OBJECTIVE:To establish a bacterial endotoxin test for monitoring pyrogen in wilsonii injection METHODS:Bacterial endotoxin test procedure and guiding principle in Chinese Phamacopoeia were used RESULTS:The test showed that the reaction with limulus agent was not interfered by wilsonii injection with a sensitivity of 0 25Eu/ml CONCLUSION:Limulus lysate method can be used to take the place of rabbit method for monitoring pyrogen in wilsonii injection

OBJECTIVE:To observe the effects of the essential conditions of storage on quality of Semen Armenniacae Am_arum (SAA).METHODS:Gravimeteric and titration method were used and orthogonal design was adopted.RESULTS:Obvious effects of baking temperature and storage duration on content of amygdalin,temperature on acid value of SAA,temperature and humidity and storage duration on peroxide value of SAA were found.CONCLUSION:The best conditions of storing SAA are as follows:A1,B1,C3 and short storage duration.

Objective To retrospectively analyse the impact of imatinib mesylate(Gleevec)on HSCT outcome in patients with Ph(+)leukemias.Methods From June 2001 to June 2005,31 CML-BC/AP patients and 8 refractory Ph(+)ALL patients were treated with imatinib mesylate to induce remission,followed by allogeneic transplantation,and we evaluated its influence on engraftment,graft versus host disease(GVHD),overall survival(OS),disease free survival(DFS),relapse rate(RR)and transplant-related mortality(TRM).Results Eighteen of 39 patients achieved complete hematologic remission(CHR),9 patients bone marrow remission(BMR)and 4 patients partial response(PR),the overall response rate being 70.96%.After HSCT,all patients achieved complete allogeneic engraftment with a median of 14 and 13.5 days for neutrophil and platelet recovery,respectively.The cumulative incidence rates of Ⅱ~Ⅳ? and Ⅲ~Ⅳ? aGVHD were 61.53% and 15.38%.The probability of OS and DFS was(73.51?9.61)% and(61.28?12.37)%,when HSCT was performed in CR,compared with(36.36?14.50)% and(31.25?13.98)% in NR.The 3-year cumulative relapse rate(RR)was 20.41% vs 75.00%.Four cases died of transplant-related complications.Conclusion Pre-transplant imatinib mesylate seems effective and safe for refractory Ph(+)leukemias;it allows transplantation to be in a more favorable status,and leads to better outcome.

Objective To investigate the influencing factors and prognosis of early and late recurrence after radical resection of primary hepatocellular carcinoma(HCC).Methods The clinical data of 117 patients who received radical resection of HCC at the Tongji Hospital of Huazhong University of Science and Technology from January 2003 to December 2006 were retrospectively analyzed.Tumor recurrence occurred within 2 years after operation was defined as early recurrence,and tumor recurrence occurred latter than 2 years after operation was defined as late recurrence.Relationship between postoperative tumor recurrence and level of alpha-fetoprotein (AFP),AFP/V,tumor diameter,tumor number,blood vessel invasion,tumor differentiation,hepatic cirrhosis,hepatic function,hepatitis B surface antigen,procedure of hepatic resection and blood transfusion was analyzed.The overall survival and disease-free survival rates were determined by Kaplan-Meier method,and the survival rate was analyzed by Log-rank test.Results Eighty-five(72.6％)patients were found with tumor recurrence,including 59(50.4％)with early tumor recurrence and 26(22.2％)with late tumor recurrence.Levels of AFP,AFP/V,tumor diameter,tumor number,blood vessel invasion,tumor differentiation and blood transfusion were the influencing factors of early recurrence(x2 = 12.78,13.40,5.79,9.98,10.26,9.48,8.32,P ＜ 0.05).Level of AFP and hepatic cirrhosis were the influencing factors of late recurrence(x2 =4.46,7.75,P ＜ 0.05).AFP/V,tumor number and blood vessel invasion were the independent risk factors of early recurrence(RR = 0.170,0.172,0.064,P ＜ 0.05).Hepatic cirrhosis was the independent risk factor of late recurrence(RR = 2.809, P ＜ 0.05).The 1-,3-,5-year overall survival rates and tumor-free survival rates were 82.6％,60.8％,54.9％ and 65.0％,38.5％,23.1％.There were significant differences in overall survival and disease-free survival rates among patients with AFP ＜20 μg/L,AFP/V ＜ 14 μg/(L · cm3)or AFP/V ≥ 14 μg/(L · cm3)(P ＜ 0.05).The 1-,3-,5-year overall survival rates of patients with early tumor recurrence were 64.9％,23.0％ and 20.5％,respectively,and the 1-,3-,5-year overall survival rates of patients with late tumor recurrence were 100.0％,88.5％ and 72.5％,respectively.A significant difference in the 1-,3-,5-year overall survival rates between patients with early or late tumor recurrence was observed(x2 = 26.918,P ＜0.05).Conclusions AFP/V,tumor number,blood vessel invasion were independent risk factors of early tumor recurrence,and hepatic cirrhosis was the independent risk factor of late tumor recurrence.There is a significant difference in the survival rate between patients with early or late tumor recurrence.

