OBJECTIVE: To investigate the pathology characteristics of congenital preauricular fistula with infection, in order to reduce the recurrence rate after surgery and improve operative technique. METHOD: Twenty-five patients diagnosed as congenital preauricular fistula with infection were analyzed. There were 14 patients in infection history group, 9 in infective stage group, and 2 in recurrence group respectively. The whole piece of fistula and scar tissue was completely excised during operation. The specimens were observed by naked eye and serial tissue sections were analyzed. RESULT: (1) Macroscopically, in infection history group, initial morphology can be maintained near the fistula orifice, but the distal tissue was dark red scar tissue. In infective stage group, the distal tissue of the specimens was granulation tissue and cicatricial tissue. The granulation tissue was crisp and bright red. In recurrence group, multicystic lesions with severe edema was observed, with a classical dumb-bell appearence. (2) Microscopically, in infection history group and recurrence group, we can see that the distal fistula tissue was discontinuous and was separated by scar tissue. In infective stage group, we can find neo-angiogenesis and infiltration of plasma cells, lymphocytes, neutrophil between interrupted fistula tissues. (3) All patients were followed up for 6-12 month, without recurrence. CONCLUSION The fistula tissue of congenital preauricular fistula with infection was divided by the scar tissue, and they did not communicate with each other. Complete delineation of fistula is hardly achieved by methylene blue staining. Radical excision of the fistula and scar tissue may help to avoid leaving viable squamous epithelial remnants and reduce the recurrence rate.
Adolescent ; Adult ; Child ; Craniofacial Abnormalities ; pathology ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Objective To compare the difference of the protein about the patient of hepatitis B who received adefovir dipivoxil(ADV)therapy,and seek the useful biomarker of effective therapy.Methods We used the two-dimensional gel electrophoresis technology to examine HBV infected serum samples aiming at searching protein's alteration after ADV therapy.Results After 1 year's treatment,haptoglobin, haptoglobin 2-alpha raised and alpha-l-antitrypsin precursor,Factor B,Chain B,transthyretin,glutathione peroxidase,alpha-2-HS-glycoprotein,retina]binding protein,retinol-binding protein precursor, apolipoprotein,apolipoprotein A-I precursor fell in viral response patients.Transthyretin raised and leucine- rich alpha-2-glyeoprotein,haptoglobin,alpha-2-actin,apolipoprotein A-I precursor fell in none viral response patients.To compare two groups:apolipoprotein A-I have the same change and haptoglobin, transthyretin have the opposite change.Conclusion Proteomics study can find the alteration of protein during the ADV treatment,and is helpful to searching the predictable biomarker to ADV.

Cell Line ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Interferon-beta ; pharmacology

OBJECTIVETo explore the correlation between pathological characteristics of target organs and excess evil syndrome in IgA nephropathy.

METHODSData were collected in multicenter cooperation. Totally 266 IgA nephropathy patients were typed into exogenous wind-heat affection syndrome (49 cases), lower energizer damp-heat syndrome (100 cases), damp-phlegm syndrome (43 cases), and blood stasis syndrome (74 cases). Meanwhile, percutaneous renal biopsy was performed in all patients for Hass classification, Oxford classification, Katafuchi integral, and Jiang's classification methods. The correlation between excess evil syndrome and pathological index was analyzed.

RESULTSFour syndrome types were correlated with their Hass levels (r = 0. 341, P <0. 01). Affection of exogenous wind-heat syndrome was correlated with segmental proliferation of endothelial cells and damaged active lesions of segmental capillary loops. Lower-energizer damp-heat syndrome was associated with Hass III level, destroying active lesions of capillary loops, segmental proliferation of endothelial cells, glomerular segmental lesions, focal interstitial infiltration of inflammatory cells, focal interstitial fibrosis and tubular atrophy. Blood stasis syndrome was associated with Hass IV level, glomerular sclerosis, segmental glomerulosclerosis (S)/adhesion, mesangial hypercellularity (M), angiohyalinosis, multi-foci interstitial infiltration of inflammatory cells, multi-foci interstitial fibrosis and tubular atrophy. Phlegm-damp syndrome had higher proportions of Hass I and III levels, but with no association with other pathological parameters.

CONCLUSIONSExcess evil syndrome was associated with partial pathological characteristics of IgA nephropathy. It could reflect pathological damage degree of target organs, activities, chronic lesions, and prognosis of IgA nephropathy to certain extent. Correlated pathological characteristics and its evolution could indicate excess evil syndrome types and their evolution rules.

Capillaries ; Fibrosis ; Glomerulonephritis, IGA ; pathology ; Glomerulosclerosis, Focal Segmental ; Humans ; Kidney Glomerulus ; Medicine, Chinese Traditional ; Prognosis ; Syndrome

OBJECTIVETo construct a eukaryotic expression vector for expressing hepatitis B virus (HBV) recombinant HBsAg-EGFP fusion protein and obtain a stable transfected Chang Liver cell line.

