It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation ; methods ; Cell Separation ; methods ; Erythrocytes ; immunology ; Humans ; Methylcellulose ; pharmacology

OBJECTIVETo investigate the effects of crypotanshinone (CPT) on the proliferation and apoptosis of DU145 prostate cancer cells as well as on the metadherin expression and the downstream PI3K/AKT signaling pathway in the DU145 cells.

METHODSWe treated DU145 prostate cancer cells with different concentrations of CPT for 24, 48, and 72 hours followed by evaluation of the proliferation and apoptosis of the cells by MTT assay and TUNEL, respectively. We determined the expressions of metadherin protein and mRNA in the DU145 cells by Western blot and RT-PCR respectively at different time points after CPT treatment. We also detected the expressions of the proteins metadherin, AKT, p-AKT, and Bcl-2 in the CPT-treated DU145 cells at 48 hours.

RESULTSCPT significantly inhibited the proliferation of the DU145 cells in a dose- and time-dependent manner (P < 0.05). After treatment with 10 µmol/L CPT for 24, 48, and 72 hours, the apoptosis rates of the DU145 cells were (29.42 ± 4.51), (55.07 ± 5.67) and (70.84 ± 4.66)%, respectively, significantly higher than (3.1 ± 2.48)% in the control group (P < 0.05). The expression of metadherin was remarkably downregulated at the transcription and translation levels (P < 0.05) and the expressions of the AKT signaling pathway and the Bcl-2 protein were markedly inhibited in the DU145 cells after treated with 10 µmol/L CPT for 48 hours (P < 0.05).

CONCLUSIONCPT can inhibit the proliferation and induce the apoptosis of DU145 prostate cancer cells, which may be associated with its suppression of the downstream PI3K/AKT signaling pathway by reducing the expression of metadherin in the DU145 cells.

Apoptosis ; drug effects ; Cell Adhesion Molecules ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Diterpenes, Abietane ; pharmacology ; Down-Regulation ; Drugs, Chinese Herbal ; pharmacology ; Humans ; In Situ Nick-End Labeling ; Male ; Neoplasm Proteins ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Signal Transduction ; drug effects ; Time Factors

OBJECTIVETo research the effects of Guizhi Fuling capsule on sex hormones levels in blood serum and breast issue morphology of hyperplasia of mammary glands model rats.

METHODThe unpregnancy SD rat models of hyperplasia of mammary glands were established by injecting 0.5 mg x kg(-1) benzoate estradiol. After five weeks doses,the effects of Guizhi Fuling capsule 2.0, 1.0, 0.5 g x kg(-1) and Rupixiao tablet 0.5 g x kg(-1) on the changes of papilla diameter, height and breast issue morphology of the naimal models were explored, and sex hormones levels in blood serum were measured.

RESULTGuizhi Fuling capsule can inhibitnipple swell, improve breast tissue morphology pathological profiles of the animal models, and decrease oestradiol (E2) level and increase progesterone (P) level in blood serum.

CONCLUSIONThese results suggested that Guizhi Fuling capsule could, improve mammary gland pathological profiles. Regulating sex hormone levels may be its important mechanism for treatment of hyperplasia of mammary glands.

Animals ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gonadal Steroid Hormones ; blood ; Hyperplasia ; Mammary Glands, Animal ; drug effects ; pathology ; Rats

Objective To observe the seasonal changes in serum levels of interleukin-1 beta (IL-1β), interleukin-6 (IL-6) and melatonin (MT) in Bizheng rat model, and explore the relationship between MT and the pathogenesis of rheumatoid arthritis.
Methods One hundred and sixty Sprague-Dawley rats were randomly divided into four groups in summer (n=80) and winter (n=80) respectively:normal group, collagen-induced arthritis (CIA) model group, operation group, and sham-operation group (n=20 in each group). The CIA model group was injected with collagen emulsion at the base of the tail to induce arthritis. The rats in the operation group received pineal gland resection, and 7 days after the first operation, underwent testectomy or oophorectomy. The rats in the sham-operation group were operated to ligature the sagittal sinus, without extracting the pineal gland. After the operations, the operation group and the sham-operation group both were immunized as the CIA group was.The serum levels of IL-1β, IL-6 and MT in different groups were measured by radioimmunoassay.
Results Compared with the normal group, the serum levels of IL-1βand IL-6 increased in the CIA model, operation, and sham-operation groups both in summer and in winter (IL-1βin summer, P=0.008, P<0.01, P=0.012; IL-1β in winter, P=0.019, P<0.01, P=0.027; IL-6 in summer, P=0.028, P<0.01, P=0.024;IL-6 in winter, P=0.006, P<0.01, P=0.008). In the operation group, the serum levels of IL-1βand IL-6 in winter were higher than in summer, but with no statistically significant differences (P=0.844, 0.679). Compared with the normal group, the serum level of MT significantly increased in summer and winter in both the CIA model group (P=0.002, 0.008) and the sham-operation group (P=0.003, 0.007), while significantly decreased in the operation group (P=0.023, 0.003). There was no significant difference in MT level in the operation group between summer and winter (P=0.947).
Conclusions The increase of serum levels of IL-1β and IL-6 may exacerbate the inflammatory reaction and cause a more severe condition in the rheumatoid arthritis. The concentrations of IL-1β, IL-6, and MT correspond with the change of seasons, confirming that there are connections between nature and human body.

