MLN4924 can inhibit the proliferation,invasion and metastasis of tumor by inducing tumor cells apoptosis,senescence and autophagy,which can inhibit tumor angiogenesis and enhance the sensitivity of radiotherapy and chemotherapy.Therefore,MLN4924 plays a good anti-tumor effect.

OBJECTIVEThe cytochrome P450 subfamily IIIA5 (CYP3A5) gene is responsible for the metabolism of many clinically used anticancer agents. So far the studies on CYP3A5 gene has only been focused on the leukemia cell lines. This study examined the polymorphism of CYP3A5 and tried to find the possible relationship between CYP3A5 gene expression and treatment outcome or prognosis in children with acute leukemia.

METHODSThe genotype distribution of CYP3A5-6986A/G gene polymorphism was detected with the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 66 children with newly diagnosed acute leukemia (AL) and 22 control individuals. Quantitative real-time RT-PCR was used to examine wt-CYP3A5 and SV1-CYP3A5 mRNA levels in the bone marrow.

RESULTSThree genotypes of CYP3A5-6986A/G polymorphisms were found: CYP3A5*1/*1, CYP3A5*1/*3 and CYP3A5*3/*3. There were significant differences in the wt-CYP3A5 mRNA expression among the AL patients with different genotypes (p<0.05). In patients with acute lymphocytic leukaemia (ALL), the complete remission (CR) rate in the group with a low expression of wt-CYP3A5 mRNA was significantly higher than that in the group with a high expression (p<0.05). A dynamic monitoring for wt-CYP3A5 mRNA expression was performed in two cases of ALL. The expression increased before ALL relapse compared with that in CR in a patient, while in the other patient, the expression was kept in a low level and the patient remained in CR CONCLUSIONS: wt-CYP3A5 mRNA expression was associated with the treatment outcome and prognosis in children with AL. Dynamic monitoring for wt-CYP3A5 mRNA expression in the bone marrow may be useful in the evaluation of the disease severity in childhood acute leukemia.

Acute Disease ; Child ; Cytochrome P-450 CYP3A ; genetics ; Genotype ; Humans ; Leukemia ; enzymology ; genetics ; Polymerase Chain Reaction ; RNA, Messenger ; analysis

Chronic myeloid leukemia (CML) is a malignant clonal disease derived from hematopoietic stem cells. CML stem cells were thought to be the root which could lead disease development and ultimately rapid change. However, a stable animal model for studying the characteristics of CML stem cells is currently lacking. This study was aimed to establish a transplanted human CML nude-mice model to further explore the biological behavior of CML stem cells in vivo, and to enrich CML stem cells in nude mice by series transplantation. The 4 - 6 weeks old BALB/c nude mice pretreated by splenectomy (S), cytoxan intraperitoneal injection (C) and sublethal irradiation (I) were transplanted intravenously with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase. Alternatively, 4 - 6 weeks old BALB/c nude mice pretreated by lethal irradiation were transplanted intravenously with 5 × 10(6) homologous bone marrow cells of BALB/c nude mice together with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase simultaneously. The leukemic cells engrafted and infiltrated in organs and bone marrow of the mice were tracked by reverse transcription-polymerase chain reaction (RT-PCR), plastic-embedded biopsy and flow cytometry. The results of these two methods were compared. The results showed that human CML cells engrafted and infiltrating into the bone marrow of two nude mice pretreated with SCI could be detected. In spite of the low successful rate, results suggested the feasibility of this method by using BALB/c nude mice as a human CML animal model. In contrast, in nude mice pretreated by the lethal dose irradiation, CML cells in the bone marrow could not be found. It is concluded that human bone marrow CML cells can results in leukemia in nude mice pretreated by SCI. Thus this study provides a new strategy for establishment of CML animal models which deserves further elaboration.
Animals ; Disease Models, Animal ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Male ; Mice ; Mice, Nude ; Mice, SCID ; Neoplasm Transplantation ; Neoplastic Stem Cells ; Transplantation, Heterologous

Objective: To investigate the clinical characteristics and risk factors of congenital choledochal cysts (CCC). Methods: This retrospective study recruited 52 cases who were antenatally diagnosed with CCC and underwent surgical treatment after birth in Guangdong Women and Children Hospital from January 2013 to August 2018, with complete clinical data. According to the enlargement of cysts during pregnancy, they were divided into two groups: progressive group (≥15 mm, 22) and stable group (<15 mm, 30). Antenatal and postpartum ultrasound and MRI features of the two groups were analyzed. Clinical manifestations and biochemical examination results before and after operation were compared between the two groups. Other data, including amylase level in cyst fluid during operation, cholangiography findings, liver biopsy results, and post-operation follow-up, were also analyzed. Chi-square test, t (t') test, and Pearson correlations tests were performed for data analysis. Results: (1) The average age of the 52 patients at operation was 46(7-822) d. The cysts of all cases were first detected during 19-21 weeks of gestation. The maximum diameter of the cyst in the progressive group was larger than that in the stable group after 34 weeks of pregnancy [31-34 weeks: (31.1±8.4) vs (23.1±6.6) mm, t=3.911; >34 weeks: (36.1±6.8) vs (27.1±7.3) mm, t=4.557; pre-operation: (51.8±18.0) vs (34.0±15.6) mm, t=3.809; all P<0.01]. (2) In the progressive group, the cysts were irregular in shape and enlarged after birth. The common hepatic duct and intrahepatic bile duct were dilated and gradually distended after birth, while the distal end of the common bile duct was narrowed, thus to form a cone-like duct. Deposits could be seen inside the cysts after delivery. Irregular cysts were also presented in the stable group, and five of them had dilatation of common hepatic duct and intrahepatic bile duct after birth. However, no cone-like formation was seen, the distal end of the common bile duct was visible, and deposits in cysts were occasionally found. (3) Twenty-five patients underwent laparotomy, and seven of them showed increased amylase level in cyst fluid including four with 2-5 times above the upper limit of normal value (one in the progressive group and three in the stable group). The other three cases were all in the stable group and their amylase levels in cyst fluid were more than ten times of the upper limit. The level of direct bilirubin in the progressive group was higher than that in the stable group before the operation [18.40(2.50-113.30) vs 8.70(0.00-16.80) μmol/L, u=2.400, P<0.05]. (4) Among the 52 cases, patients with type Ⅰ, Ⅳ and Ⅴ cyst accounted for 71.1% (37/52), 26.9% (14/52) and 2.0% (1/52), respectively. All cases were followed up regularly six months to one year after the operation. Liver function and bilirubin became normal and the growth and development of the babies were similar to those of the same age. (5) Different degrees of liver fibrosis and inflammation were shown in 46(88.5%) cases and more severe in older babies among those who underwent surgery in the progressive group. The time at operation was not associated with the severity of liver fibrosis and inflammation in the stable group. Hepatic fibrosis and inflammation were more serious in the progressive group than in the stable group (fibrosis grading: χ²=14.260, P=0.006; inflammatory activity grading: χ²=9.904, P=0.019). Conclusions Larger diameter (≥30 mm) in the initial prenatal examination or a significant increase in cystic diameter (≥15 mm) during pregnancy are risk factors for early stenosis or occlusion in the distal end of common bile duct requiring close follow-up after birth. When jaundice or abnormal liver function occur and stool color becomes light, early surgical treatment (one to two months after birth, generally within three months) for CCC is recommended to rule out the anomalous union of the pancreaticobiliary duct and hepatic disorders, as well as the cystic biliary atresia.

OBJECTIVETo explore the effects of anti-CD44 monoclonal antibody-IM7 on the in vitro adhesion and migration of chronic myeloid leukemia stem cell (CML-LSC) and its mechanism.