The value of color Doppler flow imaging (CDFI) and intravenous contrast-enhanced ultrasound (CEUS) for assessing the transplanted liver and early diagnosing complications by examining hemodynamic changes was discussed. Seventy-five patients with orthotopic liver transplantation (OLT) underwent CDFI. The following parameters were measured: peak systolic velocity (PS), resistance index (RI) and Doppler perfusion index (DPI) of the hepatic artery (HA), time average velocity (TAV) of portal vein (PV) and velocity of hepatic vein (HV) in different stages postoperation. And 11 patients of them received CEUS. Thirty healthy subjects were enrolled as controls. The results showed that: (1) In 23 patients without obvious complications, TAV of PV within 15 days post-operation was significantly higher than in controls (P<0.05), PS and DPI of HA within 7 days postoperation were lower, but RI was higher than in controls (P<0.05); (2) When the hepatic artery thrombosis (HAT) occurred, PS and DPI of HA were obviously decreased, but TAV of PV significantly increased like a high saw-tooth wave; (3) While rejection occurred, both TAV of PV and PS of HA were decreased with the increase in RI of HA, and the triphasic wave of HV disappeared and displayed as saw-tooth wave; (4) The incidence of biliary complications in liver transplantation was increased when DPI was reduced; (5) Seven cases of hepatic carcinoma relapse after OLT demonstrated hyperecho in the arterial phase and hypoecho in the portal and later phase on CEUS; (6) In 2 cases of HA thrombus, there was no visualized enhancement in arterial phase of CEUS, but enhancement during the portal vein and parenchymal phase. It was concluded that the hemodynamic changes of PV, HA and HV in the transplanted liver are valuable for assessing the transplanted liver and early diagnosing complications on CDFI and CEUS.
Bile Ducts/pathology ; Contrast Media/*administration & dosage ; Hemodynamics ; Hepatic Artery/pathology ; Infusions, Intravenous/*methods ; Liver Transplantation/*adverse effects ; Liver Transplantation/*methods ; Liver Transplantation/ultrasonography ; Perfusion ; Postoperative Complications ; Ultrasonography/*methods ; Ultrasonography, Doppler/*methods

Objective To study the outcome of HLA-identical hematopoietic stem cell transplantation ( HSCT) for severe aplastic anemia (SAA). Methods Twenty patients diagnosed with SAA received allogeneic HSCT from HLA-identical donors ( 17 from siblings and 3 from unrelated donors) between January 2000 and November 2008. Conditioning regimen consisted of cyclophosphamide ( Cy) and anti-thymocyte immunoglobulin (ATG). The patients were administrated with G-CSF-primed bone marrow (G-BM) and mobilized peripheral blood (G-PB) as grafts from the sibling donors or only G-PB from the unrelated donors. Results The median infused number of mononuclear cells and CD_(34)~+ cells were 7. 89 (4-14.21) × 10~8/kg and 2.60 (0.81-4.45) × 10~6/kg. All the patients got engraftment with 100% donor chimerism. The median time of neutrophil and platelet engraftment were 14 ( 11-20) d and 12 (8-108 )d respectively. The cumulative incidence rate of acute GVHD at 100 d was 16% (grade I : 3 cases,grade II :3 cases). Chronic GVHD occurred in 7 of the 19 evaluable cases (4 limited, 3 extensive). Till February 28, 2009, with a median follow-up of 18 months, 17 patients were alive and the overall survival rate was 82. 5%. Conclusion The study confirms that using G-PB with or without G-BM as graft after Cy + ATG conditioning results in excellent outcome of HLA-identical HSCT in patients with SAA.

