internal urethrotomy and dilation and open urethroplasty including end-to-end urethroplasty and the substitude urethroplasty etc.The ratios of using various techniques in total number of patients were obviously different by time.The most application technique for treatment of urethral stricture was endourological surgery ( 709 ) during 2004 -2006 and occupied 52.67％ in total number of patients.It was gradually decreased during 2007 -2009 (726) and only occupied 34.42％ (P ＜0.01 ).Open urethroplasty gradually increased during 2007 -2009 ( 1243,58.94％ ) compared with the first three years (563,41.83％ ) (P ＜ 0.01 ). Conclusions During the recent years there was an increase in the incidence of urethral stricture being trauma and iatrogenic causes.The main treatments of urethral strictures were endourological surgery and open urethroplasty.Endourological surgery was significantly decreased in total number of patients,while open urethroplasty were significantly increased during the late three years.

Objective To study the correlation between serum hepatitis B virus large surface protein (HBV-LP)and hepatitis B virus DNA (HBV-DNA) load in hepatitis B patients treated with lamivudine, and the application of serum HBV-LP measurement in evaluation of lamivudine therapeutic effect and end point judgement.Methods HBV DNA was detected by fluorescent quantitative PCR (FQ-PCR), serum HBV-LP was detected by enzyme linked immunosorbent assay (ELISA). Results Both HBV-DNA and HBV-LP decreased in patients responding to lamivudine therapy, and HBV-LP turned negative 3 months later than HBV-DNA. In patients resistant to lamivudine therapy, neither HBV-DNA nor HBV-LP turned negative. In patients whose symptoms relapsed, HBV-LP didn't turn negative, while HBV-DNA was undetectable only transitorily. Conclusion Measuring serum HBV-LP dynamically can be useful in evaluation of lamivudine therapeutic effect.

OBJECTIVE: To screen for proteins interacting with ataxin-3 by yeast two-hybrid system 3, and to discuss the function of ataxin-3 and pathogenesis of spinocerebellar ataxia type 3 and Machado-Joseph disease (SCA3/MJD). METHODS: First we sub-cloned the full reading frame of both wild-type and mutant ataxin-3 into carrier pGBKT7 (ataxin-3-bait), and then screened human brain cDNA library with ataxin-3-bait. RESULTS: We found five positive clones in 6.5 x 10(6) transformers. After sequencing, we knew all of them were novel ataxin-3 interacting proteins. Three were corresponded to the known sequences coding the known proteins, which were human Rho GDP dissociation inhibitor alpha, small ubiquitin-like modifier 1, and human neuronal amiloride-sensitive cation channel 2. Another two of the five were unknown. CONCLUSION Small ubiquitin-like modifier 1 probably interacted with ataxin-3, suggesting that the sumoylation probably participated in post-translation modifying of ataxin-3 and pathogenesis of SCA3/MJD.
Ataxin-3 ; Brain ; metabolism ; Gene Library ; Humans ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Protein Interaction Mapping ; Repressor Proteins ; genetics ; metabolism ; Spinocerebellar Degenerations ; genetics ; metabolism ; Two-Hybrid System Techniques ; Yeasts ; genetics

AIMIn order to explore the neurobiological mechanism of morphine addiction and treatment methods, the acute and chronic effects of morphine on the intracellular free calcium concentration ([Ca2+]i) in cultured hippocampal neurons were investigated.

METHODSChanges of [Ca2+]i induced by morphine in primarily cultured hippocampal neurons were measured by confocal laser scanning microscopy using Ca(2+) -sensitive dye fluo-4 as the calcium fluorescent probe.

RESULTSMorphine actually induced the increase in [Ca2+]i of hippocampal neurons. This process could be blocked by naltrindole (delta opioid receptor antagonist) pretreatment, but not by CTOP (micro opioid receptor antagonist) pretreatment. Pretreatment of the cells with thapsigargin almost completely blocked morphine-evoked response; while pretreatment of the cells with verapamil partially inhibited this response. After exposure to 100 micromol/L morphine for 24 h, intracellular [Ca2+]i increased and the increase could be intensified after adding 10 micromol/L naloxone to the medium.

