The fermentation broth of one actinomycete strain ACMA006 strongly inhibited growth of many tumor cells and some microorganisms, but its cytotoxicity to human normal cells were weak. Strain ACMA006 grow well on most tested media, producing exuberant vegetative hyphae and aerial hyphae. Its optimization temperature is 28?C. Phyloge-netic analysis based on 16S rDNA sequence showed that strain ACMA006 was closely related to one of the genus Streptomycetes (S.cavourensis subsp. washingtonensis) with 16S rDNA sequence similarity values of 100%, but had many differences in other features including its morphology, physiological and biochemical characteristics. The pre-liminary study supported the view that the strain ACMA006 represented a new strain of the S.cavourensis subsp. wash-ingtonensis.

BACKGROUND: The gut is capable of inducing multiple organ dysfunction syndrome (MODS). In the diagnosis and treatment of critical ill patients, doctors should pay particular attention to the protection or recovery of intestinal barrier function. However, no reliable diagnostic criteria are available clinically. This study aimed to assess the changes of intestinal mucosal barrier function in surgically critical ill patients as well as their significance. METHODS: Thirty-eight surgically critical ill patients were enrolled as a study group (APACHE II>8 scores), and 15 non-critical ill patients without intestinal dysfunction were selected as a control group (APACHE II<6). General information, symptoms, physical signs, and APACHE II scores of the patients were recorded. The patients in the study group were subdivided into an intestinal dysfunction group (n=26) and a non-intestinal dysfunction group (n=12). Three ml venous blood was collected from the control group on admission and the same volume of plasma was collected from the study group both on admission and in the period of recovery. The plasma concentrations of endotoxin, diamine oxidase (DAO), D-lactate, and intestinal fatty-acid binding protein (iFABP) were detected respectively. The data collected were analyzed by the SPSS 17.0 software for Windows. RESULTS: The levels of variables were significantly higher in the study group than in the control group (P<0.01). They were higher in the intestinal dysfunction group than in the non-intestinal dysfunction group (DAO P<0.05, endotoxin, D-lactate, iFABP P<0.01). In the non-intestinal dysfunction group compared with the control group, the level of endotoxin was not significant (P>0.05), but the levels of DAO, D-lactate and iFABP were statistically significant (P<0.05). The levels of variables in acute stage were higher than those in recovery stage (P<0.01).The death group showed higher levels of variables than the survival group (endotoxin and D-lactate P<0.01, DAO and iFABP P<0.05). CONCLUSION: The plasma concentrations of endotoxin, DAO, D-lactate, and intestinal fatty-acid binding protein (iFABP) could reflect a better function of the intestinal mucosa barrier in surgically critical ill patients.

BACKGROUNDAngiogenesis is a prerequisite for tumor growth and plays an important role in rapidly growing tumors, such as malignant gliomas. A variety of factors controlling the angiogenic balance have been described, and among these, the endogenous inhibitor of angiogenesis, tumstatin, has drawn considerable attention. The current study investigated whether expression of tumstatin by glioma cells could alter this balance and prevent tumor formation.

METHODSWe engineered stable transfectants from human glioma cell line U251 to constitutively secrete a human tumstatin protein with c-myc and polyhistidine tags. Production and secretion of the tumstatin-c-myc-His fusion protein by tumstatin-transfected cells were confirmed by Western blotting analysis. In the present study, we identify the anti-angiogenic capacity of tumstatin using several in vitro and in vivo assays. Student's t-test and one-way analysis of variance (ANOVA) test were used to determine the statistical significance in this study.

RESULTSThe tumstatin transfectants and control transfectants (stably transfected with a control plasmid) had similar in vitro growth rates compared to their parental cell lines. However, the conditioned medium from the tumstatin transfected tumor cells significantly inhibits proliferation and causes apoptosis of endothelial cells. It also inhibits tube formation of endothelial cells on Matrigel. Examination of armpit tumors arising from cells overexpressing tumstatin repress the growth of tumor, accompanying the decreased density of CD31 positive vessels in tumors ((5.62 ± 1.32)/HP), compared to the control-transfectants group ((23.84 + 1.71)/HP) and wild type U251 glioma cells group ((29.33 + 4.45)/HP).