METHODSCD34(+)CD38(-)CD123(+) leukemic stem cells (LSC) from 20 newly-diagnosed chronic myeloid leukemia (CML) patients BM cells and CD34(+)CD38(-) hematopoietic stem cells (HSC) from 20 full-term newborn cord blood cells were isolated with EasySep(TM) magnet beads. The CD44 expression of the LSC and HSC was detected by flow cytometry (FCM), and the adhesion and migration ability of the LSC and HSC pre- and post-incubated with IM7 in vitro by MTT assay and transendothelial migration assay, respectively.

RESULTS(1) After incubated with IM7, the LSC and HSC CD44 expression rates were (86.60 ± 2.10)% vs. (25.40 ± 1.70)% (P < 0.05), respectively. (2) The adhesive ability of the LSC to endothelial cells was decreased markedly after incubated with IM7, the OD value (A(570)) changing from pre-incubation of (0.62 ± 0.11) to post-incubation of (0.34 ± 0.07), while there was little change of A(570) in the HSC group. (3) The migration ability of the LSC group was inhibited evidently after incubated with IM7, the inhibition rate being 46% ∼ 63%, while little change of that in HSC group was detected. (4) The adhesive ability of the LSC group to marrow stromal cells was decreased markedly after incubated with IM7, while little change was found in that of HSC group.

CONCLUSIONThe anti-CD44 monoclonal antibody-IM7 can effectively inhibit the adhesion and migration abilities of the LSC in vitro, which might provide a theoretical evidence for targeting therapy.

Antibodies, Monoclonal ; pharmacology ; Antigens, CD34 ; metabolism ; Bone Marrow ; drug effects ; Flow Cytometry ; Hematopoietic Stem Cells ; drug effects ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; metabolism

This study was purposed to investigate the short-term effects of citrate administration on bone metabolism in the healthy blood donor volunteers. A crossover, placebo-controlled trial were conducted on 22 healthy blood donor volunteers. The volunteers received either a standardized infusion of citrate at 1.5 mg/(kg.min) or the equal volume of placebo normal saline, were washout for 2-3 weeks. During washout serial blood samples were collected and analyzed for bone biochemical markers and electrolytes, such as bone formation marker osteocalcin (OC), bone resorption marker carboxyterminal telopeptide of type I collagen (CTX), intact parathyroid hormone ((i)PTH), ionized calcium ((i)Ca(2+)) and phosphorus (P(i)). Serial urine samples were collected and analyzed for Ca(2+), P(i) and creatinine concentration. The results showed that compared with placebo group, infusion of citrate increased serum levels of OC and CTX (p < 0.0001). The greatest increase of OC and CTX levels occurred at the completion of the intervention. The increment of CTX was higher than OC (p = 0.02), and the OC/CTX ratio decreased (p < 0.01). Infusion of citrate also induced profound increase in serum (i)PTH level (p < 0.0001) and urinary calcium excretion (p < 0.0001), and decrease in serum (i)Ca(2+) (p < 0.0001) and P(i) (p < 0.01) levels. The decrease of (i)Ca(2+) level in female was higher than that in male (p = 0.007), but the changes of (i)PTH, OC, and CTX levels showed no differences between female and male. Changes of OC and CTX levels were closely related to each other (r = 0.56, p < 0.0001) and changes of both markers were negatively correlated with the change of serum (i)Ca(2+) concentration during the citrate intervention(r(OC) = -0.44, r(CTX) = -0.44, p < 0.0001). Increased levels of (i)PTH showed positively correlation with OC (r = 0.34, p = 0.02) and borderline correlation with CTX (r = 0.29, p = 0.06) in male. No such relationship was observed in female. All bone markers and electrolyte levels returned to baseline within 24 hours. It is concluded that the citrate load at the dose as a single platelet apheresis results in profound increase of bone turnover, which is characterized by a short-term increase of bone resorption and excretion of calcium. The possible effect of citrate on bone mass of long-term frequent platelet apheresis donor is worth concerning.
Adult ; Blood Donors ; Bone Remodeling ; drug effects ; Bone and Bones ; drug effects ; metabolism ; Citric Acid ; pharmacology ; Cross-Over Studies ; Female ; Humans ; Male ; Osteocalcin ; blood

