Abdominal Neoplasms ; complications ; metabolism ; pathology ; surgery ; Adult ; Dendritic Cell Sarcoma, Follicular ; complications ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Ki-1 Antigen ; metabolism ; Leukocytosis ; complications ; metabolism ; pathology ; surgery ; Receptors, Complement 3b ; metabolism ; Receptors, Complement 3d ; metabolism ; Young Adult

AIMTo elucidate the location and effects of transcription factor-nuclear factor-kappaB (NF-kappaB) in lung tissues of rats with chronic obstructive pulmonary disease (COPD).

METHODSFourteen male Wistar rats were randomly divided into COPD model and control groups equally. The COPD model was established by intratracheal instillation of lipopolysaccharide twice and exposure to cigarette smoke daily. We detected the NF-kappaB p65 protein in lung by immunohistochemical method, and the expression of NF-kappaB p65 mRNA in lung by in situ hybridization.

RESULTSImmunohistochemistry, the expression of NF-kappaB p65 protein in alveolar, bronchiolar epithelium and arteriolar endothelium was significantly higher in the COPD group (0.426 +/- 0.007, 0.434 +/- 0.012 and 0.313 +/- 0.007, respectively) than those of the control group (0.115 +/- 0.006, 0.116 +/- 0.005 and 0.095 +/- 0.007, respectively, all P < 0.01). In situ hybridization showed that the expressions of NF-kappaB p65 mRNA in alveolar epithelium (0.203 +/- 0.008), bronchiolar and arteriolar smooth muscle cell (0.208 + 0. 010 and 0.206 + 0.007) of rats in the COPD group were stronger than those in the control group (0.100 +/- 0.006, 0.102 +/- 0.002 and 0.103 +/- 0.003 respectively) by semiquantitative analysis (all P < 0.01).

CONCLUSIONThe expression and nuclear translocation of NF-kappaB may be the basis event of gene expression of many cytokines and inflammatory mediators, which may positively regulate gene expression of many cytokines and inflammatory mediators in various cell lines.

Animals ; Lung ; metabolism ; pathology ; Male ; Pulmonary Disease, Chronic Obstructive ; metabolism ; pathology ; Rats ; Rats, Wistar ; Transcription Factor RelA ; metabolism

OBJECTIVETo study the effect of multiglycosides of Tripterygium wilfordii (MTW) for treatment of proteinuria in kidney transplant recipients.

METHODForty-five kidney transplant recipients with proternuria were randomized into 3 groups (n=15) and received full daily dose (1 mg/kg) MTW, half dose (0.5 mg/kg) MTW or no MTW (control) in addition to immunosuppressant therapy. The 24-hour urinary protein (24 h Upro), blood urea nitrogen (BUN), serum creatinine (Scr), dose of ciclosporin and the adverse effects of MTW were recorded.

RESULTSMTW at both the full dose and half dose significantly reduced the 24 h Upro as compared to exclusive immunosuppressant therapy (P<0.05). The therapeutic dose of ciclosporin in patients with full and half dose of MTW was significantly lower than that in the control group (P<0.05), and the patients receiving full dose MTW showed greater adverse effects than those having half dose MTW (P<0.05).

CONCLUSIONSMTW can significantly ameliorate proteinuria, reduce the therapeutic dose of ciclosporin and protect the renal function in kidney transplant recipients. While producing similar therapeutic effect to routine full dose, long-term use of half dose MTW may reduce the adverse effect associated with MTW.

Adult ; Aged ; Female ; Glycosides ; therapeutic use ; Graft Survival ; immunology ; Humans ; Kidney Transplantation ; adverse effects ; immunology ; Male ; Middle Aged ; Proteinuria ; drug therapy ; etiology ; Tripterygium ; chemistry ; Young Adult

OBJECTIVETo investigate the effect of fosinopril (Fos) on regulating klotho gene expression and elucidate the mechanism of Fos regulating the Angiotensin II (AngII) -induced down-expression of klotho gene.

METHODSCulture cells, NRK-52E, were incubated with media either AngII or Fos or both of all. Experimental groups incubated with Fos (10(-5) mol/L) were divided according to variant points of time for 0 (control), 3, 6, 12, 24 h. Different concentration of Fos was selected to incubated with culture cells for 0 (control), 10(-9) 10(-8), 10(-7), 10(-6), 10(-5) mol/L at the optimal time point (24 h). Five groups, which were A: control; B: AngII (10(-7) mol/L); C: Fos(10(-5) mol/L); D: AngII (10(-7) mol/L) + Fos(10(-5) mol/L) and E: Cells pretreated with Fos(10(-5) mol/L)12 h incubated with AngII (10(-7) mol/L) were divided to observe the effect of Fos on expression of klotho induced by AngII. RT-PCR and immunohistochemistry (IHC) were applied to evaluate the klotho mRNA and protein expression, respectively.

