Factor VII Deficiency ; diagnosis ; genetics ; Female ; Humans ; Middle Aged ; Pedigree ; Phenotype

This study was aimed to explore the expression of microRNA-21 (miR-21) in multiple myeloma (MM), and its correlation with plasma β2-microglobulin (β2-MG) and staging of MM by Durie-Salmon (D-S) classification. The expression level of miR-21 in bone marrow mono-nuclear cells (BMMNC) of 43 patients with MM and 20 healthy individuals was examined by real-time polymerase chain reaction (real-time PCR), and the correlations among the expression level of miR-21 and related clinical pathologic features, plasma β2-MG and staging of MM by D-S classification were analyzed. The results showed that the expression of miR-21 in BMMNC of MM patients was obviously higher than that in normal controls (P < 0.05). The expression of miR-21 in BMMNC of relapsed/refractory MM patients was obviously higher than that in newly diagnosed MM patients. The expression of miR-21 in MM patients decreased after chemical therapy, especially in effective group (P < 0.05), there was no significant change of miR-21 expression level in ineffective/progressive group before and after chemical therapy (P > 0.05). The expression of miR-21 was related with staging of MM and plasma β2-MG level. It is concluded that the expression levels of miR-21 in BMMNC of MM patients are significantly higher than in normal bone marrow, these data indicated that miR-21 may play an important role in the development of MM. Super expression of miR-21 is positively correlated with level of plasma β2-MG and staging of MM by D-S classification.
Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Male ; MicroRNAs ; metabolism ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology ; beta 2-Microglobulin ; blood

Objective To investigate the effects of DL-3-n-Butylphthalide(NBP)on proliferation and apoptosis of 1-methyl-4-phenyl-pyridinium (MPP +)-induced SH-SY5Y cells, and mechanisms via mixed lineage kinase 3 (MLK3) signaling pathway. Methods The SH-SY5Y cells were divided into control group,MPP+group,NBP group and URMC-099 group,that cultured normally,with 1 mmol/L MPP+for 24 hours,with 10μmol/L NBP for 3 hours and then with MPP+for 24 hours,and with 200 nmol/L MLK3 inhibitor URMC-099 for 3 hours and then with MPP+for 24 hours,respectively.The morphology of SH-SY5Y cells was observed under inverted phase contrast mi-croscope and the survival rate was measured with 3-(4,5-Cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assays.The apoptosis was quantified under flow cytometry with Annexin V/PI fluorescence staining,and the nuclear morphology was observed with Hoechst 33342 staining.The expression of phosphorylated protein of MLK3(p-MLK3),c-Jun N-terminal kinase(p-JNK),extra cellular regulated protein ki-nases(p-ERK1/2)were detected with Western blotting.Results Compared with the control group,the survival rate reduced and apoptosis in-creased in MPP+group(P<0.05),with the increase of p-MLK3 and p-JNK and decrease of p-ERK1/2 d(P<0.05).Compared with MPP+group,the survival rate increased and apoptosis reduced in both NBP and URMC-099 groups(P<0.05),with the decrease of p-MLK3 and p-JNK and increase of p-ERK1/2(P<0.05).Conclusion NBP can decrease the apoptosis and promote the proliferation of SH-SY5Y cells in-duced by MPP+,which may be associated with inhibiting MLK3 signaling pathway,and regulating the downstream p-JNK and p-ERK1/2.

OBJECTIVETo study the expression of transcription factor LYL1 in leukemia and its possible role in leukemogenesis.

METHODSFluorescence real time quantitative polymerase chain reaction was used to detect the expression levels of LYL1 in leukemias. Specific siRNA was used to silence the expression of LYL1 in K562 cells.

RESULTSCompared to CD34 positive cells from normal bone marrow, the expression of LYL1 was significantly elevated in patients with acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). LYL1 expression was higher in chronic myeloid leukemia (CML) in blastic crisis than that in chronic phase (7.831 vs 1.672, P < 0.01). LYL1 expression in AML in complete remission (CR) was down-regulated as compared with that of un-remission patients (1.400 vs 9.985, P < 0.01). Down-regulation of endogenous expression of LYL1 in K562 cells by a combination of three specific siRNA could inhibit cellular growth and clonogenicity to some extent.