The aim of this study was to investigate the apoptosis-inducing effect of anti-CD44 monoclonal antibody IM7 on chronic myeloid leukemia (CML) stem/progenitor cells in vitro and to explore its possible mechanism. Leukemic stem/progenitor cells (LSPCs) expressing CD34(+), CD38(-) and CD123(+) were isolated from bone marrow (BM) cells of 20 patients with newly-diagnosed chronic myeloid leukemia by using EasySep(TM) magnetic beads. The percentage of apoptotic CML-LSPCs was assayed by Annexin-V/PI staining; the expression changes of c-myc and NF-kappaB mRNA were detected by real-time quantitative PCR (RQ-PCR) and RT-PCR; the NF-kappaB activity was detected by NF-kappaB Activation Nuclear Translocation Assay Kit; the BCL-2 protein expression was determined in the Western blot method. The results showed that the IM7 effectively induced apoptosis of CML-LSPCs; the mean percentage of early apoptotic cells significantly increased, as compared with the untreated control CML-LSPCs cells 12.58 +/- 2.84% vs 5.42 +/- 1.84% (p < 0.05). The c-myc, NF-kappaB mRNA expressions were down-regulated as compared with the control group (0.65 +/- 0.10 vs 1.00, 0.42 +/- 0.21 vs 1.00, respectively) (p < 0.01) by RQ-PCR and (0.49 +/- 0.09 vs 0.60 +/- 0.12, 0.47 +/- 0.11 vs 0.67 +/- 0.08, respectively)(p < 0.01) by RT-PCR. The BCL-2 protein level in CML-LSPCs treated with IM7 also decreased as compared with the control group (p < 0.01). In addition, the depression of NF-kappaB activity was observed through fluorescence microscope. It is concluded that the anti-CD44 monoclonal antibody IM7 effectively induces apoptosis of CML-LSPCs through down-regulating c-myc and bcl-2 mRNA expression, and decreasing NF-kappaB activity in CML-LSPCs.
Antibodies, Monoclonal ; pharmacology ; Apoptosis ; drug effects ; genetics ; Humans ; Hyaluronan Receptors ; immunology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; metabolism ; NF-kappa B ; metabolism ; Neoplastic Stem Cells ; drug effects ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo investigate device implanted complications and corresponding therapeutic strategies of Coflex interspinous dynamic stabilization system for lumbar spine intraoperatively and postoperatively.

METHODSFrom September 2008 to August 2010, 133 cases of degenerative disease of lumbar spine including 62 males and 71 females, ranging from 35 to 81 years of age (mean 60.8 years), underwent or planed to be underwent decompression with Coflex interspinous dynamic stabilization system were reviewed retrospectively, and 13 cases including 6 males and 7 females, ranging from 41 to 71 years of age (mean 58.6 years), occurred device implanted complications. The Coflex implanted complications were analyzed, and therapeutic strategies according to different character were carried out, scores of visual analogue scale (VAS), Oswestry disability index(ODI) and effect-related data preoperatively, postoperatively, after conservative treatment and in final follow-up were evaluated with paired-samples t test.

