With reverse transcriptase PCR, the transcripton of copper homeostasis relative gene Afe0329 in Acidithiobacillus ferrooxians standard strain ATCC23270 was investigated. The further analysis of genes in this transcripton was analyzed employed by Vector NTI, Blast, TMHMM Server, PSORTb software and so on. From the DNA of different strains, the transcripton of Afe0329 was amplified using special primer pairs to identify the universality of it in the genome of A.ferrooxidans strains. The results showed that gene Afe0330 and Afe0331 were cotranscribed with Afe0329, and they were in a single transcripton. Gene Afe0329 was supported to express a P1b3-type ATPase which is a heavy metal ion pumping transmembrane protein, protein AFE0330 which expressed by gene Afe0330 was a cytoplasmic protein, no significant ho- mologous sequences of Afe0330 or Afe0331 had been obtained by Blast analysis. And the transcripton of Afe0329 was universal in genome of A. ferrooxidans strains.

OBJECTIVETo measure the stability of Evans procedure and Chrisman-Snook technique in the treatment of II degree lateral collateral ligament of ankle joint, and provide basis for treatment and prognosis.

METHODSFrom July 2008 to June 2009,18 frozen corpes were collected, including 10 males and 8 females, with an average age of fresh 39.3 +/- 11.2 years. The frozen corpes were randomly divided into three group, including normal controls(group A), Evans procedure (group B) and Chrisman-Snook technique ( group C), 6 specimens in each group. Anterior talofibular ligament and calcaneofibular ligament were cut off to cause II degree lateral collateral ligament in group B and C. Evans procedure or Chrisman-Snook technique were applied to restore lateral collateral ligament, and measure biomechnics. The displacement of tibiotalar joint and subtalar joint were observed.

RESULTS(1) The lateral stress results of tibiotalar joint showed the displacement by Evans procedure (group B) was greater than other groups (P < 0.0001). There were no significant differences between group A and C (P > 0.05). (2) The lateral stress results of subtalar joint showed the displacement by Evans procedure (group B) was greater than other groups (P< 0.0001). There were no significant differences between group A and C (P > 0.05).

CONCLUSIONAnkle instability is caused by ankle joint lateral collateral ligament injury. Chrisman-Snook technique is better than Evans procedure in stability on the early stage of ankle joint restoration, and conform to principle of biomechanics.

Adult ; Ankle Joint ; Biomechanical Phenomena ; Female ; Humans ; Lateral Ligament, Ankle ; diagnostic imaging ; injuries ; surgery ; Male ; Mechanical Phenomena ; Prognosis ; Radiography ; Reconstructive Surgical Procedures ; methods

OBJECTIVETo gain insight into the role of Toll-like receptor 2 (TLR2) in graft rejection following penetrating keratoplasty, and investigate the expression of TLR2 mRNA in the corneal graft.

METHODSPenetrating keratoplasty was performed in 3 groups of rats for orthotopic autologous corneal transplantation (group A), allograft corneal transplantation (group B), or allograft corneal transplantation with hormone treatment (C). The transparency and neovascularization of the cornea were observed using a slit-lamp microscope and scored according to the rejection index, with normal cornea serving as the control. The corneal tissues were sampled at 5, 7, and 9 days after the transplantation for histopathological examination and detection of TLR2 mRNA expression using RT-PCR.

RESULTSWith the passage of time, edema, opacities and neovascularization of the corneal graft occurred after the operation in all the groups. Seven days after the operation, the rejection index of group B, but not that of groups A and C, met the diagnostic criteria for graft rejection with also support by histopathological evidence. The expression of TLR2 mRNA was detected in normal corneas and augmented in the corneal grafts in the 3 transplantation groups. TLR2 mRNA expression in group B was significantly higher than that of group A, and the expression in group C decreased significantly in comparison with that in group B (P<0.05).

CONCLUSIONAs the recognition receptors of native immune system, TLR2 in the rejected corneal grafts may recognize the allograft antigen and play a role in acute graft rejection after penetrating keratoplasty.

Animals ; Cornea ; metabolism ; Female ; Graft Rejection ; immunology ; Keratoplasty, Penetrating ; Postoperative Complications ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 2 ; genetics ; metabolism

BACKGROUNDStathmin was identified as an endometriosis-related protein by comparative proteomics in our previous study. As a microtubule-destabilizing factor, stathmin was shown to participate in the relay and integration of diverse intracellular signaling pathways involved in cell proliferation, differentiation, and many other cellular activities. To investigate whether stathmin is involved in the pathogenesis of endometriosis, we examined the expression of stathmin in eutopic endometrium of women with or without endometriosis.

METHODSEutopic endometrium samples were collected from thirty-six patients who were diagnosed as endometriosis and the nineteen age-matched patients who were confirmed to be free of endometriosis surgically and histologically. The expression of stathmin mRNA was detected by real-time PCR, and its protein was detected by Western blotting and immunohistochemistry.

RESULTSStathmin was overexpressed in eutopic endometrium of women with endometriosis detected by real-time PCR in mRNA levels and by Western blotting in protein levels, without significant difference between proliferative and secretory phase. Immunohistochemistry showed that stathmin protein was localized in both endometrial glandular and stromal cells throughout the menstrual cycle.

