Objective To investigate the apoptosis response to mechanical stretch in human alveolar type Ⅱ epithelial cells A549.Methods First,A549 cells were given different stretch(0%,5%,15%,30%) at frequency of 0.5 Hz for 4 h.Then,A549 cells were stretched for different periods(0 h,2 h,4 h,8 h) at stretch 15%,0.5 Hz.Last,A549 cells were stretched for different frequency(0 Hz,0.2 Hz,0.5 Hz and 1 Hz) at stretch 15% for 4 h.The early apoptosis was detected by flow cytometry. Results A549 cells submitted to cyclic stretch caused the early apoptosis in a time-,stretch-and frequency-dependent manner.In details,at 0.5 Hz and 4 h stretch-dependence occurred,at 15% stretch and 0.5 Hz time-dependence occurred,and at 15% stretch and 4 h frequency-dependence occurred. Conclusion Mechanical stretch can induce the early apoptosis in human alveolar type Ⅱ epithelial cells A549,which may be one of the mechanisms of ventilator-induced lung injury.

Discharge against medical advice (DAMA) conflicts with the purpose of disease treatment in children. Some research has shown that there are high proportions of extremely preterm infants and infants with asphyxia or congenital malformation in neonates with DAMA. This suggests that the sustainable development of neonatology needs cooperation and co-development with obstetrics, neonatal surgery, and radiology to reduce the rate of DAMA. With reference to the current status of research in both China and other countries, this article reviews the causes for DAMA and the strategies for reducing the rate of DAMA, in order to provide a theoretical basis for effectively reducing the rate of DAMA from the neonatal intensive care unit, improving treatment outcomes of the neonates, and increasing hospitals' comprehensive benefits.
Ethics, Medical ; Health Services Needs and Demand ; Humans ; Infant, Newborn ; Insurance, Health ; Intensive Care Units, Neonatal ; Patient Discharge ; Prenatal Care ; Treatment Refusal

OBJECTIVETo investigate the contents of lead, cadmium, zinc and manganese in the follicular fluid and semen of infertile couples that are not professionally exposed to the four heavy metallic elements.

METHODSIn vitro fertilization pre-embryo transfer (IVF-ET) was carried out in wives, and follicular fluid collected after routine oocyte retrieval. Semen was obtained from husbands by masturbation. The contents of zinc in the follicular fluid and semen were determined by the flame atom absorption method and those of lead, cadmium and manganese by graphite furnace atomic absorption spectrometry.

RESULTSThe average concentrations of lead, cad- mium, zinc and manganese were 151.06 microg/L, 2.02 microg/L, 0.54 mg/L and 28.54 microg/L in the follicular fluid, and 250.23 microg/L, 7.42 microg/L, 189.11 mg/L and 82.15 microg/L in the semen. The follicular fluid samples in which lead, cadmium, zinc or manganese was detected constituted 43.8% (21/48), 22.9% (11/48), 75.0% (36/48) and 50.0% (24/48), and the semen samples accounted for 70.2% (33/47), 31.9% (15/47), 100.0% (47/47) and 46.8% (22/47), respectively.

CONCLUSIONThe results suggest that the average contents of lead, cadmium, zinc and manganese are higher in the semen than in the follicular fluid in the non-professionally exposed infertile couples, and so is the percentage of the samples containing each of the elements, with the exception of manganese.

Adult ; Cadmium ; analysis ; Environmental Exposure ; analysis ; Female ; Follicular Fluid ; chemistry ; Humans ; Infertility, Female ; metabolism ; pathology ; Infertility, Male ; metabolism ; pathology ; Lead ; analysis ; Male ; Manganese ; analysis ; Metals, Heavy ; analysis ; Semen ; chemistry ; Spectrophotometry, Atomic ; Zinc ; analysis

Acquired Immunodeficiency Syndrome ; Blood Donors ; China ; HIV Infections ; Humans ; Prejudice ; Rural Population

