This study examined the effect of self-microemulsiflying drug delivery system (SMEDDS) containing Cremophor RH40 or Tween 80 at various dilutions on cytochrome P450 3A (CYP3A) enzymes in rat hepatocytes, with midazolam serving as a CYP3A substrate. The particle size and zeta potential of microemulsions were evaluated upon dilution with aqueous medium. In vitro release was detected by a dialysis method in reverse. The effects of SMEDDS at different dilutions and surfactants at different concentrations on the metabolism of MDZ were investigated in murine hepatocytes. The cytotoxicity of SMEDDS at different dilutions was measured by LDH release and MTT technique. The effects of SMEDDS on the CYP3A enzymes activity were determined by Western blotting. Our results showed that dilution had less effect on the particle size and zeta potential in the range from 1:25 to 1:500. The MDZ was completely released in 10 h. A significant decrease in the formation of 1'-OH-MDZ in rat hepatocytes was observed after treatment with both SMEDDS at dilutions ranging from 1:50 to 1:250 and Cremophor RH 40 or Tween 80 at concentrations ranging from 0.1% to 1% (w/v), with no cytotoxicity observed. A significant decrease in CYP3A protein expression was observed in cells by Western blotting in the presence of either Cremophor RH40 or Tween 80-based SMEDDS at the dilutions ranging from 1:50 to 1:250. This study suggested that the excipient inhibitor-based formulation is a potential protective platform for decreasing metabolism of sensitive drugs that are CYP3A substrates.

The aim of this study was to explore the mechanism underlying the blast crisis of chronic myeloid leukemia. Analysis of gene expression profiles of chronic myeloid leukemia patients in chronic phase and blast crisis were analyzed by using cDNA microarray representing 4096 genes for finding the differential expression genes. The results indicated that 74 differential expression genes were identified in at least 3 gene chips in blast crisis compared with chronic phase, among them 52 genes were down-regulated and 22 genes were up-regulated in blast crisis. The differential expression genes were involved in these genes including genes related to cell structure/mobility, signal transduction, transcription factor, related immunity, metabolism, cell cycle, oncogene/anti-oncogene, cell receptor, protein translation/synthesis and some unknown functions. It is concluded that the blast crisis of CML is resulted from abnormality and interaction of multigene, among them functional abnormal genes related to signal transduction, cell cycle, cell differentiation and immunity may be the critical genes for chronic myeloid leukemia blast crisis.
Adult ; Aged ; Female ; Gene Expression Profiling ; Gene Expression Regulation, Leukemic ; Genes, Neoplasm ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; Male ; Middle Aged ; Oligonucleotide Array Sequence Analysis ; Signal Transduction

To investigate the state and significance of bone marrow angiogenesis in hematological diseases, bone marrow microvascular density (BM-MVD) in plastic-embedded section was examined using acetone-fixed bone marrow tissues embedded in glycol-methacrylate (GMA) resin and by the method of immunohistochemistry. The results showed that bone marrow MVD increased greatly in newly diagnosed hematological malignancies before treatment. BM-MVD in patients with acute leukemia decreased down to the normal range as the controls at the time of complete remission. In the non-remission group, BM-MVD decreased less, but when relapsed it increased again up to the same range as the newly diagnosed hematological malignancies, significant increase of BM-MVD was found in patients with anemia, but in less degree than that in hematological malignancies. It is concluded that bone marrow angiogenesis plays a key role in the pathogenesis and development of hematological malignancy. Antiangiogenic therapy may be able to constitute a novel strategy for the treatment of hematological malignancies including leukemia.
Acute Disease ; Bone Marrow ; blood supply ; pathology ; Hematologic Diseases ; blood ; pathology ; Humans ; Leukemia ; blood ; pathology ; Microcirculation ; Neovascularization, Pathologic ; blood ; pathology

OBJECTIVETo observe the effect of activation of aldehyde dehydrogenase 2 (ALDH2) by ethanol on the expression of c-Jun N-terminal kinase (JNK) in the kidney of diabetic rats.

