Objective To investigate the effects of ketamine on the expression of NMDA receptor-1(NRⅠ)in a rat model of focal cerebral ischemia and the possible mechanism of the neuroprotection.Methods Forty healthy male SD rats weighing 250-290g were randomly divided into 2 group(n=20 each):groupⅠketamine and groupⅡpentobarbital.The aminals were anesthetized with intraperitoneal ketamine 60 mg?kg~(-1) in groupⅠor pentobarbital 40 mg?kg~(-1) in groupⅡ.Focal cerebral ischemia was produced by permanent middle cerebral artery occludion(MCAO).The animals were killed at 24 h and 72 h of MCAO and their brains removed for determination of infarct size,the number of living neurons in the penumbra and the expression of NRⅠprotein(immuno- histochemistry).Results The infarct size was significantly smaller;the number of living neurons in penumbra significantly larger and NRⅠexpression significantly down-regulated in ketamine group than in pentobarbital group.Conclusion Ketamine can protect the brain against ischemia through downregulation of NMDA receptor-1.

A xylanase producing strain was screened with xylan as the only carbon source. The strain was identified as Streptomyces cirratus. The effects of different factore on the enzyme production were studied. Corncobs xylan (water insoluble) and tryptone were the best C and N sources, respectively. The enzyme activity was increased to about 2.5 times by addition of 0.5% Tween 80 in the medium. The highest xylanase activity was up to 623u/mL.

Radical retropubic prostatectomy (RRP) has been one of the most effective treatments for prostate cancer.This study is designed to identify the related predictive risk factors for complications in patients following RRP.Between 2000 and 2012 in Department of Urology,Fudan University Shanghai Cancer Center,421 cases undergoing RRP for localized prostate cancer by one surgeon were included in this retrospective analysis.We reviewed various risk factors that were correlated with perioperative complications,including patient characteristics [age,body mass index (BMI),co-morbidities],clinical findings (preoperative PSA level,Gleason score,clinical stage,pathological grade),and surgeon's own clinical practice.Charlson comorbidity index (CCI) was used to explain comorbidities.The total rate of perioperative complications was 23.2％ (98/421).There were 45/421 (10.7％),28/421 (6.6％),24/421 (5.7％) and 1/421 (0.2％) in grade Ⅰ,Ⅱ,Ⅲ,Ⅳ respectively,and 323/421 (76.8％) cases had none of these complications.Statistical analysis of multiple potential risk factors revealed that BMI ＞30 (P=0.014),Charlson score ≥1 (P＜0.001) and surgical experience (P=0.0252) were predictors of perioperative complications.Age,PSA level,Gleason score,TNM stage,operation time,blood loss,and blood transfusion were not correlated with perioperative complications (P＞0.05).It was concluded that patients' own factors and surgeons' technical factors are related with an increased risk of development of perioperative complications following radical prostatectomy.Knowing these predictors can both favor risk stratification of patients undergoing RRP and help surgeons make treatment decisions.

OBJECTIVETo explore the biological effect on NB4 cells proliferation, all-trans retinoic acid (ATRA) inducing differentiation and resistance to apoptosis by vascular endothelial growth factor (VEGF)-C cDNA transfection.

METHODSThe recombinant eukaryotic expression plasmid pcDNA3.1-VEGF-C and the vacant pcDNA3.1 vector were introduced separately into NB4 cells by lipofectamine mediation. The positive clones were screened by G418 and identified by reverse transcriptase-PCR (RT-PCR) and Western blotting. The proliferation capacity of NB4/VEGF-C cells was analysed by MTT assay and colony forming assay in vitro. After NB4/VEGF-C cells were induced by ATRA, the expression level of C/EBPalpha gene, CD11b on cells surface and morphological alteration were analysed by real-time quantitative PCR (RQ-PCR), flow cytometry (FCM), and Wright-Giemsa staining, respectively. FCM Annexin V-FITC/PI dual labeling technique was performed to investigate the etoposide (Vp16) induced NB4/VEGF-C cells apoptosis and bcl-2 gene expression level in these cells was analysed by RQ-PCR. The NB4/pcDNA3.1 cells was used as control in the above experiments.

