BackgroundGlaucoma associates with apoptosis of retinal ganglion cells ( RGCs).Research showed that ciliary neurotrophic factor (CNTF) can repair optic nerve trauma,but it has not reported whether CNTF has a protective effect on glaucomatous optic neuropathy.ObjectiveThis experimental study was to explore the protective effect of CNTF on RGCs in rat eyes with acute ocular hypertension.MethodsOcular acute hypertension models were induced in bilateral eyes of 24 clean Wistar rats by forced perfusion of a balanced salt solution into the anterior chamber.Two days before molding,0.5 μg recombinant human CNTF (5 μl) was injected intravitreously in the left eyes,and 5 mmol/L sodium phosphate solution (5 μl) was injected in the same way as the control group.Three other Wistar rats were used as the normal control group.The animals were sacrificed by excessive anesthesia and the retinal sections were prepared 1,3,7,14 days after molding.The morphology of the retina was examined and RGCs counting was performed by hematoxylin & eosin staining and observed under a light microscope.The expression of glutamic acid in RGCs was assessed by immunochemistry.ResultsRegular retinal structure with clearly defined cell layers were observed in normal control rats.Changes in vacuoles in RGCs were seen in the model control group.The number of degenerative RGCs decreased in the CNTF group.Compared with the model control group,number of normal RGCs increased from 1 day to 14 days after molding with a significant difference between the two groups (all P=0.000).Immunochemistry assay showed that the numbers of positive cells for glutamic acid were 5.50±1.04 and 6.00±1.41 for the average of 3 fields at 3 days or 7 days in the CNTF group,and those in the model control group were 9.00±2.91 and 10.83 ± 1.94,respectively,showing significant differences between them ( all P =0.000 ).However,no comparable differences were found in the numbers of positive cells for glutamic acid at 1 day and 14 days between the two groups( P=0.578,0.180).ConclusionsCNTF down-regulates the expression of glutamic acid in RGCs and offers neuroprotection for RGCs in an acute glaucoma rat model.

To investigate the antitumor immune responses induced by MAGE-3 DNA vaccine, the recombinant mammalian expression plasmid pcDNA3.1/MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR, and the pcD-NA3.1 + vector. The recombinant plasmids were transfected into B16 cells by liposome, the expression of MAGE-3 was checked by RT-PCR, immunocytochemistry and Western blot. Then, 100 ug recombinant plasmids were injected intramuscularly per C57BL/6 mouse on 0, 10 and 20 days, with pcDNA3.1 + plasmid and PBS as controls. Splenocytes CTLs, the level of antibodies against MAGE-3 the changes of the T lymphocyte subsets and the levels of cytokines were checked after 3 times immunization. As a result, the mice immunized with pcDNA3.1/MAGE-3 plasmid can produce MAGE-3 specific immune response. The CTLs kill activities against B16/MAGE-3 cells was 51.08 +/- 7.41%, and had significant difference (P < 0.01) compared with that of pcDNA3.1 + group (8.44 +/- 1.89%) and PBS group (5.76 +/- 1.75%). The titre of antibody against MAGE-3 was 1:15, while controls were negtive. The number of CD4 + CD8 + and the levels of IFN-gamma IL-2 increased significantly after immunization with pcDNA3.1/MAGE-3 plasmid as compared with those of control groups (P < 0.01). It is concluded that the pcDNA3.1-MAGE-3 DNA vaccine are able to induce both cellular and humoral immune responses in vivo.
Animals ; Antibodies, Neoplasm ; blood ; Antigens, Neoplasm ; biosynthesis ; genetics ; immunology ; Cancer Vaccines ; biosynthesis ; immunology ; Female ; Melanoma, Experimental ; prevention & control ; Mice ; Mice, Inbred C57BL ; Neoplasm Proteins ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; T-Lymphocyte Subsets ; immunology ; Vaccination ; Vaccines, DNA ; biosynthesis ; genetics ; immunology

To express the GST-MAGE-3 protein in E. coli, and investigate the antitumor immune responses induced by Dendritic cells (DCs) pulsed with GST-MAGE-3 protein, the recombinant expression plasmid pGEX-MAGE-3 was constructed by ligating MAGE-3 gene, which was amplified by RT-PCR and confirmed by sequencing, and the pGEX-4T-2 vector. The recombinant plasmid was transformed into BL-21 E. coli. The expression of GST-MAGE-3 was induced with IPTG. The GST-MAGE-3 protein expressed as high as 32% of the total cellular protein. After purification with Glutathione Sepharose 4B, the purity of the protein was more than 90%, and 3mg GST-MAGE-3 was obtained from 100 mL BL-21 lysate. Dendritic cells from gastric carcinoma patients were pulsed with GST-MAGE-3 protein, and these DCs were used to stimulate the autologous T. lymphocytes. After 7 days, the T. lymphocytes cocultured with DCs pulsed with GST-MAGE-3 antigen exhibited specific cytotoxicity against MAGE-3 positive SGC-7901 cells. It is concluded that the GST-MAGE-3 protein are able to present antigen to T. lymphocytes, activate antigen-specific CTLs and induce special antitumor immune responses in vitro. Our results lay the groundwork for further research of the MAGE-3 vaccine.
Antigens, Neoplasm ; genetics ; isolation & purification ; metabolism ; pharmacology ; Blotting, Western ; Cell Line, Tumor ; Cells, Cultured ; Dendritic Cells ; drug effects ; immunology ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Neoplasm Proteins ; genetics ; isolation & purification ; metabolism ; pharmacology ; Plasmids ; genetics ; Stomach Neoplasms ; immunology ; T-Lymphocytes ; immunology

Most patients with trauma-hemorragic shock, prior to ICU admission, have been resuscitated and stabilized in the emergency room (ER) and/or operation room (OR). Many of them suffer from systemic edema. This extra-vascular fluid is caused by massive infusion of fluid and blood for the maintenance of blood pressure. During the recovery stage, the patients exhibit spontaneous diuresis followed by negative fluid balance. Urine volumes of some patients are more than 10000 ml/d. Do we need to maintain a balance between daily input and output of water at this situation? There are many references in the medical literature and textbooks about fluid resuscitation and the principles in maintaining the balance between input and output of water, but rarely about when and how to restrict it, that is, when and how to permit a negative balance. In this retrospective review, we examined the resuscitation records of 205 patients with systemic edema after trauma-hemorragic shock.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cause of Death ; Child ; Critical Care ; methods ; Diuresis ; physiology ; Female ; Fluid Therapy ; methods ; Humans ; Injury Severity Score ; Intensive Care Units ; Male ; Middle Aged ; Multiple Trauma ; diagnosis ; mortality ; therapy ; Predictive Value of Tests ; Probability ; Prognosis ; Retrospective Studies ; Risk Assessment ; Shock, Hemorrhagic ; diagnosis ; mortality ; therapy ; Survival Rate ; Water-Electrolyte Balance

OBJECTIVETo investigate the angiographic feature and its diagnostic value in gastrointestinal stromal tumor (GIST).

METHODSTwelve patients with pathologically proved GIST underwent angiography using PUCK and DSA one week before operation. The origin, size, morphology and angiographic appearance of the lesion were retrospectively analyzed.

RESULTSIt was found that two tumors originated from stomach, eight from jejunum, and two from ileum. Seven cases were benign but five malignant. Obviously thickened and enlarged feeding artery was detected in eight tumors, and early-developed vein was found in three. Two types of angiographic changes of GIST were observed: 1) twisted irregular neoplastic vessels with partially coarse and indistinct margin were found in four cases, which were all malignant; 2) ball-like neoplastic vessels with homogeneous stain in tumor were found in eight cases, seven of them were benign but one was malignant.

CONCLUSIONAngiography may be helpful in localization and diagnosis as well as defining the size of GIST, and especially in patients with melena of unknown origin.

Adult ; Aged ; Angiography ; methods ; Female ; Gastrointestinal Stromal Tumors ; blood supply ; diagnostic imaging ; Humans ; Jejunal Neoplasms ; blood supply ; diagnostic imaging ; Male ; Mesenteric Arteries ; diagnostic imaging ; Middle Aged ; Stomach Neoplasms ; blood supply ; diagnostic imaging

OBJECTIVETo explore the management for blunt abdominal trauma victims with probable positive diagnostic peritoneal lavage (DPL) findings.

METHODSData of 76 patients with probable positive DPL findings accepted to ICU in previous 10 years were reviewed. After admission, the patients were evaluated in a settled time according to the protocols of Advanced Trauma Life Support (ATLS). Vital signs were continuously monitored and DPL, ultrasound and/or CT scan were repeated when necessary.

