Objective Delayed xenograft rejection (DXR) is a major barrier to the long-term xenograft survial.This study evaluated the interaction between human peripheral blood mononuelear cells (PBMC) and porcine endothelial cells (PEC),and the effects of new generation of rabbit antihuman leukocyte polyclonal antibody (newRALG) inhibiting xenogeneic cell-mediated immune responses.Methods newRALG was obtained from rabbits after immunization with activated lymphocytes and monoeytes.PEC were isolated from aorta,and human PBMC were isolated from peripheral blood.Co-cultures of PKH-26 labeled PEC with PBMC were established,newRALG,thymoglobulin,isotype Ig and scavenger receptor (SR) ligand poly G were added into the co-cultures.Cells were collected,then FACS analysis was carried out to detect the up-take of PEC membrane by monocytes and the expression of costimulatory molecules.Lymphocyte proliferative responses to PEC with or without antibody were evaluated by a xenogeneie mixed lymphocyte-endothelial cell reaction (xMLER).Results FACS analysis revealed that monocytes from PBMC-PEC co-cultures became positive for PKH-26 following their interaction with PKH-26 labeled PEC,indicating that they engulfed PEC membranes during activation.PKH-26 positive monocytes up-regulated the CD40 and CD80 expression.Furthermore,SR blockade with poly-G prevented PEC membrane up-take by monocytes,newRALG greatly reduced SR-mediated PEC membrane up-take.The effects of thymoglobulin in inhibiting PEC membrane uptake were limited.xMLER demonstrated strong lymphocyte proliferation in response to PEC,and lymphocyte proliferation was dramatically inhibited by newRALG but not isotype Ig at a dosmdependent manner.Conclusions Monocytes play an important role in xenogeneic immune responses.SR ligand poly G inhibits PEC membrane up-take.newRALG inhibits PEC membrane up-take by monocytes,suggesting that newRALG blocks SR.Additionally,newRALG inhibits lymphocyte proliferation in response to PEC.These results suggest that this new polyclonal preparation may thus impair the initiation of xeno-specific immune responses and prevent xenograft rejection.

Anti-Arrhythmia Agents ; therapeutic use ; Cardiomyopathies ; drug therapy ; etiology ; Child, Preschool ; Female ; Humans ; Moricizine ; therapeutic use ; Tachycardia ; complications ; drug therapy ; Treatment Outcome

Objective To study the expression of Adiponectin (AD) and its receptors Adiponectin receptor 1 (Adipo R1) and Adipo R2 in the synovial fluids and the synovium of rheumatoid arthritis (RA). Methods ELISA was used to determine the levels of AD in 23 RA and 23 osteoarthritis (OA) patients. Real-time PCR and Western blot techniques were employed to study the expression of AD, AdipoR1 and AdipoR2 in the synovium of 10 RA and OA patients. Results It was observed that approximately twice more adiponeetin in the synovial fluids of patients with RA than with OA. Adiponectin and AdipoR1, but not AdipoR2 mRNA, were significantly expressed in synovium of RA patients in comparison with OA. Adiponectin and AdipoR1 protein were wuch more expressed in synovium from RA than those from OA. Conclusion High expression of Adiponectin and AdipoRl is likely to contribute to the formation of inflammatory joints in RA.

From air bacteria capable of decomposing creatinine, three single independent strains K9510、K9511 and K9512 have been isolated. The highest creatinine amidohydrolase (EC 3. 5. 2. 10; creatininase) producing strain K9510 was screened out. The strain K9510 was identified as Pseudomonas sp. The results of culture condition for creatininase formation by strain K9510 were obtained as follows: creatinine and creatine were found to be the effective inducers for enzyme formation; the solution of mixed metallic salts could stimulate cell growth and enzyme formation. The suitable medium for creatininase formation was consisted of 0. 9% creatinine、0. 15% yeast extract、 0. 09% malt extract、0. 05% NH4Cl and some amount of the solution of mixed metallic salts at pH5 5. When the bacterium was grown in 250mL conic flask containing 50mL of the medium mentioned above on the rotary shaker(250r/min) at 35℃ for 33 h, about 50 u creatininase was obtained.

Objective To evaluate the effect of ulinastatin on γ-aminobutyric acid (GABA) signal pathway in mice with ventilator-induced lung injury (VILI).Methods Thirty-six male Wister mice were randomly divided into 3 groups using a random number table:control group (group C), ventilator-induced lung injury group (group VILI),and ventilator-induced lung injury+ ulinastatin group (group UTI),n =12 in each group.VILI was induced by 4 h mechanical ventilation with tidal volume 40 ml/kg in groups VILI and UTI.Ulinastatin 1×10 5 U/kg was injected intraperitoneally 1 h before ventilation in group UTI,while the equal volume of normal saline was given in groups C and VILI.The mice were then sacrificed,the left lung was lavaged,and broncho-alveolar lavage fluid (BALF)was collected for determination of concentrations of protein,tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and intercellular adhesion molecule-1 (ICAM-1).The lung tissues were re-moved for determination of the wet to dry lung weight (W/D)ratio,the mRNA expression level of IL-1β,TNF-αand ICAM-1.The pathological changes of the lungs were determined under light micro-scope and the lung injury scores were also determined.Immunohistochemistry and Western blot were used to detected the protein expression level of GAD and GABAA R.Results The W/D ratio (6.7 ± 2.4 vs.8.5±2.3)and lung scores [(6.9±2.3)scores vs.(1 1.8±2.7)scores]were significantly de-creased in group UTI than those in group VILI.The concentrations of IL-1β[(56±1 1)ng/L vs.(77 ±1 5)ng/L],TNF-α[(105±29)ng/L vs.(1 58±37)ng/L]and ICAM-1 [(205±46)ng/L vs.(293 ±61)ng/L]in BALF in group UTI were significantly decreased than those in group VILI.The mRNA ex-pression levels of IL-1β(1.81±0.26 vs.2.58±0.34),TNF-α(1.61±0.15 vs.2.94±0.27)and ICAM-1 (1.74±0.27 vs.2.79±0.31)were significantly decreased in group UTI than those in group VILI.The protein expression levels of GAD (0.44±0.08 vs.0.18 ±0.04)and GABAA R (0.30 ±0.09 vs.0.15 ± 0.04)were significantly increased in group UTI than those in group VILI.Conclusion Ulinastatin can at-tenuate VILI probably through activating GABA signaling pathway.

Background The pathogenesis and development of cataract is associated with oxidative stress-induced apoptosis of human lens epithelial cells(LECs).BH3-only protein is a factor that can initiate apoptosis,and thus the apoptotic process is probably related to the activation of the c-Jun N-terminal kinase(JNK).However,the relationship between oxidative stress-induced apoptosis of human LECs and the JNK pathway remains to be illuminated.Objective This study was to investigate the effects of the JNK/c-Jun pathway and its target gene,Bim (Bcl-2 interacting mediator of cell death)and PU M A(p53 up-regulated modulator of apoptosis),on oxidative stressinduced apoptosis of human LECs.Methods The human LECs cell line(HLEC-B3)was cultured and passaged in DMEM with 10％ fetal bovine serum in vitro.Confluent cells were incubated in 24 well plates and divided into 4 groups.Hydrogen peroxide(H2O2)(50 μmol/L)was used to treat the cells for 4,8 or 12 hours,and cells without H2O2 treatment served as the control group.Apoptosis was detected using Hoechst 33258 staining and quantified by counting the number of cells with pyknotic nuclei.In addition,confluent cells were seeded in 6 well plates,and Western blot and RT-PCR were used to detect the expression of the caspase-3,c-Jun,Bim and PUMA proteins and their mRNA in HLEC-B3,respectively.The JNK/c-Jun pathway inhibitors,CEP11004 or SP600125,were added into cultured media with H2O2,and cells treated with DMSO or H2O2 only served as negative and positive control groups.The expression of the p-JNK,JNK,p-c-Jun,c-Jun,Bim,PUMA proteins was detected by Western blot and apoptosis was assayed using Hoechst 33258 staining.200 pmoL/L of Bim or PUMA small interference RNA(siBim or siPUMA)fragments were transfected into the cells for 24 hours,respectively,and H2O2 was then used to treat the cells for 8 hours.The expression of the Bim and PUMA protein and their mRNA in the cells was detected by Western blot and RT-PCR,respectively.Results After H2O2 treatment in HLEC-B3 cells for 4,8,or 12 hours,the rates of apoptosis were 4.30％±1.15％,27.08％±0.74％ and 46.59％±0.91％,showing a significant difference among them (F=1909.433,P=0.000),and those of the 4,8,12 hour groups were significantly increased in comparison to the control group(P =0.049,0.000,0.000).Compared to untreated cells,the levels of expression of the JNK,Bim,PUMA proteins and their mRNA in HLEC-B3 cells were significantly elevated.After the addition of CEP11004 or SP600125,the expression of these protein and mRNA in HLEC-B3 cells in the presence of H2O2 was significantly weaker than that in the DMSO control group(P =0.000,0.000).After the tranfection of siBim or siPUMA,the apoptosis rates of the H2O2 treated groups were significantly higher than those in the Bim-/-or PIMA-/-group (P＜0.05).Conclusions H2O2 can activate the JNK/c-Jun pathway and up-regulate the expression of its target genes Bim and PUMA in human LECs in a time-dependent manner.Inhibiting the JNK/c-Jun pathway and interfering with the expression of Bim and PUMA can protect human LECs against oxidative stress-induced apoptosis.

