Objective To investigate the changes of the expression of GDNF and GDNFR?1 of long-term denervation of posterior cricoarytenoid muscles(PCAMs).Methods 38 patients with vocal paralysis were grouped into four according to their denervated period of time while 12 normal PCAMs as control group.Using double immunofluorescence stain,changes of GDNF and GDNFR1 expression were observed in myofibers at different time points after denervation.Results Double immunofluorescence stain with antibodies against GDNF and GDNFR1 showed no staining in the control group,and study groups.However,after the muscle denervation lasted for 6-12 months and 1-2 years,noted was a significant accumulation of GDNF and GDNFR1 protein in cytolemma and endochylema of myofiber.The mean grey scales and positive region ratios were compared using the image analysis system.The results revealed the levels of GDNF and GDNFR1 protein expression in 6-12 months group,1-2yr group changed significantly(P0.05).Conclusion The changes in expression of GDNF and the acceptor GDNFR-?1,a powerful neurotrophic factor,implied that a good nervous regenerated microenvironment in PCAMs within 2 years.This experiment indicated that denervated posterior cricoarytenoid muscles are able to regain their functions through reinneration within 2 years.

Cystectomy ; Fatal Outcome ; Female ; Glial Fibrillary Acidic Protein ; metabolism ; Granular Cell Tumor ; pathology ; secondary ; surgery ; Humans ; Immunohistochemistry ; Middle Aged ; S100 Proteins ; metabolism ; Urinary Bladder ; chemistry ; pathology ; surgery ; Urinary Bladder Neoplasms ; metabolism ; pathology ; surgery ; Vaginal Neoplasms ; metabolism ; secondary ; surgery

To counteract the immune system in parasitic hosts, some viruses encode proteins to suppress the RNA interference (RNAi) effect. In this report, we established two RNAi systems to be easily observed with strong and obvious effect. The function of the P19 of tomato bushy stunt virus, which suppresses RNAi in mammal cells, was then studied using these two systems. Short hairpin RNAs targeting green fluorescence protein (pshRNA-GFP) and firefly luciferase (pshRNA-luc) were designed and inserted into a eukaryotic transcriptional vector pTZU6+1, respectively. The shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without P19. The GFP expression level was assayed by fluorescence microscopy, Western blotting and RT-PCR. The luciferase expression level was analyzed by the dual-luciferase assay system. pshRNA designed in this study down-regulated the target gene specifically and efficiently, with a decrease of expression of both genes of about 70%, respectively. When P19 was introduced into the RNAi systems, the expression of both GFP and the luciferase were mostly recovered compared with the control groups. The RNAi systems of GFP and luciferase were constructed successfully, demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNAi effect induced by shRNA in mammal cells.

To discuss the effective method of decreasing the postoperative recurrence rate of recurrent pterygium.
●METHODS:Totally 126 cases (126 eyes) with recurrent pterygium were randomly divided into A group (56 cases) and B group ( 70 cases ). Group A was treated by pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation, group B was treated by amniotic membrane transplantation. The followed-up time after surgery was 6-24mo.
●RESULTS:ln group A, postoperative 5-7d (average 5. 62± 1. 38d), cornea epithelium was repaired. ln group B, postoperative 7- 10d ( average 7. 38 ± 1. 12d), the corneal wound was healed. There was statistical significant difference between two groups (t = 4. 307,P<0. 05). Three cases recurrence were noted in A therapeutic group (56 cases), the recurrent rate was 5. 4%; Twelve cases recurrence were noted in B compared group (70 cases), the recurrent rate was 17. 1%. There was statistical significant difference between two groups(P<0. 05).
●CONCLUSlON: lt is suggested that pterygium conjunctive reverse transplantation combined with amniotic membrane transplantation is effective in the treatment of recurrent pterygium.

OBJECTIVETo search all studies that had been published in the world with regarding to the effectiveness of the extent of intralesional curettage and wide excision for recurrence rate and complications and comparative functional outcomes in patients with giant cell tumours (GCT) of the distal radius and analyze them which were in high quality by means of Meta analysis, in order to give some evidences for the choice of method dealing with giant cell tumors GCT in surgery.

METHODSCochrane central register of controlled trials(Issue 8 2014), PubMed(1970-01-01/2013-01-01), Ovid (1970-01-01/2013- 01-01), Elsevier (1970-01-01/2013-01-01), CNKI (1970-01-01/2013-01-01) were searched. Including intralesional curettage and wide excision were performed to treat giant cell tumors (GCTs) of the distal radius in the literatures, selecting on meet eligibility in the standard literatures underwent strict quality assessment. The Meta-analysis was performed with software RevMan5.0 from the Cochrane collaboration. Additionally, the analysis checked the heterogeneity of data. The effectiveness of the extent of intralesional curettage and wide excision for recurrence rate and complication in patients with giant cell tumours of the distal radius were evaluated and Odds Ratio was calculated.

RESULTSSeven relevant articles were identified involving total 163 cases. Among them, 92 cases were intralesional curettage (PMMA, n = 54; bone graft, n = 33; no PMMA or bone grafts, n = 5) and 71 cases were wide excision. The patients in the intralesional curettage group had a higher recurrence rate [OR = 3.87, 95% CI (1.42, 10.53)],especially for Campanacci grade 3 GCTs [OR = 10.12, 95% CI (1.57, 65.27)], yet fewer major complications [OR = 0.13, 95% CI (0.04, 0.40)] than the wide excision group. The use of PMMA versus bone graft did not affect the recur- rence rate [OR = 0.96, 95% CI (0.26, 3.56)]. By selecting the system evaluation of MSTS, the VAS and dynamometer, the result showed that the intralesional curettage group was equivalent or preferable to wide excision in terms of function rehabilitation.

