To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.
Calorimetry, Differential Scanning ; Curcumin ; chemistry ; Drug Liberation ; Lactic Acid ; Microscopy, Electron, Scanning ; Microspheres ; Polyglycolic Acid ; X-Ray Diffraction

BACKGROUNDControversies on the safety of the cement application between cemented and uncemented total hip arthroplasty (THA) have been existing for decades. The purpose of this study was to observe the changes in mean arterial pressure (MAP), heart rate (HR) and oxygen pressure (PaO(2)) during cemented THA, and to evaluate the intraoperative safety of using the third-generation cementing technique and investigate whether the intraoperative risk is higher in acute femoral neck fracture patients than non-traumatic patients.

METHODSForty-two patients who underwent cemented THA between November 2005 and September 2007 were prospectively included in this study. The third-generation cementing technique as vacuum mixing and pulsatile lavage was used strictly. The MAP and HR were monitored and documented during each operation. Blood gas analysis was performed at exposure, cup implantation, stem implantation and wound closure. MAP, HR and PaO(2) were compared between pre- and post-cement application. Comparisons of MAP, HR and PaO(2) between patients with acute femoral neck fracture and non-traumatic patients were performed as well.

RESULTSNo intraoperative cardiopulmonary complication occurred in these cases. No obvious changes were observed in MAP, HR and PaO(2) after cement application. There was no significant difference in MAP, HR and PaO(2) between acute femoral fracture patients (18 patients) and non-traumatic patients (24 patients).

CONCLUSIONSThe results of this study suggested that the invasive blood pressure monitoring and blood gas analysis are essential for patients undergoing cemented THA, especially for patients with femoral neck fracture. The third-generation cementing technique is safe to use in THA.

Adult ; Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; adverse effects ; methods ; Cementation ; methods ; Female ; Humans ; Male ; Middle Aged ; Monitoring, Intraoperative ; methods ; Prospective Studies

OBJECTIVETo study the application of multi-slice spiral CT angiography (MSCTA) after endoluminal exclusion of aortic diseases.

METHODS16-slice CT angiography was performed in 15 patients with aortic dissection and 4 patients with aortic aneurysm after endovascular exclusion. Two observers analysed the images and interpreted the outcomes and complications after endovascular exclusions of aortic dissection and aortic aneurysm.

RESULTSIn 19 patients, thrombus was found in all the false lumens of aortic dissection and the outer-stent cavity of aortic aneurysm. However, one patient with aortic aneurysm graft thrombosis; 4 patients had endo-leak (3 with type I endo-leak, 1 with type III endo-leak complicating graft deformation); one achieved perfusion recovery, and one experienced thrombolysis of superior mesenteric artery.

CONCLUSIONMSCTA can be an objective tool for the post-operative evaluation of endovascular exclusion of aortic diseases.

Adult ; Aged ; Aneurysm, Dissecting ; diagnostic imaging ; surgery ; Angiography ; Aortic Aneurysm, Abdominal ; diagnostic imaging ; surgery ; Female ; Humans ; Male ; Middle Aged ; Postoperative Period ; Tomography, Spiral Computed ; methods

OBJECTIVETo identify the distribution and differentiation of ABL kinase domain mutation in the Chinese Han nationality imatinib resistant chronic myeloid leukemia (CML) and Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph(+)ALL).

METHODSBone marrow or peripheral blood samples of 112 imatinib resistant CML patients and 21 Ph(+)ALL patients were obtained from the first affiliated hospital of Soochow university according to local law. Total RNA was extracted from the mononuclear cells using a TRIzol reagent. ABL kinase domain (KD) mutation was detected by direct sequencing.

RESULTSOf the 112 imatinib resistant CML patients, 54.46%(61 cases) had ABL KD mutation. Twenty-three mutants were identified in 20 amino acid sites and 23.21% (26 cases) ABL KD mutations were in P-loop region. ABL KD mutations were also detected in 71.43% (15 cases) imatinib resistant Ph(+)ALL patients, with 10 mutations in 8 amino acid sites. The most frequent mutation was T315I (28.57%), followed by E255K/V (19.05%) and Y253F/H (14.29%). The frequency of T315I was much higher in imatinib resistant Ph(+) ALL than that in imatinib resistant CML (P = 0.001). Ph(+)ALL with additional chromosomal aberrations also had a higher rate of ABL KD mutation than that of CML (P = 0.010). Ph(+)ALL gained ABL KD mutation faster than CML (P < 0.010).

