Objective To investigate the clinical application value of the laparoscopic intraoperative cholan-giography in laparoscopic cholecystectomy,and summarize the experience.Methods The clinical data of 169 cases of laparoscopic cholecystectomy intraoperative cholangiography were analyzed retrospectively.Results 169 patients were successfully completed,131 cases recovered well and no complications occurred after operation.29 patients showed hyperamylasemia,of which 3 patients had intractable hyperamylasemia,8 patients complicated with secondary acute pancreatitis,with fasting,gastrointestinal decompression,enzyme inhibition(plus somatostatin)and acid,analgesic, anti infection,rehydration treatment after remission.Conclusion Laparoscopic cholecystectomy intraoperative cholan-giography is a safe and reliable diagnostic technique,on the occurrence of biliary residual stones in prevention of post-operative prevention and timely detection of bile duct injury during operation and improves the success rate of repair of bile duct injury has important clinical value;control adaptation of intraoperative cholangiography has important clinical significance of reasonable application license.

Objective To observe the therapeutic effect of Shenmai injection on rats with acute edematous pancreatitis (AEP) and its mechanism.Methods The model of AEP in rats was induced by intraperitoneal injection of caerulein every one hour for 7 times.One hour after last injection,intravenous Shenmai injection (5 ml · kg-1 · d-1) was given to rats in Shenmai injection treatment (SI) group,while same amount of normal saline was given to rats in control (AEP) group.One hour and 1,3,5,7 d after AEP induction,the blood was taken,and the method of double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of amylase,platelet activating factor (PAF),vascular endothelial growth factor (VEGF).The rats were sacrificed at 7 and 14 d,and pancreatic tissue samples were harvested.RT-PCR and Western Blot were used to determine the expression of NF-κB mRNA and protein,and normal pancreatic tissue samples were used as control.The positive expression of CD31 in pancreatic tissue was determined by immunohistochemical method to calculate microvascular density (MVD).Results The serum levels of amylase and PAF in both groups were increased and gradually decreased 3 days later,while the decrement in SI group was greater than that of AEP group.At the 5th day after AEP induction,the serum levels of amylase in SI group and AEP group were (4569 ± 158),(5056 ± 198) U/L,and the serum levels of PAF were (25.30 ± 4.76),(36.06 ± 5.57)μg/L,and the difference between the two groups was statistically significant (P ＜ 0.05).The serum levels of VEGF were increased 1 d after AEP induction,and the increase in SI group was greater than that of AEP group 3 d later,and the serum levels of VEGF were (139.78 ±24.30),(118.51 ±21.70)pg/ml at 5th day,and the difference between the two groups was statistically significant (P ＜ 0.05).The expression levels of NF-κB mRNA and protein of pancreatic tissue in SI group were O.834 ±0.031 and 0.49±0.24,and MVD was 6.41 ± 1.14,while the corresponding values in AEP group were 1.000 ± 0.059,0.93 ± 0.45,3.62 ± 0.89,and the difference between the two groups was statistically significant (P ＜ 0.05 or P ＜ 0.01).Conclusions In the course of acute pancreatitis,Shenmai injection has the therapeutic effects of promoting new angiogenesis,improving the microcirculation,reducing the inflammatory cascade.

OBJECTIVETo evaluate the clinical value of (89)SrCl(2) (Ke xing Inc, Shanghai) as a palliative therapy modality for cancer patients with bone metastasis.

METHODSIn 504 cancer patients with painful limitation of movement due to bony metastasis, a dose of 1.48-2.22 MBq/kg (40-60 uCi/kg) iv infusion of (89)SrCl(2) was given.

RESULTSIn 97 patients (19.2%) there was no improvement in pain and life quality, 298 patients (59.1%) showed mild to moderate improvement (moderately effective), 109 patients (21.6%) became free of pain and were subsequently fully ambulatory (markedly effective). The pain relief appeared from D1-D46 after (89)SrCl(2) administration, most frequently from D5-D14. The palliative effect could last for about 56 days to 13 months. Repeated bone scans of some patients showed that the metastatic foci in the bone became smaller or even disappeared gradually after the administration of (89)SrCl(2). Approximately 55% of patients experienced grade I approximately III bone marrow depression attributable to (89)SrCl(2), which would return to the pre-treatment level within 3 approximately 9 months.