METHODSEightheen healthy male SD rats were randomly divided into 3 groups (n = 6): normal control group, diabetes group and ethanol + diabetes group. After 8 weeks, 24 h urine samples from rats were collected to detect urinary protein content. The kidney was isolated and the ratio of kidney weight/body weight (index of kidney weight) was detected. The levels of fasting blood glucose, glycosylated hemoglobin serum urea nitrogen and serum creatinine were measured. Morphological changes of renal tissue were observed by optical microscope. The protein expressions of ALDH2 and JNK in renal tissue were detected by Western blot.

RESULTSCompared with the normal control rats, the levels of fasting blood glucose, glycosylated hemoglobin, serum urea nitrogen, serum creatinine and the index of kidney weight were increased markedly in diabetic rats. The expression of ALDH2 protein was decreased, while p-JNK, JNK protein expressions and the ratio of p-JNK/JNK were increased. The morphological observation was shown that the amount of glomerular mesangial matrix were increased, basement membrane were thickened and capillary lumen were narrowed. However,in ethanol + diabetes group, renal function was improved and the damage of renal structure was attenuated. The expression of ALDH2 protein was increased, while p-JNK, JNK and the ratio of p-JNK/JNK were decreased.

CONCLUSIONEnhanced ALDH2 expression can protect kidney in diabetic rats, which may be relevant with inhibitting the activity of JNK pathway.

Aldehyde Dehydrogenase ; metabolism ; physiology ; Aldehyde Dehydrogenase, Mitochondrial ; Animals ; Diabetes Mellitus, Experimental ; enzymology ; Ethanol ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Kidney ; enzymology ; Male ; Mitochondrial Proteins ; metabolism ; physiology ; Rats ; Rats, Sprague-Dawley

Objective: To prepare an adenovirus, Ad-CD, expressing bacterial cytosine deaminase(CD),and use it for tumor suicide gene therapy. Methods: We constructed a recomb ined adenoviral vector carrying CD or LacZ. 293 packaging cell was trans fected by the newly-constructed plasmid and generated the infectious virus. The n the adenovirus was used to infect C26 cell in vitro. Results: The growth of Ad-CD-infected cell was obviously surpressed after admini stration of 5-FC. Conclusion: CD gene carried by adenoviral vec tor is effective for tumor cell suicide gene therapy in vitro.

This study was aimed to explore the anti-leukemic effect of scutellaria extract SBX in human leukemia cell lines and its mechanism. The leukemia cell lines, including HL-60, NB4, U937, K562 and Jurkat, were cultured in vitro and proliferative inhibition of these cell lines was detected by CellTiter-Glo Luminescent Cell Viability Assay in order to screen the most sensitive cell line. The effect of SBX on cell cycle was analyzed by flow cytometry and the protein expressions determined by Protein Pathway Array respectively. The results indicated that SBX (10 - 200 µmol/L, for 72 h) significantly inhibited the proliferation of different leukemia cell lines in a dose-dependent manner (r value was 0.86, 0.88, 0.95, 0.94, 0.96, respectively), the HL-60 was the most sensitive cell line. Flow cytometric analysis showed that SBX (50, 10 µmol/L, for 48 h) arrested HL-60 cells in the G(0)/G(1) phase. In addition, protein expression of p-PKC α/βII, p-p38, Cdc25B, XIAP of HL-60 cells increased, and p-AKT, p-SAPK/JNK, Notch4, Cdk4, Cdc2, cyclin E, Akt, Bcl-2, Bax, cdc42, TNF-α, p27, CaMKKa decreased after exposure to SBX (50 µmol/L, for 48 h). It is concluded that SBX can inhibit the proliferation of different leukemia cell lines, and HL-60 is a sensitive cell line. SBX significantly influences EGFR, Ras/Raf/MAPK and Notch signaling pathway, through which effects the expression of cell cycle-related proteins resulting in arrest of HL-60 cells in G(0)/G(1).
Cell Cycle ; Cell Cycle Proteins ; metabolism ; Cell Line, Tumor ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Leukemia ; drug therapy ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Scutellaria ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo describe the daily consumption of plain water and beverages of primary and middle school students in four cities of China.

