Aim To study on the diuretic effect of a phenylphthalazines compound PU1424 and its influence on electrolyte balance, glucose and lipid metabolism, hepatic and renal functions.Methods Male Sprague Dawley rats were randomly divided into solvent control group,PU1424 treated group and HCTZ treated group.Urine was collected per 6 h and blood samples were collected at the end of drug administration.Urinary osmolality was measured by Freezing Point Osmometer;urea concentration was measured by Urea Detection Kit;ion level, blood glucose level, blood lipid level, hepatic and renal function were analyzed by Automatic Biochemical Analyzer.Results Compared to the solvent control group, and urine output of rats treated with PU1424 and HCTZ was increased as 1.52 times and 1.78 times and water intake increased as 1.42 times and 1.56 times respectively.Urine osmolalities were decreased as 61.5% and 50.4% of the control group, and urine urea concentration was decreased as 57.1% and 56.8% of the control group.Urinary electrolytes were decreased by administration of PU1424 and HCTZ compared to the intact plasma electrolytes.The blood glucose levels and blood lipid levels of rats treated with PU1424 had no changes, while the blood glucose and total cholesterol were increased by administration of HCTZ.The urea nitrogen, creatinine, alkaline phosphatase and total protein were intact by administration of PU1424 and HCTZ except the alanine/straw ration increased by HCTZ.Conclusion New diuretic candidate compound PU1424 displays significant diuretic effect with electrolyte balance, blood glucose level, blood lipid level, hepatic and renal function intact.

Aim To investigate the effect of ribonucleic acidⅡon apoptosis in human leukemia cell lines K562 and KG1 a.Methods Cell counting kit-8(CCK-8)as-say was performed to detect proliferation activity of K562 and KG1 a cells treated with ribonucleic acidⅡ. Apoptosis index was assessed by flow cytometry(FCM) and fluorescent Hoechst 33258 staining was used for observing morphologic changes of apoptosis.Expres-sion levels of p53,Bax,Bcl-2 and cleaved caspase-3 were analyzed by Western blot.Results The prolifera-tion of K562 and KG1 a cells was significantly inhibited by ribonucleic acid Ⅱ treatment for 12 h,24 h,48 h at concentrations of 100~300 mg·L-1 ,which indica-ted the inhibitory effect of ribonucleic acid Ⅱ was in dose-dependent and time-dependent manners.FCM re-sults displayed a dose-dependent increase in cell apop-totic rate.Hoechst 33258 staining showed the typical apoptotic morphology in some leukemic cells treated with ribonucleic acid Ⅱ,including increased nuclear chromatin concentration and edge accumulation.West-ern blot analysis showed the increased expression of p53,Bax,cleaved caspase-3 and decreased expression of Bcl-2 in K562 and KG1 a cells treated with ribonu-cleic acid Ⅱ.Conclusions Ribonucleic acid Ⅱ can induce apoptosis of leukemia K562 and KG1 a cells by up-regulating p53,which mediates Bcl-2/Bax balance and activates caspase-3 .

Cardiac Catheterization ; Heart Septal Defects, Ventricular ; therapy ; Humans ; Prosthesis Implantation ; instrumentation ; Septal Occluder Device

OBJECTIVETo explore the role of stem cell mobilization on regeneration of partially grafted livers.

METHODSRats models with cross-sex 50% PLTx (partial liver transplantation) were established. The rats were divided into three groups: PLTx, WLTx (whole liver transplantation) and sham operation groups. Bone marrow and liver samples were collected on days 1, 3, 5, 7 postoperatively (each n = 6). The quantitative variations of the cells with stem cell markers in the bone marrow, including beta2m-/Thy-1.1+, CD45+/CD34+, Flt2/3+ and c-kit+ markers, were detected using flow cytometry. Sry gene positive cells in donor livers were detected by fluorescent in situ hybridization (FISH), and the expressions of CD34, c-kit and Thy-1.1 were detected by immunohistochemistry technique.