CONCLUSIONMorphine induced the release of Ca2+ is mainly from inositol 1, 4, 5-trisphosphate (IP3) sensitive stores in hippocampal neuron of rats through activation of delta2 subtype opioid receptor.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Hippocampus ; cytology ; Male ; Microscopy, Confocal ; Morphine ; pharmacology ; Neurons ; cytology ; drug effects ; Rats ; Rats, Wistar

OBJECTIVETo observe the experience and the outcome of pharyngo-colonic anastomosis for esophageal reconstruction in diffuse corrosive esophageal stricture involving hypopharynx.

METHODSThis retrospective report reviews the experience and results of 14 patients who underwent esophageal reconstruction by pharyngo-colonic anastomosis without resection of intra thoracic stricture esophagus. The left half colonic segment was pulled up to the neck through the substernal space in all patients.

RESULTSThere was no operative or hospital death. Postoperative complications include cervical anastomotic fistula in four patients, rupture of the abdominal incision in 1. The length of follow-up ranged from half year to 10 years with an average of 4 years. Anastomotic stenosis occurred in 2 patients. One case improved after dilatation and the other one healed by plastic operation. One patient began to vomit after diet in seven months later with barium swallowing the abdominal colon graft was redundant and this patient was cured with side by side between the colon and the stomach.

CONCLUSIONThe successful reconstruction for hypo-pharyngo-esophageal stricture requires a correct and larger hypopharyngeal opening and a good anastomotic technique. From our experience this procedure is shown to be safe and effective.

Adolescent ; Adult ; Anastomosis, Surgical ; Burns ; complications ; Child ; Child, Preschool ; Colon ; surgery ; Esophageal Stenosis ; etiology ; surgery ; Esophagoplasty ; methods ; Female ; Humans ; Male ; Pharynx ; surgery ; Retrospective Studies

OBJECTIVETo explore the prognostic values of Raf-1 kinase (Raf-1), phosphorylated mitogen extracellular kinase 1 (pMEK1), and phosphorylated extracellular signal-regulated protein kinase 1/2(pERK1/2) in hepatocellular carcinoma (HCC) patients.

METHODSWe assessed the expressions of Raf-1, pMEK1, and pERK1/2 in HCC using immunohistochemical techniques. The relationships between the expressions of Raf-1, pMEK1, and pERK1/2 and the prognosis were explored.

RESULTSThe over-expression rates of Raf-1, pMEK1, and pERK1/2 in HCC were 38.3%, 46.7%, and 38.3%, respectively. The over-expressions of Raf-1, pMEK1, and pERK1/2 were positively correlated with each other (P>0.05), but had no significant correlation with sex, age, α-fetoprotein, hepatitis B surface antigen status, the TNM stage, size,differentiation and vascular invasion of tumor, and liver cirrhosis (P>0.05). Univariate survival analysis and COX proportional hazard regression model showed that Raf-1 over-expression was an independent prognostic factor of poor survival (P<0.05).

CONCLUSIONRaf-1 over-expression is an independent marker for the patients of HCC, which may provide new clue in the future targeted therapy.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnosis ; enzymology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Female ; Humans ; Liver Neoplasms ; diagnosis ; enzymology ; MAP Kinase Kinase 1 ; metabolism ; Male ; Middle Aged ; Phosphorylation ; Prognosis ; Proto-Oncogene Proteins c-raf ; metabolism

OBJECTIVETo identify gene expression differences between abdominal aorta aneurysm (AAA) and normal abdominal artery.

METHODSFive AAA specimens and five normal abdominal artery specimens were collected. Total RNA was extracted and purified from the samples independently. Then isolated RNA was labeled and hybridized onto the Affymetrix Genechip.

RESULTSOne thousand nine hundreds and sixty-two differently expressed genes were identified in AAA compared with normal abdominal artery, of which 554 genes were up-regulated and 1408 genes were down-regulated. Functional analysis revealed that genes involved in immune response, inflammatory response and chemokines were significantly up-regulated in AAA.

CONCLUSIONThe differently expressed genes may be useful in providing new insights and foster a better understanding of the mechanism of AAA.

Aorta, Abdominal ; metabolism ; Aortic Aneurysm, Abdominal ; genetics ; Gene Expression Profiling ; Humans ; Oligonucleotide Array Sequence Analysis