To explore the regulation of eIF4E, we screened the protein interacting with eIF4E from human cDNA library by using yeast two-hybrid system. Several clones interacting with eIF4E were identified. One of them was homologous with HUWE1 (HECT, UBA and WWE domain containing 1, also named as ARF-BP1, HECTH9 or HUWE1). Cell co-immunoprecipitation showed that eIF4E could bind to HUWE1 in mammalian cells. We also found that HUWE1 bearing the HECT domain is necessary for its association with eIF4E.
Animals ; Eukaryotic Initiation Factor-4E ; metabolism ; Humans ; Ubiquitin-Protein Ligases ; metabolism

Objective To analyse the clinical efficacy of methotrexate (MTX) combined with intrauterine embryo sac garrotte injection in the treatment of cesarean scar pregnancy (CSP), and discuss its clinical significance. Methods A total of 77 patients with CSP treated in our hospital during June 2013 to December 2016 were selected in this study. Forty patients treated with embryo sac destruction and methotrexate injection were included in the observation group, while 37 cases treated by uterine artery embolization combined with curettage were used as the control group. The time of vaginal bleeding, the time of postoperative blood level of human chorionic gonadotropin (HCG) returned to the normal level, average hospitalization cost and the curative rate were recorded in two groups. All patients were followed up by the outpatient visit. Results In the observation group, the vaginal bleeding time [(22.1±6.7) days vs. (29.5±10.8) days] and treatment cost [(8774.2 ± 714.5) yuan vs. (15258.3 ± 1084.2) yuan] were less than those of the control group (P<0.001). There were no significant differences in the recovery time of HCG [(26.4±9.0) days vs. (25.1±10.4) days] and treatment success rate (87.5%vs. 91.9%) between the two groups (P>0.05). No bleeding or threatened rupture of scar were found in two groups of patients. Conclusion In this study, we take the embryo sac puncture combined with methotrexate injection in the treatment of scar pregnancy. This method has the advantages of low operative difficulty, definite clinical curative effect and low cost

High dose methotrexate is a cornerstone of therapy for acute lymphoblastic leukemia and osteosarcoma.However MTX levers exhibit significant inter-individual variability in clinical,and acute toxicity after high-does MTX is often unexpected,thus it needs individual administration.

Viruses (e.g. Human immunodeficiency virus, Human simplex virus and Prototype foamy virus) are obligate intracellular parasites and therefore depend on the cellular machinery for cellular trafficking. Bovine foamy virus (BFV) is a member of the Spumaretrovirinae subfamily of Retroviruses, however, details of its cellular trafficking remain unknown. In this study, we cloned the BFV gag gene into prokaryotic expression vector pET28a and purified the denaturalized Gag protein. The protein was used to immunize BALB/c mouse to produce antiserum, which could specifically recognize the BFV Gag protein in BFV-infected cells through western blot assay. Additionally, these results demonstrated that both the optimal and suboptimal cleavage of Gag protein occur in BFV-infected cells. Subsequently, the Gag antiserum was used to investigate subcellular localization of BFV. In immunofluorescence microscopy assays, colocalization microtubules (MTs) and assembling viral particles were clearly observed, which implied that BFV may transport along cellular MTs in host cells. Furthermore, MTs-depolymerizing assay indicated MTs were required for the efficient replication of BFV. In conclusion, our study suggests that BFV has evolved the mechanism to hijack the cellular cytoskeleton for its replication.

Objective To investigate the changes of lead,zinc,copper,iron and calcium in blood of chronic poisoned infantal mice.(Methods) Forty-eight 21 day-old kunzea mice were randomly divided into 4 groups,each having 12 mice.Distilled water group was as control group and other three lead acetate poisoning groups had a dose of 10,20,40 mg/kg,respectively.The poisoning was carried out by lavage once a day,and consecutively for 46 days.Eyeballs of mice were picked then for blood sampling,and BS trace element analysis grapher was used to determine level of lead,zinc,copper and iron.Level of calcium was measured by Dimentional-RXL auto-biochemistry analysis meter.Results The lead and zinc levels in poisoned mice blood were increased with increasing lead acetate level administration,while zinc level changed inversely with lead acetate level.Significant differences were shown among control group and poisoning groups in terms of lead(P0.05).Conclusion Lead posioning can lead to zinc decreasing and copper(increa)-sing,which suggests that zinc works as a poential antidote of lead poisoning.

