BACKGROUNDNR2B containing N-methyl-D-aspartate (NMDA) receptor plays an important role in the facilitation and maintenance of neuropathic pain. The discrete distribution of NR2B subunit in the central nervous system (CNS) may support reduced side effects of agents that act selectively at this site. Therefore, we investigated the hypothesis that a humoral autoimmune response targeting the NR2B subunit of NMDA receptor relieves pain like behaviours produced by peripheral injury.

METHODSRats were immunized subcutaneously with NR2B-Keyhole Limpet Hemocyanin (NR2B-KLH) three times at two-week intervals. NR2B specific IgG titres in sera and cerebrospinal fluid were determined by indirect ELISA. Seven days after the third immunization, 2 of the 3 terminal branches of the sciatic nerve (tibial and common peroneal nerves) were tightly ligated. Behavioural testing was carried out on every other day after surgery, until 7 days after surgery. The lumbar spinal cord (L4-6) was removed on day 7 after ligation. The expression of NR2B protein in the lumbar spinal cord was determined using Western blotting.

RESULTSAfter the second vaccination, NR2B specific IgG in sera was detected to be > 0.5 microg/ml in six of nine rats. After the third vaccination, all the immunized rats had > 2.2 microg/ml. Titres of NR2B specific IgG in sera peaked 42 days post initial immunization and persisted for over 70 days. No NR2B specific IgG was detected in sera from PBS or KLH group. The behavioural thresholds in NR2B group were significantly higher than those in PBS and KLH groups on day 7 after ligation. NR2B specific IgG in CSF in NR2B group could not be detected on day 1 before spinal dissection; but could be detected on day 7 after surgery. The expression of NR2B protein in group NR2B was significantly lower than in PBS and KLH groups on day 7 after surgery.

CONCLUSIONThe NR2B peptide could be used as a vaccine against neuropathic pain, which could be associated with reduction of NR2B protein in the lumbar spinal cord.

Adjuvants, Immunologic ; Animals ; Blotting, Western ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Female ; Hemocyanins ; immunology ; Immunoglobulin G ; immunology ; Neuralgia ; immunology ; physiopathology ; prevention & control ; Pain Measurement ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; immunology ; metabolism ; Recombinant Fusion Proteins ; administration & dosage ; immunology ; Spinal Cord ; drug effects ; metabolism ; physiopathology ; Time Factors ; Vaccines ; administration & dosage ; immunology

OBJECTIVETo assess the feasibility and the significance of surgical resection of small intrahepatic lesions adjacent to the major vasculature.

METHODSThe results of treatment were retrospectively reviewed in 40 patients who received operation for intrahepatic lesions less than 3 cm in diameter between Jan. 2003 and Dec. 2005. The lesions were all adjacent to the major vasculature in the liver.

RESULTSIn the 40 patients, a total of 44 small intrahepatic lesions were successfully resected with minimal morbidity and blood loss (mean 163 ml). A second lesion was found in 4 patients (10%) during intraoperative exploration. Histologically the lesion was malignant in 29 cases (including 4 cases with two lesions) and benign in 11 cases, with correct preoperative diagnosis in 62.5% of all cases. For 26 patients with hepatocellular carcinoma, the 1-, 2-, and 3-year postoperative survival rates were 90.1%, 83.2% and 64.7%, respectively, while the patients with benign lesions were cured with the operation.

CONCLUSIONSSurgical resection of small intrahepatic lesions adjacent to the major vasculature is demanding but feasible and with satisfying effect. The significance of surgical management of these small lesions is not only excising the lesions but also making definite diagnosis and finding new lesions in some patients.

Adult ; Aged ; Blood Vessels ; pathology ; Feasibility Studies ; Female ; Follow-Up Studies ; Hepatectomy ; Humans ; Liver ; blood supply ; pathology ; surgery ; Liver Neoplasms ; pathology ; surgery ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo investigate the protective effects of morphine postconditioning on myocardial ischemia-reperfusion (I/R)injury and the potential mechanisms in rabbits.

METHODSThirty-two New Zealand male white rabbits were randomly assigned into 4 groups: Group 1 (Sham), Group 2 (I/R), Group 3 (ischemic postconditioning), Group 4 (ischemia and morphine postconditioning). Group 1 was perfused for 160 min; Group 2 underwent 40 min ischemia and 120 min reperfusion; Group 3 underwent three cycles of 30 s reperfusion and 30 s left anterior descending coronary artery re-occlusion immediately after 40 min ischemia and before 120 min reperfusion; Group 4 was given morphine 1.0 mg/kg immediately after 40 min ischemia in 1 min and before 120 min reperfusion. Blood samples were taken from arterial line at 20 min before occlusion, 20 min after occlusion, 40 min after occlusion, 1 h after reperfusion and 2 h after reperfusion for determination of the plasma levels of cardiac troponin I (cTnI). At the end of the reperfusion, infarct size (IS) and area at risk were defined by Evans and TTC staining. Plasma SOD activity and MDA were determined at the end of reperfusion.

RESULTThe levels of cTnI were significantly lower during reperfusion in the two postconditioning groups than those in I/R group. The plasma MDA content was significantly lower and SOD activity was significantly higher in the two postconditioning groups than those in I/R group, but there was no difference between two postconditioning groups. Morphine significantly reduced infarct size of the left ventricular area at risk as compared with I/R group (P<0.05).

CONCLUSIONMorphine postconditioning is as effective as ischemic postconditioning in the protection of myocardium against I/R injury in rabbits. Decrease in oxygen free radicals and increased antioxidant activity might be involved in its mechanism.

Animals ; Coronary Artery Disease ; drug therapy ; Ischemic Preconditioning, Myocardial ; Male ; Malondialdehyde ; blood ; Morphine ; therapeutic use ; Rabbits ; Random Allocation ; Reperfusion Injury ; prevention & control ; Superoxide Dismutase ; blood

Rapid discrimination of compounds with similar structures and properties is a hot topic in analytical chemistry. In this study, a cyclic chemiluminescence (CCL) system was designed by using nano-magnesium oxide as catalyst. The effects of reactant concentration, reaction temperature and detection wavelength on CCL analysis were studied. It was found that the CCL signal satisfied the first order exponential decay law. Each reaction had an exponential decay equation (EDE) describing its signal change law. The initial variable A was proportional to the reactant concentration. The decay-coefficient k was a characteristic constant that was independent of the reactant concentration. For iso-butanol, it's A-values versus concentration was linear in 0. 89-14. 24 mg / L, the average of k-value in this range was 32. 0 with a RSD of 2. 2% . Thus, qualitative and quantitative analysis could be conducted according to the A and k values. The system was used to analyze eight kinds of alcohol compounds, and it was found that there were significant differences in the k-values for different alcohols. For example, the k-values for n-butanol, iso-butanol and sec-butanol were 27. 2 ±0. 2, 32. 0±0. 8 and 19. 5±0. 1, respectively.

Objective To etiologically diagnose and analyze a patient with suspected cases of brucellosis,and to provide a experimental basis for the confirmation of the first case of human brucellosis in Guizhou province.Methods Conventional and molecular techniques [genus specific Brucella surface protein 31 PCR (BCSP31-PCR)and Brucella suis species-specific PCR (AMOS-PCR)] were used to identify suspicious bacteria strains isolated from the suspected patient of brucellosis.Results The results showed that the Brucella suspicious colonies were identified as Brucella melitensis biotype 3 using conventional tests and were further identified as Brucella spp.by genus specific Brucella surface protein 31 PCR (BCSP31-PCR) and classified as Brucella melitensis with Brucella abortus,Brucella melitensis,Brucella ovis,Brucella suis species-specific PCR(AMOS-PCR).Conclusions Laboratory diagnostic results show that the bacteria strain isolated from the suspected patient of brucellosis is Brucella melitensis biotype 3.It is the first case of human brucellosis in Guizhou province.

