OBJECTIVETo predict and screen the efficient antigenic epitopes in genus-specific envelope protein LipL41 of Leptospira interrogans and to determine the immunoreactive diversity of LipL41s from different genotypes.

METHODSBioinformatic methods were applied to predict the T/B combined epitope candidates in LipL41/1 and LipL41/2 molecules. The nucleotide fragments encoding epitopes were amplified by PCR. Phage display system with SDS-PAGE was performed to obtain the recombinant PIIIs containing different T/B combined epitopes. Western Blot assays were performed to determine the immunoreactivity of recombinant PIIIs to various antisera including antiserum against rLipL41/1, rLipL41/2 and whole cell of L.interrogans strain Lai, and serum from patients with leptospirosis.

RESULTBased on the predicting data, eight common or differential combined epitopes in LipL41s were selected. The nucleotide fragments encoding the epitopes were obtained by PCR. All the T/B combined epitope fragments were correctly inserted into the N end of phage PIII protein and then successfully expressed. All the antisera were able to recognize each of the epitopes but the hybridization signal intensity was different. Among these epitopes, the common T/B combined epitopes LipL41/1-30 and LipL41/1-233 showed a stronger and stable hybridization signals.

CONCLUSIONAll 8 selected T/B combined epitopes in the study are the efficient antigenic epitopes. The common T/B epitopes LipL41/1-30 and LipL41/1-233 can be first used in development of leptospiral MAP vaccine. The cross immunoreaction is between the differential T/B epitopes LipL41s-89,LipL41s-299 and the different antisera.

Amino Acid Sequence ; Antigens, Bacterial ; genetics ; immunology ; Bacterial Vaccines ; genetics ; immunology ; Cloning, Molecular ; Epitopes, B-Lymphocyte ; genetics ; immunology ; Epitopes, T-Lymphocyte ; genetics ; immunology ; Genotype ; Molecular Sequence Data ; Peptide Library

Chlamydia trachomatis ; genetics ; Chlamydiaceae Infections ; diagnosis ; genetics ; Genes, Bacterial ; genetics ; Genes, Viral ; genetics ; Gonorrhea ; diagnosis ; genetics ; Herpes Simplex ; diagnosis ; genetics ; Herpesvirus 2, Human ; genetics ; Humans ; Multiplex Polymerase Chain Reaction ; Neisseria gonorrhoeae ; genetics ; Ureaplasma Infections ; diagnosis ; genetics ; Ureaplasma urealyticum ; genetics

BACKGROUNDHemorrhagic shock induces immune dysfunction. Regulatory T cells (Tregs), T-helper (Th) cells, and cytotoxic T-lymphocytes (CTLs) can execute many crucial actions in immune and inflammatory responses. This study was conducted to investigate the early pathophysiological changes of CD4(+)CD25(+)Foxp3(+) Treg and Th1/Th2, Tc1/Tc2 profiles in the peripheral blood of rats with controlled hemorrhagic shock and no fluid resuscitation.

METHODSA rat model of controlled hemorrhagic shock with no fluid resuscitation was established. Peripheral blood samples were taken before and four hours after hemorrhagic shock with no fluid resuscitation. Three color flow cytometry was used to detect Tregs, Th1, Th2, Tc1 and Tc2 cells in the samples.

RESULTSIn the peripheral blood of rats, the percentage of Tregs four hours after hemorrhagic shock was significantly lower than before hemorrhagic shock (P = 0.001). The ratios of Th1/Th2 and Tc1/Tc2 were changed from (23.08 ± 8.98)% to (23.91 ± 15.36)%, and from (40.40 ± 21.56)% to (65.48 ± 23.88)%, respectively.

CONCLUSIONSAt an early stage, the advent of hemorrhagic shock is related to an early decrease of Tregs, and a mild shift in the Th1/Th2, Tc1/Tc2 balance toward Th1 and Tc1 dominance. These changes are part of a hyper-inflammatory state of the host, and will deteriorate the maintenance of immune balance. Further influences and detailed mechanisms need to be investigated.