Objective To explore the effects and the mechanisms of curcumin on in vitro viability,migration and invasion of human breast cancer cell line.Methods The test group (the concentrations of curcumin were 5,10,20 μg · mL-1,respectively)and control group were set in experiment.The inhibition effect of curcumin on the viability of MDA-MB-231 was determined with methyl thiazolyl tetrazolium (MTT).The apoptosis rate was determined with TUNEL assay.Themigration rate was determined with wounding assay.The invasion number of MDA-MB-231 was determined with tranwell assay.The expression of phosphorP38 was determined with Western blot.Results Curcumin suppressed MDA-MB-231 cell viability in a concentration-dependent manner when compared to the control.In high-dose group,cell viability after 48 h of curcumin treatment was decreased 49.17％ compared to control group;In medium-dose group,cell viability after 48 h of treatment was decreased 67.63％.In medium-dose group,the apoptosis rate after 48 h of curcumin treatment was increased 6.44％ compared to control group (0.38％);the apoptosis rate 48 h of curcumin treatment was increased 24.22％ in high-dose group.In medium-dose group,the migration rate after 48 h of curcumin treatment was decreased 55.33％ compared to control group;the migration rate 48 h of curcumin treatment was decreased 40.67％ in high-dose group.In medium-dose group,the invasion number after 48 h of curcumin treatment was decreased (37 ± 7) cell,compared to control group (104 ± 8) cell;the invasion number 48 h of curcumin treatment was decreased (25± 9) cell in high-dose group.In medium-dose group,the expression of phosphor-P38 after 48 h of curcumin treatment was decreased 57.63％ compared to control group;the expression of phosphor-P38 was decreased 42.51％ in high-dose group.Conclusion Curcumin suppresses human liver cancer cell MDA-MB-231 viability,migration and invasion by suppressing the expression of phosphorylation P38.

OBJECTIVETo construct the prokaryotic and eukaryotic expression vectors pET32/E5 and pcDNA3.1/E5 for transformation into E. coli BL21 and NIH(3)T(3) cells respectively to observe the expression of human papillomavirus type 16 E5 protein (HPV16 E5).

METHODSHPV16 E5 gene was amplified by PCR from clinical isolates of HPV 16 and inserted into the plasmid pET32a(+) followed by digestion with BamH I and Hind III. The recombinant plasmid pET32/E5 was transformed into E. coli JM109 and selected with ampicillin. The positive clones containing the recombinant plasmid pET32/E5 were verified by restriction endonucleases BamH I and Xho and sequence analysis. The expression of HPV16 E5-TRX fusion protein in E. coli BL21(ED3) was identified by SDS-PAGE and Western blotting. The digestion product of BamH I and Xho was purified and inserted into the eukaryotic expression vector pcDNA3.1(+) to construct pcDNA3.1/E5, which was identified by sequencing and transfected into NIH3T3 cells. The NIH(3)T(3) cells with stable expression of HPV16 E5 were selected by G418 and confirmed by RT-PCR.

RESULTSThe pET32/E5 and pcDNA3.1/E5 vectors were constructed successfully. E.coli BL21(DE3) transformed by the recombinant plasmid pET32/E5 expressed HPV16 E5-TRX fusion protein efficiently. In the presence of 1 mmol/L IPTG at 28 degrees C, HPV16 E5-TRX recombinant protein accounted for about 10% of the total bacterial proteins. NIH3T3 cells stably expressing HPV16 E5 were harvested by selection with 250 g/ml of G418. HPV16 E5 gene from pcDNA3.1/E5-transfected NIH(3)T(3) cells was amplified by RT-PCR, and sequence analysis demonstrated the acquisition of the full-length gene fragment.

CONCLUSIONSThe prokaryotic and eukaryotic vectors for the HPV16 E5 gene have been successfully constructed. The acquisition of E .coli and NIH(3)T(3) cells with stable expression of HPV16 E5 protein may facilitate subsequent research of the biological properties and the transformation mechanism of HPV16 E5 protein on specific cells.

3T3 Cells ; Animals ; Escherichia coli ; genetics ; metabolism ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; Humans ; Mice ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; virology ; Plasmids ; genetics ; Prokaryotic Cells ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics

OBJECTIVETo evaluate clinical therapeutic effect of post-stroke shoulder pain treated by acupuncture combined with Tuina.

