Objective To construct eukaryotic expression vector of wild type E 4F1 and the mutant deleting amino acid region 32-81, and to detect the interaction between wild type or mutant E 4F1 and p53 and to study the effect of E4F1 on the expression level of p21.Methods Wild type and mutant sequences of E 4F1 were amplified from the mammary library using standard PCR and recombinant PCR .The sequences were cloned into pXJ 40-MYC vector to generate the MYC-E4F1 and MYC-E4F1(Δ32-81) recombinant plasmids that were transfected into 293T cells and identified by Western blotting . FLAG-p53 and MYC-E4F1 or MYC-E4F1(Δ32-81) were co-transfected into 293T cells and immunoprecipitation assay was performed to detect the interaction of wild type or mutant E 4F1 with p53.Wild type and mutant E4F1 expressing vec-tors were co-transfected into osteosarcoma U2OS cells and the expression of p21was detected.Results Recombinant plas-mids of MYC-E4F1 and MYC-E4F1(Δ32-81) were successfully constructed.Both wild type and mutant E4F1 interacted with p53.Deletion of amino acid region 32-81 of E4F1 increased the interaction .The expression level of p21 was in-creased by wild-type E4F1, but not by mutant E4F1.Conclusion The eukaryotic expression vector of wild type E4F1 and its deletion mutant is successfully constructed .Both of them interact with p53.Deletion of amino acid region 32-81 of E4F1 increases the interaction .This study contributes to further studies on the regulation and mechanism of E 4F1 on p53.

To study the chemical constituents from the root of Berchemia lineata (L.) DC., nine compounds were isolated from the EtOAc extract by using silica gel, RP-C18 silica gel column chromatography and preparative HPLC. Based on the spectroscopic analysis, their structures were identified as 5-hydroxy-7-(2'-hydroxypropyl)-2-methyl-chromone (1), (-)-(1'R, 2'S)-erythro-5-hydroxy-7-(1', 2'-dihydroxypropyl)-2-methyl-chromone (2), naringenin (3), eriodictyol (4), (+)-aromadendrin (5), (+)-taxifolin (6), (+)-catechin (7), (+)-epigallocatechin (8) and quercetin (9). Among them, compound 2 is a new chromone derivative. Compound 1 is a known chromone derivative and isolated from this genus for the first time. Compounds 3-9 are known flavonoids and isolated from this plant for the first time.
Catechin ; analogs & derivatives ; chemistry ; isolation & purification ; Chromones ; chemistry ; isolation & purification ; Flavanones ; chemistry ; isolation & purification ; Flavonoids ; chemistry ; isolation & purification ; Molecular Structure ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; analogs & derivatives ; chemistry ; isolation & purification ; Rhamnaceae ; chemistry

Adult ; Aged ; Cell Cycle Proteins ; analysis ; Coal Mining ; Cyclin-Dependent Kinase Inhibitor p27 ; Female ; Humans ; Immunohistochemistry ; Lung ; chemistry ; pathology ; Male ; Middle Aged ; Occupational Diseases ; metabolism ; Pneumoconiosis ; metabolism ; Tumor Suppressor Proteins ; analysis

OBJECTIVETo examine the differences in the expression of HSP27 and c-kit between benign prostatic hyperplasia (BPH) and prostatic cancer (PCa) tissues and to analyse the relationship between their expression and BPH and PCa, especially the relationship with the occurrence, development, prognosis and treatment of PCa.

METHODSAn immunohistochemical staining (SP method) for HSP27 and c-kit was undertaken on 40 BPH and 40 PCa tissues samples.

RESULTSConsistent patterns of cytoplasmic staining for HSP27 were seen in all sections of tissue from BPH. The glandular epithelium stained very strongly positively and the stroma stained positively. The staining for HSP27 in PCa tissues was located in the cytoplasm of glandular epithelia, but the expression of HSP27 in PCa was higher than BPH (P < 0.05). The staining for c-kit in BPH tissues was located in the cytoplasm of smooth muscle cells, and in PCa tissues was located in epithelial cells. The expression of c-kit in PCa tissues was lower than BPH (P < 0.05). The expression level of both HSP27 and c-kit were decreased with the development of grade of PCa (P < 0.05); HSP27 was increased with the development of clinical stage of PCa (P < 0.05 ); c-kit was decreased with the development of clinical stage of PCa (P < 0.05).

CONCLUSIONThe expression level of HSP27 and c-kit was highly correlated with the process of the development from BPH to PCa, and also correlated with tumor grades and stages. The expression of HSP27 and c-kit may be used as an important pathological index and may be helpful for the treatment of PCa.