Animals ; CD4 Antigens ; metabolism ; Forkhead Transcription Factors ; metabolism ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Resuscitation ; Shock, Hemorrhagic ; immunology ; metabolism ; T-Lymphocytes, Cytotoxic ; metabolism ; T-Lymphocytes, Regulatory ; metabolism ; Th1 Cells ; metabolism ; Th2 Cells ; metabolism

OBJECTIVETo investigate the changes and effects of arginine vasopressin (AVP) in patients with acute traumatic subarachnoid hemorrhage (tSAH).

METHODSThe plasma and cerebrospinal fluid (CSF) level of AVP, and intracranial pressure (ICP) were measured in a total of 21 patients within 24 hours after tSAH. The neurological status of the patients was evaluated by Glasgow Coma Scale (GCS). Correlation between AVP and ICP, GCS was analyzed respectively. Meanwhile, 18 healthy volunteers were recruited as control group.

RESULTSCompared with control group, the levels (pg/ml) of AVP in plasma and CSF (x+/-s) in tSAH group were significantly increased within 24 hours (38.72+/-24.71 vs 4.54+/-1.38 and 34.61+/-21.43 vs 4.13+/-.26, P less than 0.01), and was remarkably higher in GCS less than or equal to 8 group than GCS larger than 8 group (50.96+/-36.81 vs 25.26+/-12.87 and 44.68+/-31.72 vs 23.53+/-10.94, P less than 0.05). The CSF AVP level was correlated with ICP (r eqaul to 0.46, P less than 0.05), but no statistically significant correlation was found between plasma AVP, CSF AVP and initial GCS (r equal to -0.29, P larger than 0.05 and r equal to -0.32, P larger than 0.05, respectively). The ICP (mm Hg) in tSAH patients was elevated and higher in GCS less than or equal to 8 group than in GCS larger than 8 group (25.9+/-9.7 vs 17.6+/-5.2, P less than 0.05).

CONCLUSIONOur research suggests that AVP is correlated with the severity of tSAH, and may be involved in the pathophysiological process of brain damage in the early stage after tSAH. It seems that compared with the plasma AVP concentration, CSF AVP is more related to the severity of tSAH.

Adult ; Arginine Vasopressin ; blood ; cerebrospinal fluid ; Female ; Glasgow Coma Scale ; Humans ; Intracranial Pressure ; Male ; Middle Aged ; Subarachnoid Hemorrhage, Traumatic ; metabolism

OBJECTIVESTo investigate the possibility and efficacy of vasoligation and deferent vein ligation in treating and preventing benign prostatic hyperplasia(BPH).

METHODS40 male pooches were divided into A, B, C and D groups randomly. Each group has 10 pooches group A and B were made into the model of BPH. Two years later, the pooches in group A and C accepted the operation of vasoligation and deferent vein ligation on both sides. Group B and D only accepted the operation of dissection and relieving its deferent duct. Then continuing to feed the 40 pooches after the operation, they were killed another two years later. After taking out their prostates and calculating their volume and weight, the sections of the prostates were checked by microscope to observe their histology and pathology changes.

RESULTSThere were significant differences of weight and volume as well as the changes of histology and pathology between group A and group C (P < 0.01). It was the same with group B and D (hyperplasia changes).

CONCLUSIONSVasoligation and deferent vein ligation before benign prostatic hyperplasia at pooches grow-up stage can reduce the extent of BPH at old age, and treatment after benign prostatic hyperplasia can make prostatic tissue appear different degrees of atrophy.

Animals ; Disease Models, Animal ; Dogs ; Ligation ; Male ; Prostatic Hyperplasia ; prevention & control ; therapy ; Vasectomy

To compare the early effects of hypertonic and isotonic saline resuscitation on heme oxygenase-1 (HO-1) expression in organs of rats with hemorrhagic shock. Rats were randomly divided into hypertonic saline resuscitation (HTS), normal saline resuscitation (NS) and sham groups. HO-1 mRNA, protein expression and apoptosis were evaluated in organs. In the HTS group, significant difference was noted in HO-1 protein in small intestinal mucosa and liver compared with the NS and sham groups, and in HO-1 mRNA in liver and kidney compared with the sham group. The apoptosis of small intestinal mucosa, liver, heart, and lung was significantly lower in the HTS group than that in the NS group. In this study, small volume resuscitation with HTS can efficiently up-regulate the expression level of HO-1 in small intestinal mucosa and liver, which may be one of the mechanisms alleviating organ damage.
Animals ; Base Sequence ; Blood Pressure ; DNA Primers ; Gene Expression Regulation, Enzymologic ; drug effects ; Heme Oxygenase-1 ; metabolism ; Intestine, Small ; enzymology ; Kidney ; enzymology ; Liver ; enzymology ; RNA, Messenger ; genetics ; Rats ; Resuscitation ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; Saline Solution, Hypertonic ; pharmacology ; Shock, Hemorrhagic ; enzymology

BACKGROUNDHemorrhagic shock is usually associated with complicated immune and inflammatory responses, which are sometimes crucial for the prognosis. As regulators of the immune and inflammatory system; proliferation, migration, distribution and activation of myeloid-derived suppressor cells (MDSCs) are intimately linked to the inflammation cascade.

METHODSIn a model of severe hemorrhagic shock, thirty-five rats were randomly divided into control, sham, normal saline resuscitation (NS), hypertonic saline resuscitation (HTS), and hydroxyethyl starch resuscitation (HES), with seven in each group. MDSCs were analyzed by flow cytometric staining of CD11b/c(+)Gra(+) in peripheral blood mononuclear cells (PBMC), spleen cell suspensions, and bone marrow nucleated cells (BMNC). Simultaneously, the expressions of arginase-1 (ARG-1) and inducible nitric oxide synthase (iNOS) mRNA in MDSCs were evaluated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR).

RESULTSIn the early stage after hemorrhagic shock, fluid resuscitation and emergency treatment, the MDSCs in the PBMC of NS, HTS and HES groups markedly increased, and MDSCs in BMNC of these groups decreased accordingly, significantly different to the control group. In hemorrhagic shock rats infused with HTS at the early resuscitation stage, MDSCs in PBMC increased about 2 and 4 folds, and MDSCs in BMNC decreased about 1.3 and 1.6 folds, as compared to the sham group respectively, with statistically significant difference. Furthermore, compared to the NS and HES groups, the MDSCs in PBMC of HTS group increased 1.6 and 1.8 folds with statistically significant differences; the MDSCs decrease in BMNC was not significant. However, there was no statistically significant difference in MDSCs of spleen among the five groups. In addition, compared to the control, sham, NS and HES groups, the ARG-1 and iNOS mRNA of MDSCs in PBMC, spleen and BMNC in the HTS group had the highest level of expression, but no statistically significant differences were noted.

CONCLUSIONSIn this model of rat with severe and controlled hemorrhagic shock, small volume resuscitation with HTS contributes to dramatically early migration and redistribution of MDSCs from bone marrow to peripheral circulation, compared to resuscitation with NS or HES.

Animals ; Arginase ; genetics ; metabolism ; Blood Pressure ; physiology ; Disease Models, Animal ; Flow Cytometry ; Fluid Therapy ; methods ; Leukocytes, Mononuclear ; metabolism ; Male ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Saline Solution, Hypertonic ; therapeutic use ; Shock, Hemorrhagic ; immunology ; metabolism ; therapy

UNLABELLEDThe purpose of this study was to evaluate the effects of cellular immunity activation on P58(+) cells expressing killer cell inhibitory receptor (KIR) and their regulatory function on cellular immunity, and provid theoretical data for preventing graft-vers-host disease (GVHD) in stem cell transplantation therapy. The mononuclear cells from human peripheral blood were incubated with IL-2, Con A and Lipostin (LP) for 72 hours. The KIR expressing cells, P58.1(+) and P58.2(+) cells, were analyzed by flow cytometry. The results showed that the percentages of CD3(+), CD4(+), CD8(+), CD16(+)CD56(+), P58.1(+) and P58.2(+) cells were greatly increased after treated with IL-2, Con A and LP, separately or in combination, and the percentages of above cells in combined treatment groups were higher than those of single stimulated groups, especially the percentage of cells in the IL-2 + LP group was significantly higher than those in IL-2 and LP singly treated groups.

IN CONCLUSIONIL-2, Con A and LP possess the ability to induce the expression of KIR and stimulate proliferation of P58.1(+) and P58.2(+) cells while to activate the celluar immunity response, the expression of P58 gene may be regulated by the activation of cellular immunity.