RESULTSA stable NB4 cell line that secrets VEGF-C and its control lines were established. The proliferation capacity of the former was stronger than that of the latter. The expression level of C/EBPalpha gene of NB4/VEGF-C cells on ATRA treatment was only 1/32 that of NB4/pcDNA3.1 cells. The CD11b level and the degree of differentiation of NB4/VEGF-C were weaker than that of NB4/pcDNA3.1 cells. The percentage of apoptotic NB4/VEGF-C cells induced by Vp16 [(7.20 +/- 2.52)%] was significantly lower than that of NB4/pcDNA3.1 cells [(16.07 +/- 3.58)%] (P = 0.005), but the bcl-2 gene expression level of NB4/VEGF-C cells is 2.28-fold that of NB4/pcDNA3.1 cells.

CONCLUSIONThe VEGF-C via VEGFR-3 signaling pathway could promote the proliferation of leukemic cells by autocrine pathway and inhibit the cell differentiation mediated by ATRA and chemotherapy-induced apoptosis. VEGF-C/VEGFR-3 signaling loops might play an important role in disease progression and be potential therapeutic target for the treatment of leukemias.

Apoptosis ; drug effects ; genetics ; physiology ; Blotting, Western ; CCAAT-Enhancer-Binding Protein-alpha ; genetics ; metabolism ; CD11b Antigen ; genetics ; metabolism ; Cell Differentiation ; genetics ; physiology ; Cell Line, Tumor ; Cell Proliferation ; DNA, Complementary ; genetics ; Drug Resistance, Neoplasm ; Flow Cytometry ; Genetic Vectors ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; pathology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Tretinoin ; pharmacology ; Vascular Endothelial Growth Factor C ; genetics ; metabolism ; physiology

OBJECTIVETo assess the diagnostic value of 8 equations using different variables for determining the estimated glomerular filtration rate (eGFR) in patients with cardiovascular diseases.

METHODSGFR was estimated in 208 patients with cardiovascular diseases by (99m)Tc-DTPA dynamic renal imaging, and the eGFR was derived from 8 equations using different variables.

RESULTSIn patients with chronic kidney disease (CKD) stages 1-3, the eGFR calculated suing serum creatinine (SCr)-based equation was better correlated to GFR estimated by (99m)Tc-DTPA renal imaging than that derived from cystatin C (Cys C)-based equations, whereas in patients with CKD stages 4 and 5, the estimates by the latter equation showed a better correlation to GFR. Compared with (99m)Tc-DTPA renal imaging, MDRD-based equation and simple MDRD equation resulted in a higher eGFR in patients with CKD stages 4 and 5, the Rule equation had a lower eGFR in CKD stages 1 and 2, the Macisaac equation yielded a higher eGFR in CKD stages 2-5, and the Tan equation showed a higher eGFR in CKD stages 2 and 3. In patients with mild renal dysfunction, the Scr-based equation had a higher AUC(ROC) than Cys C-based equation, which was reversed in patients with severe renal dysfunction; the AUC(ROC) of the two equations were comparable in patients with moderate renal dysfunction. Compared with (99m)Tc-DTPA renal imaging, the modified MDRD equation and Arnal-Dade equation showed no significant difference in the eGFR in patients with CKD stages 1-5.

CONCLUSIONModified MDRD equation (or simple MDRD equation) and Arnal-Dade equation are superior to other calculation methods for estimating the GFR in Chinese patients with cardiovascular disease.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cardiovascular Diseases ; physiopathology ; Child ; Chronic Disease ; Creatinine ; blood ; Cystatin C ; blood ; Female ; Glomerular Filtration Rate ; physiology ; Humans ; Kidney Diseases ; physiopathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo detect the IgH-MMSET fusion gene resulted from t (4;14) translocation in multiple myeloma and illuminate its significance.

METHODSIgH-MMSET fusion gene was detected in bone marrow specimens of 25 multiple myeloma (MM) patients and MM cell line NCI-H929 using reverse-transcription PCR (RT-PCR) assay followed by nested PCR to increase the sensitivity. The purified PCR products were cloned into pGEM-T vector and then sequenced using M13 forward primers. The fragment sequences were compared with that in GenBank to find matched sequences.