METHODSA total of 5914 students from Beijing, Shanghai, Guangzhou and Chengdu were selected using multiple-stage random sampling method, and 5868 students completed the study from September to October 2011. The information on amounts and types of drinking water was recorded using a 24 hour measurement for seven consecutive days. The amount of plain water and beverages was analyzed for subjects in different gender, grades and cities.

RESULTSThe daily consumption of plain water of subjects was (744 ± 484) ml (68.3% of total drinking water) with statistically significant difference among the Guangzhou, Beijing, Shanghai and Chengdu ((869 ± 528), (818 ± 518), (702 ± 471), and (573 ± 333) ml; F = 113.74, P < 0.05). The amount of plain water in boys (809 ± 534) ml was significantly higher than in girls (683 ± 436) ml (Z = 9.58, P < 0.05) while higher in urban (792 ± 531) ml than in rural (695 ± 427) ml (Z = -6.09, P < 0.05). The consumption of plain water in high school students was the highest (829 ± 513) ml, and that in primary students was the lowest (672 ± 426) ml (F = 55.23, P < 0.05). The average daily consumption of beverages was (345 ± 287) ml (31.7% of total drinking water) and the highest in Shanghai (424 ± 304) ml, then in Beijing (347 ± 303) ml and in Guangzhou (316 ± 267) ml, the lowest in Chengdu (293 ± 255) ml (F = 58.94, P < 0.05). The consumption of beverages for students in urban areas (394 ± 301) ml was higher than that in rural areas (296 ± 264) ml (Z = -14.48, P < 0.05), but no significant difference between boys (348 ± 306) ml and girls (342 ± 269) ml (Z = -1.44, P > 0.05). The consumption of beverages of high school students (356 ± 309) ml and middle school students (360 ± 301) ml were higher than primary school students (328 ± 263) ml (F = 8.37, P < 0.05).

CONCLUSIONThe major drinking water of primary and middle school students in four cities of China was plain water. The amounts of consumption of plain water and beverages varied in different cities, urban and rural and levels of education.

Beverages ; Child ; China ; Diet Surveys ; Drinking ; Drinking Water ; Feeding Behavior ; Female ; Humans ; Male ; Students ; Urban Population

OBJECTIVETo investigate the effect of bone mesenchymal stem cell (BMSC) transplantation on repair of glomerular podocytes and on the Nephrin expression in rats with puromycin aminonucleoside (PAN) -induced nephrosis.

METHODSForty-five Sprague-Dawley rats were randomly divided into three groups (n=15 each): a nephrosis model group that received a single intraperitoneal injection of PAN (0.15 mg/g); a BMSC transplantation group that received a single intraperitoneal injection of PAN (0.15 mg/g) followed by BMSC transfusion; a control group that received a single intraperitoneal injection of normal saline. Ten days after injection, the rats were sacrificed. The 24 hrs urinary protein content and serum albumin and cholesterol levels were measured 24 hrs before sacrifice. Changes of glomerular podocytes were observed under an electron microscope. Brdu labeled positive cells in kidneys were measured by immunohistochemical technology. RT-PCR and Western blot were used to assess the expression of mRNA and protein of Nephrin.

RESULTSIn the nephrosis model group, urinary protein and blood cholesterol contents increased, plasma albumin content decreased compared with those in the control group. Extensive fusion of podocyte foot processes was observed in the nephrosis model group. The BMSC transplantation group had decreased urinary protein and blood cholesterol contents and increased plasma albumin content compared with the nephrosis model group. Fusion of podocyte foot processes was also improved. Brdu labeled positive cells were seen in kidneys in the BMSC transplantation group, but not in the nephrosis model and the control groups. Nephrin mRNA and protein expression decreased significantly in the nephrosis model group compared with that in the control group. The BMSC transplantation group had increased Nephrin mRNA and protein expression compared with the nephrosis model group.