Objective To explore the expression and clinical significance of caudal homeobox gene CDX2 in acute myeloid leukemia(AML) patients .Methods Bone marrow (BM )and peripheral blood (PB)samples were colleted in 114 cases of donor AML patients and 56 patients undergoing chemotherapy .The CDX2 gene expression in every patient′s mononuclera cells were de‐tected by RT‐PCR .Among these patients ,19 cases were detected the gene continuous every three months .Eight healthy PB and five patients with iron deficiency anemia BM as control .Results CDX2 gene transcript levels were detectable in bone marrow mononu‐clear cells from 114 AML patients and 13 healthy donors ,but the level of gene expression was higher in AML patients(90/114 , 78 .9% ) .There was a statistically significant difference between the AML patients and normal donor (P< 0 .01) .The higher or lower expression of CDX2 gene showed no correlation with CR rate .CDX2 gene expression level had a positive correlation in BM and PB mononuclera cells(the correlation coefficient r= 0 .656 ,P< 0 .01) .The expression of CDX2 in patients with CR was 10 .3% -86 .2% of pre‐chemotherapy ,wihch decreased with the treatment course ,while elevated in recurrence .19 cases of patients underwent half a year of follow‐up ,there was no significant difference of the rate of early recurrence in two groups(P>0 .05) .Con‐clusion Higher expression level of CDX2 gene is mostly in AML patients ,but its expression has no relation with CR rate .CDX2 gene may be a prognostic molecular marker in AML patients ,and can be used to monitor the minimal residual disease of Normal chromosome karyotype AML .

Objective The aim of the study was to explore the difference between the expression of CDg7 in peripheral blood and bone marrow on acute myeloid leukemia (AML n=30). Methods A flow cytometric quantitative analysis of expression levels for CD87 was performed on fresh blast cells in peripheral blood and bone marrow from patients with acute myeloid leukaemia using CD87, monoclonal antibodies. Analysis the difference between the expression of CD87 using matched t -test. Results The values of CD87 expression in bone marrow of 14 M5 cases are from 9.47 %～80.32 %, and from 11.49 %～87.46 % in peripheral blood. The values of CD87, expression in bone marrow of 8 M4 cases are from 14.27 %～46.28 %,and from 14.79 %～47.19 % in peripheral blood. The values of CD87 expression in bone marrow of 6 M2 cases are from 4.67 %～34.26 %, and from 8.96 %～39.78 % in peripheral blood. The values of CDs, expression in bone marrow of 2 MI cases are from 3.56 %～7.69 %, and from 5.21 %～8.96 % in peripheral blood.The expression of CD87 in peripheral blood and bone marrow from patients with acute myeloid leukaemia had statistical difference (t =3.13, P<0.05). Conclusion The levels of CD87 expression had difference between peripheral blood and bone marrow. The level in peripheral blood was higher than bone marrow. So when we performed quantitative analysis of expression levels for CD87, peripheral blood instead of bone marrow was commended.

Objective To investigate the inhibitory effects of overexpression of PTEN on renal epithelial-mesenchymal trans-differentiation induced by TGF-β1,and the signaling transduction mechanism.Methods HKC cells were transfected with GFP-PTEN via lipofectAMINE2000.The efficiency of transfection was detected by fluorescence microscope.The expression of PTEN protein and mRNA in the translected cells was detected by Western blot and RT-PCR respectively.The experiment was divided into four groups:normal group,TGF-β1 stimulation group,GFP-PTEN+TGF-β1 group and empty vector+TGF-β1 group.The expression of E-cadherin,a-SMA,Akt and p-Akt was detected by Western blot.Results Most ceils transfected with GFP-PTEN expressed GFP.The expression of PTEN protein and mRNA was strongly increased when HKC cells were transfected with GFP-PTEN(all P<0.05).In both TGF-β1 stimulation group and empty vector+TGF-β1 group,the expression level of E-cadherin was lower(all P<0.05),while that of p-Akt and a-SMA was higher than in normal group(both P<0.05).The expression level of p-Akt and a-SMA in GFP-PTEN+TGF-β1 group was Iower(both P<0.05),while that of E-cadherin was higher than in TGF-β1 stimulation group and empty vector+TGF-β1 group(both P<0.05).The expression of Akt was similar in the four groups.Conclusion Overexpression of PTEN can inhibit renal epithelial-mesenehymal trans-differentiation induced by TGF-β1 through suppressing the activation of PI3K/Akt signal pathway.