Aged ; Aged, 80 and over ; HSP27 Heat-Shock Proteins ; Heat-Shock Proteins ; biosynthesis ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-kit ; biosynthesis

OBJECTIVETo investigate the expression variation and significance of Skp2 and p27(kip1) during the proliferation of lymphoma cell line Jurkat cells.

METHODSThe binding of p27(kip1) and Skp2 in Jurkat cells were detected by immunoprecipitation. Jurkat cells were treated with serum starvation and release synchronization. The expression variation and subcellular localization of p27(kip1) and Skp2 were detected by subcellular fractionation, Western blot and double immunofluorescence labelling.

RESULTSThe results of immunoprecipitation suggested that p27(kip1) and Skp2 could bind each other in Jurkat cells. During the proliferation of Jurkat cells, the protein expression of p27(kip1) decreased and intranuclear p27(kip1) decreased significantly, while the Skp2 protein increased and cytoplasmic Skp2 increased significantly.

CONCLUSIONDuring the proliferation of Jurkat cells, the increased cytoplasmic synthesis of Skp2 may speed up p27(kip1) degradation via the ubiquitin-proteasome pathway, then intranuclear p27(kip1) decreases significantly, leading to an increased cell cycling activity.

Cell Nucleus ; metabolism ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; metabolism ; Cytoplasm ; metabolism ; Humans ; Jurkat Cells ; Lymphoma, B-Cell ; metabolism ; pathology ; Protein Binding ; S-Phase Kinase-Associated Proteins ; metabolism

AIMTo analyse the impurities of gatifloxacin.

METHODSThe impurity of gatifloxacin were analysized and determinated by RP-HPLC/electrospray ionization mass spectrometry with a Zorbax SB-C18(4.6 mm x 150 mm ID, 5 microns). The mobile phase was 3% acetic acid/acetonitrile-3% acetic acid/water (15:85). The two compounds were synthesized: 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-methoxy-7-(1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMP) and 1-cyclopropyl-6-fluoro-1, 4-dihydro-8-hydro-7-(3-methy-1-piperazinyl)-4-oxo-3-quinolinecarboxylic acid (DMO). Their liquid chromatogram, UV, MS were compared with those of the impurity of gatifloxacin.

RESULTSThe mass of the impurity was 14 less than that of gatifloxacin. It means the impurity was CH2 less than gatifloxacin. The tR (HPLC), UV and MS of DMP were the same as those of the impurity of gatifloxacin.

CONCLUSIONBased on the tR (HPLC), UV and MS, the impurity of gatifloxacin is confirmed as DMP.

Anti-Infective Agents ; analysis ; chemistry ; Chromatography, High Pressure Liquid ; Drug Contamination ; Fluoroquinolones ; analysis ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective To explore the clinical distribution and significant of 23 kinds of human papillomavir-us(HPV)genetypes in normal cells and atypical squamous cells(ASC-US),meanwhile analysis result of cervi-cal histological pathology diagnosis in cases of ASC-US.Methods A total of 1 000 women with normal cells specimens were recruited into control group,and 236 women with ASC-US were selected into the ASC-US group.Polymerase chain reaction(PCR)and gene-chips technology were utilized for the detection of 23 kinds of HPV genetypes,all cases of ASC-US diagnosis of cervical pathological histology.Results A total of 106 ca-ses of HPV infection were detected in the control group,as the total HPV infection rate was 10.6%,in which the single genotypes infection rate was 9.3% and the multiple genotypes infection rate was 1.3%.A total of 139 cases of HPV infection were detected in ASC-US group,as the total HPV infection rate was 58.9%,in which the single genotypes infection rate was 38.1%,and the multiple genotypes infection rate was 20.8%. There were significant differences on the total HPV infection rate,the infection rates of type 1 and multiple geontypes between the control group and ASC-US group(P<0.05).The top six of constituent ratio in the control group were type 43,16,58,33,52,18,42,those in the ASC-US group were type 16,18,52,58,33,51,66.Conclusion PCR combined with the gene-chip technology could be used in the HPV genotypes detection in cervical cells,which has important clinical significance on the further distribution management of ASC-US,and should be draw great attention.