RESULTSFos up-regulated klotho mRNA in time-dependent manner, and independent of dose-dependent manner; AngII obviously decreased the levels of kloltho mRNA and protein expression in NRK-52E as compared to the control (P < 0.05), the down-regulating effect was reversed by incubating both with AngII and Fos (P < 0.05), and Fos could inhibit the down-regulated expression of klotho gene induced by Ang II in NRK-52E.

CONCLUSIONFosinopril up-regulates klotho mRNA in time-dependent manner, and inhibits the down-regulated expression of klotho gene induced by Ang II.

Angiotensin II ; pharmacology ; Animals ; Cells, Cultured ; Down-Regulation ; Epithelial Cells ; drug effects ; metabolism ; Fosinopril ; pharmacology ; Gene Expression ; drug effects ; Glucuronidase ; genetics ; metabolism ; Kidney Tubules ; cytology ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Rats

OBJECTIVETo develop a screening system for more rapid and sensitive mutation detection of autosomal dominant polycystic kidney disease (ADPKD) gene 1 (PKD1) by using denaturing high-performance liquid chromatography (DHPLC) protocol.

METHODSUsing genomic DNA as templates extracted from blood samples of 19 Han pedigrees with 67 family members, the complete codon areas were amplified by long-range PCR and nested PCR in succession, and then the PCR products were analyzed by DHPLC. The mutations from screened abnormal PCR products were confirmed by DNA sequencing, and then compared with the mutations identified by single strand conformation polymorphism (SSCP) before.

RESULTSThere were 14 mutations found in this study, including 10 missense, 1 insertion, 1 deletion and 2 nonsense mutations. Besides 12 mutations identified before, mutations nt32819G>A and nt37137T>C were the novel mutations found. The mutation detection ratio was 73.7%.

CONCLUSIONThis developed system via DHPLC can be used as a more effective approach for mutation detection of autosomal dominant polycystic kidney disease PKD1 in Hans.

Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Chromatography, High Pressure Liquid ; methods ; DNA Mutational Analysis ; Family Health ; Female ; Humans ; Male ; Middle Aged ; Mutation ; Pedigree ; Polycystic Kidney, Autosomal Dominant ; diagnosis ; ethnology ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; TRPP Cation Channels ; genetics

Objective To investigate the imaging findings of the lacerating injury of the lung. Methods Ten patients of lung lacerating injury were examined by X-ray and CT within 1—5 h after injury. X-ray(2—5 times)and CT(3—5 times)examinations were repeated for 7 patients.Results The lung lacerating injury involved 10 sides and 14 lung lobes(21 lesions in total)in the 10 cases,among which 1 case involved the right upper lobe with 1 lesion,2 cases in the right lower lobe with 2 lesions,1 case in the right upper and lower lobes with 2 lesions for each lobe,3 cases in the left lower lobe with 9 lesions,and 3 cases in both the left upper and the lower lobes with 7 lesions.The X-ray findings were cavity-like shadows with smooth margin in 9 lesions(9/21),and patchy shadows of fogging margin in 12 lesions(12/21).The CT imaging findings included 6 pulmonary hematomas(6/21),and 15 cavitary lesions with air-fluid levels (15/21).In the 15 cavitary lesions,CT revealed 14 single cavities and 2 small cavities within a big cavity. On dynamic follow-up observation,the cavity was the biggest in 1—5 h after injury,but the hematoma was the biggest in 2—3 days after injury.Hematomas tended to absorb slower than the cavities.After 16— 32 days,all lesions revolved into small patchy or stripe-like shadows with slightly increased density. Conclusion Cavitary lesion with air-fluid level is the characteristic imaging finding of lung lacerating injury.CT surpasses X-ray plain film in revealing the details of lung lacerating injury.

OBJECTIVETo detect the mutations of autosomal dominant polycystic kidney disease gene 2(PKD2)in Chinese.

METHODSThe white blood cell genomic DNA from patients of 94 Chinese autosomal dominant polycystic kidney disease(ADPKD) pedigrees was isolated and amplified by polymerase chain reaction(PCR). The PCR products were analyzed by denaturing high-performance liquid chromatography(DHPLC). The samples with abnormal profiles were sequenced.

RESULTSEight mutations were identified, including 2 nonsense mutations, 2 deletion mutations,1 insertion mutation and 3 missense mutations. Two nonsense mutations occurred in exon 5(1249C-->T) and exon 13(2407C-->T),both resulted in a stop codon. The insertion was in exon 2(636-637 ins T),and the deletion mutations were in exons 12(2348-2351 del AGAA) and 13(2401 delete A),resulting in the reading frame shift. Three missense mutations were in exons 1(G568-->A),4(C964-->T),and 5(G1168-->A), which caused amino acid changes (190Ala-->Thr,322Arg-->Trp,390Gly-->Ser).