RESULTSCompared with the WLTx and sham operation groups, beta2m-/Thy-1.1+, CD45+/CD34+ cells in bone marrows in the PLTx group increased on the first postoperative day and decreased on the following days. The CD34, c-kit and Thy-1.1 positive cells detected in portal tract areas peaked during the 3-5 postoperative days. CD34+/CD45+ positive cells could be detected. The expressions of CD34, c-kit and Thy-1.1 positive cells were rare in the WLTx and sham operation groups. Sry+ cells could be detected in portal tract areas and few Sry+/CD34+ and Sry+/Thy-1.1+cells were detected.

CONCLUSIONIn the PLTx group, the stem cells in the bone marrow were mobilized and stem cells in the liver were activated.

Animals ; Antigens, CD34 ; immunology ; Bone Marrow Cells ; cytology ; immunology ; Female ; Hematopoietic Stem Cell Mobilization ; Leukocyte Common Antigens ; immunology ; Liver Transplantation ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; immunology ; Thy-1 Antigens ; immunology

Antibody-drug conjugate (ADC) is a new class of therapeutics composed of a monoclonal antibody and small cytotoxin moieties conjugated through a chemical linker. ADC molecules bind to the target antigens expressed on the tumor cell surfaces guided by the monoclonal antibody component. The binding ADC molecules can be internalized and subsequently the toxin moieties can be released within the tumor cells via chemical and/or enzymatic reactions to kill the target cells. The conjugation combines the merits of both components, i.e., the high target specificity of the monoclonal antibody and the highly potent cell killing activity of the cytotoxin moieties. However, such complexities make the pharmacokinetic and metabolic studies of ADCs highly challenging. The major challenges should include characterization of absorption, distribution, metabolism and excretion, investigation of underlying mechanisms, assessment of pharmacokinetic- pharmacodynamic relationship, and analytical method development of ADC drugs. This review will discuss common pharmacokinetic issues and considerations, as well as tools and strategies that can be utilized to characterize the pharmacokinetic and metabolic properties of ADCs.
Antibodies, Monoclonal ; pharmacokinetics ; Cytotoxins ; pharmacokinetics ; Humans ; Immunoconjugates ; pharmacokinetics ; Neoplasms ; drug therapy

AIM:To study the influence of bone marrow mesenchymol stem cell-drived exosomes(BMSC-exo-somes)on hindlimb activity,and the numbers of reactive astrocytes and residual neurons in spinal cord injury(SCI)rats. METHODS:BMSCs were cultured using the whole bone marrow adherent culture method and surface markers CD 90 and CD34 were verified by flow cytometry.Exosomes were isolated by ultracentrifugation and the morphology of exosomes was observed under transmission electron microscope.The protein markers CD63 and CD9 were verified by Western blot.After exosomes were applied to SCI rats,the Basso,Beattie and Bresnahan locomotor rating scale score,the Nissl staining of the lesion site,and the numbers of reactive astrocytes and residual neurons were assessed at various time points.RESULTS:Transmission electron microscopic observation revealed the presence of saucer -shaped vesicles.BMSC-exosomes were found to express high levels of CD63 and CD9.Compared with injury group,significant improvement of hindlimb activity scores from day 14 after injury in treatment group was observed(P<0.05),and less reactive astrocytes and more residual neu-rons from day 7 after injury were also observed(P<0.05).CONCLUSION:BMSC-exosomes inhibit reactive astrocytes and death of neurons,and improve hindlimb activity in the rats after SCI.

OBJECTIVETo prepare cytochrome (CYP)2E1-silenced hepatocytes by lentivirus-mediated RNA interference technology and to investigate the hepatotoxicity of trichloroethylene (TCE) in CYP2E1-silenced hepatocytes.

