Objective To discuss the critical effect of studying and producing the practical cleaning stomach machine of once for all. Methods The process of the cleaning stomach by using the center loaded pressure system equipment was imitated, and the time of the liquid inside and outside, the whole cleaning time and the problems appeared during the cleaning stomach process was recorded. Results After finishing designing the equipment,the mimic stomach was used for critical experiment, all kinds of connecting tubes were airtight and no leaking air and water phenomena, the cleaning process was successful and it could save time to achieve the designing goal.Conclusion The cleaning stomach machine of once for all produced by using the center loaded pressure system had well solved the tube sterilized problems appeared by cleaning machine on hand, and could reduce the risk of hospital inflecting, the cleaning stomach effect was comparatively good,it had practical value and deserve to critical expanding.

OBJECTIVETo observe the expression of phospholipid scramblase 1 (PLSCR1) in matrine (MAT) induced differentiation of all-trans retinoic acid (ATRA) resistant acute promyelocytic leukemia (APL) cells, and to explore its correlation to cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) signal pathway.

METHODSNB4 (an APL cell line sensitive to ATRA) and NB4-R1 (a resistant strain of ATRA) were observed as subjects in this study. Effects of combined treatment of 0.1 mmol/L MAT and 1 [mol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium (NBT) reduction test and flow cytometry (CD11b). Expressions of PML/RARot and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay. Meanwhile, H89, PKA antagonist, was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.

RESULTSMAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells, and significantly down-regulate the expression of PML/RARapha-fusion protein/gene (P < 0.05, P < 0.01). ATRA used alone could obviously enhance the expression of PLSCRI in NB4 cells at protein and mRNA levels (P < 0.01). But the expression of PLSCR1 was up-regulated in NB4-R1 cells, but with statistical.difference only at the protein level (P <0. 01). In combination of MAT, PLSCR1 protein expression was further elevated in the two cell lines (P < 0.01). Besides, there was statistical difference in mRNA expressions in NB4-R1 cells (P < 0.05). All these actions could be reversed by treatment of 10 micromol/L H89 (P < 0.05, P < 0.01).

CONCLUSIONMAT combined ATRA could significantly induce the differentiation of NB4-R1 cells, and inhibit the expression of PML/RARalpha fusion gene/protein, which might be associated with up-regulating PLSCR1 expression.

Alkaloids ; Antineoplastic Agents ; Cell Differentiation ; Cell Line, Tumor ; Down-Regulation ; Humans ; Leukemia, Promyelocytic, Acute ; metabolism ; Phospholipid Transfer Proteins ; metabolism ; Quinolizines ; RNA, Messenger ; Signal Transduction ; Tretinoin ; Tumor Cells, Cultured ; Up-Regulation

Objective To explore the clinical feature,suitable treatment and prognosis of soft tissue leiomyosarcoma of the trunk and extremities.Methods Clinical data of 18 cases of pathologically confirmed soft tissue leiomyosarcoma of the trunk and extremities from January 1999 to December 2012 were analyzed retrospectively.Primary tumors in 7 cases were marginally excised before admitted to our hospital,2 had open biopsy before admission to our hospital,and 8 cases had local relapse at admission; Only one patient took our institute as the first visit.Seventeen cases were performed extended excision of tumors and 1 case underwent marginal resection.All the patients were followed up and the follow-up period was from 16 to 158 months.Results During follow-up period,9 patients developed lung metastasis,and local recurrence occurred in 5 patients.Ten patients died and 8 survived.Of the ten dead cases,seven died of tumor progression and 3 died from non-tumor factors.In the 8 patients survived,2 survived with tumor.The 5-year overall survival rate was 59.2％.Conclusion Soft tissue leiomyosarcoma of the trunk and extremities are rare malignant tumors,mostly occurred in elder patients and presenting soft tissue mass.Local recurrence and distant metastasis are common and associated with a poor prognosis.Surgical excision combined with adjuvant radiation is the common treatment strategy.

Objective To search for the changes of T cells subpopulations and immunoglobulin and their relation-ship in children patients with simple nephrotic syndrome. Design Case-control research. Patients aud Participants 39 patients with simple nephrotic syndrome were divided into two groups:the incipient group and relapse group (6 cases were determined at the incipient and relapse time) .Thereare 28 patients in incipient group, 19 males and 9 females, at the age of 2 to 10 years old. There are20 patients in relapse group, 12 males and 8 females, at the age of 3 to 13 years old. There are 35health children in control group, 21 males and 14 females, 2～13 years old. Interventions T cells subpopulations were determined by indirect immunofluorescence of OKT linesmonoclonal antibodies. The serum IgG was determined by routine simple agar immunodiffusion tests. Results and Conclusions The CD_3~+ and CD_4~+ cells are of no change in the children patients withsimple nephrotic syndrome, and the CD_8~+ and CD_(10)~+ cells are obviously increased, the Values of CD_4~+/CD_8~+ are obviously lower than those in the control qroup, there are no difference between the incipientand relapse groups. The levels of serum IgG were decreased in the 85.3% children patients, IgM were inc-reased in 29.4% of that. The values of CD_4~+/CD_8~+ have positive correlation and negative correlationwith the levels of serum IgG and IgM respectively.