CONCLUSIONBased on data obtained from the limited number of studies available, intralesional curettage appears to be moreappropriate for the treatment of local lesions (Grade 1 and 2) than Grade 3 GCTs of the distal radius. Moreover, PMMA was not additionally effective as an adjuvant, the intralesional curettage group was found to be equivalent or preferable to wide excision in terms of function rehabilitation.

Bone Neoplasms ; surgery ; Curettage ; methods ; Giant Cell Tumor of Bone ; surgery ; Humans ; Radius ; surgery

Objective To compare the curative effect of high ligation+exfoliation and subfascial endoscopic perforator surgery(SEPS)for superficial varicose veins in calf+invagination spot-striping surgery in great venous varicosity.Methods Study group(42 patients)accepted SEPS+invagination spot-striping surgery and control group (42 patients)accepted traditional surgeries.Operation duration,bleeding volume in operation,the time of beginning movement away from bed after operation,hospitalization duration,the degree of pain,the scar,the recrudescence af- ter operation and the instance of the ulcer heals of two groups were compared.Results Operation duration,bleeding volume in operation,the time of begin movement away from bed after operation and hospitalization durations of study group were significantly lower than those of control group(P0.05).All of the patients in study group recovered without severe syndromes such as venous thrombosis,skin necrosis,lower limb functional disorder etc.They had no recrudesce after 4～16 months and were satisfied with the curative effect.Con- elusions The clinical curative effect of SEPS+invagination spot-striping surgery in great venous varicosity is superi- or to that of traditional operation and it has the advantages such as minor wound,few scars,light pains,short hospi- talization duration,without recrudescence,the ulcer heals quickly and so on.

OBJECTIVETo study the effect of HCV NS5A protein on HCV IRES-dependent translation in HepG2 cells.

METHODSHepG2 cells were co-transfected with a plasmid vector containing a bicistronic transcript carrying Renilla luciferase and firefly luciferase genes separated by HCV IRES sequences, and an expressing vector producing the NS5A protein. The luciferase activity and the mRNA of the luciferase gene were then detected. The NS5A expression was confirmed by fluorescence microscopy.

RESULTSHCV NS5A protein was detected in the cytoplasm of the HepG2 cells transfected with pcDNA-NS5A, and the luciferase activity was up-regulated in the presence of the HCV NS5A protein while the expression of luciferase mRNA showed no difference.

CONCLUSIONHCV NS5A protein can upregulate the HCV IRES activity and this effect is dose-dependent with NS5A.

Hep G2 Cells ; Hepacivirus ; genetics ; metabolism ; Humans ; Plasmids ; Protein Biosynthesis ; Protein Structure, Secondary ; Ribosomes ; metabolism ; Transfection ; Viral Nonstructural Proteins ; metabolism

Airway remodeling is an important pathological feature of asthma and the basis of severe asthma. Proliferation of airway smooth muscle cells (ASMCs) is a major contributor to airway remodeling. As an important Ca(2+) channel, transient receptor potential vanilloid 1 (TRPV1) plays the key role in the cell pathological and physiological processes. This study investigated the expression and activity of TRPV1 channel, and further clarified the effect of TRPV1 channel on the ASMCs proliferation and apoptosis in order to provide the scientific basis to treat asthmatic airway remodeling in clinical practice. Immunofluorescence staining and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of TRPV1 in rat ASMCs. Intracellular Ca(2+) was detected using the single cell confocal fluorescence microscopy measurement loaded with Fluo-4/AM. The cell cycles were observed by flow cytometry. MTT assay and Hoechst 33258 staining were used to detect the proliferation and apoptosis of ASMCs in rats respectively. The data showed that: (1) TRPV1 channel was present in rat ASMCs. (2) TRPV1 channel agonist, capsaicin, increased the Ca(2+) influx in a concentration-dependent manner (EC50=284.3±58 nmol/L). TRPV1 channel antagonist, capsazepine, inhibited Ca(2+) influx in rat ASMCs. (3) Capsaicin significantly increased the percentage of S+G2M ASMCs and the absorbance of MTT assay. Capsazepine had the opposite effect. (4) Capsaicin significantly inhibited the apoptosis, whereas capsazepine had the opposite effect. These results suggest that TRPV1 is present and mediates Ca(2+) influx in rat ASMCs. TRPV1 activity stimulates proliferation of ASMCs in rats.

Aged ; Castleman Disease ; immunology ; pathology ; Diagnosis, Differential ; Humans ; Immunoglobulin G ; metabolism ; Lymphatic Diseases ; immunology ; pathology ; surgery ; Lymphoma ; pathology ; Male ; Plasma Cells ; immunology ; Pseudolymphoma ; immunology ; pathology

Humanin(HN,its analogue [Gly14]-Humanin,HNG)was originally identified as an endogenous peptide that protects neuronal cells from apoptosis induced by various types of Alzheimer's disease-related insults.But the relative low content of this peptide in its natural sources limits its further characterization.An expression vector pET32a/HNG was corstructed and transformed it into E.coli BL21 trxB(DE3).HNG was expressed as a fusion protein in the soluble fraction and was purified by nickel affinity chromatography.Subsequently,the purified fusion protein was cleaved by enterokinase and was further purified by reverse-phase HPLC.A 23 mg recombinant HNG(rHNG)from 1 L bacterial culture was purified.The molecular weight of rHNG determined by ESI-MS was 2876.5 Da which was the expected size for correctly processed peptide.The N-terminal amino acid sequence of rHNG determined by Edman degradation method is identical to the theoretical sequence.Neuroprotective bioassay studies of rHNG exhibited its potential neuroprotective effect comparable to that of the natural HNG peptide.