Objective To explore the effects of donor dendritic cells(DC)treated with Ad-IL- 12p35siRNA on the survival of allogragft recipients.Methods The recombinant adenoviral vectors carrying IL-12p35siRNA and HKsiRNA were transfected into bone marrow derived DC of BALB/C murine.C57BL/6 recipients were infused with DG(Ad-IL-12p35siRNA DC,Ad-HKsiRNA DC and control DC)from BALB/C donors 7 days before cardiac allograft,the survival time of murine and the change of T_H 1 and T_H2(IL-2,IL-4,IL-10 and IFN-?)cytokine were observed.Results The survival time of p35 group(20.17?2.71)days was longer than that of control group(7.81?1.61)days and HK group(7.17?1.60)days.The concentration of IL-2 and IFN-?in p35 group were significantly lower than those of control group and HK group,otherwise were the concentration of IL-4 and IL-10. Conclusion Pretreatment of dondor dendritic ceils with Ad-IL-12p35siRNA could prolonged cardiac allograft survival in recipicents.

Terminal restriction fragment length polymorphism (T_RFLP) analysis is a culture- independent approach for analyzing microbial community in environment. It bases on PCR technology, and its process includes DNA extraction of environmental samples, amplification of genes encoding the 16S rRNA, 18S rRNA or enzymes with fluorescently labeled primers, the restriction enzyme digestion of PCR products, cap- illary electrophoresis and the analysis of T_RFLP profile. It has been proved to be powerful applied on mi- crobial community in environment since developed in 1997. Currently, T_RFLP rarely applied in China, and it has no applications on microbial community analysis of nitrifying bacteria. In this article, the fundamental principle of this technique and the recent applications of T_RFLP on microbial community are summarized; in addition, it illustrates the confinements of conventional culture-dependent of nitrifying bacteria and the foreground of T_RFLP applying on microbial community structure analysis of nitrifying bacteria.

Objective To prepare controlled release microspheres of vascular endothelial growth factor(VEGF)& calcium alginate and observe their effect on proliferation of human umbilical vein endo- thelial cells(HUVEC)in order to provide theoretical basis for controlled release of VEGF facilitating an- giogenesis of tissue engineering bone.Methods VEGF-calcium alginate microspheres were prepared by using the needle extrusion/external gelation method to investigate physicochemical character and in vitro release of VEGF.According to the different ingredients added into the culture medium,the seconda- ry cultured HUVEC were divided into four groups,ie,control group,microsphere group,VEGF group and VEGF-calcium alginate microsphere group,in which the proliferation of the cultured HUVEC was ob- served with cell counting method,MTT method and flow cytometry.Results The calcium alginate mi- crospheres were revealed as spherical shape and evenly distributed,with mean grain diameter of(560?50)?m,carrying capacity of 0.72 ng/mg and the encapsulation efficiency of 54%.Smooth controlled re- lease in VEGF-alginate microspheres lasted for more than 10 days.Proliferation of the cultured HUVEC was accelerated the most in VEGF group at the beginning but in EGF-calcium alginate microsphere group at midanaphase compared with other groups,with statistical difference(P＜0.05).There was no statis- tical difference upon cell counting,cell activity and time point of cell cycle between control group and mi- crosphere group.Conclusion VEGF-sodium alginate microapheres can continue activity of VEGF,re- lease VEGF for over 10 days and promote proliferation cultured HUVEC for a long time.

Dimethyl sulfide (DMS) has been historically recognized as a metabolite of the marine microorganism or a disgusting component for the smell of halitosis patients. In our recent study, DMS has been identified as a cytoprotectant that protects against oxidative-stress induced cell death and aging. We found that at near- physiological concentrations, DMS reduced reactive oxygen species (ROS) in cultured PC12 cells and alleviated oxidative stress. The radical-scavenging capacity of DMS at near-physiological concentration was equivalent to endogenous methionine(Met)-centered antioxidant defense. Methionine sulfoxidereductase A (MsrA), the key antioxidant enzyme in Met-centered defense, bound to DMS and promoted its antioxidant capacity via facilitating the reaction of DMS with ROS through a sulfonium intermediate at residues Cys72, Tyr103, Glu115, followed by the release of dimethyl sulfoxide (DMSO). MTT assay and trypan blue test indicated that supplement of DMS exhibited cytopro?tection against 6-hydroxydopamine and MPP + induced cell apoptosis. Furthermore, MsrA knockdown abolished the cytoprotective effect of DMS at near- physiological concentrations. The present study reveals new insight into the potential therapeutic value of DMS in Parkinson disease.