METHODSThree hundred cases of post-stroke shoulder pain were randomly divided into an acupuncture and Tuina group and a rehabilitation group by double-center randomized controlled clinical trial method. In acupuncture and Tuina group, normalized electroacupuncture and Tuina therapy were applied, that was electroacupuncture at main points, such as Chize (LU 5), Quze (PC 3), Shaohai (HT 3), Jianyu (LI 15), Jianliao (TE 14) and Jianjing (GB 21),etc., combined with traditional Tuina manipulations; in rehabilitation group, the rehabilitation methods such as the electrostimulation through nervus cutaneus and the squeezing and stabilizing manipulations of Proprioceptive Neuromuscular Facilitation (PNF), etc. were applied. The treatment courses of both groups were 6 weeks. The main therapeutic effect indices were the Assessment Face Scale (AFS) for pain when shoulder was in passive motion and the Fugl-Meyer Motor Assessment for upper limbs active function; the secondary indices were the moditied Rankin Scale (mRS) and the clinical incidences of shoulder-hand syndrome of hemiplegia and shoulder joint subluxation of hemiplegia.

RESULTSAfter 6 weeks treatment and 12 weeks follow-up, AFS score, Fugl-Meyer motor assessment of upper limbs active function and mRS evaluation in acupuncture and Tuina group were more obviously improved than those in rehabilitation group (P < 0.05, P < 0.01). Although the clinical incidences of shoulder-hand syndrome of hemiplegia and shoulder joint subluxation of hemiplegia in acupuncture and Tuina group was equal to those in rehabilitation group [3.55% (5/141) vs 8.45% (12/142), 1.42% (2/141) vs 5.63% (8/142), both P > 0.05], the data indicated that there was a superiority tendency in acupuncture and Tuina group.

CONCLUSIONThe combined therapy of electroacupuncture and Tuina is a normative manipulation, and the therapeutic effect is satisfying for post-stroke shoulder pain, superior to that of comprehensive rehabilitation treatment.

Aged ; Amobarbital ; Combined Modality Therapy ; Drug Combinations ; Electroacupuncture ; Female ; Humans ; Male ; Middle Aged ; Range of Motion, Articular ; Secobarbital ; Shoulder Pain ; etiology ; physiopathology ; therapy ; Stroke ; complications

OBJECTIVETo evaluate the prevalence and distribution of C-kit, NPM1 and FLT3 gene mutations in patients with acute myeloid leukemia (AML), and to analyze the relationship between the gene mutations and their prognosis.

METHODSMutations in exon 8 and 17 of C-kit gene, exon 12 of NPM1 gene, exon 20 of FLT3-TKD gene, and exon 14/15 of FLT3-ITD gene were detected by direct sequencing. Clinical data was collected and followed up if the patient had accepted treatment in our hospital.

RESULTSAmong the 656 AML patients, mutations in C-kit exon 8 were found in 6 patients (0.9%), C-kit exon 17 in 33 (5.0%), NPM1 in 169 (25.8%), FLT3-TKD in 46 (7.1%), and FLT3-ITD in 178 (27.1%). Six subtypes of mutations were detected in C-kit exon 8, 8 in C-kit exon 17, 11 in FLT3-TKD, 15 in NPM1, of which 5 were not reported before. C-kit exon 17 mutations were more frequently detected in patients with t(8;21) and exon 8 in patients with inv(16) cytogenetic abnormality. No other gene mutations except FLT3 were detected in M(3) patients. NPM1 and ITD mutations were often detected in individuals with normal cytogenetics or M(5) and M(1) of FAB classification, and accompanied with high white blood cell counts in peripheral blood, high blast counts in bone marrow and low CD34 expression. The older the patients were when diagnosed, the more gene mutations and the higher white blood cell count were detected. More mutations were found in individuals with normal karyotype than that with other karyotypes. It appeared that FLT3-ITD was significantly associated with shorter overall survival (OS) (P = 0.004), NPM1 was not significantly associated with OS, but NPM1(+)/ITD(-) patients had the longest OS.