CONCLUSIONChinese imatinib resistant CML and Ph(+)ALL patients had different characteristics in ABL KD mutation. The rate of ABL KD mutation in Ph(+)ALL with additional chromosomal aberrations was much higher than that of CML with additional chromosomal aberrations.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Benzamides ; pharmacology ; Chromosome Aberrations ; Drug Resistance, Neoplasm ; genetics ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Middle Aged ; Mutation ; Philadelphia Chromosome ; Piperazines ; pharmacology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Protein-Tyrosine Kinases ; genetics ; Proto-Oncogene Proteins c-abl ; genetics ; Pyrimidines ; pharmacology ; Young Adult

Objective To investigate the cause of differences in confluent growth between hepatic(L02) and hepatoma carcinoma(HCCLM3) cells by comparing responses of the two cells to different substrate stiffness (0.5, 4 kPa and glass). MethodsThe real-time photomicrography, immunofluorescence staining, flow cytometry, and Western Blotting techniques were respectively employed to observe the morphological characteristics, the cytoskeleton conformation and the distribution of E-cad of confluent L02 and HCCLM3 cells on different substrates, and test the changes in expression of E-cad, Integrinβ1 and p-Src. Results (1) Confluent L02 cells displayed a round or cubic shape, while HCCLM3 cells showed a polygon shape. The morphology of HCCLM3 cells were spread and polarized more obviously than that of L02 cells. With the increase of substrate stiffness, the variation of L02 cells with time was smaller than that of HCCLM3 cells. (2) The cytoskeleton of confluent L02 cells showed a ring-like conformation under the cortex, and E-cad was located at the cell-cell contact sites. However, the ring-like cytoskeleton of HCCLM3 cells was incomplete and distributed radially along the basement, while E-cad was dispersed in cytoplasm. (3) As the substrate stiffness increased, expression of E-cadherin in both L02 and HCCLM3 cells was significantly decreased (P<0.01), while the level of p-Src and integrinβ1 was increased significantly, with greater changes in HCCLM3 cells than in L02 cells. Conclusions The assembling of cortical ring-like cytoskeleton was positively correlated with the location of E-cad at the cell-cell contact sites. The substrate stiffness showed a more obvious impact on the balanced regulation between cadherin and integrin mediated adhesion system of hepatocarcinoma cells than that of hepatic cells.

OBJECTIVETo investigate the in vitro maturation of human oocytes (IVM) from pregnant late term, natural cycles and Gn stimulating cycles and the effect of granulose cells on IVM from pregnant late term.

METHODSA total of 1086 immature oocytes were obtained including 633 oocyte-cumulus complexes (OCCs) and 453 denuded oocytes (DOs). OCCs were divided into pregnant late term group, natural cycle group and IVM group, and DOs were divided into pregnant late term group, natural cycle group and controlled ovarian hyperstimulation (COH) group. All the oocytes were matured in IVM culture system and fertilized by ICSI. The embryos were cultured to blastocyst stage except that those in IVM group were transferred into the uterus. The main outcomes were assessed including maturation rate (MR), fertilization rate (FR), cleavage rate (CR), and blastulation rate (BR) (natural cycle group, pregnant late term group and COH group) and pregnancy rate per transfer cycle (PR) of IVM group.

RESULTSMR of OCCs in pregnant late term group, natural cycle group and COH group was 74.3%, 76.9% and 82.2%, respectively, showing statistical difference between pregnant late term cycle group and IVM group. No statistical difference was observed in FR, CR or BR between the three groups. For IVM cycle group, clinical pregnancy rate of 20% per aspiration was achieved. For DOs, MR of COH group (86.0%) was significantly higher than that of the natural cycle group (72.5%) and pregnant late term group (72.7%) (P<0.01). FR, CR and BR showed no statistical difference among the 3 groups. No difference was found in MR, FR, CR and BR between OCCs group and DOs group from pregnant late term.

CONCLUSIONSThe oocytes from pregnant late term have the same development potential as those from natural cycles or Gn stimulating cycles in vitro, and provide a new source of donor oocytes. Granulose cells do not affect the IVM from pregnant late term.

Cell Differentiation ; Cells, Cultured ; Female ; Fertilization in Vitro ; Humans ; Oocytes ; cytology ; Pregnancy ; Pregnancy Trimester, Third

Adult ; Aged ; Alcohol Drinking ; China ; ethnology ; Fatty Liver, Alcoholic ; etiology ; Female ; Humans ; Male ; Middle Aged ; Surveys and Questionnaires

OBJECTIVETo investigate the feasibility of multi-modalities in the reconstruction of circumferential defects after resection of cancers in pharyngoesophageal regions, and to compare the pros and cons between different surgical procedures.