Adult ; CD3 Complex ; analysis ; CD4 Antigens ; analysis ; CD56 Antigen ; analysis ; CD8 Antigens ; analysis ; Cell Count ; Cell Division ; drug effects ; Concanavalin A ; pharmacology ; Flow Cytometry ; Humans ; Interleukin-2 ; pharmacology ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Receptors, IgG ; analysis ; Receptors, Immunologic ; analysis ; Receptors, KIR ; Receptors, KIR2DL3

BACKGROUNDParaquat (PQ), an effective and widely used herbicide, has been proven to be safe when appropriately applied to eliminate weeds. However, PQ poisoning is an extremely frustrating clinical condition with a high mortality and with a lack of effective treatments in humans. PQ mainly accumulates in the lung, and the main molecular mechanism of PQ toxicity is based on redox cycling and intracellular oxidative stress generation. The aim of this study was to evaluate whether lysine acetylsalicylate (LAS) could protect the lung from the damage of PQ poisoning and to study the mechanisms of protection.

METHODSA model of PQ poisoning was established in 75 Sprague-Dawley rats by intragastric administration of 50 mg/kg PQ, followed by treatment with 200 mg/kg of LAS. The rats were randomly divided into sham, PQ, and PQ + LAS groups, with 25 in each group. We assessed and compared the malonaldehyde (MDA) content, superoxide dismutase activity (SOD), glutathion peroxidase (GSH-Px), and catalase (CAT) in serum and lung and the hydroxyproline (HYP) content, pathological changes, apoptosis and expression of Bcl-2/Bax protein in lung of rats on days 1, 3, 7, 14 and 21 after PQ poisoning and LAS treatment.

RESULTSCompared to the PQ group rats, early treatment with LAS reduced the MDA and HYP contents, and increased the SOD, GSH-Px, and CAT activities in the serum and lung on days 1, 3, 7, 14, and 21 after PQ poisoning (all P < 0.05). After early LAS treatment, the apoptotic rate and Bax expression of lung decreased, the Bcl-2 expression increased, and the Bcl-2/Bax ratio increased, compared to the PQ group rats. Furthermore, the pathological results of lungs revealed that after LAS treatment, early manifestations of PQ poisoning, such as hemorrhage, edema and inflammatory-cell infiltration, were improved to some degree, and collagen fibers in the pulmonary interstitium were also obviously reduced.

CONCLUSIONIn this rat model of PQ poisoning, LAS effectively ameliorated the lung injury induced by PQ, possibly through antioxidation, anti-fibrosis, anti-apoptosis, and anticoagulation.

Animals ; Antioxidants ; metabolism ; Aspirin ; analogs & derivatives ; standards ; therapeutic use ; Catalase ; metabolism ; Glutathione Peroxidase ; metabolism ; Lung ; drug effects ; metabolism ; Lung Injury ; chemically induced ; drug therapy ; metabolism ; Lysine ; analogs & derivatives ; standards ; therapeutic use ; Male ; Malondialdehyde ; metabolism ; Paraquat ; toxicity ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

2,4-dinitrophenol (DNP), an organic compound which frequently used in industry, is considered to have high toxicity. This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning. Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as control were enrolled. The patients received immediately comprehensive supportive treatments, including large-dose glucocorticoid and repeated hemoperfusion (HP). The ratio of CD4+/CD8+ T cells were significantly higher in patients upon admission compared to healthy controls (P < 0.01); however, counts of total lymphocytes, CD3+, CD3+CD4+, CD3+CD8+, B (CD19+), and natural killer (NK) cells (CD16+CD56+) were significantly reduced (all P < 0.001). The NK cell count was negatively correlated with initial plasma 2,4-DNP concentration (r = -0.750, P = 0.026). Thus, acute occupational 2,4-DNP poisoning was accompanied by immediate complex immune cell reactions, especially NK cells might play important role in severe 2,4-DNP poisoning.
2,4-Dinitrophenol ; poisoning ; toxicity ; Adult ; China ; Coloring Agents ; poisoning ; toxicity ; Female ; Humans ; Killer Cells, Natural ; drug effects ; Lymphocyte Subsets ; drug effects ; Male ; Middle Aged ; Occupational Diseases ; chemically induced ; T-Lymphocytes ; drug effects