CONCLUSIONOver-expression of LYL1 is highly associated with AML as well as ALL. RNA interference targeting specific oncogenes such as LYL1 is potentially useful in the treatment of hematological malignancies.

Adult ; Aged ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Differentiation ; genetics ; Cell Proliferation ; Down-Regulation ; Female ; Gene Expression Regulation, Leukemic ; Humans ; K562 Cells ; Leukemia ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Proteins ; genetics ; metabolism ; RNA Interference ; Young Adult

OBJECTIVETo investigate the effects of early enteral nutrition supplemented with immune nutrient on intestine immune function in mice with severe burn.

METHODSTwenty-four BALB/c mice were inflicted with 20% TBSA full-thickness scald, then they were randomly divided into EN(with oral administration of common enteral nutrition after 2 hours) and EIN (with oral administration of common enteral nutrition and glutamine, arginine after 2 hours) groups. Another 10 mice were used as the normal control (NC) group. The supplied energy ratio( carbohydrate: fat: protein)in former 2 groups was 82:3:15, and the ratio of energy to nitrogen was 150: 1. The energy requirement of each mouse was calculated according to 732.2 kJ x kg(-1) x d(-1), one third of the requirement was administrated on 1st day, and one half of it on 2nd day, and full energy requirement was started on the 3rd day,and the requirement was divided into 4-6 portions every day. The feed was isocaloric, isonitrogenous, and isovolumic for the 2 experimental groups. All mice were sacrificed and entire small intestine was harvested for determination of intestinal IgA level by ELISA, total Peyer's patches (PP) lymphocytes and their apoptosis ratio, and changes in PP lymphocytes (CD3+, CD4+, CD19+) on 7th day of the experiment.

RESULTSCompared with those in NC group [(4.5 +/- 0.6) x 10(6), (42 +/- 7) microg/cm, respectively], total PP lymphocytes and intestinal IgA levels in EN and EIN groups obviously decreased [(2.3 +/- 0.4) x 10(6), (35 +/- 6) microg/cm, (3.8 +/- 0. 5) x 10(6), (38 +/- 6), microg/cm, respectively, P < 0.05] , among which the values in EIN group were higher than EN group (P < 0.05). The changes in PP lymphocytes were similar to that of total PP lymphocytes. Compared with that in NC group [(4.8 +/- 2.1)%], the apoptosis ratio of PP lymphocytes in EN and EIN groups significantly increased [(12.7 +/- 2.4)%, (8.0 +/- 1.7)%, respectively, P < 0.05], however the ratio in EIN group was lower than that of EN group (P < 0.05).

CONCLUSIONSEarly enteral nutrition supplemented with immune nutrient can improve intestinal immune function in mice with severe burn.

Animals ; Burns ; immunology ; physiopathology ; therapy ; Enteral Nutrition ; Intestine, Small ; immunology ; Intestines ; immunology ; Lymphocyte Subsets ; Lymphocytes ; immunology ; Male ; Mice ; Mice, Inbred BALB C

Aim To explore the therapeutic effects of main active compounds of panaxadiol ( PD ) in on Alzheimer’s disease ( AD) via network pharmacologi-cal analysis and Mmolecular docking. Methods A to-tal of 107 prescriptions for AD treatment were screened by using network pharmacology, screening for the high-est frequency of ginseng and its target for AD. Use mo-lecular docking technology was used to find components with the highest score for non-receptor tyrosine kinase ( FYN) docking. Then we successfully estimatedestab-lished AD cell model with overexpressinged APP pro-teins in vitro. Next,the cell viability was detected by MTT assay,the cell damage was detected by LDH as-say,the apoptosis and intracellular Ca2+concentration were detected by flow cytometry, and phosphorylated FYN protein expression was detected by Western blot detection of . phosphorylated FYN protein expression. Results Eighteen active components of Gginseng and 29 AD-related targets were screened by the method of network pharmacology. The results of molecular doc-king showed that PD had strong binding effects with FYN. The results showed that PD could increase the survival rate of cells,reduce the release of LDH,reduce apoptosis,and improve AD cells’ intracellular Ca2+o-verload and reduce the expression of FYN-Y416 pro-tein. Conclusion The experimental results of network pharmacology were are verified and the protective effect of PD on AD may be related to inhibition of FYN signa-ling pathway.