Abstract: Objective　To analyze the distribution and drug resistance evolution characteristics of pathogenic bacteria of bloodstream infection in nine tertiary hospitals in Yunnan Province from 2017 to 2021, so as to provide reliable basis for rational selection of antibiotics in clinic. Methods Using the drug sensitive paper method or instrument method, the bacteria identification and drug sensitivity test were carried out in nine tertiary hospitals in different regions according to the unified technical scheme. The results were judged according to the Clinical and Laboratory Standards Institute (CLSI) breakpoint standard in 2021, and use WHONET5.6 for data statistical analysis. Results A total of 12 003 strains of pathogenic bacteria were isolated from bloodstream infection samples in the past five years, including 7 442 strains of Gram-negative bacteria (62.0%) and 4562 strains of Gram-positive bacteria (38.0%), with an increasing trend in the number of isolated strains; of these, 163 strains (1.4%) were isolated from outpatients and 11 840 strains (98.6%) were isolated from inpatients. The top three gram-negative bacteria were Escherichia coli, Klebsiella pneumoniae and Acinetobacter baumannii, of which 309 strains (4.2%) were carbapenem-resistant Klebsiella pneumoniae (CR-KPN), 29 strains (0.4%) carbapenem-resistant Escherichia coli and 19 strains (0.3%) carbapenem-resistant Enterobacter cloacae, and the number of CR-KPN was on the rise year by year. The top three Gram-positive bacteria were coagulase-negative staphylococci, Staphylococcus aureus and Enterococcus faecium, of which methicillin-resistant Staphylococcus aureus (MRSA) was detected for 213 strains, accounting for 27.7%, and decreased from 40.0% in 2017 to 23.4% in 2021, showing a downward trend year by year. No vancomycin-resistant staphylococci and enterococci were found. Conclusions The detection and composition of bloodstream infection pathogenic bacteria in multicenter have not changed much in the past five years, but each hospital has its own characteristics. The number of carbapenem resistant Enterobacteriaceae increased year by year, which should be paid more attention.

OBJECTIVETo observe the influence of H102 on the expression of amyloid protein and amyloid precursor protein in the hippocampus of APP695 transgenic mice.

METHODSThe 9-month-old APP695 transgenic mice were randomly divided into the model group and the H102 group; C57BL/6J mice were adopted as normal control group. The H102 group were injected with H102 in a dose of 3 microl/per mouse in lateral ventricle, once a day, for ten days; while the model group and the control group were injected with saline. The hippocampus and temporal cortex of the brain sections from transgenic mice and wild type female mice were subjected to immunohistochemistry and Congo red histological staining, and observed the difference of the protein expression under microscope. The expression of the APP protein was detected by Western blot.

RESULTSAbeta and APP immunohistochemistry showed density of positive cell in the CA1 region of hippocampus of control group were less than model group. H102 peptide reduced the area, and density of positive cells. Congo red staining showed there were lots of amyloid plagues in the brains of model mice but not in the brains of normal control. And the Western blot showed the content of the APP protein of the model group was much higher than the H102 group. H102 significantly decreased the amyloid plagues.

CONCLUSIONThe expression of APP, Abeta are increased in APP695 transgenic mice, and H102 can decrease the level of APP, Abeta in transgenic mice.

Amyloid beta-Protein Precursor ; metabolism ; Amyloidogenic Proteins ; metabolism ; Animals ; Brain ; metabolism ; Female ; Gene Expression ; Hippocampus ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic