AIM:To construct adenovirus vector expressing mice B7-H1 gene, transfect dendritic cells ( DCs ) , and to study the therapeutic effect of modified DC on thyroid-associated ophthalmopathy ( TAO) in mice.
METHODS: We designed and constructed B7-H1 gene adenovirus expression vector, and transfected DCs from mouse bone marrow, tested the phenotype and function of modified DCs, identificated its negative regulation to immune responses. The modified DCs were infected the sicked mice. And then the immunotherapeutic effect of modified DCs to TAO were tested.
RESULTS: B7 - H1 gene adenovirus vector was constructed and transfected DCs from bone marrow. The titer of the recombinant adenovirus was 1. 8í109 PFU/mL. B7-H1 gene modified DCs characteristics of regulatory DCs, could inhibit positive immune responses. The inhibition proceeding of TAO into mice infected modified DCs, was obviously prior to the control mice. The gene modified DCs, maybe become the new immunotherapy biological agent to thy TAO.
CONCLUSION: We constructed the expression of mouse B7 - H1 gene adenovirus expressed vector successfully, transfected DCs, by vector have properties of regulatory DCs, inhibiting positive immune response and the occurrence and development of thyroid eye disease. Gene modified DCs, reveal potent to the treatment of thyroid eye disease.

Objective To provide a practicable method for the complete display and localization of the posterolateral structures (PLS) of the normal knee through MRI study. Methods 30 tibial bone specimens were observed to establish the bony landmark for localizing the knee. In 50 cadaver knees, the angles between lateral tibial plateau and the long axis of the individual structure of PLS were measured. Then the scan methods of the oblique MR images were determined based on above results. The routine and oblique scans of T 1WI were performed in 40 normal knees. The display effect and appearance of the PLS were observed on MRI. Results The lateral tibial plateau was a stable bony landmark for measuring and localizing of the knee. In the 40 normal knees, The fibular collateral ligament could be intactly displayed on 70? posterior coronal oblique images in 34 cases (85%). The popliteus could be better seen on either 45? medial sagittal oblique in 34 cases (85%) or 60? posterior coronal oblique planes in 36 cases (90%). The popliteofibular ligament could be intactly appreciated on both 60? posterior coronal oblique in 32 cases (80%) and 70? lateral sagittal oblique images in 34 cases (85%). Although the arcuate ligament and the fabellofibular ligament could occasionally be seen on routine and oblique images, but the display rate was lower. Conclusion The oblique MR imaging can intactly display the main structures of PLS, and can be useful in diagnosing the injuries in those structures.

A complete aniline dioxygenase gene cluster cloned from an Acinetobacter sp. strain, which could utilize aniline as the sole carbon, nitrogen and energy, was sequenced. Sequence analysis showed that the gene cluster had six intact ORFs, and the whole sequence had high similarity with that of Acinetobacter sp. YAA at amino acid level. A recombinant strain was formed with the gene cluster ligated to vector pLAFR6 and transferred to E.coli. After optimizing the fermentation conditions of this strain for producing catechol, LB was confirmed as the final medium, pH7.0, aniline concentration 0.5mg/ml, E.coli DH5?as the host, incubation temperature 37℃, amount of inoculum 3%. Under above conditions, the yield of catechol could get to 0.546mg/ml, and the converting rate of substrate at molecule level could get to 92.4%.

AIMTo investigate the effect of ciliary neurotrophic factor (CNTF) on the nuclear translocation of protein kinase C (PKC) following NMDA administration in the primary cultured hippocampal neurons.

METHODS(1) PKCGAMMA or PKCepsilon- immunocytochemistry staining method was used after treating neurons with NMDA or CNTF. (2) The gray of the nucleus of the PKC-positive neurons were measured under the image pattern analysis system.

RESULTS(1) After NMDA administration of different concentration and time, Nucleus appear PKCgamma and PKCepsilon activities, especially the 100 micromol/L NMDA 30 min group. (2) The gray of nucleus in CNTF + 500 micromol/L NMDA group is similar to control group.

CONCLUSIONNMDA can induce nuclear translocation of PKC in the primary cultured hippocampal neurons, and CNTF can inhibit the translocation. It suggests that the inhibition of PKC translocation induced by NMDA is one of the important reasons for the neuro-protective effects of CNTF.