CONCLUSIONThe method of DHPLC was used in detecting mutations successfully and 8 mutations in PKD2 were identified. It will be useful in the molecular diagnosis of ADPKD in advance of the cysts formation and birth.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chromatography, High Pressure Liquid ; Female ; Humans ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Mutation ; Nucleic Acid Denaturation ; Polycystic Kidney, Autosomal Dominant ; genetics ; TRPP Cation Channels

OBJECTIVETo identify the overall anatomical characteristics and the clinical value of the lumbar nerves under CT multiplanar reconstruction.

METHODSFifty normal subjects and 30 patients with LN diseases (51 sides) were selected, including 10 patients with lumber intervertebral disk hernia, eight patients with spinal stenosis, 5 patients with spondylolisthesis, 1 patient with tethered cord syndrome, 1 patient with lumbar scoliosis, and 5 patients with spinal trauma The 16-slice helical CT (Light Speed, GE Company) was used for scanning the lumbar vertebra with multiplanar reconstruction in Workstation (ADW4.1) with UNIX System in DICOM format. The image was set on the same slice for the overall anatomy and manifestations of the lesions.

RESULTSThe same-slice imaging showed the strip-like LN slightly lower than the surrounding muscle in density. Each LN went out of the dural sac at an acute angle. The course of the lumbar plexus and its major branches, including the obturator nerve, femoral nerve and reproductive nerve, and their relations to the adjacent structures were clearly revealed. The percentage of the segments displayed was well associated with the reconstruction angle, with the order being oblique coronal > outward-rotated oblique coronal > oblique sagittal > coronal > sagittal section. The major manifestations of abnormal LN included compression and displacement (50 sides, 98.0%), morphological changes (49 sides, 96.1%), adhesion (41 sides, 80.4%).

CONCLUSIONSThe CT multiplanar reconstruction is ideal for the imaging of the overall size, shape, running and tension of the LN root; it is valuable in clinical diagnosis.

Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Image Processing, Computer-Assisted ; Lumbosacral Plexus ; anatomy & histology ; diagnostic imaging ; Male ; Middle Aged ; Spinal Nerve Roots ; anatomy & histology ; diagnostic imaging ; Tomography, Spiral Computed ; Young Adult

Epithelial cell adhesion molecule (EpCAM) is a popular target for cancer therapy. In this research, 3 nanobodies with high specificity and endocytosis activity against EpCAM were developed, which provides a basis for the study of immunotoxin based on EpCAM. In our preliminary experiments, we have immunized a camel with EpCAM-Fc antigen and constructed a high-quality phage display library. Seventeen nanobodies with different complementarity determining region (CDR) 3 sequences have been screened after 3 rounds of biopanning by phage display technology. The animal procedures were approved by the Institutional Animal Care and Use Committee (IACUC) of Fudan University School of Pharmacy. After purification, 7 nanobodies showed high cell binding activity by fluorescent activated cell sorting (FACS) identification. Furthermore, 3 nanobodies presented high endocytosis activity based on FACS and laser confocal microscopy, which also showed high affinity to EpCAM measured by ForteBio. According to this study, we aimed to provide a novel alternative approach to the EpCAM-targeted therapy and to provide guidance for the study of nanobody based immunotoxins for other targets.

ObjectiveTo investigate the effect of LncRNA GAPLINC on the cell proliferation of RA-FLSs. MethodsRA-FLSs were cultured from synovial specimens. The expression of LncRNA GAPLINC in RA-FLSs and trauma-FLSs groups was detected by qRCR. GAPLINC suppression was transfected by siRNA and the inhibition efficiency was detected by qRCR. Flow cytometry was adopted to determine the change of cell growth and cell cycle distribution. 【ResμLts】 The expression of LncRNA GAPLINC was significantly higher in RA-FLSs than that of the trauma-FLSs （P<0.05）.Transfection of GAPLINC-siRNA significantly decreased the expression of LncRNA GAPLINC. GAPLINC silence in RA-FLSs revealed significant inhibition in cell proliferation which was showed by the reduced cell number in S phase（P<0.05）. Moreover， flow cytometry assay showed GAPIINC-siRNA treatment group had an accumμLation of cells in the G0/G1 phase and decreased RA-FLSs in the S and G2/M phase（P<0.05）. After GAPLINC knockdown， mRNA and protein levels of Cyclin D1 and PCNA， which were positively correlated with proliferative phenotype， were decreased （P<0.05）， while p21， which was negatively correlated with proliferative phenotype， was up-regμLated （P<0.05）. ConclusionsThe mRNA expression of GAPLINC was higher in RA-FLSs compared with trauma-FLSs ，which was statistically significant（P<0.05）. The silence of LncRNA GAPLINC coμLd significantly inhibit RA-FLSs cell growth and suppress the cell cycle transformation， which suggests that GAPLINC may play a role in the regμLation of proliferation of RA-FLSs， leading to synovial hyperplasia and contributing to RA progression.