METHODSShort hairpin RNA fragments were designed and synthesized and were then ligated into the lentiviral vector; single colonies were screened; the plasmid was extracted after PCR and sequence identification and then transferred into L02 hepatocytes; the CYP2E1-silenced hepatocytes were selected; real-time quantitative PCR and Western blot were used to evaluate the interference effects. The obtained CYP2E1-silenced hepatocytes, as well as normal L02 hepatocytes, were treated with TCE (0, 0.25, 0.50, 1.00, 2.00, and 4.00 mmol/L). The cell viability and half maximal inhibitory concentration (IC50) of TCE were measured; the apoptotic rate of cells was measured by flow cytometry; the mRNA expression levels of apoptosis genes and oncogenes were measured by real-time quantitative PCR.

RESULTSThe IC50s of TCE for L02 hepatocytes and CYP2E1-silenced hepatocytes were 15.1 mmol/L and 23.6 mmol/L, respectively. The apoptotic rate increased as the dose of TCE rose in the two types of cells; the CYP2E1-silenced hepatocytes hada significantly lower apoptotic rate than L02 hepatocytes when they were exposed to 2.0 and 4.0 mmol/L TCE (P < 0.05 or P < 0.01). The mRNA expression level of bcl-2 (anti-apoptosis gene) in CYP2E1-silenced hepatocytes was 15% ∼ 60% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of caspase-3 and caspase-9 (apoptosis genes) in CYP2E1-silenced hepatocytes were 30% ∼ 60% lower than those in L02 hepatocytes (P < 0.01). The mRNA expression level of p53 (cancer suppressor gene) in CYP2E1-silenced hepatocytes was 81 - 278% higher than that in L02 hepatocytes (P < 0.01), while the mRNA expression levels of c-fos and k-ras (oncogenes) in CYP2E1-silenced hepatocytes were 20-68% lower than those in L02 hepatocytes (P < 0.01).

CONCLUSIONCYP2E1-silenced cells can be successfully prepared by lentivirus-mediated RNA interference technology. Silencing CYP2E1 gene can reduce the hepatotoxicity of TCE and inhibit the expression of some apoptosis genes and oncogenes, suggesting that CYP2E1 gene plays an important role in TCE metabolism and is related to the hepatotoxicity of TCE.

Apoptosis ; drug effects ; genetics ; Cell Line ; Cell Survival ; drug effects ; genetics ; Cytochrome P-450 CYP2E1 ; genetics ; metabolism ; Genetic Vectors ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lentivirus ; genetics ; RNA Interference ; Trichloroethylene ; toxicity

OBJECTIVETo identify the survival prognostic factors and clinical outcome of the patients with spinal metastatic tumors and to discuss the surgical treatment strategy of spinal metastatic tumors.

METHODSThe patients with spinal metastatic tumors who received surgeries during January 2003 to June 2012 were enrolled. The survival was analyzed by Kaplan-Meier survival curve. The prognostic factors, divided into patient-related factors, tumor-related factors and therapy-related factors, were analyzed univariately and multivariately by Cox comparative hazard model.

RESULTSThere were 453 patients were enrolled in research including 263 male and 190 female patients with an average age of (56 ± 13) years (10-86 years). The median postoperative survival was 9 months. Local recurrences and peri-operative complications were found in 78 (17.2%) and 72 (15.9%) patients, respectively. Univariate analysis showed the significant prognostic factors for postoperative survival included poor preoperative general condition (χ(2) = 4.16), severe preoperative neurologic deficit(χ(2) = 10.23), not receiving bisphosphonate therapy(χ(2) = 10.47), short disease-free interval before spinal metastasis (χ(2) = 23.31), spinal metastasis as the first manifestation (χ(2) = 10.94), rapid-growth primary tumor(χ(2) = 15.45), visceral metastasis (χ(2) = 4.10), not receiving postoperative radiotherapy(χ(2) = 18.10) and not receiving post-operative sensitive systemic therapy(χ(2) = 11.20) (P < 0.05). Multivariate analysis showed the independent prognostic factors include severe preoperative neurologic deficit (P = 0.012, 95%CI: 1.11-2.30), short disease-free interval before spinal metastasis (P = 0.023, 95%CI:1.05-1.83), rapid-growth primary tumor (P = 0.000, 95%CI:1.74-3.06), visceral metastasis (P = 0.008, 95%CI: 1.08-1.68), not receiving postoperative radiotherapy (P = 0.000, 95%CI:1.38-2.35) and not receiving post-operative sensitive systemic therapy (P = 0.045, 95%CI:1.01-1.58).

