Objective To investigate the effect of porous silk fibroin scaffolds (PSFSs) combined with chondroitinase ABC (ChABC)for treatment of rats with spinal cord injury (SCI).Methods After exposed to T9 spinal cord transection injury,96 SD rats were divided into control group,PSFSs group,ChABC group,and PSFSs plus ChABC group according to random number table.BBB scoring system was used to evaluate hindlimb motor function in rats.Immunohistochemistry and Western blot analysis were performed to detect expression levels of neurofilament-200 (NF-200),glial fibrillary acidic protein (GFAP),and growth associated protein-43 (GAP-43) of the injured spinal cord.Immuno-fluorescence staining was carried out to evaluate regeneration of nerve fiber.Results BBB score improved in PSFSs group (8.1 ± 0.8),ChABC group (9.0 ± 1.1),and PSFSs plus ChABC group (13.7 ± 1.3) compared with control group 4 weeks after injury (5.3 ±0.7,P ＜0.05).Immunohistochemistry showed higher integral absorbance (IA) values of NF-200 and GAP-43 in those treatment groups,but smaller GFAP-positive area was observed compared with control group (P ＜ 0.05).Immuno-fluorescence staining indicated more GAP-43 growth at injury sites in PSFSs plus ChABC group in contrast with other 3 groups.Western blotting showed levels of NF-200,GFAP,and GAP-43 differed among groups (P ＜ 0.05).Conclusion PSFSs combined with chondroitinase ABC transplantation can enhance axonal regeneration,inhibit glial scar proliferation and hence promote motor function recovery.

Objective To study the effect of lipopolysaccharide(LPS)on the endo-pulmonary natrium channel(ENaC)expression in the lung of rats with acute lung injured.Method Sixteen rats were randomly divided into normal control group and LPS-group.Rats of normal control group and LPS-group were killed at 6 hours after intravenous injection of normal saline(8 ml/kg)or LPS(8 mg/kg).The extent of lung injury was assessed by arterial blood gas analysis and histological examination.At the same time,?-ENaC protein and???- ENaC mRNA expression in the lung tissue were analyzed by immunohistochemistry and RT-PCR.Results PaO_2 in LPS-group was noticeably lower than in normal control group(P

OBJECTIVEIn order to fully understand hepatitis c virus (HCV) genotype 3b, 1a, 2b and 6a infection in China, We built HCV 5'-noncoding region (5'-NCR) of different genotypes and subtypes.

METHODSThe classification HCV into variable genotypes (subtypes) was carried on by programs A, B and C A. Using a combination of three restriction endonuclease BHH' (BsrB I, Hae II, Hinf I) digestions at the same time. The distinct genotypes were classified into 5 groups: genotype 1 (1a, 1b), 6a, 2 (2a, 2b), genotype 3 (3a, 3b), genotype4 (4a). B. With regard to genotype 1, we could distinguish subtype 1a from 1b using BstU I digestion. C. Using restriction endonuclease Hae III, genotype 2a, 2b, 3b, 4a, 6a are differentiated respectively.

RESULTS(1) HCV genotype 1a, 1b, 2a, 2b, 3a, 3b, 4a, 6a are fully discriminated by comparison with the genotypes regular samples. (2) Of the 93 patients, HCV genotype distribution in China was 66.67% for 1b, 18.28% for 2a, 3.23% for 1b/2b, 3b, 2b respectively. 2.15% for 2a/2b, 1b/2a respectively. 1.08% for 1a.

CONCLUSIONThis research indicated that adoption of HCV 5'-NCR A B C restriction endonuclease digestions techniques, might be sensitive and efficient to detect HCV and discriminate HCV genotype (subtypes) 1a to 6a.

5' Untranslated Regions ; chemistry ; DNA Restriction Enzymes ; Genotype ; Hepacivirus ; classification ; genetics ; RNA, Viral ; analysis

OBJECTIVETo assess the prognosis and reproductive outcomes of laparoscopic intracapsular myomectomy.

METHODSA total of 673 women received subserosal and intramural intracapsular laparoscopic myomectomy between March, 2007 and March, 2012, and their post-operative complications, the need for subsequent surgery, symptomatic relief and reproductive outcomes were analyzed.

RESULTSOf these patients, 42.4% had subserosal myomas and 57.6% had intramural myomas. The mean total operative time was 96∓41 min with a mean blood loss of 128∓46.2 ml, and 82.3% of the patients were discharged 48 h after the operation without early complications. A small fraction (2.3%) of the patients had a second laparoscopic myomectomy for recurrent fibroids. Of the fertility-demanding women who underwent myomectomy, 71% achieved pregnancy, 49.8% underwent caesarean section, 8% had operative vaginal deliveries, and 42.2% had spontaneous deliveries; uterine rupture occurred in none of the cases.