Background Some aboard studies showed that polyatomic pathway play an important role in the microsvasculopathy of type 2 diabetes,and sorbitol dehydrogenase (SDH) gene is involved in the pathogenesis of diabetic retinopathy(DR).There is no relevant research at home up to now.Objective This study was to investigate the correlation of sorbitol dehydrogenase (SDH) G-888C gene polymorphism with diabetic retinopathy (DR) in type 2 diabetes mellitus.Methods The SDH genotypes were determined by polymerase chain reactionreaction fragment length polymorphism (PCR-RFLP) in 187 patients with diabetes and 123 normal contrels.Patients were divided into two groups depending on the presence or absence of DR(DR:n=118;NDR:n=69).The frequencies of SDH genotype and allele were assayed and compared among these groups.This study was approved by Ethic Committee of Guangdong General Hospital.Written informed consent was obtained from each individual prior to any relevant medical procedure.Results The disease course in the DR group was significantly longer than that in NDR group(t=2.070,P=0.042),and other clinical features in both groups were non-significant (all P>0.05).The genotype frequencies in the DR group,NDR group and normal control group were 24.6%,8.7% and 8.1%,respectively,and frequencies of the G allele were 42.4%,25.4% and 29.7%,showing statistically significant differences among these three groups.The GG genotype and G allele frequencies were significantly higher in the DR group than those in the NDR group and normal control group (GG:P=0.007,P=0.001;G:P=0.001,P=0.004).There were no significant differences in the frequencies of the GG genotype and G allele between the NDB group and normal control group ( P>0. 05) as well as the proliferative DR group and non-proliferative DR group (P>0.05).Conclusion SDH G-888C gene polymorphism is associated with the development of diabetic retinopathy in southern Chinese.

BACKGROUNDWe presented the technique and outcomes of laparoscopic radical cystectomy performed in 28 patients with bladder cancer to evaluate its clinical efficacy and feasibility.

METHODSAmong the 28 patients, aged from 58 to 73 years (mean 68.5 years), 26 were transitional cell carcinoma grads II - III and 2 squamous carcinoma. Laparoscopic radical cystectomy plus bladder reconstruction was performed in all cases, among them 15 with ileum, 10 with rectum pouch, and 3 with ureterostomy. The operating time, the blood loss, the intestine function and the complications were observed.

RESULTSAll procedures were successful. The operating times were 7 - 10 hours, the blood loss was 400 - 1500 ml (mean 850 ml), the intestine function recovered at 72 hours after operation, and all ureteral catheters were removed at 2 weeks after surgery. The results of intravenous urography were normal at 3-month, 1-year, and 2-year follow-up after surgery.

CONCLUSIONSLaparoscopic radical cystectomy for invasive bladder cancer is safe and efficient, with good operating field, reliable hemostasis, mini-invasion, less celiac complications, and rapid recovery. This surgery is worth being extended.

Aged ; Carcinoma, Transitional Cell ; surgery ; Cystectomy ; methods ; Humans ; Ileum ; surgery ; Laparoscopy ; methods ; Male ; Middle Aged ; Urinary Bladder Neoplasms ; surgery ; Urinary Diversion ; methods ; Urinary Reservoirs, Continent

Adult ; Aged ; Chyle ; Female ; Humans ; Kidney ; surgery ; Laparoscopy ; methods ; Lymphatic Diseases ; surgery ; Lymphatic System ; surgery ; Male ; Middle Aged ; Retroperitoneal Space ; Urine ; Urologic Surgical Procedures ; methods

A strain KX6,producing thermotolerant endoglucanase,was isolated from compost. The morpholo-gical identification and 16S rRNA sequence analysis showed it belongs to Streptomyces xylophagus. The production and characterization of endoglucanase from Streptomyces xylophagus KX6 was studied. Maximum endoglucanase yield of 0.538 IU/ml was achieved with medium pH8.0,containing CMC2Na 1.0% as carbon resource,soybean meal 1% as nitrogen resource,2% inoculating volume,30% 250 ml triangle flask bulk for medium volume at 40℃ 200r/min shaker for 48h. The endoglucanase exhibited optimum catalytic activity at pH7.0 and 50℃. The enzyme was stable at 50℃,and able to retain 60% of the full activity,when it was incubated at 60℃ for 1h.The enzyme was stable at pH6.0~7.0. All these findings suggest that the enzyme is a thermotolerant neutral endoglucanase.