Objective To study the difference in therapeutic effects and side effects of ibuprofen versus indomethacin for symptomatic patent ductus arteriosus (PDA) in the premature infants.Methods Meta analysis was used to qualitatively and quantitatively evaluate the data extracted from 6 public papers about comparative study of ibuprofen and indomethacin.Results The rate of ductal closure was similar with the two treatment regimes (intravenous ibuprofen and indomethacin).In side effects on PDA,the incidence of oliguria induced by ibuprofen was significantly lower than that of indomethacin though there were no difference in other side effects.Conclusions The efficacy of ibuprofen for the early treatment of PDA in preterm infants is similar with indomethacin,and has low incidence of oliguria.

OBJECTIVETo evaluate the clinical efficacy of treating myelodysplastic syndrome (MDS) by hematopoietic stem cell transplantation (HSCT) combined with Chinese medical syndrome typing.

METHODSFrom July 2009 to July 2013, 6 MDS patients were treated with allo-HSCT combined with Chinese medical syndrome typing from HLA-identical sibling donors at Department of Hematology, Zhejiang Provincial Hospital of Chinese Medicine. Patients were classified as refractory anemia (RA, 2 cases), refractory anemia with ringed sideroblast (RARS, 1 case), refractory cytopenia with multilineage dysplasia (RCMD, 2 cases), and RA with excess blasts-I (RAEB-I , 1 case). Modified BuCy conditioning regimen was used in all 6 cases. Two patients received bone marrow transplantation (BMT), 1 patient received peripheral blood stem cell transplantation (PBSCT), and 3 patients received BMT + PBSCT. In order to prevent the occurrence of graft-versus-host disease (GVHD), all patients were treated with cyclosporine + methotrexate + mycophenolate mofetil. Different Chinese medical treatment methods (by syndrome typing) were given to patients according to different criticality of international prognostic scoring system (IPSS, 5 at moderate risk and 1 at high risk).

RESULTSAll 6 patients successfully reconstructed their hematopoietic system. The time from transplantation to ANC ≥ 0.5 x 10(9)/L and platelet (PLT) ≥ 20 x10(9)/L were 13 (9-15) days and 11 (9-22) days respectively. Main complications were GVHD. Acute GVHD (aGVHD) occurred in 4 cases, 3 cases of grade I and 1 case of grade II, and local chronic GVHD (cGVHD) occurred in 1 patient. All cases survived with median follow-up of 18 (11-58) months. The overall survival (OS) and disease-free survival (DFS) rate were 100%.

CONCLUSIONSHSCT combined with Chinese medical syndrome typing could improve clinical symptoms, reduce transplant as- sociated complications. So it was an effective treatment choice for MDS.

Biomedical Research ; Blood Platelets ; Bone Marrow Transplantation ; Cyclosporine ; therapeutic use ; Disease-Free Survival ; Drugs, Chinese Herbal ; therapeutic use ; Graft vs Host Disease ; prevention & control ; Hematopoietic Stem Cell Transplantation ; Humans ; Medicine, Chinese Traditional ; Methotrexate ; therapeutic use ; Myelodysplastic Syndromes ; therapy ; Transplantation Conditioning ; Transplantation, Homologous ; Treatment Outcome

Objective To evaluate the optical quality in patients with meibomian gland dysfunction (MGD) by using double-pass optical quality analysis system (OQAS-Ⅱ) for offering the reference data for clinical practices.Methods A total of 50 patients with a diagnosis of MGD,who were divided into MGD with dry eye disease (DED) group (n =22) and MGD without DED group (n =28) according to the results of tear film break-up time (BUT) and Schirmer I test (SIT),and another 25 healthy subjects were enrolled in this study.Then several parameters,including modulation transfer function cutoff (MTF cutoff),Strehl ratio (SR) and objective scattering index (OSI),were measured to evaluate the optical quality by using OQAS-Ⅱ.Additional clinical examination,including meibomian gland and tear film assessment were per formed and compared between the three groups.In addition,a correlation analysis was conducted among OSI,MTF cutoff,SR,BUT,SIT and meibomian gland assessment.Results Statistical differences were approached in MTF cutoff,SR,OSI of the three groups (all P ＜ 0.05).The MTF cutoff and SR in MGD without DED group were significantly lower than those in the control group [(31.36 + 1.83) c · deg-1 vs.(35.87 ± 1.59)c·deg-1,(0.21 +0.02) vs.(0.23 ±0.03)],but the OSI got higher [(0.57+ 0.06) vs.(0.45 ±± 0.06)] (all P ＜ 0.05).The SR and MTF cutoff in MGD patients with DED was significantly lower than those in MGD patients without DED [(27.87 ±± 3.08),c·deg-1 vs.(31.36±±1.83)c· deg-1,(0.16 ±±0.02) vs.(0.21 ± 0.02)],but OSIgot significantly higher [(0.72 ± 0.10) vs.(O.57 ± 0.06)] (all P ＜ 0.05).Among the three groups,OSI,MTF cutoff and SR showed no significant correlation with BUT,SIT and meibomian gland assessment (all P ＞ 0.05).Conclusion There is changes in visual quality parameters in MGD patients by OQAS-Ⅱ,and MGD with DED patients has significant changes than MGD patients without DED.