CONCLUSIONLaparoscopic intracapsular myomectomy, by preserving the fibroid pseudocapsule and myometrial integrity, has no early postoperative complications and ensures good fertility rates and reproductive outcomes.

Adult ; Female ; Fertility ; Humans ; Laparoscopy ; Leiomyoma ; surgery ; Prognosis ; Retrospective Studies ; Uterine Myomectomy ; Uterine Neoplasms ; surgery

Objective To study the suppressive effect of knockdown of miR-21 on the U87 human giioma xenograft growth and the possible mechanism. Methods Nude mice bearing U87 human glioblastoma subcutaneously were treated with miRNA-21 anfisense oligonucleotides(AS-miR-21)intratumomlly every 3 d until the observation peded ended.The tumor volume of the mice treated withAS-miR-21 was measured regularly as compared with that in the control untreated mice and in the mice treated with scramble oligonucelotides(ODN).Finally,the tumors were removed from nude mice for the examination.In-sire hybridization and real-time PCR were conducted to detect the miRNA expression of miR-21.The biological charaetedsties of the tumors were evaluated by HE and immunohistochemieal staining, and the cell apoptosis was detected by TUNEL method. Resulls During the observation period,the tumor growth was delayed and the final tumor volume of AS-miR-21 heated group was smaller than that in the control and scramble ODN treatedg roup(F=6-056,P=0.007).The expression of miRNA precursor was knocked down in As-miRNA treated tunlors compared with that in untreated or scramble ODN treated tumors.Histopathological examination exhibited the appearance of degraded malignancy.The expressions of PCNA and MMP-9 were down-regulated while Septin-7 and P21 were up-regulated and apoptotic index was increased significantly (F=141.021,P=000) as well.Conclusion The suppressive effect of anti-miR-21 ODNs on the growth of U87 human glioma xenogratts is significant and miR-21 Call be taken as a candidate for gene therapy ofhuman glioma.

OBJECTIVETo screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients.

METHODSSerum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software. The differential protein spots were further identified by MALDI-TOF MS/MS.

RESULTSComparing the results using 12.5% SDS-PAGE gel, there were more protein bands (between 3 x 10(3) and 20 x 10(3)) and low molecular weight (MW) protein spots (less than 20 x 10(3)) were clearly shown in the 16% SDS-PAGE gel. Fifteen differential protein spots representing 5 proteins were found in the 3 groups by inter-class comparison and they were then identified. Compared with those in the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were all down regulated in HCC groups and haptoglobin-2 was over expressed. All 5 proteins decreased more in the PVTT group than in the non-PVTT group.

CONCLUSIONThe expression of low MW serum protein obviously changes in the beginning and in the progressive stage of HCC, and differentially expressed low MW proteins might be potential biomarkers in an early prognostic prediction and surveillance in the treatment for HCC and PVTT.

Adult ; Blood Proteins ; analysis ; Carcinoma, Hepatocellular ; blood ; pathology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Portal Vein ; pathology ; Proteome ; analysis

OBJECTIVETo assess the feasibility and efficiency of eliciting leukemia-specific T cell responses in acute myeloid leukemia patients in complete remission (AML-CR) in vitro by dendritic cells (DC) pulsed with the leukemia cells total RNA.

METHODSThe immature DCs were generated from the adherent bone marrow mononuclear cell in vitro in the presence of combined cytokines (GM-CSF 100 ng/ml, IL-4 500 U/ml), and pulsed with total RNA isolated from autologous leukemic cells by cationic lipid 1,2-dioleoyloxy-3-trimethyl ammonium propane (DOTAP) at day 5 of culture. Then the cells were incubated for another 24 h in a medium containing 10 ng/ml of TNF-alpha for maturation of DC. After the total 7 days culture, the cells were harvested as the mRNA-DC and the expression of mature DC markers were determined by FACS. The proliferative capacity of T cell activated by mRNA-DC was determined by MTT assay. Meanwhile, the mRNA-DC was co-cultured with T lymphocytes at a ratio of 1:3 for 7 days. The activated T lymphocytes were harvested, the secretion of IFN-gamma was determined by ELISPOT assay, and the cytotoxicity was analyzed in vitro by LDH release assay.

RESULTSAfter culture, the BMMNC from 14 AML-CR patients developed morphologic and phenotypic characteristics of mature DC. At a stimulator/reactor ratio of 1:16, auto-T lymphocytes primed with mRNA-DC exhibited significant proliferative activity compared with T lymphocyte primed with non-pulsed DC [(36.84 +/- 5.68)% vs (12.20 +/- 3.16)%, (P < 0.05)]. An expansion of mRNA reacted T cell secreting IFN-gamma could be observed on ELISPOT assay. At an effector/target ratio of 20:1, the auto-T lymphocytes primed with mRNA-DC exhibited significant killing activity to auto-AML cells (45.46 +/- 6.34 )% as compared with that stimulated by IL-2 alone (13.26 +/- 2.28)% or primed by non-pulsed DC (12.32 +/- 1.32)% (P < 0.05).