BACKGROUND:Cycloserine with low hepatotoxicity exhibits no cross-resistance with the existing anti-tuberculosis drugs,and has been commonly used for the treatment of drug-resistant tuberculosis.However,its oral administration or injection leads to a certain degree of neurotoxicity.OBJECTIVE:To prepare poly(lactic-co-glycolic acid) (PLGA)-cycloserine sustained-release microspheres which are expected to reduce the neurotoxicity and adverse reactions,and maintain the drug concentration in the bone tuberculosis region for a long time,and to observe the in vitro drug release of the microspheresMETHODS:Double emulsion solvent evaporation method was used to prepare PLGA-cycloserine microspheres that were bonded into sponge implant by Bletilla striata polysaccharide extract.Then,morphology,particle size,encapsulation efficiency and in vitro performance of the microspheres were observed.The drug loading,burst release,appearance and dispersion of the microspheres were observed at 0,1,2 months after the microspheres were placed in room temperature (25 ℃),high temperature (60 ℃) and high humidity (93％),respectively.RESULTS AND CONCLUSION:The PLGA-cycloserine microspheres that were round and spherical presented with the mean particle size of (143±38) μm,the drug loading of 38.38％ and the encapsulation efficiency of 67.54％.No burst release occurred,and the cumulative release of drug within 50 days was 65.62％ After being stored at room temperature,high temperature and high humidity for 1 and 2 months,the microspheres were intact in the appearance and morphology,and showed insignificant changes in drug loading and burst release.To conclude,the time of degradation and the release of drug accord with the biological requirements of bone restoration.

Objective To investigate the changes and characteristics of body mass index (BMI) of low birth weight infants during catch-up growth within 24 months of life. Methods Using the birth cohort method, 126 low birth weight children (birth weight less than 2 500 g) among the registered and permanent born in Jiading District from January 2016 to December 2016, were enrolled in the study voluntarily.According to the calculation of birth weight and gestational week, 73 children were included in the preterm appropriate for gestational age group and 53 in the full-term small for gestational age group.105 children with gestational age of 37-41 weeks and birth weight of 2 500-3 999 g were included as the control group.The differences of BMI mean and standard deviation were compared between 0-24 months old in three groups, and the changes of BMI curve analyzed between 0-24 months old in boys and girls. Results ① There were 231 infants investigated, who were composed by 111 boys and 120 girls; ② The BMI of the two groups of low birth weight infants at birth and at 2 months old were lower than those of the control group.There was no significant difference between the BMI of preterm appropriate for gestational age group and the control group since the age of 4 months.The BMI of the term small for gestational age group was less than the other two groups between 4 and 18 months of age, the difference was statistically significant (P < 0.05);③ The BMI index of the three groups showed a rapid rise after birth.It peaked at 4-6 months of age, and the BMI value of 7-9 months of age began to fall.Preterm appropriate for gestational age group infants caught up with the BMI of normal-weight infants at 6 months of age.Until the age of 24 months, the BMI of small for gestational age group was still different from normal weight infants, but the difference between the three groups decreased.The rising curves of BMI between boys and girls were similar, but the peak of preterm appropriate for gestational age group girls was lengthened. Conclusion There is a significant catch-up growth for low birth weight infants aged 0-24 months, having a similar trend of normal infants in the late stage.It is necessary to deliver proper breeding education and intervention to the low birth weight infants in their early stages.

BACKGROUNDPopulation based epidemiologic study on the main diseases and birth status of liveborn neonates remains scarce in China, especially in rural areas where a large number of neonates are born. The aim of this study was to establish an epidemiological basis of live births in Julu County, a representative of the northern and mid-western parts of China in terms of demography, disease pattern and women and children's health care infrastructure.

METHODSThe perinatal data of all live births were prospectively collected in three participating county-level hospitals from September 1, 2007 to August 30, 2008.

RESULTSThere were 5822 live births in these hospitals. Among all live births, 53.7% were male and 4.5% were born prematurely. Mean (SD) birth weight (BW) was (3348 ± 503) g. The low (< 2500 g) and very low BW (< 1500 g) infants accounted for 3.8% and 0.5% of the total births, with 6.5% as small for gestational age and 2.8% as multi-births. Cesarean section rate was 30.2%, of which 68.6% were elective. There were 745 infants (12.8% of the live births) admitted to local neonatal wards within 7 days of postnatal life, in which 48.3% and 19.3% were due to perinatal asphyxia and prematurity, respectively. The incidences of perinatal aspiration syndrome, transient tachypnea and respiratory distress syndrome were 4.9%, 0.6% and 0.5%, respectively. Neonatal mortality was 7.6‰ (44/5822), with 16 in delivery room and 28 in neonatal ward before discharge.

CONCLUSIONSThis study provided a population-based perinatal data of live births and neonatal mortality in a northern China county with limited resources. Neonatal disorders related to perinatal asphyxia remain a serious clinical problem, which calls for sustained education of advanced neonatal resuscitation and improvement in the quality of perinatal-neonatal care.

Asphyxia Neonatorum ; epidemiology ; Birth Weight ; China ; epidemiology ; Female ; Humans ; Infant Mortality ; Infant, Newborn ; Infant, Newborn, Diseases ; epidemiology ; therapy ; Male ; Prospective Studies ; Respiratory Distress Syndrome, Newborn ; epidemiology ; therapy