Objective To discuss and analyze the diagnosis,management and surgical techniques for cranioplasty of skull vault defects and skull base reconstruction in order to raise the therapeutic effect. Methods The clinical data of 169 cases of skull vault and skull base defects treated with cranioplasty of skull vault defect and skull base reconstruction were retrospectively analyzed.Results Overlay tech- nique or inlay technique with imported titanium alloy mesh was used in 160 cases,and homologous bone was used in 9 cases.The surgical time ranged from 3 months to 8 years after injury.Eight cases presented with complications such as hematoma,subcutaneous effusion,infection and epilepsy postoperatively,but no operative death occurred.Conclusion For patients with skull vault defect with the diameter≥3 cm the best operative time is 3 months after injury,and for patients with intracranial and extracranial communica- ting tumors,skull base reconstruction can be performed when tumors are removed.Much attention should be paid to perioperative management and surgical skills.

Parkinson's disease (PD) is characterized pathologically by the relatively preferential loss of dopaminergic neurons with resultant depletion of striatal dopamine and presence of Lewy bodies mainly composed by alpha-synuclein (alpha-SYN) in the remaining neurons in the substantia nigra. A lot of evidence suggests that the aggregation of alpha-SYN play an essential role in the pathogenesis of PD and formation of Lewy body. Increasing findings have implicated that some proteins, including parkin, synphilin-1,14-3-3, agrin and tau, interact with alpha-SYN and are involved in the abnormal aggregation of alpha-SYN.
Animals ; Carrier Proteins ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Lewy Bodies ; metabolism ; pathology ; Nerve Tissue Proteins ; metabolism ; Parkinson Disease ; etiology ; metabolism ; Protein Interaction Domains and Motifs ; Substantia Nigra ; pathology ; alpha-Synuclein ; metabolism

OBJECTIVETo investigate the binding characteristics of endothelial cell (EC) with LPS free from the participation of serum factors.

METHODSLaser confocal microscope was employed in the observation of the binding of EC with FITC-LPS. The KD and the binding sites of each EC were calculated by radioligand binding assay of receptors (RBA) using [(3)H]-LPS.

RESULTSThe binding of EC with LPS was saturable, time and concentration dependent and it could be competed with overdosed LPS of the same type. The fluorescence mainly distributed in cytoplasm, especially near the nucleus, which could also be stained.

CONCLUSIONSThere might be some specific LPS binding sites existing on ECs and LPS could function intracellularily.

Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; metabolism ; Fluorescein-5-isothiocyanate ; Fluorescent Dyes ; Humans ; Lipopolysaccharides ; metabolism ; Microscopy, Confocal ; Radioligand Assay ; Umbilical Veins ; cytology

OBJECTIVETo identify whether SH3GL1 gene serves as a disease associated gene of adolescent idiopathic scoliosis (AIS).

METHODSPositioning candidate cloning: "case-sibling or case-family control design" research scheme based on family constellation was designed. Fifty-six AIS patients (15 male and 41 female, mean age 15 years old, ranged from 8 to 22 years old, Cobb angle from 25 degrees to 110 degrees , average Cobb angle of 67.5 degrees ) from November 2007 to December 2008 were recruited. In all patients, blood preparation was collected, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed. Sequence comparative analysis for exon sequencing result between sib pairs or family pairs, and that between sib pair or family pairs and NCBI (National Center for Biotechnology Information) were conducted through Vector NTI Advance 10.3 software to judge whether basic group mutation occurred or not. Amino acid sequence comparative analysis for prediction was made.

RESULTSTen exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair and family pairs, and the sequencing obtained positive results. Twelve basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (3 mutations), the fourth exon (1 mutations), the fifth exon (4 mutations), the sixth exon (1 mutations), the eighth exon (1 mutations), and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutated to T, termination codon(TAG) came into being and open reading frame was altered. The sequence of protein showed brachytmema protein was encoded, which could cause changes of primary structure.