CONCLUSIONAnti-angiogenic gene therapy using human tumstatin gene may be an effective strategy for the treatment of glioma.

Animals ; Autoantigens ; genetics ; Brain Neoplasms ; blood supply ; therapy ; Cell Line, Tumor ; Cell Proliferation ; Collagen Type IV ; genetics ; Genetic Therapy ; Glioma ; blood supply ; pathology ; therapy ; Humans ; Mice ; Mice, Inbred BALB C ; Neovascularization, Pathologic ; prevention & control ; Platelet Endothelial Cell Adhesion Molecule-1 ; analysis ; Transfection

Objective To explore the activation and function of Janus protein-tyrosine kinase (JAK)/ signal transducer and activator of transcription (STAT1) signal transduction pathway in kidney,lung and brain of MRL/lpr mice.Methods MRL/lpr mice with systemic lupus erythematosus (SLE) were studied at the age of 12 weeks up.Non-SLE MRL/lpr mice were used as controls.We used phosphospecific antibodies to detect STAT1 activation in kidney,lung and brain by immunohistochemistry and Western blots.Gene expression of the STAT induced feedback inhibitors of cytokine signaling 1 (SOCS-1) was investigated by SYBR green I real-time reverse transcriptase polymerase chain reaction (PCR).Results Phosphorylation of STAT1 protein was markedly activated in these three organs,although renal and pulmonary STAT1 activation were much more evidently activated.SOCS-1 gene expression increased in all three organs,while renal SOCS-1 gene expres- sion increased less than lung and brain.Conclusion The activation of JAK/STATI signal transduction path- way may be pathogenic in the organ involvement and progression of SLE.The pathogenesis of lupus nephritis may also be associated with the down-regulation of SOCS-1 feedback inhibition.

OBJECTIVE: To determine the mRNA and protein expressions of RCAS1 in human astrocytic tumors, and to explore the relation between their expression and the genesis and development of tumor. METHODS: The RCAS1 mRNA expression in human astrocytic tumors was evaluated by RT-PCR, and the RCAS1 protein expression was studied by immunohistochemical staining. RESULTS: The quantities of RCAS1 mRNA expression between diffusive astrocytoma(Grade II) and anaplastic astrocytoma(Grade III), anaplastic astrocytoma and glioblastoma(Grade IV) were significantly different(P<0.05), while the expression scores of RCAS1 protein were different only between the anaplastic astrocytoma and glioblastoma(P<0.01). RCAS1 protein expression was positively correlated with the tumor grade (r=0.573,P<0.001). The RCAS1 protein was not detected in normal brain tissues by immunohistochemical staining. CONCLUSION The RCAS1 expression is related to the histological grade of astrocytic tumor. In astrocytic tumors, the RCAS1 expression is regulated transcriptionally and posttranscriptionally.
Adolescent ; Adult ; Aged ; Antigens, Neoplasm ; genetics ; metabolism ; Astrocytoma ; genetics ; metabolism ; Brain Neoplasms ; genetics ; metabolism ; Child ; Child, Preschool ; Female ; Humans ; Male ; Middle Aged ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVETo investigate the relationship between the helicobacter pylori (HP) infection and the genetic instability of mitochondrial DNA (mtDNA) in human gastric adenocarcinoma epithelial cells (AGS).

METHODSAfter treated with extracts of HP11638 (CagA+, VacA+) or Hp11638 mutant strain (CagA+, VacA-), AGS cells were collected, and mitochondrial DNA was extracted and Cox-I, Cox-II, Cox-III, ATPase6, ATPase8 and Cytb genes and the D-Loop region were amplified by PCR and then sequenced.

RESULTSThe mutation rates of the mtDNA in AGS cells were correlated with the extracts of the two HP strains in a concentration- and time-dependent manner. But the mtDNA mutation rate in AGS cells treated with the HP11638 extract was higher than that treated with the Hp11638 mutant extract. Total of 616 mutations in D-Loop region were detected, including 489 point mutations, 81 insertions and 46 deletions. Among them, 70.9% (437/616) belonged to GC to AT and AT to GC transition. Seventeen out of 20 (85%) AGS cells treated with extract of HP had mutations in 303PolyC, 16184PolyC and 514CA regions of mtDNA D-Loop. No mutation was detected in Cox-I, Cox-II, Cox-III, ATPase6 and ATPase8 genes, three point mutations were found in the Cytb gene.

CONCLUSIONHP can cause the accumulation of mutations in mtDNA, in particular, in the D-Loop region, and the VacA participated in the process.

Antigens, Bacterial ; pharmacology ; Base Sequence ; DNA, Mitochondrial ; drug effects ; genetics ; Endothelial Cells ; drug effects ; pathology ; Helicobacter Infections ; complications ; Helicobacter pylori ; chemistry ; Humans ; Mutation ; Stomach ; pathology

OBJECTIVETo assess the left ventricular longitudinal rotation (LR) in patients with dilated cardiomyopathy (DCM).

METHODSConventional echocardiography (GE-Vivid7) was performed in 35 healthy subjects and 42 DCM patients. Left atrial diameter was measured by M-mode echocardiography, left ventricular end-systolic, end-diastolic volume and left ventricular ejection fraction (LVEF) were calculated by bi-plane simpson's method. The peak velocity during early diastole (Ve) and late diastole (Va) of anterior mitral valve were measured by pulse-waved doppler, and the ratio Ve/Va was calculated. The peak radial systolic strain, strain rate in systolic, early and late diastolic periods were measured. Segmental LR and global LR were assessed using two-dimensional speckle tracking imaging (2D-STI).

RESULTSThe peak radial systolic strain, strain rate in systolic, early and late diastolic periods in DCM group were significantly lower than in healthy subjects, the rotation degrees of the middle and base lateral, the apex and the base septum walls were significantly lower than those of the healthy subjects. A prominent counterclockwise LR (0.76° ± 2.63°) was shown in healthy subjects while prominent clockwise LR (-1.58° ± 3.42°) was present in DCM patients. The time delay between the left ventricular lateral wall and the base septum wall in DCM patients significantly correlated with the peak LR of the left ventricular (r = 0.409, P < 0.01; r = 0.396, P < 0.01).

CONCLUSIONS2D-STI can be used to assess the LR in DCM patients and a clockwise LR is present in DCM patients which might be caused by the time delay between the left ventricular lateral wall and the base-septum wall.

Cardiomyopathy, Dilated ; physiopathology ; Case-Control Studies ; Diagnostic Imaging ; Diastole ; Echocardiography ; Echocardiography, Doppler ; Heart Atria ; Heart Ventricles ; Humans ; Rotation ; Systole ; Ventricular Function, Left

Chromogranin A (CGA) is a soluble protein existed in most secreted cells and neurons. It was recently found that the bovine CGA N terminal region has vasoinhibitory, antibacterial and antifungal activities. Since the need for effective antifungal agents increases in parallel with the expanding number of immunocompromised patients at risk for fungal infections, it becomes imperative to find antifungal compounds with low toxicity toward mammalian cells. To study the antifungal activity of CGA N terminal region, the DNA fragment encoding for the N terminal 1-76 amino acid sequence (CGA1-76) of human CGA was amplified by PCR technique. After DNA sequence analysis, the amplified DNA fragment was cloned into the Bacillus subtilis inducible and expression vector pSBPTQ constructed in this study and the resultant plasmid pSVTQ was then transformed into triple-protease deficient Bacillus subtilis strain DB403 competent cells. The transformants was screened on LB plates containing 10 microg/mL kanamycin. The positive transformant DB403 (pSVTQ) was grown on kanmycin containing 2 x MSR medium and sucrose was added to 2% final concentration for induction after 2h cultivation. The culture supernatant was used to run SDS-PAGE. The result of SDS-PAGE showed that the CGA1-76 was expressed by sucrose induction and the expressed product secreted into the medium with a yield of 5 mg/L. The expressed product reacts specifically with mouse anti CGA47-68 monoclonal antibody. The antifungal activity of the expressed product was examined by adding the culture supernatant to the fungal spore or Candida albican suspensions at appropriate proportion and found that the recombinant human CGA1-76 produced in Bacillus subtilis inhibits the growth of Fusarium sp. Alternaria sp. and Candida albican at the concerntration of 4 micromol/L. These results demonstrate that human CGA1-76 has expressed in Bacillus subtilis and the expressed product is immunogenic and has the antifungal activity.
Amino Acid Sequence ; Animals ; Antifungal Agents ; pharmacology ; Bacillus subtilis ; genetics ; metabolism ; Base Sequence ; Chromogranin A ; biosynthesis ; genetics ; pharmacology ; Cloning, Molecular ; DNA ; genetics ; Humans ; Mice ; Molecular Sequence Data ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology

This study was purposed to investigate the serum levels of soluble intracellular adhesion molecule (sVCAM-1), interleukin 18 (IL-18) and vascular endothelial growth factor (VEGF) in patients with aplastic anemia (AA) and their clinical significance. Enzyme linked immunosorbent assay (ELISA) was used to detect sVCAM-1, IL-18 and VEGF in serums of 30 patients with AA and 25 normal controls. The results showed that the serum levels of sVCAM-1 and IL-18 in patients with AA [(839.08 +/- 173.97) ng/ml, (380.35 +/- 47.76) pg/ml] were significantly higher than those in normal controls [(538.16 +/- 91.21) ng/ml, (256.39 +/- 59.52) pg/ml] (p < 0.01; p < 0.01). The levels of sVCAM-1 and IL-18 in severe AA patients [(969.94 +/- 182.54) ng/ml, (388.96 +/- 46.06) pg/ml] were higher than those in chronic AA patients [(709.26 +/- 165.32) ng/ml, IL-18 (352.21 +/- 47.08) pg/ml] (p < 0.01; p < 0.05), but the level of VEGF in AA patients [(69.63 +/- 27.42) pg/ml] was lower than that in the normal controls [(125.62 +/- 32.15) pg/ml] (p < 0.01)]. The level of VEGF in severe AA patients [(51.30 +/- 29.86) pg/ml] was significantly lower than that in chronic AA patients [(80.02 +/- 25.14) pg/ml] (p < 0.01). The levels of sVCAM-1 and IL-18 in AA patients after treatment were lower than those before treatment (p < 0.01; p < 0.05), but the level of VEGF after treatment was significantly higher than that before treatment (p < 0.05). It is concluded that the high levels of sVCAM-1, IL-18 and low level of VEGF in serum may be involved in the pathogenesis and progress of AA.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; blood ; Case-Control Studies ; Female ; Humans ; Interleukin-18 ; blood ; Male ; Middle Aged ; Vascular Cell Adhesion Molecule-1 ; blood ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

OBJECTIVESTo evaluate the long-term facial nerve function of patients following microsurgical removal of large and huge acoustic neuroma, and to identify the factors that influence these outcomes.

METHODSA retrospective review was performed which included 176 consecutive patients with a large acoustic neuroma (≥ 30 mm) underwent a retrosigmoid craniotomy for tumor resection between January 2002 to November 2009. House-Brackmann (HB) Scale was used preoperatively and in a long-term follow-up after surgery. Test for linear trend was applied for statistic analysis.

RESULTSComplete resection was achieved in 168 (95.5%) of these 176 patients with a mortality of 1.7%. Anatomic preservation of the facial nerve was attained in 96.0% of the patients. In the series of 96 patients who had at least 1-year follow-up (mean 3.0 years) the facial nerve function preservation (HB grade 1 - 2) was totally attained in 79 patients (82.3%), and 40 of 55 patients (72.7%) who presented huge tumors (diameter > 40 mm) among the 96 patients had facial nerve function preserved. Analysis showed that facial nerve function correlated linearly with tumor sizes (χ(2) = 14.114, ν = 1, P < 0.05).

CONCLUSIONSComplete removal of large and giant acoustic neuroma may be obtained via retrosigmoid approach with facial nerve preservation. Excellent long-term facial function can be expected in the majority of patients who undergo microsurgical removal of vestibular schwannoma via the suboccipital retrosigmoid approach. Tumor size is a significant prognostic parameter for facial nerve function following vestibular schwannoma surgery.

Adolescent ; Adult ; Aged ; Facial Nerve ; physiopathology ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; Middle Aged ; Neuroma, Acoustic ; surgery ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Young Adult