METHODSThe coding region of HBsAg gene of HBV was amplified by PCR and was digested by BamH I/EcoR I . This fragment was inserted into pEGFPN1 with T4 ligase and transformed E-coli TG1. The positive recombinant plasmid was selected, then the recombinant plasmid was transfected into Chang Liver cell by Lipofectamine 2000 cells containing stable transformants were selected by the ability of resistance to G418 and isolated with a limited dilution. The stable transfected cell line expressing high level HBsAg-EGFP fusion protein was obtained.

RESULTSThe eukaryotic expression vector named pEGFPN1-HBsAg was successfully constructed and the stable transfected Chang Liver cell line expressing pEGFPN1-HBsAg fusion protein was obtained.

CONCLUSIONThe stable transfected Chang Liver cell line could express pEGFPN1-HBsAg fusion protein, could be used to screen the proteins differentially expressed in HBsAg expression Chang Liver cells, which brought some new clues for studying the potential molecular mechanism of HBsAg protein.

Cell Line ; Gene Expression ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Hepatitis B Surface Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Liver ; cytology ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection ; methods

OBJECTIVETo develop a novel gene delivery vector TAT-PEI-beta-CyD.

METHODSbeta-cyclodextrin (beta-CyD) was linked by low molecular weight (PEI 600) via 1, 1-carbonyldiimidazole (CDI), and TAT peptide (RRRQRRKKRC) was coupled to PEI 600 by [N-succinimidy-3-(2-pyridyldithio) propionate, SPDP]. The copolymer was characterized by (1)H-NMR and FT-IR. Physiochemical characteristics of TAT-PEI-beta-CyD/DNA complexes were tested by agarose gel electrophoresis and particle size measurements. Cell viability and transfection efficiency were evaluated in A293 and B16 cells using PEI 25 kDa as a control.

RESULTTAT peptide was successfully coupled to PEI-beta-CyD. The result of gel electrophoresis showed that the TAT-PEI-beta-CyD was able to condense DNA efficiently at N/P ratio of 4. The particle size of TAT-PEI-beta-CyD/DNA complexes was around 100 nm. The cytotoxicity of TAT-PEI-beta-CyD was lower than that of PEI 25 kDa. The transfection efficiency of TAT-PEI-beta-CyD was higher than that of PEI 25 kDa in A293 and B16 cells at N/P ratio of 30.

CONCLUSIONThe novel vector TAT-PEI-beta-CyD has been developed successfully with low cytotoxicity and high transfection efficiency.

Cell Line ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Humans ; Peptide Fragments ; chemistry ; Polyethyleneimine ; chemistry ; beta-Cyclodextrins ; chemistry ; tat Gene Products, Human Immunodeficiency Virus ; chemistry

OBJECTIVETo study the genome sequence of hepatitis A virus L-A-1 strain which has been applied for live attenuated vaccine production in China, to compare with other HAV strains, to understand some characteristics of L-A-1 strain, and to find the mechanism of attenuation and cell adaptation.

METHODSGenome fragments were prepared by antigen-capture PCR from infected cell (2BS), PCR products were cloned into T vector, sequenced and analyzed by using bioinformatics program.

RESULTSAnalysis of the genomic sequences(nt 25-7,418) showed that the open reading frame contains 6,675 nucleotides in length encoding 2,225 amino acids. Sequence homology comparison showed 98.00% and 94.00% homology at nucleotide level, and 98.51% and 98.65% homology at amino acid level with international strains MBB and HM 175, respectively. Through comparison with other attenuated, cell adapted and cytopathic effect (CPE) strains, L-A-1 strain had mutation at nt 152, 591, 646, 687 and insertion at nt 180-181 in 5?NTR and had mutation at nt 3,889 (aa 1 052-Val) in 2B region, these mutations and insertion are molecular basis for cell adaptation; mutation at nt 4,185 (aa 1 152-Lys) in 2C region should be attenuated marker; deletion in 3A region (nt 5,020-5,025) that caused two amino acids deletion is virus fast growth basis.

CONCLUSIONThrough analyzing L-A-1 strain genomic sequence, certain sites related to cell adaptation and attenuation were found.

Adaptation, Biological ; genetics ; Amino Acid Sequence ; Base Sequence ; Gene Deletion ; Genome, Viral ; Hepatitis A Vaccines ; genetics ; Hepatitis A virus ; genetics ; growth & development ; Mutation ; Open Reading Frames ; genetics ; Sequence Homology ; Vaccines, Attenuated ; genetics

OBJECTIVETo further study the mechanism of the inhibitory effect of interferon beta-1a (IFN beta-1a) on the activation of human hepatic stellate cell (HSC) LX-2, and to analyze the differences on the protein expression in LX-2 induced by I IFN beta-1a.