CONCLUSIONSStathmin is overexpressed in endometrium of patients with endometriosis and may play a role in the pathogenesis of endometriosis.

Adult ; Blotting, Western ; Endometriosis ; metabolism ; Endometrium ; metabolism ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Polymerase Chain Reaction ; Stathmin ; genetics ; metabolism

Allergic diseases have become global social health problems. The binding of IgE with its high affinity receptor FcepsilonRI plays a key step in I-type allergy. Recently, more and more key molecules on the IgE/FcepsilonRI signaling transduction pathway were to be the drug candidates against allergic diseases, with in-depth study of FcepsilonRI signal pathway gradually. The main drugs include molecule antibodies, peptides, vaccines, fusion proteins, small molecules, and other drugs related to IgE/FcepsilonRI. The recent progress in the study of mechanisms of representative drugs targeting on IgE/FcepsilonRI signaling pathway was reviewed in this article.
Aminophenols ; pharmacology ; therapeutic use ; Animals ; Anti-Allergic Agents ; pharmacology ; therapeutic use ; Antibodies, Anti-Idiotypic ; pharmacology ; therapeutic use ; Antibodies, Monoclonal, Humanized ; pharmacology ; therapeutic use ; Humans ; Hypersensitivity ; drug therapy ; immunology ; Immunoglobulin E ; metabolism ; Intracellular Signaling Peptides and Proteins ; antagonists & inhibitors ; Molecular Targeted Therapy ; Omalizumab ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; therapeutic use ; Receptors, IgE ; metabolism ; Signal Transduction ; Syk Kinase

OBJECTIVETo investigate the clinical features and management of ureteral endometriosis.

METHODSPatients surgically and histologically diagnosed as ureteral endometriosis from January 2001 to January 2007 in Peking Union Medical College Hospital were retrospectively reviewed.

RESULTSTen patients were diagnosed as ureteral endometriosis among 7561 cases with surgically and histologically proved diagnosis of endometriosis, with an incidence of 0.132%. Nine out of 10 patients were extrinsic ureteral endometriosis and concomitant with severe pelvic endometriosis, and the other was intrinsic ureteral endometriosis. Hormone therapy failed in 2 patients with urinary tract obstruction. Ureterolysis was performed in 6 patients and ureterectomy was performed in 4 patients. One case of ureteral recurrence was observed in a postmenopausal woman without hormonal replacement therapy who received laparoscopic ureterolysis and hysterectomy with bilateral adnexectomy. No relapse was observed in the other 9 patients.

CONCLUSIONSUreteral endometriosis is a rare entity. The upper urinary tract should be evaluated in patients with severe endometriosis, even in postmenopausal women. The treatment of ureteral endometriosis usually requires surgery, while ureterolysis should not be performed in patients with extensive disease. As a form of adjuvant therapy of surgery, hormonal therapy is an appropriate option.

Adult ; Diagnosis, Differential ; Endometriosis ; complications ; pathology ; therapy ; Female ; Humans ; Middle Aged ; Retrospective Studies ; Ureter ; pathology ; Ureteral Obstruction ; etiology

Objective: To investigate the dynamic expressions of tenascin-C (TN-C) at different phases of atherosclerosis in ApoE-/-mice. Methods: Our research included in 2 groups: ApoE-/- group, n=50 SPF male mice with ApoE-/- and Control group, n=50 wild male C57BL/6 mice. Atherosclerosis model was established by high fat diet in both groups. The mice were sacrificed at 4, 8, 16, 24 and 32 weeks, blood lipids were examined, pathologic changes of plaque were observed by microscope for quantitative analysis and TN-C expressions in atherosclerosis plaque were measured by immunohistochemistry. Results: Compared with Control group, ApoE-/- group had elevated blood levels of total cholesterol (TC) and low density lipoprotein cholesterol (LDL-C), both P<0.05. In ApoE-/- group, plaque area and the ratio of plaque area/lumen area were increasing upon prolonged modeling time, all P<0.05; TN-C expressions were increasing by progress of atherosclerosis, the highest TN-C expression was found at 32 weeks of modeling (0.49±0.07) which was higher than it was at 8 weeks (0.04±0.02), 16 weeks (0.12±0.03) and 24 weeks (0.21±0.04), all P<0.05. Conclusion: TN-C expression was increasing with plaque progress which might be related to the development of atherosclerosis and plaque instability.

OBJECTIVETo prepare cytochrome (CYP)2E1-silenced hepatocytes by lentivirus-mediated RNA interference technology and to investigate the hepatotoxicity of trichloroethylene (TCE) in CYP2E1-silenced hepatocytes.

METHODSShort hairpin RNA fragments were designed and synthesized and were then ligated into the lentiviral vector; single colonies were screened; the plasmid was extracted after PCR and sequence identification and then transferred into L02 hepatocytes; the CYP2E1-silenced hepatocytes were selected; real-time quantitative PCR and Western blot were used to evaluate the interference effects. The obtained CYP2E1-silenced hepatocytes, as well as normal L02 hepatocytes, were treated with TCE (0, 0.25, 0.50, 1.00, 2.00, and 4.00 mmol/L). The cell viability and half maximal inhibitory concentration (IC50) of TCE were measured; the apoptotic rate of cells was measured by flow cytometry; the mRNA expression levels of apoptosis genes and oncogenes were measured by real-time quantitative PCR.