Objective To investigate the effect of movie enjoyment on emotion of male patients with chronic schizophrenia.Methods A total of 62 standard patients with chronic schizophrenia from September to November 2011were selected and randomly divided into the experimental group(30) and the control group(32) by using a random number table.Patients in experimental group received movie enjoyment and routine rehabilitation project,while those in the control group received only routine rehabilitation project.Before the intervention and 12 weeks after the intervention,the patients were investigated with the Positive and Negative Affect Scale for efficacy assessments.Results After the nursing intervention,the scores of positive and negative affect were (30.79 ± 7.54) and (18.07 ± 5.97) respectively in the experimental group in contrast to (22.29 ± 8.14) and (18.00 ± 7.79) before the intervention,the difference was statistically significant (t =3.97,4.85,respectively;P ＜0.01).After the nursing intervention,the scores of positive and negative affect were(21.41 ±7.40) and (17.72 ± 7.03)respectively in the control group while were (23.25 ± 6.37)and (16.50 ± 6.08) before the intervention,the difference was statistically significant (t =2.67,1.98,respectively; P ＜ 0.05).There was statistically significant between the two groups in the scores of positive and negative affect after intervention(t =5.69,6.78 ;P ＜ 0.01).Conclusions The movie enjoyment can help improve positive emotions affect of patients with chronic schizophrenia.

OBJECTIVETo investigate the distribution of γ-glutamyl hydrolase gene (GGH) 452C/T genotype and allele frequency in children with acute leukemia (AL) and healthy children.

METHODSBone marrow samples from 92 children with AL and peripheral blood samples from 124 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for a GGH 452C/T polymorphism by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis (RT-PCR-DGGE) and direct sequencing.

RESULTSThe frequencies of the AL patients with TT, CT and CC genotypes were 2.2%, 13.0% and 84.8%, and the frequencies of the control children were 1.6%, 16.9% and 81.5%, respectively. There was no significant difference in GGH genotype or T allele frequency between the two groups (P> 0.05). However, the T allele frequency in Han Chinese children was significantly different from those reported in Japanese, Mexican and African-American populations.

CONCLUSIONThe frequency of 452C/T polymorphism of GGH gene in Han Chinese children has been determined. The results suggested that an ethnic difference may exist.

Acute Disease ; Base Sequence ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genotype ; Humans ; Infant ; Infant, Newborn ; Leukemia ; enzymology ; genetics ; Male ; Polymorphism, Single Nucleotide ; gamma-Glutamyl Hydrolase ; genetics

To clarify the important functional residues in the active site of N-myristoyltransferase (NMT), a novel antifungal drug target, and to guide the design of specific inhibitors, multiple sequence alignments were performed on the NMT family and thus evolutionary trace was constructed. The important functional residues in myristoyl CoA binding site, catalytic center and inhibitor binding site of NMT family were identified by ET analysis. The trace residues were mapped onto the active site of CaNMT. Trpl26, Asn175 and Thr211 are highly conserved trace residues and do not interact with current NMT inhibitors, which are potential novel drug binding sites for the novel inhibitor design. Pro338, Leu350, Ile352 and Ala353 are class-specific trace residues, which are important for the optimization of current NMT inhibitors. The trace residues identified by ET analysis are of great importance to study the structure-function relationship and also to guide the design of specific inhibitors.
Acyl Coenzyme A ; metabolism ; Acyltransferases ; chemistry ; genetics ; metabolism ; Amino Acid Sequence ; Animals ; Binding Sites ; Conserved Sequence ; Enzyme Inhibitors ; chemistry ; pharmacology ; Evolution, Molecular ; Humans ; Imidazoles ; chemistry ; pharmacology ; Models, Molecular ; Molecular Sequence Data ; Oligopeptides ; chemistry ; pharmacology ; Phylogeny ; Protein Structure, Tertiary ; Sequence Homology, Amino Acid

OBJECTIVETo assess whether polymorphisms of methylenetetrahydrofolate reductase (MTHFR) gene is associated with susceptibility to acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML) in Chinese Han children.

METHODSThe study has included 87 patients with ALL, 22 patients with AML and 120 healthy controls. All subjects were analyzed with reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis and sequencing.

RESULTSA 677CT genotype of the MTHFR gene was associated with decreased risk of ALL (OR=0.23, 95%CI: 0.07-0.79). However, MTHFR A1298C genotypes were not associated with the risk of either disease. 677TT/1298AA and 677CC/1298AC genotypes were associated with increased risk of ALL(OR=3.78, 95% CI: 1.38-10.40; OR=3.17, 95% CI: 1.18-8.53, respectively), whereas the genotype 677CT/1298AA was associated with susceptibility to AML (OR=0.23, 95% CI: 0.06-0.97).

CONCLUSIONOur data suggested that C677T polymorphism of MTHFR gene may increase the risk of childhood AML.

Acute Disease ; Base Sequence ; Child ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Leukemia ; diagnosis ; enzymology ; genetics ; Male ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Molecular Sequence Data ; Polymorphism, Single Nucleotide

This study was aimed to clone the gene coding mouse CXC chemokine receptor 4 (CXCR4), to construct the recombinant lentiviral vector carrying enhanced green fluorescence protein (EGFP) and to explore its expression in eukaryotic cells (293FT cells). The full length CXCR4 gene was cloned by RT-PCR using bone marrow cells from C57BL/6 mouse as template and inserted into PCR-Blunt vector. CXCR4 fragment was generated by digestion with restriction endonuclease and subcloned into a lentiviral vector to generate recombinant lentiviral vector LV-IRES-EGFP-CXCR4, which was co-transfected into 293FT cells together with envelope plasmid and packaging plasmid by lipofectamine 2000. Viruses were gathered and concentrated using ultracentrifuge, and then transfected into 293FT cells. Expression of EGFP was detected by fluorescent microscopy and flow cytometry (FCM). And the expression of CXCR4 protein was detected by Western blot. The results demonstrated that mouse CXCR4 gene was cloned and the lentiviral vector was successfully constructed. The lentiviral particles were correctly packaged, and the virus titers were above 10(8) TU/ml in the supernatant after concentration. Expression of EGFP was detected by fluorescent microscopy in the transfected 293FT cells, and the transfection efficacy > 95% was determined by FCM. Expression of CXCR4 protein detected by FCM and Western blot was significantly higher than that in control group. It is concluded that the CXCR4 gene along with the gene coding EGFP is successfully inserted into a lentiviral vector to construct a recombinant lentiviral vector, which can be expressed in eukaryotic cells.
Animals ; Cell Line ; Gene Expression ; Genetic Vectors ; Green Fluorescent Proteins ; genetics ; Humans ; Lentivirus ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; Receptors, CXCR4 ; genetics ; Transfection

BACKGROUNDInflammation plays a pivotal role in cardiac remodeling, especially in myocardial fibrosis. Abnormal growth of cardiac fibroblasts is critically involved in the pathophysiology of cardiac hypertrophy/remodeling. Previous study has demonstrated that many inflammation stimulating factors trigger transforming growth factor-β (TGF-β) induction and reactive myocardial fibrosis. Activin A (ACT A) is a member of TGF-β superfamily, and follistatin (FS) is an activin-binding protein, i.e. an antagonist of ACT A. Our previous studies have shown that ACT A-FS imbalance occurs in rats with heart failure (HF), and overexpression of ACT A can lead to ventricular remodeling, and resultant HF. Low expression of FS after myocardial infarction further exacerbated HF. The pathogenic change resulting from overexpression of ACT A is consistent with that of overexpression of angiotensin II (AngII). Ventricular remodeling includes cardiocyte remodeling and myocardial interstitial collagen deposition and fibrosis. Therefore, the present study was designed to investigate the effects of inflammatory factors on the ACT A-FS and the secretions of cardiac fibroblasts in order to explore in depth the mechanism of myocardial fibrosis.

METHODSA rat model with HF was established, and the results showed that there was a greater degree of cardiac fibrosis in HF rats. In addition, we found that there was an imbalance of the ACT A/FS system in HF rats, which was characterized by increased levels of ACT A. Further, primary rat cardiac fibroblasts were cultured and the MTT assay was performed to determine the effect of the inflammatory factor-bacterial endotoxin lipopolysaccharide (LPS) on cardiac fibroblast proliferation.

RESULTSThe results showed that LPS can stimulate the cardiac fibroblasts to proliferate in a dose-dependent manner. Cellular immunohistochemical staining showed that the rat cardiac fibroblasts themselves could express ACT A and FS proteins, and stimulation by LPS could apparently promote the cultured primary rat cardiac fibroblasts to secrete ACT A, but inhibit the secretion of FS. The results also showed that ACT A promoted, in a dose-dependent manner, the proliferation of the cultured primary rat cardiac fibroblasts, and the expression of collagen types I and III. Moreover, ACT A promoted, in a dose dependent manner, the cardiac fibroblasts to secrete nitric oxide (NO), and unregulated the expression of inducible nitric oxide synthase (iNOS) mRNA.

CONCLUSIONSThese results suggest that the inflammatory mediator LPS can promote ACT A-FS imbalance in cardiac fibroblasts, mainly overexpression of ACT A. Overexpression of ACT A promotes the proliferation and the secretion of collagens in cardiac fibroblasts through autocrine/paracrine stimulation of NO, and is involved in the pathological process of myocardial fibrosis.

Activins ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Fibroblasts ; cytology ; drug effects ; Follistatin ; genetics ; metabolism ; Immunohistochemistry ; Lipopolysaccharides ; pharmacology ; Myocardium ; cytology ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Real-Time Polymerase Chain Reaction ; Ventricular Remodeling ; drug effects