Objective To assess the efficacy and safety of recombinant human granulocyte colony stimulating factor (rhG-CSF) primed donor peripheral blood stem cell (PBSC) on the treatment of poor graft function (PGF) after allogeneic stem cell transplantation(allo-HSCT).Methods The patients diagnosed as PGF after allo-HSCT and transfused with rhG-CSF primed PBSC from January 2003 to November 2012 were retrospectively analyzed.Hematological response was assessed at day 30 after transfusion.Graft versus host disease (GVHD) was assessed until 6 months after transfusion.Results There were 28 patients including 21 men and 7 women with a median age of 28 (12-50) years old.Of these patients,16 were diagnosed as primary PGF.The median number of transfused mononuclear cells was 2.0 (1.0-5.8) ×108/kg.Totally 42.9％ (12/28) patients achieved good response.Eight patients (28.6％) developed GVHD.Sixteen patients (57.1％) survived.Age (≤/＞ 28 years),gender,donor type (matched sibling/mismatched related),additional conditioning regimen prior to transfusion,time of neutrophil engraftment (≤/＞ 18 days) time of transfusion (≤/＞ 100 days after allo-HSCT) and number of mononuclear cells (≤/＞ 2.0 × 108/kg) did not impact hematological response.However,response rate of primary PGF (4/16) was significantly lower than that of secondary PGF (8/12) (P =0.022).Conclusion Transfusion of PBSC mobilized by rhG-CSF could be considered as an option to treat secondary PGF after allogeneic stem cell transplantation.

To examine the role of ultrasound in gene delivery in vitro, three cells lines were exposed to the low-frequency ultrasound of varying intensities and for different durations to evaluate their effect on gene transfection and cell viability of the cells. Microbubble (MB), Optison (10%), was also used to observe the role of the microbubbles in gene transfection. The results demonstrated that as the ultrasound intensity and the exposure time increased, the gene transfer rate increased and the cell viability decreased, but at high energy intensities, the cell viability decreased dramatically, which caused the transfer rate to decrease. The most efficient ultrasound intensity for inducing gene transfer was 1 W/cm(2) with duration being 20 s. At the same energy intensity, higher ultrasound intensity could achieve maximal gene transfer rate earlier. Microbubbles could increase ultrasound-induced cell gene transfer rate by about 2 to 3 times mainly at lower energy intensities. Moreover, microbubbles could raise the maximum gene transfer rate mediated by ultrasound. It is concluded that the low-frequency ultrasound can induce cell gene transfer and the cell gene transfer rate and viability are correlated with not only the ultrasound energy intensity but also the ultrasound intensity, the higher ultrasound intensity achieves its maximal transfer rate more quickly and the ultrasound intensity that can induce optimal gene transfer is 1 W/cm(2) with duration being 20 s, and microbubbles can significantly increase the maximal gene transfer rate in vitro.
3T3 Cells ; CHO Cells ; Cell Line ; Cell Survival/*genetics ; Contrast Media/metabolism ; Cricetinae ; Cricetulus ; Microbubbles ; Transfection/*methods ; Ultrasonics

Liver resection is the most effective treatment for hepatocellular carcinoma (HCC). The Barcelona Clinic Liver Cancer (BCLC) staging system is commonly accepted as a guideline for HCC treatment, but it only recommends liver resection for the patients with HCC at stage 0 to A1. The surgical indications of the BCLC staging system need to be re-evaluated. 120 HCC patients undergoing curative liver resection were retrospectively stratified to the BCLC staging system, and the survival of the patients at stages A, B and C was analyzed. The justification of the BCLC staging system was re-evaluated. Fifty-two patients were classified at stage A, 51 at stage B and 17 at stage C respectively. The hospital mortality of this cohort was zero and the morbidity was 24.1%. The 1-, 2-, 3-year overall survival rate of this cohort was 81.6%, 68.3%, and 57.5% respectively. There was no significant difference in the survival rate between the patients at stage A and B (P>0.05). If the treatment guidelines of BCLC staging system were followed, the majority of the patients at stages A and B (77.7%, 80/103) would not have been treated surgically. Our data suggest that the surgical indications of the BCLC staging system are not justified for HCC treatment. More studies may be needed as for how to further broaden the surgical indications of the BCLC staging system in the future.

Objective To investigate the efficacy of chimeric anti-CD2 monoclonal antibody (basiliximab) on acute graft-versus-host disease (GVHD) in patients following aliogeneic hematopoietic stem cell transplantation. Methods Thirty-six patients who were suffered from acute GVHD from March 2005 to July 2007 were studied. All of them were treated with steroid first and got no response, then began basiliximab therapy. Results Thirty of 36 patients showed response to basiliximab therapy including 25 complete responses and 5 partial responses. The efficacy was associated with the degree of GVHD and the source of donor. Conclusion Patients suffered from steroid-resistant acute GVHD can be successfully treated with basiliximab.