CONCLUSIONSOur results showed that the mutation types and amounts had particular distribution in MICM subtypes, and were associated with white blood cell counts in peripheral blood, blast counts in bone marrow and prognosis. Especially for patients with normal karyotype, the genetic mutations could be new molecule marker.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group ; genetics ; DNA Mutational Analysis ; Female ; Humans ; Karyotyping ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Male ; Middle Aged ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

A two-dimensional liquid chromatography method was developed for the analysis of rice leaves proteomics based on the coupling of hydrophilic interaction liquid chromatography-reversed-phase liquid chromatography with online tandem mass spectrometry. The influence of pH value of chromatographic mobile phase on the orthogonality of the hydrophilic interaction-reversed-phase two-dimensional liquid chromatography was evaluated by the changes of standard peptide retention. The results indicated that the better orthogonality (R2=0.34113) was achieved from the system with hydrophilic interaction columns(pH 9.3) in the first and C18columns(pH 3.3) in the second LC dimension. Coupled with multiple fraction concatenation strategy,the orthogonality of two-dimensional liquid chromatography was further evaluated in the analysis of complex rice leaf proteins. The results showed that more than 50% of the total peptides were identified less than two times, and the peptides obtained from first-dimension were well distributed across the elution window,indicating that the method showed significant orthogonality in the identification of complex rice leaf proteins. Based on the proteome discoverer software,207345 peptides belonged to 2930 protein clusters were identified.

OBJECTIVETo determine the sero-prevalence of and the risk factors for human immunodeficiency virus infection among residents in a former commercial blood donating community, Shanxi Province.

METHODSA community-based cross-sectional survey was conducted among all residents aged 18 - 64 years in 4 villages in a rural county of Shanxi Province. A standardized structured questionnaire was administered to collect socio-demographic, medical and risk behavioral information. Venous blood was collected for HIV antibody testing. Generalized estimating equation with logistic regression was used to analyze the risk factors for HIV infection. Population attributable fraction was used to evaluate the role of public health in the risk factors for the incidence of HIV.

RESULTSA total of 3062 villagers participated in the study and provided their blood samples for HIV antibody testing. 29.5% of the participants had a history of selling blood and/or plasma. Among the subgroup of former blood donors (FBDs), 71.9% sold whole blood only, 4.1% sold plasma only and 24.0% sold both. Forty HIV infected persons were identified in the four villages. HIV prevalence was 1.3%, overall, 4.1% in FBDs and 0.1% in non-FBDs in the community. Amongst the subgroup of FBDs, HIV prevalence was 1.5% for whole blood only, 21.6% for plasma only and 8.8% for both. Univariate analysis showed that village of residence, education, marital status, gender, plasma/blood donation were statistically associated with HIV seropositivity in the villagers. Other variables, such as gender, age, occupation, number of sexual partners, drug use, histories of tooth extraction, acupuncture, surgical operation and medical injection were not found to be statistically associated with HIV seropositivity. Multivariate analysis found that selling whole blood only (adjusted OR = 13.25, 95% CI: 3.02 - 58.25) and selling plasma or both (adjusted OR = 87.85, 95% CI: 22.22 - 347.27) were significantly associated with HIV seropositivity, and the additive population attributable fraction (PAF) was 89.8%. Villagers in village 03 were more likely to be HIV positive than those in village 04 (adjusted OR = 12.26, 95% CI: 1.46 - 103.00).

CONCLUSIONAt present, HIV epidemic in the former commercial blood donating community has a low prevalence in Shanxi Province and the majority of HIV infections should be related to the blood and plasma donation. This study reveals the existence of HIV second-generation transmission from the infected blood donors to the general population. However, HIV had not been evidently extended from the blood/plasma donors to general population.

Adult ; Blood Donors ; China ; Cross-Sectional Studies ; Female ; HIV Infections ; epidemiology ; Humans ; Logistic Models ; Male ; Middle Aged ; Risk Factors ; Rural Population ; Serologic Tests ; Surveys and Questionnaires