CONCLUSION(89)SrCl(2) is effective and safe for the relief of bone pain and improvement of quality of life in cancer patients with painful bony metastasis.

Adult ; Aged ; Aged, 80 and over ; Bone Neoplasms ; complications ; radiotherapy ; secondary ; Breast Neoplasms ; pathology ; Female ; Humans ; Lung Neoplasms ; pathology ; Male ; Middle Aged ; Pain Measurement ; Pain, Intractable ; etiology ; radiotherapy ; Quality of Life ; Strontium Radioisotopes ; therapeutic use

To explore the feasibility of real-time quantitative PCR (QRT-PCR) for selecting cell strains which overexpress a certain transgene, expression level of RbAp46 was detected in transfected cell strains by using optimal real-time PCR with SYBR Green I. Meanwhile, semi-quantitative RT-PCR and Western blot were performed to compare with the QRT-PCR. The results showed that values of RbAp46(N) were 2064.42 +/- 253.47, 860.94 +/- 291.07, 234.456 +/- 31.08, 18.17 +/- 5.14 and 1.46 +/- 0.54 in K562/RbAp46, K562/CMV, SHG44/RbAp46 monoclone, SHG44/RbAp46 multiclone and SHG44/CMV, respectively. The results were consistent with that determined by semi-quantitative RT-PCR and Western blot. It is concluded that QRT-PCR provides a highly efficient and reproducible method for selection of transfected cell subclones at different level of transgene expression.
Blotting, Western ; Carrier Proteins ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; K562 Cells ; Nuclear Proteins ; genetics ; metabolism ; Organic Chemicals ; chemistry ; RNA, Neoplasm ; metabolism ; Retinoblastoma-Binding Protein 7 ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transfection ; Transgenes ; genetics

cDNA microarray, recently applied to analyze gene expression profile of cancers, was difficult to be utilized in myelodysplastic syndrome (MDS) for a special need of excessive mRNA hardly provided by ordinary bone marrow aspiration in MDS patients. The aim of this study was to investigate the feasibility of exploring the molecular events underlying MDS by using cDNA microarray and mixing mRNA from multiple patients. A commercially purchased BioStar H141 cDNA microarray containing 14,110 clones of cDNA or EST was employed to analyze the gene expression profile of bone marrow mononuclear cells from two cases of MDS. Equal amount of total RNA from each patient was mixed, reversely transcribed to cDNA and labeled with Cy5. Mixture of Cy5-labeled cDNA and Cy3-labeled cDNA from normal bone marrow cells was concomitantly hybridized to H141 microarray in duplicate. In H141 chips, 1,064 cDNAs were spotted at least twice targeting different fragments of a single gene cDNA. The results showed that among these 1,064 cDNA clones, the expression level of 625 (58.7%) and 630 (59.2%) ones was consistent within these two chips, respectively, 297 (27.9%) and 191 (18.0%) inconsistent, 21 (2.0%) and 11 (1.0%) in opposite. Among 411 duplicately spotted cDNAs with complete data, expression levels of 400 (97.3%) was consistent between two chips. 488 genes with known function were identified as differentially expressed in MDS, among which 101 genes were involved in hematopoiesis regulation, including those encoding transcription factors, cell cycle-regulating proteins, metabolism-relating enzymes, and adhesive molecules. It is concluded that cDNA microarray can be used for profiling gene expression of mixed MDS samples and replication shall be necessary for reducing the data bias caused by experimental operation.
Bone Marrow Cells ; metabolism ; Gene Expression Profiling ; Humans ; Leukocytes, Mononuclear ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; Oligonucleotide Array Sequence Analysis ; Reproducibility of Results

OBJECTIVETo evaluate the significance of quantitative detection of CCAAT/enhancer binding protein zeta (C/EBP zeta) transcripts in patients with chronic myeloid leukemia (CML).

METHODSReal-time quantitative polymerase chain reaction (RQ-PCR) assay was established and performed to measure the transcript level of C/EBP zeta in the bone marrow mononuclear cells from 76 patients with CML at different stages.

RESULTSC/EBP zeta transcripts were significantly decreased in CML patients in chronic phage, accelerated phage, and blastic crisis (median 2.5, 3.31, and 2.22, respectively), compared to that in normal controls (median 12.20). Further down-regulation of C/EBP zeta was observed in the patients resistant to interferon (median 1.56); however, its expression level returned to normal in the patients who obtained complete cytogenetic remission (median 15.43).