CONCLUSIONThe prognostic factors for survival are useful for determining the indication of operation and improving survival and clinical outcome for patients with spinal metastatic tumors.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Spinal Neoplasms ; diagnosis ; secondary ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo identify the differentially expressed genes associated with hypersplenism in patients with portal hypertension.

METHODSThe total RNA were extracted from the macrophages isolated from normal spleen and the spleen of patients with portal hypertension and reversely transcribed to cDNA with the incorporation of fluorescent (cy3 and cy5)-labeled dCTP to prepare the hybridization probes. After hybridization of Biostar-H140s chip containing 14,112 spots of cDNAs with the prepared probes, the gene chip was scanned for fluorescence intensity to screen the differently expressed genes. Three gene chips were used for hybridization and only the genes with differential expression in all the three chips were considered to associate with hypersplenism in patients with portal hypertension.

RESULTSTotaling 896, 1330 and 898 genes were identified to be differentially expressed by the three chips, respectively, and 121 genes (0.86%) showed differential expression in all the three chips, including 21 up-regulated known genes and 73 down-regulated known genes. The differently expressed genes were functionally related with ion channels and transport proteins, cyclins, cytoskeleton, cell receptors, cell signal transduction, metabolism, immunity, and so forth. These genes might be involved in hypersplenism in the condition of portal hypertension.

CONCLUSIONcDNA microarray-based screening of differentially expressed genes in the macrophages in the spleen may provide new insights into the pathogenesis of hypersplenism in patients with portal hypertension.

Female ; Gene Expression Profiling ; Humans ; Hypersplenism ; etiology ; genetics ; Hypertension, Portal ; complications ; genetics ; Macrophages ; metabolism ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Spleen ; metabolism ; pathology

OBJECTIVETo explore Chinese medicine syndrome distribution laws of asymptomatic HIV infection patients.

METHODSUsing Chi-square test, Chinese medicine syndrome distribution laws were compared and analyzed in 1 156 asymptomatic HIV infection patients from March 2009 to October 2011 from four aspects, i.e., age, possible infection time, disease duration, and different routes of infection.

RESULTSQi deficiency syndrome (QDS) and internal dampness-heat accumulation syndrome (IDHAS) were dominant in all syndrome types. Along with aging, QDS showed a growing tendency, while IDHAS showed obvious declining tendency. There was no obvious change in other syndrome types. There was statistical difference in the distribution of each syndrome type among each age period (P < 0.01). Within 15 years, along with the increase of infection time, QDS showed a growing tendency, while IDHAS ratio showed an obvious declining tendency. No obvious laws were found in other syndrome types. There was statistical difference in the distribution of each syndrome type (P < 0.01). Along with the prolongation of disease duration, the case number of each syndrome showed a decreasing trend, but QDS and IDHAS still accounted for higher ratios in each stage. There was statistical difference in the distribution of each syndrome type (P < 0.01). As for infection routes, QDS was predominant in paid blood donation, blood transfusion infection, intravenous drugs. IDHAS was predominant in sexual transmit. No obvious laws were found in other syndrome types. There was statistical difference in the distribution of each syndrome type (P < 0.01).

CONCLUSIONSDIS, IDHAS, and no confirmable syndrome typing were dominant in asymptomatic HIV infection patients. Deficiency and dampness were important pathological factors for them.

Adolescent ; Adult ; Aged ; HIV Infections ; diagnosis ; Humans ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Young Adult