METHODSCultured LX-2 cells were treated with 2000 U/ml IFN beta-1a for 48 h. Two-dimensional gel electrophoresis (2-DE) was performed to compare protein patterns of the control (untreated) and IFN beta-1a treated LX-2 and for quantitative and qualitative analyses of protein expression. A rat liver fibrosis model was established and the rats were sacrificed and their various tissues were obtained for the same analyses. Western blotting and RT-PCR were used to validate the expression of the changed proteins after treatment of IFN beta-1a in LX-2 cells and of various tissues of the rats.

RESULTS708 +/- 25 spots were detected in control LX-2 cells and 804 +/- 32 spots in IFN beta-1a-treated LX-2 cells. A match rate of 73%-82% was achieved. The results also showed that 31 protein spots displayed quantitative changes in expression after IFN beta-1a treatment. Of the 31 spots, 21 proteins were identified, of which, one was newly found, two were enhanced in abundance and 18 showed lower expressions. The newly found protein was glia maturation factor beta (GMF beta). The treatment of LX-2 with IFN beta-1a increased the production of GMF beta(GMF beta) protein in comparison with the untreated cells (t=1.81, P < 0.01). The expression of GMF beta protein (1.81 vs 0.10) and mRNA (0.85 vs 0.12) were more in the normal liver tissues than in the cirrhotic liver tissues (t=2.53, 2.13 respectively, P < 0.01). The expressions of GMF beta protein and mRNA were weak in rat heart and lung tissues, however, they were strong in rat liver, kidney, spleen and brain tissues (t=1.91, 1.94 respectively, P < 0.01).

CONCLUSIONThere is a significant difference of protein expression levels between IFN beta-1a untreated and treated LX-2 cells. These proteins, especially GMF beta, may be involved in an inhibition process of IFN beta-1a on activation and apoptosis of LX-2 cells. This proteome study may be useful in further studies of the relationship of IFN beta-1a treatment and human liver diseases.

Animals ; Cell Line ; Female ; Glia Maturation Factor ; metabolism ; Hepatic Stellate Cells ; metabolism ; Humans ; Interferon beta-1a ; Interferon-beta ; pharmacology ; Liver ; cytology ; Liver Cirrhosis ; metabolism ; Proteome ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the expression and clinical implication of tissue inhibitor of metalloproteinase-1 (TIMP-1) in colorectal carcinoma.

METHODSTIMP-1 expression in 54 colorectal carcinoma was observed by SP immunohistochemical method, and the results were analyzed in relation to the clinical data of patients.

RESULTSTIMP-1 was localized on the membrane and in the cytoplasm of the enteric epithelial cells, and its expression rate was 100% in normal tissue but only 59.6% (31/52) in colorectal carcinoma tissues. In addition, the expression rate of TIMP-1 was higher in the tumor tissues without lymph node metastasis than in tissues with lymph node metastasis (P<0.05).

CONCLUSIONThe expression of TIMP-1 is inversely correlated to lymph node metastasis of colorectal carcinoma, and decreased TIMP-1 expression may play a role in the progression of colorectal carcinoma.

Adult ; Aged ; Colorectal Neoplasms ; metabolism ; pathology ; Epithelial Cells ; chemistry ; pathology ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; biosynthesis

BACKGROUNDAtypical meningioma is one of the rare subtypes of meningioma, which is lacking of optimal consensus on treatment strategies. This study aimed to investigate the radical treatment strategies to improve the long-term outcome of recurrent atypical meningiomas.

METHODSThe prognostic factors including the age and gender of patients; the location, histology, recurrence pattern and mitotic cell rate of the tumors; and the resection extents, surgical strategies and adjuvant therapies of 15 cases of recurrent atypical meningiomas were analyzed retrospectively.

RESULTSThe age and gender of patients were not associated with tumor recurrence. However, high recurrence rates and poor prognosis for atypical meningiomas were associated with the high mitotic cell rate, failure to achieve Simpson grade I-II resection, and without the dura and bone flap replacement intraoperatively. Post-operative radiotherapy improved the outcomes of tumors in patients after the second surgery.

CONCLUSIONRadical treatment strategies such as dura and bone flap replacements and radiotherapy should be considered in patients diagnosed with atypical meningiomas.

Adult ; Aged ; Female ; Humans ; Male ; Meningeal Neoplasms ; radiotherapy ; surgery ; Meningioma ; radiotherapy ; surgery ; Middle Aged ; Neoplasm Recurrence, Local ; radiotherapy ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult