OBJECTIVETo construct a recombinant lentiviral vector for p38 MAPK and establish a human prostatic carcinoma cell line that stably expresses p38 MAPK.

METHODSEGFP/p38 fusion gene was subcloned into the lentiviral vector pTYF- EF1α-IRES-EGFP. The recombinant lentiviral vector pTYF-EF1α-EGFP/p38 was indentified by restriction enzyme digestion, and packaged in HEK 293T cells using lipofectamintm2000 with the packaging plasmid psPAX2 and envelope plasmid pMD2.G. The viral titer was tested according to the expression level of GFP. The resulting recombinant lentiviral vector was transduced into human prostatic carcinoma DU145 cells, and stably transduced cells were selected by limiting dilution analysis. The intracellular expression level of total p38 was detected by Western blotting and the cell growth curve was drawn.

RESULTSDNA restriction enzyme digestion demonstrated that the recombinant lentiviral vector of the fusion gene EGFP/p38 (pTYF-EF1α-EGFP/p38) was constructed successfully. The recombinant lentiviral vector was packaged in 293T with a viral titer of 4.7×10(6) TU/ml. A stable cell line, EGFP/p38-DU145, was established, which stably expressed exogenous EGFP/p38 MAPK fusion protein as detected by Western blotting and showed a lowered growth rate compared to the control cells.

CONCLUSIONWe have successfully constructed a recombinant lentiviral vector of the fusion gene EGFP/p38 and established a stable cell line EGFP/p38-DU145. Overexpression of p38 has a significant inhibitory effect on the proliferation of DU145 cells in vitro.

Cell Line, Tumor ; Cloning, Molecular ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; HEK293 Cells ; Humans ; Lentivirus ; genetics ; metabolism ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection ; p38 Mitogen-Activated Protein Kinases ; biosynthesis ; genetics

OBJECTIVETo investigate the cerebral distribution of propofol during continued infusion at a constant rate when the cerebral propofol uptake reaches equilibrium in dogs.

METHODSSix healthy 1-year-old male dogs were used in this study. The venous channel was established in the great saphenous vein of the right posterior limb. Anesthesia was induced with a single bolus injection of propofol (7 mg/kg), followed by propofol infusion at a constant rate of 70 mg/(kg.h) using a microinfusion pump. The blood samples were taken from the right internal carotid and internal jugular vein at 30 min (T30) and 50 min (T50) during propofol infusion for measurement of plasma propofol concentrations with high performance liquid chromatography (HPLC). At T50, the frontal lobe, parietal lobe, temporal lobe, hippocampus, cingulate gyrus, thalamus, midbrain, pons, and cerebellum were dissected respectively for determination of propofol concentrations.

RESULTSPropofol concentrations in the internal carotid artery and internal jugular vein blood plasma were 3.107-/+1.067, 3.095-/+1.085 microg/ml at T30 and 3.091-/+1.101, 3.117-/+1.091 microg/ml at T50, respectively, showing no significant differences (P>0.05). Propofol concentrations in the frontal lobe, parietal lobe, temporal lobe, hippocampus, cingulate gyrus, thalamus, midbrain, pons, cerebellum at T50 were 3.085-/+1.123, 3.116-/+1.125, 3.073-/+1.159, 3.117-/+1.090, 3.075-/+1.178, 3.073-/+1.146, 3.075-/+1.151, 3.102-/+1.174, and 3.072-/+1.192 microg/g respectively, suggesting homogeneous propofol distribution in these cerebral tissues (P>0.05).

CONCLUSIONAt T50, the cerebral uptake of propofol reached equilibrium when propofol is distributed homogeneously in the cerebral tissues in dogs.

Anesthetics, Intravenous ; administration & dosage ; blood ; pharmacokinetics ; Animals ; Brain ; metabolism ; Carotid Artery, Internal ; metabolism ; Chromatography, High Pressure Liquid ; Dogs ; Infusions, Intravenous ; Jugular Veins ; metabolism ; Male ; Propofol ; administration & dosage ; blood ; pharmacokinetics ; Tissue Distribution

The purpose of this study is to investigate the effects of formulation on the swelling behavior of choline fenofibrate hydrogel matrix tablets and reveal the relation between swelling property and release profile using dynamic image analysis. The volume swelling ratio (SR) and height/width (k) could evaluate the swelling behavior of matrix tablets well. The mount of hydroxypropyl methylcellulose (HPMC) and the ratio between K15M and K4M affected the volume swelling ratio, while PVP didn't. The three factors all impacted k, which was an indicator of the strength of the gel formed by HPMC. The accumulative release ratio and SR, the rate of swelling and the rate of release were compared. The proper model equations were established for the results with an excellent correlation. The results prove that there is a strong relevance between the swelling behavior and release property. This study provides a guideline in the study design for hydrogel matrix tablets.

OBJECTIVE: To investigate the efficacy, safety and the risk factors affecting prognosis of high-risk acute myeloid leukemia (AML) patients treated by cladribine-based intensified conditioning regimen. METHODS: The clinical data of 28 patients with high-risk AML treated by cladribine in combination with busulfan plus cyclophosphamide (BuCy) intensified conditioning regimen before allogeneic hematopoietic stem cell transplantation (allo-HSCT) in Zhujiang Hospital, Southern Medical University from October 2016 to June 2020 were analyzed retrospectively. The overall survival (OS) rate, cumulative progression-free survival (PFS) rate, relapse rate, non-relapse mortality (NRM), regimen related toxicity (RRT) and risk factors affecting prognosis of the patients were analyzed. RESULTS: The 1-year OS and PFS of the patients after implantation was (78.8±8.6)% and (79.8±8.1)%, while the 1-year cumulative relapse rate and NRM of the patients was 9.3% and 22.0%, respectively. The 1-year expected OS of MRD^- high-risk patients before HSCT was 100%. The 1-year expected OS and PFS of the patients in pre-transplant relapse group was (46.9±18.7)% and (50.0±17.7)%, respectively. The incidence of I/II grade RRT was 39.3%. NO III/IV grade RRT were found in 28 patients. Multivariate analysis showed that pre-transplant relapse was the independent risk factor affecting OS and PFS of the patients. CONCLUSION The intensified conditioning regimen of cladribine in combination with BuCy can reduce the relapse rate of high-risk AML transplantation, and its RRT is mild, exhibiting good safety. MRD^- high-risk patients before HSCT can achieve better transplant benefits, but the prognosis of patients with relapse before transplantation is not significantly improved. Therefore, for non-relapsed high-risk AML patients, this intensified conditioning regimen deserves to be considered.
Busulfan ; Cladribine ; Graft vs Host Disease ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute/therapy* ; Retrospective Studies ; Transplantation Conditioning