CONCLUSIONImmunization with DC-leukemia cell RNA vaccines may be a simple, rapid and potent approach to elicitation of T cell-mediated anti-leukemia immunity.

Adolescent ; Adult ; Cell Communication ; Cells, Cultured ; Coculture Techniques ; Dendritic Cells ; drug effects ; immunology ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; immunology ; Male ; Middle Aged ; RNA ; pharmacology ; T-Lymphocytes ; immunology

Objective To explore the efficacy and safety of autologous peripheral blood stem cell transplantation (APBSCT) in the treatment of systemic lupus erythematosus.Methods Nine patients with systemic lupus erythematosus were enrolled in this study.Patients were given cyclophosphamide and granu- locyte colony-stimulating factor(G-CSF)as the mobilization regimen.Urine was alkalinized and hydrolyzed to protect the function of the heart,liver and kidney of the patients.A CS3000 Plus blood cell separator was used to collect peripheral blood stem cells,which were preserved in liquid nitrogen.Two to five days before the administration of the stem cells,the patients were pretreated with intravenous injection of cyclophos- phamide (50 mg?kg~(-1)?day~1) for 4 consecutive days and antithymocyte globulin (ATG,2.5 mg?kg~(-1)?day~1) for 3 consecutive days.Granulocytes were recoverd by G-CSF stimulation.Then,the peripheral blood stem cells were reinfused.Therapeutic effect was evaluated by assessment of alteration of clinical manifestation (skin erythema),levels of proteinuria and antoantibodies,hematopoietic reconstitution and occurrence of transplantation related complications.Results After transplantation,all patients had been successfully en- grafted.The time for peripheral leucocyte count to reach 1.0?10~9/L was 7～15d;the time for platelets to reach 20?10~9/L was 0～21 d.The skin erythema resolved in all patients;proteinuria decreased to normal level and the autoantibodies became negative in most of the patients.Serum sickness-like response occurred in all patients,renal and heart failure in 1 patient,hemorrhagic cystitis in 3 patients,psychiatric disorders in 1 patient,candidal infection in 1 patient.Conclusion One-year follow up suggests that autologous stem cell transplantation is markedly effective and relatively safe for systemic lupus erythematosus.However,the duration of remission remains to be investigated in a long-term follow up study.

Adult ; Female ; Humans ; Laparoscopy ; methods ; Liver Neoplasms ; diagnostic imaging ; pathology ; surgery ; Magnetic Resonance Imaging ; Neoplasms, Multiple Primary ; diagnostic imaging ; pathology ; surgery ; Tomography, X-Ray Computed

OBJECTIVETo study the feasibility of binding pancreatic duct to mucosa anastomosis (BDM)-a complementary procedure to both binding pancreaticojejunostomy and binding pancreaticogastrostomy.

METHODS(1) Animal experimental study:gastrostomy and jejunostomy were performed on six adult New Zealand rabbits. The gastrostomy and jejunostomy shared a same stent (rubber urethral catheter, silicone tube or plastic infusion tube). Both ends of the stent were placed in gastric and enteric cavity. Purse-string suture was performed around the stent before the jejunum and the stomach were brought together for fixation by few stitches. And to observe whether the purse-string suture around a plastic tube, rubber tube or silicon tube inserted into jejunum and/or stomach can prevent leaking out of the jejunal or gastric content to cause peritonitis. (2) Clinically 7 patients were performed with BDM anastomosis. The procedure was consisted of five steps: preparation of the pancreatic stump;preparation of the jejunum; preparation of the fixing sutures between the pancreatic stump and the jejunum; implementation of the anastomosis; lastly, fixation of the jejunum beside the pancreas stump. Post-operative periodic examination of the blood amylase and the amylase in the abdominal drainage. Pancreatic fistula was classified in to two categories: parenchymal fistula (pancreatic cut surface fistula) and anastomotic leakage.

RESULTSAnimal experiment did not show any leakage around the plastic tube or silicon tube inserted into jejunum and(or) stomach. There was no anastomotic leak in all the patients. There was transient increase of amylase in two cases, but the volume of drainage did not exceed 50 ml/d and the recovery of the patients was not affected.

CONCLUSIONSBDM is a simple, safe and easy procedure to perform. It provides to the surgeons with a new option in different situations to achieve the most ideal surgical result.

Anastomosis, Surgical ; methods ; Animals ; Gastric Mucosa ; surgery ; Intestinal Mucosa ; surgery ; Pancreatic Ducts ; surgery ; Pancreaticoduodenectomy ; methods ; Pancreaticojejunostomy ; methods ; Rabbits