This study aimed to evaluate the vasorelaxant effect and mechanisms of compound reserpine and triamterene tablets (CRTTs) and its component triamterene on isolated rat thoracic aorta rings. Isolated rat thoracic aorta rings pre-contracted by high potassium or norepinephrine (NE) were used to evaluate the vasodilatory effect of CRTTs and its component triamterene. The mechanisms concerning endothelium, potassium channels and calcium channels were studied through the interventions of several tool drugs. Animal welfare and experimental procedures followed the requirements of the Laboratory Animal Management and Animal Welfare Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College. The results showed that both CRTTs and triamterene had potent relaxant effect on KCl and NE pre-contracted vessels. Triamterene showed partial endothelium dependency, and N-nitro-L-argininemethyl ester hydrochloride (L-NAME) could influence the relaxant effect, which may be related to the opening of calcium-activated potassium channels. Meanwhile, CRTTs could inhibit intracellular Ca²⁺ release and extracellular Ca²⁺ influx. Overall, CRTTs and its component triamterene have vasorelaxant effects on vessels in vitro, and the vasorelaxant effect of CRTTs may be related to intracellular Ca²⁺ release and extracellular Ca²⁺ influx, while this effect of triamterene may depend on vascular endothelial function and may also be related to the opening of calcium-activated potassium channels.

This study analyzed the clinical features of 5 children with hereditary spherocytosis (HS) and the characteristics of ANK1 and SPTB gene mutations. All 5 children were confirmed with HS by peripheral blood genetic detection. Anemia, jaundice and splenomegaly were observed in all 5 children. Three children had an increase in erythrocyte osmotic fragility. All 5 children had negative results of the Coombs test, glucose 6 phosphate dehydrogenase test, sucrose hemolysis test, acidified-serum hemolysis test and thalassemia gene test. Peripheral blood smear showed an increase in spherocyte count in one child. High-throughput sequencing revealed ANK1 gene mutations in patients 1 to 3, namely c.3398(exon29)delA, c.4306C>T and c.957(exon9)_c.961(exon9)delAATCT, among which c.3398(exon29)delA had not been reported before. Patient 4 had c.318delGExon3 mutation in the SPTB gene. Patient 5 had mutations in the SPTB and SLC4A1 genes, among which c.3484delC in the SPTB gene was a spontaneous mutation; the mutation site of the SLCA4A1 gene was inherited from the father and was a non-pathogenic gene. This study suggests that anemia, jaundice and splenomegaly are major clinical manifestations of HS children. Most children with HS do not have the typical spherocytic changes. Genetic detection may help with the accurate diagnosis of HS.
Ankyrins ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Spectrin ; genetics ; Spherocytosis, Hereditary ; genetics

OBJECTIVETo evaluate the feasibility and the effect of surgical resection of hepatic tumor originated from segment IXb.

METHODSThe cases with hepatic tumors in segment IXb who had been operated on between March 2003 and January 2007 were retrospectively reviewed.

RESULTSA total of 15 tumors in segment IXb, including 13 primary liver cancers and 2 benign tumors with a mean diameter of (4.3 +/- 1.6) cm, were successfully resected by anterior transhepatic approach under sequential occlusions of portal tride and total hepatic vascularity or portal tride clamping only. There was no operative mortality,with a mean operative time of (190.3 +/- 37.6) min and a mean operative blood lose of (376.7 +/- 252.7) ml. All the patients had uneventful postoperative course except one who suffered from ascites and edema of the low body, which was successfully managed medically. The mean postoperative hospital stay was (13.3 +/- 6.0) d. During the follow-up of 1-47 months, two patients with benign tumor enjoyed a normal life. Among the 13 patients with primary live cancers, 1 patient died of recurrence, 2 patients remained alive with intrahepatic recurrence and 10 patients survived without any sign of relapse, with a median tumor-free survival time of 23.5 months.

CONCLUSIONSSurgical resection of hepatic tumor in segment IXb, despite their sophisticated anatomic position, is feasible in technique with high safety. The local resection can provide the patients with potential to cure.