RESULTSThe IC50s of TCE for L02 hepatocytes and CYP2E1-silenced hepatocytes were 15.1 mmol/L and 23.6 mmol/L, respectively. The apoptotic rate increased as the dose of TCE rose in the two types of cells; the CYP2E1-silenced hepatocytes hada significantly lower apoptotic rate than L02 hepatocytes when they were exposed to 2.0 and 4.0 mmol/L TCE (P < 0.05 or P < 0.01). The mRNA expression level of bcl-2 (anti-apoptosis gene) in CYP2E1-silenced hepatocytes was 15% ∼ 60% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of caspase-3 and caspase-9 (apoptosis genes) in CYP2E1-silenced hepatocytes were 30% ∼ 60% lower than those in L02 hepatocytes (P < 0.01). The mRNA expression level of p53 (cancer suppressor gene) in CYP2E1-silenced hepatocytes was 81 - 278% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of c-fos and k-ras (oncogenes) in CYP2E1-silenced hepatocytes were 20-68% lower than those in L02 hepatocytes (P < 0.01).

CONCLUSIONCYP2E1-silenced cells can be successfully prepared by lentivirus-mediated RNA interference technology. Silencing CYP2E1 gene can reduce the hepatotoxicity of TCE and inhibit the expression of some apoptosis genes and oncogenes, suggesting that CYP2E1 gene plays an important role in TCE metabolism and is related to the hepatotoxicity of TCE.

Apoptosis ; drug effects ; genetics ; Cell Line ; Cell Survival ; drug effects ; genetics ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Genetic Vectors ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lentivirus ; genetics ; RNA Interference ; Trichloroethylene ; toxicity

In order to obtain nucleotides aptamers bind to IgE, 80 bp nucleotides single-stranded DNA library containing 40 random nucleotides was designed and synthesized. Oligonucleotides that bind to human Cepsilon3-Cepsilon4 protein were isolated from ssDNA pools by the systematic evolution of ligands by exponential enrichment (SELEX) method using nitrocellulose filters as screening medium. Through the optimization of critical PCR and asymmetric PCR parameters including annealing temperature, cycles, and molar ratios of target protein and ssDNA etc, a suitable screening system was established. The aptamers of Cepsilon3-Cepsilon4 protein with high affinity and high specificity were identified by ELISA with biotin-streptavidin-horseradish peroxidase system, and its primary sequence and second structure were analyzed by DNAMAN package and DNA folding sever after being cloned and sequenced. Moreover, target protein was bound to one aptamer and another aptamer modified with biotion together forming a sandwich-like complex, which was captured in microwell to detect IgE concentration using the optimal combination in the sandwich method named enzyme-linked aptamers sorption assay (ELASA). The method could be used for the quantitative detection of human IgE, and whose sensitivity reached to 120 ng x mL(-1).
Aptamers, Nucleotide ; chemistry ; genetics ; isolation & purification ; Base Sequence ; DNA, Single-Stranded ; chemistry ; Humans ; Immunoglobulin epsilon-Chains ; chemistry ; genetics ; Oligonucleotides ; chemistry ; SELEX Aptamer Technique ; methods ; Sensitivity and Specificity

BACKGROUNDAnnexin-1 was identified as an endometriosis-related protein by comparative proteomics in previous study. As an endogenous anti-inflammatory mediator, Annexin-1 has been shown to regulate the immune response, cell proliferation and apoptosis. To investigate whether Annexin-1 is involved in the pathogenesis of endometriosis, we examined the expression of Annexin-1 in eutopic endometrium of women with or without endometriosis, and detected its expression in peritoneal fluids of those with endometriosis.

METHODSEutopic endometrium samples from twenty-five women with endometriosis and those from sixteen age-matched women without endometriosis were collected. Peritoneal fluids were obtained from ten patients with endometriosis. The expression of Annexin-1 protein in eutopic endometrium was detected by immunohistochemistry and Western blotting, and mRNA detected by real-time PCR. Annexin-1 protein in the peritoneal fluids was detected by Western blotting.

RESULTSAnnexin-1 mRNA and protein were overexpressed in eutopic endometrium of endometriosis without significant differences between the proliferative and secretory phase. Immunohistochemistry showed that Annexin-1 protein was expressed mainly in endometrial glandular cells throughout the menstrual cycle. Annexin-1 protein was detected in the peritoneal fluids of all the ten patients with endometriosis.

CONCLUSIONSAnnexin-1 is overexpressed in eutopic endometrium and presents in the peritoneal fluids of patients with endometriosis, and may play a role in the pathogenesis of endometriosis.

Adult ; Annexin A1 ; genetics ; metabolism ; Ascitic Fluid ; metabolism ; Blotting, Western ; Endometriosis ; genetics ; metabolism ; pathology ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction