Adolescent ; Anti-Glomerular Basement Membrane Disease ; pathology ; Diagnosis, Differential ; Granulomatosis with Polyangiitis ; pathology ; Humans ; Kidney ; pathology ; Lung ; pathology ; Lupus Erythematosus, Systemic ; pathology ; Male

OBJECTIVETo observe the effects of three fluid resuscitation methods on apoptosis of visceral organs in rats with hemorrhagic shock.

METHODSA model of rat with severe hemorrhagic shock and active bleeding was established in 32 SD (Sprague-Dawley) rats. The rats were randomly divided into control group, no fluid resuscitation group (NF group), controlled fluid resuscitation group (NS40 group) and rapid large scale fluid resuscitation group (NS80 group). Each group contained 8 rats. The curative effects were compared. At the same time, the apoptosis in liver, kidney, lung and small intestinal mucosa of survivors after hemorrhage and resuscitation was detected by light microscopy in HE (hematoxylin and eosin) stained tissue sections, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL).

RESULTSThe survival rate of early fluid resuscitation (14/16) was markedly higher than that of NF group (3/8). There was some apoptosis in liver, kidney, lung and small intestinal mucosa of all survivors. Compared with NF and NS40 groups, the apoptosis of liver, kidney and small intestinal mucosa of NS80 group was obviously increased.

CONCLUSIONSAmong three fluid resuscitation methods, controlled fluid resuscitation can obviously improve the early survival rate and the apoptosis of liver, kidney and small intestinal mucosa in rats with severe and uncontrolled hemorrhagic shock, and may benefit improvement of prognosis.

Animals ; Apoptosis ; Blood Pressure ; Flow Cytometry ; Fluid Therapy ; methods ; In Situ Nick-End Labeling ; Intestine, Small ; pathology ; Kidney ; pathology ; Lactic Acid ; blood ; Liver ; pathology ; Lung ; pathology ; Male ; Organ Specificity ; Rats ; Rats, Sprague-Dawley ; Resuscitation ; methods ; Shock, Hemorrhagic ; pathology ; physiopathology ; therapy

OBJECTIVETo investigate the effect of intravesical instillation of antifibrinolytic agents with bacillus Calmette-Guerin (BCG) on preventing recurrence of superficial bladder transitional cell carcinoma (BTCC) after surgical management.

METHODSA total of 326 cases of superficial BTCC undergoing transurethral resection of bladder tumor (TURBT) or partial cystectomy were divided into 5 groups. Then the different dosage BCG with or without antifibrinolytic agents was regular instilled into bladders (once a week, then once a month after 6 times). Group A including 66 cases received intravesical instillation of 100-120 mg BCG plus 100 mg para-aminomethyl benzoic acid (PAMBA). Group B including 64 cases: instillation of 50-60 mg BCG plus 100 mg PAMBA; Group C including 65 cases: 100-120 mg BCG plus 2.0 g epsilon-aminocaproic acid (EACA); Group D including 64 cases: 50-60 mg BCG plus 2.0 g EACA; Group E (control group) including 67 cases: 100-120 mg BCG. All the cases had been followed up for 4 to 69 months (mean, 28.5 months). Not only was cystoscopy performed every 3 months, but also biopsy was carried out to identify recurrence when necessary. Side effect was recorded after instillation.

RESULTSThe rate of tumor recurrence of Group A, Group B, Group C and Group D was 12%, 10%, 9%, 9% respectively, which was significantly lower than that of Group E (30%) (chi(2) = 5.699, 6.818, 7.380, 7.867, P = 0.017, 0.009, 0.007, 0.005). And there was no significant difference of tumor recurrence rate between Group A and Group B or between Group C and Group D (Group A and Group C: high dosage BCG plus antifibrinolytic agents, while Group B and Group D: low dosage BCG plus antifibrinolytic agents) (P > 0.05). But the side effects developing in Group B and Group D after BCG instillation were less than those in Group A and Group C.

CONCLUSIONSThe efficacy of BCG on prevention the recurrence of superficial BTCC can be enhanced when combined with antifibrinolytic agents. Even if the dosage of BCG was reduced by half, the efficacy didn't changed. A new approach of low dosage BCG plus antifibrinolytic agents is recommended in the prophylaxis of recurrence of bladder cancer.