OBJECTIVETo investigate the molecular mechanism of haemophilia B caused by the novel mutation of Arg327Ile (R327I) in FIX gene.

METHODSThe R327I, R327Ala(A), R327Lys(K), R327Asn(N) and a replacement mutant (FIXβFVII), in which FIX β strand 324-329 was replaced by that of FVII 298-303, expression plasmids were constructed with site-directed mutagenesis method based on the wild-type (WT) FIX expression plasmid. The HEK293 cell was transiently transfected, then the activity of FIX (FIX:C) was assayed by one stage method in the conditioned medium, while the FIX:Ag in both the conditioned media and the cell lysates was measured by ELISA. The molecular weight and the semi-quantity of expressed FIX were analyzed by Western blot. Fluorescent protein expression plasmid was constructed to investigate the synthesis and secretion of the FIX R327I mutation in the viable cells.

RESULTSFIX:C of the R327I mutant protein was 4.49% of the level of the WT in the conditioned medium, and the FIX:Ag of the R327I mutant protein in the conditioned medium and the cell lysates was 31.02% and 129.29% compared to that of WT, respectively. The mutation was characterized as cross-reaction material reduced (CRMR). The viable cell fluorescent assays showed that the R327I protein was more in both the viable cells and in lysosome than that of WT. The FIX:C of the R327A, R327K, R327N and FIXβFVII mutants was reduced compared to that of WT, the reduction of FIX:C of FIXβFVII was the most significantly amount among all the mutants in medium. FIX:Ag of all the mutants in the medium, except that the R327K increased, was reduced. The result of Western blot showed that the molecular weight of R327I protein was the same as that of WT, but the amount of the protein was much less compared with WT in the conditioned medium.

CONCLUSIONThe abnormal synthesis and secretion as well as the abnormal function of the R327I mutant protein causes haemophilia B. The residue of R327 as well as the β strand domain of R327 located play important roles of the specific function of FIX.

Factor IX ; genetics ; HEK293 Cells ; Hemophilia B ; genetics ; pathology ; Humans ; Mutagenesis, Site-Directed ; Mutation ; Transfection

OBJECTIVETo study the role of Ezh2 in the all-trans retinoic acid RA induced P19 neural differentiation.

METHODSThe expression of Ngn1 in the RA induced P19 cells was detected at the mRNA and protein levels using real time RT-PCR and Western blot assays. The binding of Ezh2 and H3K27me3 on the Ngn 1 promoter was analyzed using chromatin immunoprecipitation assay.

RESULTIn the RA induced P19 cells, the recruitment of Ezh2 and its methylated substrate H3K27me3 on the promoter of Ngn 1 gene elevated in the first 2 days, and then declined rapidly, followed by the initiation of neuronal differentiation.

CONCLUSIONSEzh2 produces a repressive histone mark H3K27me3 in the early stage of RA induced P12 cells. By avoiding the premature expression of Ngn1 gene, Ezh2 can ensure the normal differentiation of P19 cells.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Differentiation ; drug effects ; physiology ; Enhancer of Zeste Homolog 2 Protein ; Histone-Lysine N-Methyltransferase ; genetics ; metabolism ; Histones ; metabolism ; Mice ; Nerve Tissue Proteins ; genetics ; metabolism ; Neurons ; cytology ; drug effects ; metabolism ; Polycomb Repressive Complex 2 ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

The identity of higher-order neurons and circuits playing an associative role to control renal function is notwell understood.We identified specific neural populations of rostral elements of brain regions that project multisynaptically to the kidneys in 3~ days after injecting a retrograde tracer pseudorabies virus (PRV)-614 into kidney of 13 adult male C57BL/6J strain mice.PRV-614 infected neurons were detected in a number of mesencephalic (e.g.central amygdala nucleus),telencephalic regions and motor cortex.These divisions included the preoptic area (POA),dorsomedial hypothalamus (DMH),lateral hypothalamus,arcuate nucleus (Arc),suprachiasmatic nucleus (SCN),periventricular hypothalamus (PeH),and rostral and caudal subdivision of the paraventricular nucleus of the hypothalamus (PVN).PRV-614/Tyrosine hydroxylase (TH) double-labeled cells were found within DMH,Arc,SCN,PeH,PVN,the anterodorsal and medial POA.A subset of neurons in PVN that participated in regulating sympathetic outflow to kidney was catecholaminergic or serotonergic.PRV-614 infected neurons within the PVN also contained arginine vasopressin or oxytocin.These data demonstrate the rostral elements of brain innervate the kidney by the neuroanatomical circuitry.