RESULTSOnly a 438 base pair long fragment was obtained after RT-PCR assay and was confirmed by sequencing to be a fusion gene product of IgH gene and MMSET gene in MM cell line NCI-H929. The breakpoints were located within the C micro region of IgH gene on chromosome 14 and intron 3 of MMSET gene on chromosome 4. IgH-MMSET hybrid transcripts were detected in 3 of 25 MM patients through nested PCR assay. The amplified fragments of the 3 patients were 237 base pairs (bp), 239 bp and 239 bp in length, respectively. The breakpoints on chromosome 4 were identical to that of NCI-H929 cell.

CONCLUSIONSThe formation of IgH-MMSET fusion gene is resulted from t (4;14) translocation in MM. The incidence rate is 12.0%. The presence of IgH-MMSET fusion gene may predict poor prognosis.

Adult ; Aged ; Base Sequence ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 4 ; Female ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; Oncogene Proteins, Fusion ; genetics ; Protein-Tyrosine Kinases ; Receptor, Fibroblast Growth Factor, Type 3 ; Receptors, Fibroblast Growth Factor ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Translocation, Genetic

OBJECTIVETo establish NB4/VEGF-C cells xenograft in nude mice model, and explore the effect of VEGF-C on hematological malignancies

METHODSNB4/VEGF-C or NB4/pcDNA3.1 cell lines were established by transfecting the recombinant pcDNA3.1-VEGF-C plasmid and the vacant pcDNA3.1 vector into NB4 cells. The recombinant VEGF-C was identified by RT-PCR and Western blotting. Eighteen male BALB/c nude mice aged 4 - 5 weeks were equally divided into two groups. Mice irradiated by 4 Gy ⁶⁰Co were subcutaneously injected with 1 × 10⁷NB4/VEGF-C or NB4/pcDNA3.1 cells into one side of axilla. The volumes of xenograft tumor was evaluated according to L × t² × 0.52. Microvessel density (MVD) on the xenograft tumor section was detected by IHC with VWF antibody.

RESULTSNB4 cell xenograft tumors were developed in all mice of both the two groups. The growth of NB4/VEGF-C cells in nude mice was faster than in controls. There were statistically significant differences in the volume and weight of xenograft tumor between NB4/VEGF-C and NB4/pcDNA3.1 cell groups \[(631.44 ± 114.42) mm³ vs (491.22 ± 70.05) mm³\] (P = 0.006) and \[(321.78 ± 27.84) mg vs (288.57 ± 40.12) mg\] (P = 0.031), respectively. MVD in xenograft tumor of NB4/VEGF-C cells \[(50.8 ± 11.7)/mm²\] was higher than that in controls \[(18.9 ± 7.0)/mm²\] (P = 0.021). The Bcl-2 protein level in NB4/VEGF-C cells xenografts was higher than that in controls.

CONCLUSIONVEGF-C could promote proliferation of NB4 cells by inducing angiogenesis and inhibit cells apoptosis by upregulating antiapoptotic Bcl-2 protein expression in NB4 cells xenograft tumor.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Xenograft Model Antitumor Assays

Peritoneal dialysis catheter surgery has been used in clinical treatment for nearly 40 years, and open surgery under local anesthesia is the conventional method. However, catheter displacement after open surgery is still the thorny issue during our clinical practice. Then the reset surgery is often required to be taken again. Nowadays, laparoscopic peritoneal dialysis catheter draws our attention due to its advantages of accurate positioning, smaller incision, and less pain, and its clinical application has been limited. While laparoscopic surgery is recognized, there are few relevant studies on whether there is difference during the catheter reset process between the two surgical approaches. In this study, we mainly discussed the rate of secondary catheter migration, the incidence of complications after catheter reset for two surgical approaches and the hospital stay as well as the total clinical cost for the two surgical approaches. In this study, we retrospectively analyzed 25 cases of end-stage renal disease, who received catheterization for peritoneal dialysis and regular peritoneal dialysis in our hospital from March 2010 to December 2013, and had a medical history of catheter migration. We collected the relevant clinical data for all patients. Fifteen patients selected laparoscopic catheter reset, and 10 patients selected the traditional surgical method for catheter reset by themselves. For all patients enrolled, we analyzed the incidence of secondary catheter migration and postoperative complications, hospitalization time, and total cost for different methods of reset. Through the studies above, we found that laparoscopic peritoneal dialysis catheter surgery offered accurate catheter location and a small incision that was easy to heal. Besides, the incidence of postoperative complications for the laparoscopic surgery was lower than that for traditional surgical approach for catheter reset. The average hospitalization time for laparoscopic surgery was shorter than that for the traditional surgical approach. The total cost of laparoscopic surgery was more than that of the traditional surgery. Therefore, the rational application of a laparoscopic peritoneal dialysis catheter and reset surgery can increase the success rate of peritoneal dialysis, reduce the complications, shorten hospitalization time of patients, and thus enhance patient's confidence to stick it out.
Humans ; Laparoscopy ; standards ; Peritoneal Dialysis