Adult ; Aged ; Feasibility Studies ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Liver Neoplasms ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo investigate the differences in semen quality at different times of reanalysis and the correlation of sperm DNA fragmentation index (DFI) with sperm motility alteration using semen samples completely liquefied and normal in initial examination.

METHODSWe analyzed 127 semen samples up to the inclusion criteria with the computer-assisted semen analysis (CASA) system at 15, 30 and 60 min after semen collection, and obtained sperm morphology parameters and DFI by Shorr staining and acridine orange test (AOT) , respectively.

RESULTSSperm concentration, and the percentages of grades a and b sperm showed no statistically significant differences at the three time points (P > 0.05). The percentages of grades a + b and a + b + c sperm were significantly higher at 15 min than at 30 and 60 min after semen collection (P < 0.05), but with no significant difference between the latter two time points (P > 0.05). The incidence of alternation from normal to abnormal in at least one index of sperm motility at different times was 25.2%, but there were no significant differences in sperm DFI and morphology between the normal and abnormal groups (P > 0.05). Among the altered parameters of sperm motility from 15 to 60 min, the percentages of grades a, a + b and a + b + c sperm were all positively correlated with sperm DFI (P < 0.05).

CONCLUSIONSemen samples completely liquefied within 15 min after collection and normal in initial examination, when reanalyzed at 30 and 60 min, showed significant decreases in the percentages of grades a + b and a + b + c sperm, but not in the percentages of grades a and b sperm, and the parameters of sperm motility might be abnormal. Thus, at least 2 sperm analyses are required for a comprehensive evaluation of fertility. Significant difference between the results of the two analyses, and particularly a markedly reduced percentage of rapidly progressive sperm, might indicate sperm DNA damage, and thus the necessity of sperm DNA damage detection.

Adult ; DNA Damage ; DNA Fragmentation ; Diagnosis, Computer-Assisted ; Fertility ; genetics ; Humans ; Male ; Middle Aged ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Young Adult

OBJECTIVETo evaluate the influence of extended hepatic pedicle occlusion (HPO) on hepatic ischemic/reperfusion (I/R) injury and intraoperative blood loss in major hepatectomy for primary liver cancer (PLC).

METHODSBetween June 2001 and December 2005, a total number of 843 patients with PLC had been operated on. Those whose hepatic pedicle were occluded continuously for or longer than 30 min during hepatectomy were retrospectively reviewed (continuous HPO group) and compared to the patients whose hepatic pedicle were occluded for the same length of time but intermittently (intermittent HPO group). The amount of intraoperative blood loss, the percentage of the patients who needed blood transfusion and postoperative liver biochemical tests were compared between the two groups.

RESULTSThere were 35 cases in continuous HPO group and 38 cases in intermittent HPO group with occlusion time between 30 min and 45 min. The two groups were matched for underlying liver disease ,preoperative liver function, tumor size and location, major intrahepatic vessel involvements and the types and extensions of the hepatectomies. The mean intraoperative blood loss in continuous HPO group was significantly less than that in intermittent HPO group (660 ml vs. 1054 ml, P < 0.05); accordingly, the percentage of patients who need blood transfusion in continuous HPO group was significantly lower than that in intermittent HPO group (48.6% vs. 78.9%, P < 0.01). Patients in both of the groups were recovered smoothly after operation, with no occurrence of liver failure.

CONCLUSIONSThe hepatic pedicle can be continuously occluded for 3045 min in cirrhotic patients with well compensated liver function, and when compared to routine intermittent HPO, continuous HPO significantly decreases the intraoperative blood loss and reduces the need for transfusion. Meanwhile it does not increase the hepatic I/R injury.

Adult ; Aged ; Blood Loss, Surgical ; prevention & control ; Female ; Hepatectomy ; methods ; Humans ; Liver ; blood supply ; physiopathology ; Liver Neoplasms ; physiopathology ; surgery ; Male ; Middle Aged ; Retrospective Studies