Animals ; Cells, Cultured ; Ciliary Neurotrophic Factor ; pharmacology ; Hippocampus ; cytology ; N-Methylaspartate ; pharmacology ; Neurons ; drug effects ; metabolism ; Protein Kinase C ; metabolism ; Protein Transport ; Rats ; Rats, Sprague-Dawley

BACKGROUNDTraditional Chinese medicine has been used widely for many years in China to treat acute pancreatitis. We have investigated the effects of Dachengqi decoction on intestinal mucosal permeability and outcome in patients with severe acute pancreatitis (SAP).

METHODSForty patients with sustained SAP that required admission to the surgical intensive care unit were enrolled prospectively in the study. All of these patients were divided randomly into the Dachengqi decoction group (n = 20) and control group (n = 20) on admission. Intestinal permeability was assessed by measuring absorption of two metabolically inert markers, lactulose (L) and mannitol (M), which were administered orally. Serum concentrations of endotoxin (lipopolysaccharide, LPS) and the ratio of lactulose to mannitol in urine (L/M) were detected in all patients.

RESULTSCompared with those in the control group, urinary L/M ratio decreased significantly in the Dachengqi decoction group on the 7th day after admission (P = 0.001). Also, serum concentrations of LPS were reduced on the 5th and 7th day after admission (P = 0.006, P = 0.008, respectively). Incidence of multiple organ dysfunction syndrome (MODS) and pancreatic infection was significantly lower in the Dachengqi decoction group compared with those in the control group (P = 0.038,P = 0.025, respectively).

CONCLUSIONDachengqi decoction may promote the recovery of intestinal mucosal permeability and decrease the incidence of MODS and pancreatic infection in patients with SAP.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-6 ; metabolism ; Intestinal Mucosa ; drug effects ; metabolism ; pathology ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Pancreatitis ; drug therapy ; metabolism ; pathology ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

OBJECTIVETo analyze the risk factors of postoperative complications in gastric carcinoma patients complicated with liver cirrhosis.

METHODSA retrospective research of 41 gastric cancer patients with liver cirrhosis who underwent surgical therapy was performed. The multivariate Logistic regression analysis was carried out to determine risk factors of postoperative complications.

RESULTSThere were no postoperative death in this cohort. Twenty-seven(65.9%) patients developed postoperative complications. As determined by the Logistic regression analysis, Child-Pugh score(OR=27.96, 95% CI:1.16-672.23), albumin level(OR=17.98, 95% CI:1.28-253.36) and intraoperative blood loss(OR=10.60, 95% CI:1.21-92.82) were independent risk factors of postoperative complications.

CONCLUSIONAs for gastric cancer patients with liver cirrhosis, positive perioperative management, adjustment of Child-Pugh classifications score, following damage control principle during operation and reducing the intraoperative bleeding may decrease the morbidity of postoperative complications.

Factor Analysis, Statistical ; Humans ; Liver Cirrhosis ; Morbidity ; Postoperative Complications ; Retrospective Studies ; Risk Factors ; Stomach Neoplasms

OBJECTIVETo evaluate the efficacy and optimal re-implantation time of two-stage revision for management of periprosthetic infection following hip arthroplasty.

METHODSWe retrospectively analyzed the clinical data of 15 patients (15 hip joints) undergoing two-stage ipsilateral total hip arthroplasty (THA) revision from January, 2006 to January, 2010. In the first stage, after surgical debridement and thorough removal of all the implants, a self-made Vancomycin-loaded cement spacer was implanted. The second stage operation was performed 3-6 months later for debridement and removal of the antibiotic-loaded spacer, followed by re-implantation of Vancomycin-loaded bone cement prosthesis in 9 cases and cementless prosthesis in 6 cases. The patients were followed up for 9-46 months (mean 25 months) after the operation.

RESULTSNo reinfection or prosthesis loosening/displacement was found in these cases after the operation. The Harris score increased from 40.3 before the operation to 54.0 after the first-stage operation, and to 88.2 at the last follow-up.