CONCLUSIONSH3GL1 is possibly one of the disease associated genes of AIS.

Adolescent ; Base Sequence ; Child ; Exons ; genetics ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Male ; Scoliosis ; genetics ; Sequence Alignment ; Young Adult

OBJECTIVETo investigate the clinical value of spiral CT with multi-planar reconstruction (MPR) and three diamensions reconstruction (3D) in the diagnosis and treatment of pelvic ring fractures.

METHODSFifty-seven patients with pelvic ring fractures were examined with digital radiography and spiral CT in our hospital from April 2004 to April 2009. According to Tile classification, there were thirty-eight cases in type A, twelve in type B and seven in type C. Expectant treatment was used in type A, while surgery, open reduction internal fixation or percutaneous lag screw internal fixation technique for sacroiliac joint due to the displacement of fractures, for type B and C. Three days to twenty-seven months after operation, spiral CT examinations were used to evaluate the location of internal fixation. Cross-check analysis of images of digital radiography and spiral CT was performed before and after operation.

RESULTSFive posterior and three anterior pelvic ring fractures were diagnosed as suspected fractures. Nine posterior and three anterior pelvic ring fractures were missdiagnosed according to plain radiographs, which were corrected by spiral CT examination. According to the postoperative imageology evaluation, the results were excellent in 15, good in 3 and bad in 1. According to clinical evaluation, 16 cases were excellent, 3 good.

CONCLUSIONSpiral CT with multi-planar reconstruction (MPR) and three diamensions reconstruction (3D) has clinical values for precise diagnosis and treatment for the complex pelvic ring fractures.

Adolescent ; Adult ; Aged ; Female ; Fracture Fixation, Internal ; Fractures, Bone ; diagnosis ; diagnostic imaging ; surgery ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Pelvic Bones ; diagnostic imaging ; injuries ; surgery ; Reconstructive Surgical Procedures ; Tomography, Spiral Computed ; methods ; Young Adult

Objective To report our experience with retroperitoneoscopic nephroureterectomy with excision of a bladder-cuff for renal pelvic and ureteral tumors.Methods Thirty-five patients (21 women and 14 men;mean age,67 years;age range,49 -82 years) with upper urinary tract tumors underwent retro- peritoneal laparoscopic nephrourcterectomy with excision of a bladder-cuff.Of the 35 cases,15 had pelvic tumors and 20 had ureteral tumors;19 cases had the tumors on the right side and 16 on the left.Two cases had ureteral tumors combined with bladder tumors.One case had bilateral ureteral tumors then concomitantly had bladder tumors.The needle electrode was used to circleround incise the bladder thoroughly 0.5 cm away from the ureterostoma.Three trocars in the waist were used for dissecting the kidney;and the ureter was dis- sected as far distally downward.Then an incision of 5-9 cm was created in the lower abdomen to allow dis- section of the distal ureter and bladder-cuff and intact specimen extraction.Results The operation was successful in all 35 patients.The mean operative time was 3.1 h ( range,1.5-6.0h).The mean estimated blood loss was 166 ml (range,20-1600 ml).Four cases received blood transfusion.The patient's activity re- covered in 20-32 h after operation.Postoperative pathology showed transitional cell carcinoma in 30 cases, poorly differentiated adenocareinoma in 2 (ureter),squamous cell carcinoma in 1 (ureter),leiomyosarcoma in 1 (ureter),xanthogranulomatous pyelonephritis in 1.Duodenal leakage occurred in 1 patient who had had dialysis a drainage catheter was placed on the third d after operation,and the patient died of heart failure af- ter 2 months.Postoperative vesical irrigation was performed to prevent tumor recurrence.The mean hospital stay was 11 d.During a mean follow-up of 14 months(range,1-32 months),1 patient developed pelvic me- tastasis and was alive with the tumor.The other 33 patients survived free of tumor to date.No patient had re- current transitional cell carcinoma of the bladder.Conclusions Our data demonstrate that retroperitoneo- scopic nephroureterectomy for renal pelvic and ureteral tumors has shorter incision and more rapid postopera- tive recovery compared with open surgery.Using resectoscope to resect the termination of ureter allows more complete excision of the ureter.