4-Aminobenzoic Acid ; administration & dosage ; Adjuvants, Immunologic ; therapeutic use ; Administration, Intravesical ; Adult ; Aged ; Aged, 80 and over ; Aminocaproic Acid ; administration & dosage ; Antifibrinolytic Agents ; therapeutic use ; BCG Vaccine ; therapeutic use ; Carcinoma, Transitional Cell ; drug therapy ; surgery ; Combined Modality Therapy ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; prevention & control ; Urinary Bladder Neoplasms ; drug therapy ; surgery ; para-Aminobenzoates

OBJECTIVETo investigate the effect of small interfering RNA on cell proliferation and apoptosis in gastric cancer cell lines with high expression of RegIα.

METHODSTotal RNA was isolated from six gastric cancer cell lines,and the expression of RegI α mRNA was detected by RT-PCR. RegI α RNAi expression vector was constructed and stably transfected into MKN45 and AGS cells with high RegI α expression, empty-vector was used as control. RegI α mRNA and protein expression was measured by RT-PCR and Western blot respectively in stable transfected cell lines. Cell proliferation and apoptosis were detected with MTT assay and flow cytometry.

RESULTRT-PCR results indicated that RegI α mRNA expression was significantly inhibited by RNAi in both cell lines compared with empty-vector. Western blot results showed that RegIα protein was down-regulated to (44 ± 4)% and (25 ± 4)% respectively in MKN45 and AGS cells compared to empty-vector. MTT results showed that cell growth was significantly inhibited in MKN45 and AGS cells. The apoptosis rate in MKN45 and AGS cells was remarkable increased compared to that of empty-vector (12.96 ± 0.50)% compared with (3.99 ± 0.30)% and (11.59 ± 1.10)% compared with (4.22 ± 0.40)% (P < 0.05).

CONCLUSIONSmall interfering RNA of RegI α gene can efficiently down-regulate RegI α expression in MKN45 and AGS cell lines, and further inhibit cell growth and induce cell apoptosis.

Animals ; Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Lithostathine ; genetics ; metabolism ; Mice ; Plasmids ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transfection

OBJECTIVETo construct RegIα over-expression vector and to evaluate the effect of RegIα on the proliferation and apoptosis of gastric cancer MKN28 cells in vitro.

METHODSFull sequence of RegIα cDNA was amplified from normal gastric tissue samples by RT-PCR and cloned into pIRES2-EGFP vector. RT-PCR and Western blot were performed to detect expression levels of RegIα in MKN28 cells. The effects of over-expression RegIα on cell proliferation was measured by MTT assay and apoptosis was detected by flow cytometry.

RESULTRegIα cDNA over-expression vector of pIRES2-RegIα-EGFP was successfully constructed. The expressions of RegIα in MKN28 cells, including mRNA and protein levels, were significantly increased after stable transfection, which resulted in cell proliferation and anti-apoptotic effect induced by H(2)O(2).

CONCLUSIONThe over-expression of RegIα can promote cell proliferation and reduce cell apoptosis when induced by H(2)O(2) in gastric cancer cells.

Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Plasmids ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Transfection

OBJECTIVETo observe the effects of antisense RNA of connective tissue growth factor (CTGF) on rat liver fibrosis.

METHODSGene recombinant techniques were used to construct a rat antisense RNA of CTGF recombinant plasmid which could be expressed in eukaryotic cells. The recombinant plasmids were encapsulated with lipofectamine and then transducted into a carbon tetrachloride (CCl4) induced rat liver fibrosis model. Expression of CTGF was assessed by RT-PCR, Western blot and immunohistochemistry. Immunohistochemistry was used to identify type I and III collagens. HE stained liver slides were used for pathological study.

RESULTSThe mRNA and protein expression of CTGF in the fibrotic liver transfected with antisense-CTGF were significantly decreased compared with those of the controls (P<0.01). The depositions of type I and type III collagens were also decreased (P<0.05). Antisense-CTGF also minimized the pathological fibrosis in the rat livers (P<0.01).

CONCLUSIONThe results demonstrate that the antisense RNA of CTGF recombinant plasmid has certain effects in preventing liver fibrosis and makes it a possible candidate for use in future gene therapy.

Animals ; Connective Tissue Growth Factor ; genetics ; Genetic Therapy ; Liver ; pathology ; Liver Cirrhosis, Experimental ; pathology ; Male ; Plasmids ; RNA, Antisense ; genetics ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo study the effects of decreased leptin expression on liver fibrosis.