RESULTSEighteen (24%) of 76 patients presented positive DPL findings after repeated DPL. Surgical findings confirmed 7 cases of spleen rupture, 3 hepatorrhexis (infra-Glisson capsule), 4 intestinal perforation, 2 gastric perforation, 1 colon perforation and 1 injured mesentery.

CONCLUSIONSPatients with probable positive DPL findings were admitted to ICU with vital signs continuously monitored. Repeated DPL with supplemental ultrasound and/or CT scan can work together to increase the sensitivity and accuracy of the diagnosis, reduce the rate of exploratory laparotomy, ensure patients' safety and provide a reliable basis for therapeutic operations.

Abdominal Injuries ; diagnosis ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Intensive Care Units ; Male ; Middle Aged ; Peritoneal Lavage ; Wounds, Nonpenetrating ; diagnosis

OBJECTIVEThis study compared the differences in clinical features between chronic aplastic anemia (CAA) and myelodysplastic syndrome (MDS) in children in order to provide a basis for the differential diagnosis of the two diseases.

METHODSA retrospective study of 23 cases of CAA and 9 cases of MDS from September 2007 to September 2010 was performed. The clinical data including routine blood test results, reticulocyte counts, serum lactate dehydrogenase level, serum ferritin level, cytological examination of bone marrow, bone marrow CD34+ cell counts, bone marrow chromosome and FISH test results were compared between the CAA and MDS groups.

RESULTSNeutrophils, reticulocytes, and serum ferritin and lactate dehydrogenase levels increased in the MDS group compared with those in the CAA group. There were significant differences in bone marrow blast cell counts and dyshematopoiesis phenomena of three lines blood cells between the CAA and MDS groups. The bone marrow CD34+ cell counts and the rate of chromosomal abnormalities detected in bone marrow cytogenetic analysis in the MDS group were significantly higher than those in the CAA group.

CONCLUSIONSThere are differences in the results of laboratory examinations and morphological and cytogenetic examinations of bone marrow between the children with CAA and MDS. The differences are useful to the differential diagnosis of the two diseases.

Anemia, Aplastic ; genetics ; pathology ; Bone Marrow Examination ; Child ; Child, Preschool ; Chromosome Aberrations ; Chronic Disease ; Female ; Humans ; Male ; Myelodysplastic Syndromes ; genetics ; pathology

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 how transient low dose of hydroperoxide pretreatment prevents cardiac ischemia/reperfusion injury.

METHODSSD rats were divided into 4 groups: sham operation (Sham), standard ischemia/reperfusion (I/R), ischemic preconditioning (IPC) and IR preceded by low H2O2 treatment. Cardiac function and injury parameter were compared among groups.

RESULTSIPC protected reperfusion injury and improved cardiac function. Low H2O2 treatment played a role in cardioprotection similar to IPC. Low H2O2 was indeed generated in the early phase of simulated ischemia and attenuated cytochrome c release induced by high Ca2+ in isolated mitochondria.

CONCLUSIONLow H2O2 plays a critical role in cardioprotection probably by inhibiting mitochondrial permeability transition.

Animals ; Hydrogen Peroxide ; administration & dosage ; Ischemic Preconditioning ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; prevention & control

OBJECTIVETo study the chemical constituents from the roots and rhizome of Cynanchum atratum.

METHODThe chemical constituents were isolated and repeatedly purified by silica gel chromatography and the structures were elucidated by the NMR spectra and physicochemical properties.

RESULTTwelve compounds were obtained and nine of them were identified as 2, 4-dihyroxyacetophe none (1), 2, 6-dihyroxyacetophenone (2), 4-hydroxybenzenemethanol (3), benzoic acid (4), beta-amyrin acetate (5), palmitic acid (6), beta-sitosterol (7), beta-daucosterol (8), glaucogenin C-3-O-alpha-D-oleandropyranosyl-(1 --> 4)-beta-D-digitoxopyranosyl-(1 -->4)-alpha-D-oleandropyranoside (9).

CONCLUSIONSeven compounds were obtained from C. atratum for the first time.

Benzoic Acid ; chemistry ; isolation & purification ; Chromatography, Gel ; Cynanchum ; chemistry ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Palmitic Acid ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Silica Gel ; Silicon Dioxide