METHODSAccording to the nature and extend of defects, five different methods including pectoralis major myocutaneous flap, laryngeal tube replacement, free jejunum, free forearm flap and gastric pull-up were used to reconstruct the circumferential pharyngoesophageal defects in 72 patients. Function of deglutition and restoration of swallowing was regularly followed up and objectively evaluated.

RESULTSA total of 35 cases of pectoralis major myocutaneous (PM) flaps, 8 cases of laryngeal tube replacement, 12 cases of free jejunum, 12 cases of free forearm flaps and 16 cases of gastric pull-up were performed. Different complications including wound infection, pharyngeal fistula, partial necrosis of PM flap, partial necrosis of gastric wall, stricture of anastomotic site were encountered in 15 cases. All patients survived the operation except one due to partial necrosis of the gastric wall. Two of 4 patients who developed anastomotic stricture can ingest half-liquid food, the remaining cases regained normal deglutition function. The mean postoperative follow-up time was 1. 6 years with 2-year survival rate of 45.3%.

CONCLUSIONSCircumferential defects resulting from resection of carcinomas in pharyngoesophageal region can be reconstructed with different operative techniques depending on the nature and extend of the defects. Once the operative indications are properly selected, the good reconstructive results are to be achieved.

Adult ; Aged ; Esophageal Neoplasms ; surgery ; Esophagus ; surgery ; Feasibility Studies ; Female ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods

OBJECTIVETo investigate the expression of AML1/ETO9a isoform in the acute myeloid leukemia (AML)-M2 patients.

METHODSExpressions of AML1/ETO fusion gene and AML1/ETO9a isoform were detected by using reverse transcriptase-polymerase chain reaction (RT-PCR) in leukemia patients, MDS patients, leukemia cell lines and healthy subjects. Karyotype was studied by R-banding technique.

RESULTIn 30 newly diagnosed AML-M2 patients 15 were found to express AML1/ETO9a isoform, while the rest including 20 AML-M2CR, 18 other subtypes of AML, 5 chronic myelogenous leukemia (CML), 3 myelodysplastic syndromes (MDS), 3 leukemia cell lines (NB4, KG-1, K562) and 5 healthy subjects were AML1/ETO9a negative. Among the 15 AML/ETO9a isoform expressing cases, 13 were demonstrated t(8;21) translocation and AML1/ETO expression.

CONCLUSIONIsoform AML1/ETO9a was correlated to AML/M2, and it may promote the development of leukemia in combination with the AML1/ETO fusion gene.

Adolescent ; Adult ; Aged ; Core Binding Factor Alpha 2 Subunit ; genetics ; metabolism ; Female ; Gene Expression ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; genetics ; metabolism ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Protein Isoforms ; genetics ; metabolism ; RUNX1 Translocation Partner 1 Protein

Objective To explore the possibility of rat adipose-derived stem cells (ADSCs) to differentiate into oligodendrocyte precursor cells(OPCs) and find an effective way to treat demyelinating disease. Methods ADSCs from the inguinal region of SD rats were isolated, digested with collagenase type I and trypsin, collagenase type I digestion method as control, counted and compared; Cultured in vitro and observed the growth characteristics. After ADSCs subcultured 3 times of passages, CD29, CD90 and CD45 were detected by flow cytometry; After differentiation into adipocyte, the cells were identified by the staining of oil red 0; After differentiation into OPCs by stem cell differentiation medium and OPCs induced differentiation medium, the expression of a-N-acetylneuraminic acid a-2, 8-sialyltransferase I (A2B5) and NG2 was detected by immunofluorescent staining. Results The number of ADSCs in the combined enzyme group was higher than the collagenase type 1 group (P < 0 . 05, re = 7); ADSCs grew in a long shuttle type and their morphology tended to be stable after passage. The surface marker CD29, CD90 were positive, and CD45 was negative. After adipogenic induction, oil red 0 staining showed red lipid droplets of varying sizes in the cells. After OPCs induction, immunofluorescence detection showed that positive reaction of cell surface fluorescence was seen with antibody to A2B5 and NG2,(87. 03±0. 94)% expressed A2B5, (90. 07±0. 96) % expressed NG2. After cultured for 3 days, immunof'luorescence detection showed that positive reaction of cell surface fluorescence was seen with antibody to myelin basic protein (MBP). Conclusion ADSCs are obtained by combined enzyme digestion and the cells are much more than collagenase alone and can be induced to OPCs in vitro.