CONCLUSIONC/EBP zeta was down-regulated in CML patients, which might play a role in the leukemogenesis.

CCAAT-Enhancer-Binding Proteins ; genetics ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Transcription, Genetic

OBJECTIVETo investigate Wilms' tumor gene (WT1) expression levels in bone marrow (BM) of acute leukemia patients (ALs).

METHODSA real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method was established for detecting WT1 and internal reference GAPDH expression levels in BM of 108 ALs and 23 non-leukemia controls by Light Cycler.

RESULTSThe median expression levels of WT1 in 70 newly diagnosed ALs and 11 relapsed ALs were statistically higher than those in 23 ALs in complete remission (CR) and 23 non-leukemic controls (75.10 and 89.56 vs 2.07 and 1.51 respectively). No statistic differences was found between the CR group and control group, nor between the newly diagnosed group and relapsed group. Of the 70 newly diagnosed ALs, median WT1 expression level of acute granulocytic leukemias was significantly higher than that of acute monocytic leukemias (M(5)), but there was no statistic differences among the M(1), M(2), M(3) and ALL subtypes. Furthermore the WT1 levels were not correlated to peripheral WBC counts, BM blast percentage and multidrug resistant gene (mdr1) expression at presentation, but correlated to chromosome karyotypes. Dynamic analysis of WT1 levels of 2 patients on treatment showed that WT1 expression levels predicted relapse.

CONCLUSIONWT1 expression levels in ALs were strikingly higher than that in non-leukemias. WT1 can be a marker for detecting MRD and evaluating therapy efficacy in leukemias.

Acute Disease ; Adolescent ; Adult ; Aged ; Bone Marrow Cells ; metabolism ; Child ; Female ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia ; blood ; genetics ; Leukemia, Monocytic, Acute ; blood ; genetics ; Leukemia, Myeloid ; blood ; genetics ; Male ; Middle Aged ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; WT1 Proteins ; genetics ; Young Adult

The aim of this study was to investigate the incidence, risk factors of acute renal failure (ARF) after allogeneic hematopoietic stem cell transplantation (allo-HSCT), and evaluate its effect on the prognosis of patients after allo-HSCT. A retrospective analysis was performed in 86 patients undergoing allo-HSCT at Peking University First Hospital from June 2003 to April 2007. ARF is defined as a doubling of baseline serum creatinine at any time during the first 100 days post-transplant. The risks of ARF and mortality after ARF were examined using univariate analysis and multivariate unconditional logistic regression. The correlation of ARF and survival was examined using Cox regression. The results indicated that 27 patients (31.40%) developed ARF at a median of 59.5 days after transplant (range 1 to 93 days). The univariate analysis showed that elevated risks were severe acute GVHD (OR 6.196; 95% CI 1.121 - 34.249, p = 0.033), sepsis or septic shock (OR 4.184; 95% CI 1.314 - 13.325, p = 0.018) and hyperbilirubinemia (OR 3.709; 95% CI 1.428 - 9.635, p = 0.006). Renal disease before transplant (OR 6.711; 95% CI 1.199 - 37.564, p = 0.027), hypertension (OR 2.067; 95% CI 0.739 - 5.782, p = 0.165), the use of vancomycin (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) or foscarnet sodium (OR 2.133; 95% CI 0.844 - 5.392, p = 0.106) may be potential risks. Multivariate logistic regression analysis showed that renal disease before transplant (OR 6.288; 95% CI 1.218 - 32.455, p = 0.028), sepsis or septic shock (OR 3.614; 95% CI 1.040 - 12.544, p = 0.043) and hyperbilirubinemia (OR 4.448; 95% CI 1.563 - 12.665, p = 0.005) appear to be independently associated with an increased risk of ARF. Age, gender, baseline serum creatinine level, advanced malignant disease, unrelated-donor, total body irradiation (TBI) and cyclosporine levels were not associated with the development of ARF. Cox regression showed that ARF (RR 2.124; 95% CI 1.016 - 4.441, p = 0.045) was independently associated with survival of patients after allo-HSCT. The mortality of patients with ARF within 6 months post-transplant was significantly higher than that of those without ARF (44.4% vs 8.47%, p < 0.001). It is concluded that the cumulative incidence of ARF after allo-HSCT remains high. Renal disease before transplant, hyperbilirubinemia and sepsis or septic shock are all related factors which can increase the risk of ARF. ARF appears to be independent factor influencing survival of patients after allo-HSCT.
Acute Kidney Injury ; etiology ; Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Incidence ; Male ; Middle Aged ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous ; Young Adult

OBJECTIVETo explore the therapeutic feasibility of allogeneic hematopoietic stem cell transplantation (allo-HSCT) from partially HLA-mismatched related donors for hematologic diseases.