OBJECTIVETo investigate the potential of siRNAs targeting sphingomyelin phosphodiesterase 1 (SMPD1) in protecting the oocytes from apoptosis, and explore new approaches to female fertility preservation.

METHODSChemically synthesized siRNA targeting SMPD1 were introduced into mouse oocytes retrieved by hyperstimulation, and the cell apoptosis was analyzed by comic assay 48 and 72 h later.

RESULTSIn the oocytes without any siRNA injection, oocyte DNA damage occurred after 24 h, and large amount of DNA fragments migrated from the cells 48 h later. In oocytes injected with siRNA003, DNA migration decreased significantly as compared with the control and the other two groups injected with siRNA001 and siRNA002 (P<0.01).

CONCLUSIONsiRNA targeting SMPD1 may protect the oocytes from apoptosis, and has the potential for use in future female fertility preservation.

Animals ; Apoptosis ; drug effects ; genetics ; Comet Assay ; Female ; Mice ; Oocytes ; cytology ; RNA Interference ; RNA, Small Interfering ; genetics ; Sphingomyelin Phosphodiesterase ; genetics ; physiology ; Transfection

OBJECTIVEThis study was aimed to investigate the risk factors of renal impairment and the predictive factors of renal function recovery so as to provide basis for its prevention and treatment.

METHODSMedical records of 161 patients with MM firstly diagnosed from January 2007 to April 2013 were analyzed retrospectively. Among them 58 cases accompanied with renal insufficiency (group A, others belong to group B) and 39 of them regain normal renal function after some treatment. The possible related renal impairment risk factors and reversible predictors were analyzed with chi-square test for significance firstly, then factors that have significant difference were entered into multivariate logistic regression analysis.

RESULTSSystolic blood pressure (SBP), hemoglobin, uric acid, blood calcium, phosphorus, serum β2-microglobulin, urine β2-microglobulin levels, M-component type, light chain type, nephrotoxic drug use, infection in group A had significant difference (P<0.05) compared with those in group B; the systolic blood pressure, diastolic blood pressure, platelet, globulin, blood calcium, and urine β2-microglobulin levels, the chemotherapy applied and the response to chemotherapy in reversed group were significantly different from no-reversed group (P<0.05). Multivariate logistic regression showed that light chain type, Hb, uric acid, Ca were the independent risk factors for the development of renal failure in MM, and Ca, chemotherapy and the response to chemotherapy were the predictors of renal function recovery.

CONCLUSIONHigh blood calcium, severe anemia, λ light chain, high uric acid are the independent risk factors of renal impairment in MM patients. Patients with high blood calcium before treatment easily regain normal renal function after effective chemotherapy. Bortezomib-based chemotherapy has higher response rate and higher reversal rate, and it may be related with its unique mechanism.

Bortezomib ; Humans ; Kidney ; Logistic Models ; Multiple Myeloma ; Renal Insufficiency ; Retrospective Studies ; Risk Factors

To validate in situ rats intestinal single pass perfusion model based on P-glycoprotein (P-gp). Firstly, phenol red perfusion was carried out to verify the close connection structure of intestinal epithelial cells, and the integrity of the intestinal epithelium, with a gravimetric method for correcting water flux. The level of phenol red was determined by high performance liquid chromatography (HPLC) both before and after perfusion. Secondly, the positive drug digoxin specified by FDA was used to validate the model. After different mass concentrations of verapamil were given in the rats, the absorption parameters of digoxin in ileum of rats were observed and compared. The results showed that the phenol red was absorbed in rats ileum segment, with an effective permeability coefficient of (1.09±0.62)×10 ⁻⁶ cm•s ⁻¹. The experiment results indicated that the close connection structure of intestinal epithelial cells was normal, and the integrity of the intestinal epithelium was maintained well. In digoxin perfusion experiment, in case no verapamil was given, digoxin showed certain degree of absorption in rat ileum, with an effective permeability coefficient (Peff) of (1.07±0.59)×10 ⁻⁵ cm•s ⁻¹; after mass concentrations of 0.01,0.1 mmol•L ⁻¹ verapamil were given, the absorption of digoxin was on the rise in rat ileum, with an effective permeability coefficient Peff of (1.58±0.69)×10 ⁻⁵, (3.28±0.95)×10 ⁻⁵ cm•s ⁻¹ respectively (P<0.05). Digoxin perfusion experiment verified that P-gp expression in small intestine epithelium was intact and can be used in the research of P-gp efflux transporter.