RESULTSThirteen cases of Coflex implanted complications and treatment applied included: 3 cases occurred fracture of spinous processes intraoperatively were treated by pedicle screws instead; 2 cases occurred fracture of spinous processes postoperatively or during follow-up, including 1 case underwent revision with pedicle screws, another 1 case treated with conservative treatment; 4 cases with degenerative coronal spondylolysis in surgical segments, 1 case with sagittal instability preoperatively, and 1 case with device dislodgment in follow-up all suffered aggravated pain and received conservative treatment; 1 case suffered implanted malposition intraoperatively was underwent internal fixation with pedicle screws instead; at length, 1 case with aggravated pain postoperatively and without definite reason received revision with internal fixation of pedicle screws demolishing the Coflex. The follow-up time of 13 cases ranged from 20 to 38 months (mean 27.6 months); and 7 cases implanted Coflex with aggravated pain of lumbar and lower limb, but the position of device can still maintained, were received conservative treatment, and whose score of VAS and ODI in the final follow-up were 1.9 ± 0.7 and 23.2 ± 3.4, and comparing to 6.1 ± 1.1 and 58.1 ± 3.0 preoperatively, evident improvement was got finally (t = 8.2 and 18.2, P < 0.01). Scores of VAS and ODI of 2 cases with Coflex implanted complications underwent revision with pedicle screws were also improved correspondingly.

CONCLUSIONSCoflex interspinous dynamic stabilization system implanted should be avoided to cases who suffered with osteoporosis, too narrow interspinous space and intervertebral coronal spondylolysis or sagittal instability; and choice of device, depth of implantation and intensity of clumping should be appropriate. For patients with symptom but device still in right position, conservative treatment can be carried out; but for patients subjected to malposition of device, failure of implantation intraoperatively or intolerance to device, revisions and salvages should be underwent with internal fixation of pedicle screws.

Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Internal Fixators ; adverse effects ; Intervertebral Disc Degeneration ; surgery ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Postoperative Complications ; Retrospective Studies ; Spinal Fusion ; adverse effects ; instrumentation ; methods ; Treatment Outcome

The HIV-1 Rev protein facilitates nuclear export of unspliced and singly spliced viral transcripts containing RRE RNA through the CRM1 export pathway. Inhibition of Rev-mediated RNA nuclear export can arrest HIV-1 transcriptional process, which clearly, reveals a target for anti-HIV drug development. In this work, a cell-based assay has been established for screening anti-HIV compounds targeting the Rev-mediated RNA nuclear export. This assay utilized a codon-optimized green fluorescent protein (GFP) as reporter gene, which expression is in a Rev-dependent manner. Any compound that inhibits the Rev-mediated RNA nuclear export is identified by reducing emission of GFP. The Z' score of this model is 0.8220. Three thousands compounds were screened and the positive rate was 9.3% with a cutoff at 50% inhibition. IMB7C7, one of the positive compounds, efficiently inhibits viral production from HIV-1 infected cells.
Active Transport, Cell Nucleus ; drug effects ; Anti-HIV Agents ; pharmacology ; Cell Nucleus ; metabolism ; Codon ; Fatty Acids, Unsaturated ; pharmacology ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; metabolism ; HEK293 Cells ; HIV-1 ; drug effects ; genetics ; High-Throughput Screening Assays ; Humans ; Karyopherins ; genetics ; metabolism ; RNA, Viral ; Receptors, Cytoplasmic and Nuclear ; genetics ; metabolism ; Transfection ; Virus Replication ; drug effects ; rev Gene Products, Human Immunodeficiency Virus ; genetics ; metabolism

Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).
Anti-HIV Agents ; pharmacology ; Benzoxazines ; pharmacology ; Bioluminescence Resonance Energy Transfer Techniques ; methods ; Fusion Proteins, gag-pol ; genetics ; metabolism ; HEK293 Cells ; HIV Protease ; metabolism ; physiology ; HIV-1 ; enzymology ; High-Throughput Screening Assays ; methods ; Humans ; Plasmids ; genetics ; Protein Precursors ; metabolism ; physiology ; Pyridazines ; pharmacology ; Transfection ; Virion ; growth & development ; Virus Assembly ; gag Gene Products, Human Immunodeficiency Virus ; genetics ; metabolism