METHODSThirty patients with hematologic diseases received allo-HSCT from 1 - 3 loci mismatched related donors conditioning regimen consisting of ATG (thymoglobulin, total dose of 10 mg/kg, intravenously on - 4 d to - 1 d), and only 5 (18%) of 28 recipients from HLA-identical sibling donors were treated with regimen containing ATG. Donors were given G-CSF prior to hematopoietic stem cell harvest and CsA, short-term MTX and mycophenolate mofetil (MMF) were used for GVHD prophylaxis in both group.

RESULTSAll patients were successfully engrafted. There was no significant difference in the incidence of grade II to IV acute graft-versus-host disease (aGVHD) and grade III to IV aGVHD between the mismatched and matched groups (34% vs 32%, and 13% vs 11%, respectively). 3-year overall survival (OS) and disease-free survival (DFS) in mismatched and matched groups were 57% vs 77% (P = 0.14) and 57% vs 69% (P = 0.28), respectively. Multivariate analysis showed that advanced disease pre-transplant (P = 0.006) and CMV infection (P = 0.04) were risk factors for OS. OS for patients with stable disease in mismatched and matched groups were 87% vs 81% (P = 0.65) respectively, and for those with advanced disease were 21% vs 71% (P = 0.02).

CONCLUSIONSIt is feasible to perform allo-HSCT from 1 -3 loci HLA-mismatched related donors for patients with stable disease who lack HLA-identical sibling donors. Nevertheless, for patients with advanced disease optimized conditioning regimen and intensive supporting therapy should be administered to obtain better clinical outcomes.

Graft vs Host Disease ; prevention & control ; HLA Antigens ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Siblings ; Tissue Donors ; Transplantation Conditioning

BACKGROUNDWide application of umbilical cord blood transplantation (UCBT) in adult patients is limited by low cell-dose available in one umbilical cord blood (UCB) unit. The aim of this study was to investigate the safety and long-term outcomes of UCBT from unrelated donors in adult and adolescent patients with leukemia.

METHODSThirteen patients with leukemia received double-unit UCBT with human leukocyte antigen (HLA) mismatched at 0 - 2 loci. We analyzed the engraftment, graft-versus-host disease (GVHD) and survival.

RESULTSTwelve evaluable patients (92.3%) had neutrophil and platelet engraftment at a median of 21 days (range, 16-38 days) and 34 days (range, 25 - 51 days), respectively. At day 30, engraftment was derived from one donor in 8 patients (66.7%, 95%CI 40.0% - 93.4%), and from both donors in 4 patients (33.3%, 95%CI 6.7% - 60.0%) with 1 unit predominated. Unit with larger nucleated cell (NC) dose would predominate in engraftment (P = 0.039), whereas CD34(+) cell dose or HLA-match failed to demonstrate any relationship with unit predominance. Only one patient developed grade II acute graft-versus-host disease (aGVHD). Chronic GVHD (cGVHD) was observed in 2 of 11 patients who survived more than 100 days, and both were limited. The median follow-up after transplantation for the 13 patients was 45 months (range 1.5 - 121.0 months) and 72 months (range 41.0 - 121.0 months) for the 8 alive and with full donor chimerism. The 5-year cumulative disease free survival (DFS) was (61.5 ± 13.5)%. Of the 13 patients, 5 patients died in 1 year and 1-year transplantation related mortality (TRM) was 23.1% (95%CI 0.2% - 46.0%).

CONCLUSIONDouble-unit UCBT from unrelated donors with HLA-mismatched at 0-2 loci may overcome the cell-dose barrier and be feasible for adults and adolescents with leukemia.

Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; adverse effects ; methods ; Disease-Free Survival ; Female ; Graft vs Host Disease ; etiology ; Humans ; Leukemia ; immunology ; mortality ; therapy ; Male ; Treatment Outcome ; Young Adult