OBJECTIVES: To study the protective effect of breviscapine against brain injury induced by intrauterine inflammation in preterm rats and its mechanism. METHODS: A preterm rat model of brain injury caused by intrauterine inflammation was prepared by intraperitoneal injections of lipopolysaccharide in pregnant rats. The pregnant rats and preterm rats were respectively randomly divided into 5 groups: control, model, low-dose breviscapine (45 mg/kg), high-dose breviscapine (90 mg/kg), and high-dose breviscapine (90 mg/kg)+ML385 [a nuclear factor erythroid 2-related factor 2 (Nrf2) inhibitor, 30 mg/kg] (n=10 each). The number and body weight of the live offspring rats were measured for each group. Hematoxylin-eosin staining was used to observe the pathological morphology of the uterus and placenta of pregnant rats and the pathological morphology of the brain tissue of offspring rats. Immunofluorescent staining was used to measure the co-expression of ionized calcium binding adaptor molecule-1 (IBA-1) and nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) in the cerebral cortex of offspring rats. ELISA was used to measure the levels of interleukin-6 (IL-6), interleukin-8 (IL-8), and interleukin-1β (IL-1β) in the brain tissue of offspring rats. Western blotting was used to measure the expression of Nrf2 pathway-related proteins in the brain tissue of offspring rats. RESULTS: Pathological injury was found in the uterus, and placenta tissue of the pregnant rats and the brain tissue of the offspring rats, and severe microglia pyroptosis occurred in the cerebral cortex of the offspring rats in the model group. Compared with the control group, the model group had significant reductions in the number and body weight of the live offspring rats and the protein expression levels of Nrf2 and heme oxygenase-1 (HO-1) in the brain tissue of the offspring rats (P<0.05), but significant increases in the relative fluorescence intensity of the co-expression of IBA-1 and NLRP3, the levels of the inflammatory factors IL-6, IL-8, and IL-1β, and the protein expression levels of NLRP3 and caspase-1 in the brain tissue of the offspring rats (P<0.05). Compared with the model group, the breviscapine administration groups showed alleviated pathological injury of the uterus and placenta tissue of the pregnant rats and the brain tissue of the offspring rats, significant increases in the number and body weight of the live offspring rats and the protein expression levels of Nrf2 and HO-1 in the brain tissue of the offspring rats (P<0.05), and significant reductions in the relative fluorescence intensity of the co-expression of IBA-1 and NLRP3, the levels of the inflammatory factors IL-6, IL-8, and IL-1β, and the protein expression levels of NLRP3 and caspase-1 in the brain tissue of the offspring rats (P<0.05). The high-dose breviscapine group had a significantly better effect than the low-dose breviscapine (P<0.05). ML385 significantly inhibited the intervention effect of high-dose breviscapine (P<0.05). CONCLUSIONS Breviscapine can inhibit inflammatory response in brain tissue of preterm rats caused by intrauterine inflammation by activating the Nrf2 pathway, and it can also inhibit microglial pyroptosis and alleviate brain injury.
Animals ; Female ; Pregnancy ; Rats ; Body Weight ; Brain Injuries/prevention & control* ; Caspase 1 ; Inflammation/drug therapy* ; Interleukin-6 ; Interleukin-8 ; NF-E2-Related Factor 2 ; NLR Family, Pyrin Domain-Containing 3 Protein ; Flavonoids/therapeutic use*