CONCLUSIONTwo-stage revision is effective for treatment of periprosthetic infection following hip arthroplasty, and 3-6 months can be the optimal interval between the two the first-stage and second-stage operation for re-implantation.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Female ; Hip Prosthesis ; Humans ; Male ; Middle Aged ; Prosthesis-Related Infections ; surgery ; Reoperation ; Retrospective Studies ; Treatment Outcome

Injury of vascular endothelial barrier function is implicated in several pathophysiological processes. The integrity of vascular endothelium is regulated by cytoskeleton and cell-cell junctions. Small guanosine triphosphatases of the Rho family (Rho GTPases) are known to play a central role in vascular endothelial barrier function. It has been reported that RhoA, Rac1, Cdc42 and RhoB are involved and they exert both positive and negative effect on endothelial barrier integrity, depending on their subcellular location. When inflammatory factors such as thrombin attack the vascular endothelial cells, GEF of RhoA will be widely distributed throughout the cells. Thus, activated RhoA causes aggregation of F-actin fibers in a short time and disrupts the vascular endothelial barrier, a process named acute cell contraction. However, RhoA may also induce the production and maturation of intercellular junctions in new cells. Rac1 and Cdc42 help to maintain the integrity of vascular endothelial barrier at the resting state. They cause the phosphorylation of LIM kinase and inhabitation of cofilin, resulting in less remodeling of cytoskeletal in the vascular endothelial cells. On the other hand, Cdc42 can translocate to the cortex rapidly after a stimulation, where Cdc42 will activate the myosin Ⅱ and promote the reorganization of adjective junction to facilitate the recovery of vascular endothelial barrier. In this review, we overviewed how Rho GTPases regulate the vascular endothelial barrier integrity.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; complications ; pathology ; physiopathology ; Liver ; pathology ; physiopathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo establish a novel model of lumbar disc degeneration on the early stage in the rhesus monkey using percutaneous needle puncture guided by CT.

METHODS(1) Thirteen rhesus monkeys aged from 4 to 7 years, female 7 and male 6 were selected for establishing a model of the early stage of lumbar disc degeneration. (2)13 monkeys, 91 discs were divided into 3 groups: 64 discs from L1/2 to L5/6 were percutaneous punctured with a needle 20G as experimental group and 1 disc with a needle 15G as puncture control group and 26 discs were not be punctured from L6,7 to L7-S1 as control group. (3) Lumbar disc localization for needle puncture was guided by CT. All discs were examined by MRI, the HE, Masson's trichrome, Safranine-O and immunohistochemical staining of type II collagen before disc puncture and after puncture at 4, 8 and 12 weeks.

RESULTSMRI: (1) Experimental group: Pfirmann's Grade I was shown at postoperation 4, 8 and 12 weeks; (2) Puncture control group: Grade III was shown at postoperation 4 weeks and Grade IV at 8 weeks; (3) CONTROL GROUP: Grade I was shown at postoperation 4, 8 and 12 weeks. Histological examination: (1) In experimental group, there was no any change at postoperation 4 weeks, and the cell population of the nucleus was decreased at 8 weeks and more decreased at 12 weeks in HE. (2) There was no any change at postoperation 4 weeks, the clefts among the lamellae of the annulus fibrosus (AF) were shown at 8 weeks and more wider of the clefts of AF at 12 weeks in Masson's trichrome. (3) No any change was shown at postoperation 4 weeks, proteoglycan were progressively decreased at 8 and 12 weeks in Safranine-O. (4) No statistically significant difference in positive rate was observed at 4 and 8 weeks compared with control group in immunohistochemical staining of type II collagen. There was statistical difference at 12 weeks compared with control group (P<0.05). In puncture control group postoperation 8 weeks, the morphology of cell of nucleus pulposus was not clear in HE. The wider clefts of lamellae of the AF were shown in Masson's trichrome. The proteoglycan was obviously decreased in Safranine-O. Immunohistochemical staining collagen II synthesized was decreased. In normal control group, no any change was shown at 4, 8 and 12 weeks.

CONCLUSIONSThe degeneration of lumbar intervertebral disc on the early stage could be induced by the percutaneous needle puncture (20G) to the annulus fibrosus. The assessment of disc degeneration on early stage is not shown on MRI and only confirmed by histological examination.

Animals ; Disease Models, Animal ; Female ; Intervertebral Disc ; metabolism ; pathology ; surgery ; Intervertebral Disc Displacement ; etiology ; metabolism ; pathology ; Lumbar Vertebrae ; surgery ; Macaca mulatta ; Male ; Minimally Invasive Surgical Procedures ; Random Allocation