METHODSThe small interfering RNA, targeting leptin gene, was designed according to the secondary structure of leptin gene. The recombinant plasmids were encapsulated with lipofectamine and then injected into carbon tetrachloride (CCl4) induced rat liver fibrosis models. Leptin and I, III collage were detected by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe mRNA and protein levels of leptin in the fibrotic liver transfected with leptin shRNA were significantly decreased compared with those in controls (P less than 0.01). The depositions of type I and type III collagens were also decreased (P less than 0.01).

CONCLUSIONDecreased leptin expression prevents liver fibrosis.

Animals ; Leptin ; genetics ; Liver Cirrhosis, Experimental ; therapy ; Male ; RNA, Messenger ; genetics ; RNA, Small Interfering ; Rats ; Rats, Sprague-Dawley

Objective To study the relationship between the initial expression level of glucocorticoid receptor (GR) and the treatment outcome in children with acute lymphoblastic leukemia (ALL). And to evaluate if the initial expression level of GR could be the prognostic factor for children with ALL.Methods Anti-GR-antibody was used to measure the GR expression level in the bone marrow samples from 48 newly diagnosed children with ALL with flow cytometry. Also the GR expression levels in the patients at complete remission were mea-sured. Fifteen randonmized samples from ALL patients in continuous complete remission (CCR) were measured in this study. The GR expre-ssion levels of 30 blood samples from children in control group were monitored. Results The initial GR expression level had no association with the results after therapy. The GR expression level in CR and CCR had no statistic difference compared with that in control group.Conclusions It is not clear yet if the initial GR expression level could be the prognostic factor in children with ALL. Monitoring dynamic changes of the GR expression level in children with ALL seems to be of no remarkable significance.

OBJECTIVETo develop a specific SARS virus-targeted antibody preparation for emergent prophylaxis and treatment of SARS virus infection.

METHODSBy using phage display technology, we constructed a naive antibody library from convalescent SARS patient lymphocytes. To obtain the neutralizing antibody to SARS virus surface proteins, the library panning procedure was performed on purified SARS virions and the specific Fab antibody clones were enriched by four rounds of repeated panning procedure and screened by highthroughput selection. The selected Fab antibodies expressed in the periplasma of E. coli were soluble and further purified and tested for their binding properties and antiviral function to SARS virus. The functional Fab antibodies were converted to full human IgG antibodies with recombinant baculovirus/insect cell systems and their neutralizing activities were further determined.

RESULTSAfter four rounds of the panning, a number of SARS-CoV virus-targeted human recombinant Fab antibodies were isolated from the SARS patient antibody library. Most of these were identified to recognize both natural and recombinant SARS spike (S) proteins, two Fab antibodies were specific for the virus membrane (M) protein, only one bound to SARS-CoV nucleocapsid protein. The SARS-CoV S and M protein-targeted Fab or IgG antibodies showed significant neutralizing activities in cytopathic effect (CPE) inhibition neutralization test, these antibodies were able to completely neutralize the SARS virus and protect the Vero cells from CPE after virus infection. However, the N protein-targeted Fab or IgG antibodies failed to neutralize the virus. In addition, the SARS N protein-targeted human Fab antibody reacted with the denatured N proteins, whereas none of the S and M protein specific neutralizing antibodies did. These results suggested that the S and M protein-specific neutralizing antibodies could recognize conformational epitopes which might be involved in the binding of virions to cellular receptors and the fusion activity of the virus.

CONCLUSIONThe SARS-CoV spike protein and membrane proteins are able to elicite efficient neutralizing antibodies in SARS patients. The neutralizing antibodies we generated in this study may be more promising candidates for prophylaxis and treatment of SARS infection.

Amino Acid Sequence ; Animals ; Antibodies, Viral ; immunology ; Cercopithecus aethiops ; Humans ; Membrane Glycoproteins ; immunology ; Neutralization Tests ; Peptide Library ; Protein Binding ; Protein Engineering ; Recombinant Proteins ; immunology ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; virology ; Spike Glycoprotein, Coronavirus ; Vero Cells ; Viral Envelope Proteins ; immunology ; Viral Matrix Proteins ; immunology

The development and metastasis of uterine tumors depend highly on tumor angiogenesis. Multiphase dynamic contrast-enhanced magnetic resonance imaging can quantitatively describe the hemodynamic changes of uterine tumors based on a variety of tracer kinetic models and time-signal curves and by simulating the distribution of contrast inside and outside the blood vessels. Functional parameters can accurately and noninvasively assess tumor angiogenesis. It provides a non-invasive functional evaluation method for the differential diagnosis,staging,response evaluation,and prognostic prediction of uterine tumors.