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.

Autologous stem cell transplantation(ASCT) significantly increased the complete response rate(CR) and long-term survival in the patients younger than 65 years with multiple myeloma(MM). ASCT in the early stage of disease is currently considered the standard therapy in younger patients with newly diagnosed MM. High-dose melphalan with 200 mg/m2 is still considered the best conditioning. Double autotransplant is recommended for the patients without having VGPR after the first ASCT. Novel pretransplant induction regimens and conditioning with novel drugs such as bortezomib could improve the effect of ASCT. The best curative approach in multiple myeloma is allogeneic transplantation (allo-HSCT) with much higher CR. The long-term survival of patients receiving conventional/myeloablative conditioning (MAC) allo-HSCT is not better than that of the ASCT because of higher transplant-related mortility (TRM). Allo-HSCT with MAC has almost universally been replaced by the dose-reduced intensity conditioning allogeneic transplantation (Allo-RIC),which has lower TRM. More patients and long-term follow-up are needed to test the efficacy of single ASCT followed by Allo-RIC in patients with multiple myeloma.

In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Cell Survival/genetics ; Green Fluorescent Proteins/genetics ; Hep G2 Cells ; Poloxalene/*pharmacology ; Transfection ; Ultrasonics

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.

0.05). Second and third generation of the implanted tumor were obtained in 8 nude mice respectively,and all transplanted tumors survived. Liver,bone and lung metastasis were found in 100% of nude mice between 10 w～14 w. [WT5”HZ] Conclusions This nude mouse HCC model holds the biologic features of human HCC and serves the purpose of in vivo study investigation .

Purpose　To study myocardial pathology and its pathogenesis in diabetes. Methods　Myocardial structure of alloxan induced diabetic rats was observed under light microscopy (LM)and transmission electron microscopy(TEM).Activity of superoxide dismutase(SOD),glutathione peroxidase (GSH-Px)，nitric oxide synthase(NOS)and content of malondialdehyde (MDA),nitric oxide(NO) were detected biochemically in myocardial homogenate. Results　Atrophy and degeneration of myocardium and interstitial fibrosis were found under LM and expansion of mitochondria and smooth endoplasmic reticulum, destruction of myofibril and interstitial proliferation of collogen fiber under TEM. Activity of SOD and GSH-Px decreased significantly, while content of NO and MDA and activity of NOS increased significantly in diabetic rats. Conclusion　Atrophy of myocardium, expansion of mitochondria,destruction of myofibril and interstitial fibrosis are the main morphological changes of diabetic cardiomyopathy, and lipid peroxidation and NO may be involved in it.

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

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

Malignant mesothelioma(MM) is a rare but highly invasive carcinoma associated with asbestos exposure.Its incidence is in rising trend and most cases occur in pleura and peritoneum.The majority of patients diagnosed at late stage with poor prognosis that the median overall survival is only 12 months.Accurate diagnosis depends on the histopathology combined with immunohistochemistry.At present, the treatment of MM is mainly based on the tumor reduction or resection surgery combined with chemotherapy and radiotherapy.The potential molecular target needs to be further investigated.In this paper, we summary the incidence, diagnosis and treatment of MM, which will benefit the diagnosis and treatment of MM in China.

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.