OBJECTIVETo investigate the effect of hydrogen sulfide-induced delayed preconditioning on glutathione S-transferase (GST) expression during myocardial ischemia-reperfusion in rats.

METHODSSprague-Dawley male rats were randomly divided into 4 groups (n= 10 in each): Group S (sham operation group), Group IR (ischemia/reperfusion group), Group H (IR+ NaHS 0.05 mg/kg iv, 24 h before ischemia) and Groups D receiving IR+NaHS 24 h before ischemia and 5-hydroxydecanoate (5-HD)15 min before ischemia. Animals in groups IR, H and D were subjected to ischemia by 30 min of coronary artery occlusion followed by 2 h of reperfusion. At the end of the reperfusion, myocardial infarct size (IS) was examined. Glutathione S-transferase (GST) was measured by Western blotting. The myocardial ultrastructures were observed under the electron microscopy.

RESULTSThe IS was significantly smaller in Group H than that in Group IR [(25.40 ± 3.54)% compared with (38.27 ± 5.64)%, P<0.05]. The GST expression in myocardium was significantly higher in Group H than that in Group IR. Microscopic examination showed less myocardial damage in Group H than in Group IR. The protective effects of delayed preconditioning by hydrogen sulfide was prevented by 5-HD pre-treatment.

CONCLUSIONThe hydrogen sulfide-induced delayed preconditioning attenuates myocardial IR injury possibly through up-regulating glutathione S-transferase expression in rats.

Animals ; Disease Models, Animal ; Glutathione Transferase ; metabolism ; Hydrogen Sulfide ; administration & dosage ; therapeutic use ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Reperfusion Injury ; enzymology ; pathology ; therapy ; Myocardium ; enzymology ; ultrastructure ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the outcomes after 2 methods of laparoscopic gastric bypass surgery for patients with type 2 diabetes mellitus(T2DM).

METHODSFrom December 2009 to June 2011, 21 patients with T2DM underwent laparoscopic gastric bypass surgery, including laparoscopic Roux-en-Y gastric bypass (LRYGB, n=11), and laparoscopic mini-gastric bypass (LMGB, n=10). Clinical data were analyzed retrospectively.

RESULTSThe clinical complete remission rate of T2DM was 64%(7/11) in LRYGB group, and 60%(6/10) in LMGB group. The clinical partial remission rate of T2DM was 36%(4/11) in LRYGB group, and 40%(4/10) in the LMGB group. There was no significant difference between the two groups(both P>0.05). The levels of BMI, waist circumference, HOMA-IR and HbA1c within the postoperative 6 months were improved in each group (all P<0.05), but there was no significant difference between the two groups(all P>0.05). There were no conversion or perioperative deaths in both groups. Compared to LMGB, the LRYGB group had longer operative time[(147.0±35.9) min vs. (110.5±39.7) min, P=0.038] and postoperative hospital stay [(8.9±2.3) d vs. (7.1±1.4) d, P=0.046). One patient suffered from ileus in LRYGB group, one patient suffered from reflux esophagitis and one suffered chronic diarrhea in LMGB group. The incidence of postoperative complication was similar between the two groups(P>0.05).

CONCLUSIONLRYGB and LMGB may result in satisfactory and safe effects for the treatment of T2DM, while the LMGB is simpler and associates with quicker recovery.

Adult ; Diabetes Mellitus, Type 2 ; surgery ; Female ; Gastric Bypass ; methods ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Treatment Outcome