CONCLUSIONSBMSCs can repair glomerular podocytes in PAN-induced nephrosis rats, and the changes of Nephrin expression may be involved in the process.

Animals ; Bromodeoxyuridine ; metabolism ; Kidney ; pathology ; Male ; Membrane Proteins ; genetics ; Mesenchymal Stem Cell Transplantation ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy ; Podocytes ; pathology ; Puromycin Aminonucleoside ; toxicity ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Objective To investigate the clinical features of recurrent epileptic seizures and perioperative management principles and methods of neurosurgical patients with recurrent epileptic seizures.Methods The clinical data,reasons for aggravated seizures,seizure characteristics,and treatment methods and results of 9 patients with recurrent epileptic seizures were analyzed retrospectively.Results Of all the 9 patients,3 were combined with glioma,1 with arachnoid cyst,1 with cavernous hemangioma and 1 with encephalomalacia; epilepsy history was noted in 7 patients; frontal lobe epilepsy was noted in 7,and temporal lobe epilepsy in 2.The reasons for seizure aggravation included drug-decrement(n=3),recent-diagnosed brain tumor(n=2)and surgical operation during intracranial electrode implantation(n=1); and unknown reason was noted in the other 3.Epileptic seizure types included partial seizure and secondary generalized seizure; the frequency of seizure ranged from 3 minutes of interval to several hours of interval.Patients given multiple antiepileptic drugs,including oral and parenteral administration,received good seizure-control; and levetiracetam showed good curative effect during the treatment.Conclusion Recurrent seizures show drug refractory,and is hard to control.The AEDs having good curative effect in partial seizure should be combined used in these patients with higher dosage than conventional initial dose; intravenous and intramuscular administration can be used; the seizures should be controlled as soon as posible.Levetiracetam has good curative effect on patients with recurrent seizures for its fast oral-absorption,rapid onset of action and good antiepileptic effect.

OBJECTIVETo investigate whether the release of nitric oxide (NO) was involved in the cardioprotection of ethanol postconditioning in isolated rat hearts.

METHODSHearts isolated from male SD rats were subjected to 30 min of regional ischemia (occlusion of left anterior descending artery) followed by 120 min of reperfusion. Ethanol postconditioning was fulfilled through perfusion of 50 mmol/L ethanol for 15 min (at the end of cardiac ischemia for 5 min and at the beginning of reperfusion for 10 min). The rats were divided into five groups: normal, ischemia and reperfusion, ethanol postconditioning, ethanol postconditioning + L-nitro-arginine-methylester (L-NAME) and ethanol postconditioning + atractyloside. The ventricular hemodynamic parameters and lactate dehydrogenase (LDH) release during reperfusion were measured. The infarct size was measured by TTC staining method and NO content was measured by nitric acid reductase method. The expressions of Bcl-2 and Bax mRNA were detected by RT-PCR analysis.

RESULTSIn contrast to ischemia and reperfusion, ethanol postconditioning improved left ventricular developed pressure, rate pressure product during reperfusion, reduced LDH release and infarct size. NO content was decreased. The ratio of Bcl-2/Bax was increased. Administration of nitric oxide synthase inhibitor L-NAME or mitochondrial permeability transition pore opener atractyloside both attenuated the role of ethanol postconditioning, which inhibited the recovery of hemodynamic parameters, the decreases of LDH and infarct size. NO content was decreased further. The ratio of Bcl-2/Bax was decreased.

CONCLUSIONThe cardioprotection of ethanol postconditioning may be associated with reducing nitric oxide release, inhibiting the opening of mitochondrial permeability transition pore and decreasing the happening of apoptosis.

Animals ; Ethanol ; therapeutic use ; In Vitro Techniques ; Ischemic Postconditioning ; Male ; Mitochondria, Heart ; metabolism ; Myocardial Ischemia ; metabolism ; prevention & control ; Myocardial Reperfusion Injury ; metabolism ; prevention & control ; Myocardium ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley