Cell Differentiation ; drug effects ; Dimethyl Sulfoxide ; pharmacology ; HL-60 Cells ; Humans ; Proteomics

Objective To characterize molecule profiling for hypoxia/hypothermia-conditioned human endothelial cells(EC),and to examine the hypothesis that the interaction between hypoxia/hy- pothermia-conditioned EC and monocytes plays a key role in early hypoxia/hypothermic reperfusion in- jury.Methods Human aortic EC were conditioned in hypoxia/hypothermia chamber at 4℃for 91) rain followed by 4 to 8 h re-warming at 37℃and then analyzed initially by RT-PCR to evaluate RNA transcripts.To examine early cellular interactions,a cellular adhesion study was performed using EC and fluorescence-conjugated DIO labeled purified human monocytes,and CD4~+ T cells as well as PB- MC.Interactions between PKH-26-labeled EC and these cells were carried out to determine EC mem- brane uptake by monocytes.Results RT-PCR analysis demonstrated that hypoxia/hypothermia-con- ditioned EC up-regulated RNA transcripts of inflammatory cytokines,ECE-1 and endothelin,co-stim- ulatory molecules,and adhesion molecules CD54.FACS showed minimal up-regulation of CD54 on hy- poxia-hypothermia-conditioned EC.Low level adhesion of purified CD4~+ T cells to resting and hypoxi- a/hypothermia-conditioned EC was noted.In contrast,high levels of monocyte adhesion to hypoxia/ hypothermia-conditioned EC were observed when compared to resting EC.EC membrane up-take by monocytes but not CD4~+ T cells was revealed by FACS,and the levels of EC membrane up-take were similar to those of the normal EC.Conclusions Hypoxia/hypothermia-conditioned EC demonstrated up-regulation of multiple RNA transcripts associated with inflammatory responses,cellular adhesion and co-stimulation.Human monocyte adhesion to hypoxia/hypothermia-conditioned EC was greater than T cells.Monocytes up-took EC membrane during their interaction.These findings suggest that the initial interaction between monocytes and hypoxia-hypothermia EC,but not T cells,may play a critical role during early reperfusion injury.

Genetic mutations are important molecular biomarkers for cancer diagnosis and surveillance.Therefore,the development of methods for mutation detection characterized with straightforward,highly specific and sensitive to low-level mutations within various sequence contexts is extremely needed.Although some of the currently available methods have shown very encouraging results,their discrimination efficiency is still very low.Herein,we demonstrate a fluorescent probe coupled with blocker and property of melting temperature discrimination,which is able to identify the presence of known or unknown single-base variations at abundances down to 0.1％ within 20 min.The discrimination factors between the perfect-match target and single-base mismatched target are determined to be 10.15-38.48.The method is sequence independent,which assures a wide range of application.The new method would be an ideal choice for high-throughput in vitro diagnosis and precise clinical treatment.

OBJECTIVE: To analyze the cases of anaphylactic death cases and explore the standards of judicial expertise of anaphylactic death for providing evidence for judicial expertise. METHODS: Fifty-nine cases death due to allergic reaction in Shanghai were collected. And details of medical history, clinical manifestation of anaphylactic reaction and postmortem examination findings were reviewed for all cases. RESULTS: In the 59 cases, there were 58 cases died from drug allergy, including 77.6% of them were antibiotics. The rates of treating in standard hospital and illegal clinic were 37.3% and 61.0%, respectively. The allergic symptoms were dyspnea and facial cyanosis. The time from contacting allergens to death ranged from 1 min to 3 d. The concentration of total serum IgE ranged from 50 to 576.92 IU/mL. The results of clinical manifestation and pathological anatomy had obviously changes. CONCLUSION Based on the exclusion of all other cause of death and synthetically analysis of details of cases, medical history, clinical manifestation and anatomy, the conclusion of anaphylactic death can reached. The details of cases including clinical history, exposure to allergens, and clinical manifestation play an important role in diagnosis of anaphylactic death.
Anaphylaxis/mortality* ; Anti-Bacterial Agents/adverse effects* ; Autopsy ; China ; Drug Hypersensitivity/mortality* ; Forensic Sciences ; Humans

This paper reported that the activation of oncogenes in human fetal esopha geal epithelium treated by alternariol (AOH). It was found that NIH/3T3 cells were transformed via transfeetion of DNA extracted from human fetal esophageal epithelium which was cultured and treated by 10?g/ml AOH in a short term in vitro. The efficiency of primary loci was 0.17 focus per ?g of DNA. In the secondary transfection, the efficiency was 0.58 focus per ?g of DNA (P

Adolescent ; Adult ; Female ; Follow-Up Studies ; Humans ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; Treatment Outcome ; Young Adult

OBJECTIVETo report the clinical characteristics and treatment of 3 patients with juvenile xanthogranuloma (JXG).

METHODSA retrospective review of the medical records of 3 patients with JXG.

RESULTSJXG was characterized by solitary or multiple yellowish cutaneous nodules, or eye involvement . It could also affect pituitary. JXG was easily misdiagnosed as Langerhans cell histiocytosis (LCH). Treatment for JXG was surgical excision of a solitary skin lesion and some cases might be, spontaneous regression. In cases with multisystem involvement, chemotherapy regimens used to treat LCH may be effective.

CONCLUSIONSJXG is one of the more common non-Langerhans histiocytic proliferations and is frequently seen in infants and children. LCH-like chemotherapy is effective for patients with symptomatic multisystem JXG.

Child, Preschool ; Female ; Humans ; Infant ; Male ; Xanthogranuloma, Juvenile ; diagnosis ; therapy

OBJECTIVETo evaluate the impact of specific siRNA on survivin gene in transfected leukemia cells.

METHODThe small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into K562 cell by Hiperfect into human leukemia cell line K562, which has high survivin expression level. The level of survivin mRNA expression was determined by quantitative reverse transcription polymerase chain reaction (RT-PCR) with SYBR GREEN I. The apoptosis index of cytotrophoblasts were determined and analyzed by FCM (Annexin V-FITC/PI staining methods). The cell proliferation was examined by MTT at 48 h and 72 h after transfection.

RESULTThe level of mRNA expression was significantly inhibited by the siRNA 48 h and 72 h after transfection, the suppression rate of survivin mRNA separately reached 85.21%, 94.35% mensurated by quantitative RT-PCR with SYBR GREEN I, cell proliferation was inhibited significantly by 45.02% and 50.88%, respectively, the apoptotic rate detected by Annexin V-FITC assay reached 12.28%and 21.55%, respectively.

CONCLUSIONThe chemosynthesized siRNA targeting survivin could significantly down-regulate survivin mRNA. Survivin siRNA was able to inhibit the proliferation of leukemia cell line K562. Survivin may become a new target for leukemia gene therapy.

Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; RNA, Small Interfering ; pharmacology ; Transfection

OBJECTIVESince most reports on bystander effect have been only concerned with radiation-induced damage, the present paper aimed at disclosing whether low dose radiation could induce a stimulatory or beneficial bystander effect.

METHODSA co-culture system containing irradiated antigen presenting cells (J774A.1) and unirradiated T lymphocytes (EL-4) was established to observe the effect of J774A.1 cells exposed to both low and high doses of X-rays on the unirradiated EL-4 cells. Incorporation of 3H-TdR was used to assess the proliferation of the EL-4 cells, expression of CD80/86 and CD48 on J774A.1 cells was measured with immunohistochemistry and flow cytometry, respectively. NO release from J774A.1 cells was estimated with nitrate reduction method.

RESULTSLow dose-irradiated J774A.1 cells could stimulate the proliferation of the unirradiated EL-4 cells while the high dose-irradiated J774A.1 cells exerted an inhibitory effect on the proliferation of the unirradiated EL-4 cells. Preliminary mechanistic studies illustrated that the differential changes in CD48 expression and NO production by the irradiated J774A.1 cells after high and low dose radiation might be important factors underlying the differential bystander effect elicited by different doses of radiation.

CONCLUSIONStimulatory bystander effect can be induced in immune cells by low dose radiation.

Animals ; Antigen-Presenting Cells ; immunology ; metabolism ; radiation effects ; Antigens, CD ; immunology ; B7-1 Antigen ; immunology ; B7-2 Antigen ; Bystander Effect ; radiation effects ; Cell Division ; immunology ; Cell Line ; Coculture Techniques ; Dose-Response Relationship, Radiation ; Membrane Glycoproteins ; immunology ; Mice ; Nitric Oxide ; biosynthesis ; T-Lymphocytes ; immunology ; X-Rays

To investigate the specific anti-leukemia effect of cytotoxic T lymphocytes (CTLs) induced by dendritic cells (DCs) activated by exosomes alone or in combination with CpG ODN in vitro and the feasibility of exosomes as remedial vaccine, the DCs induced from normal volunteer PBMNCs were divided into 7 groups. Three groups of them were added with the exosomes: Kexo (exosomes derived from K562 cells) or DCexo (exosomes derived from DCs induced from K562 cells) or FTexo (exosomes derived from DCs induced from K562 cells and pulsed by freeze-thawing antigen of K562 cells) as experimental groups (Kexo, DCexo and FTexo). The other three groups were added with CPG ODN while added the exosomes (Kexo, DCexo and FTexo), and were used as experimental groups also (Kexo+CpG, DCexo+CpG and FTexo+CpG). The seventh group DCs was added with nothing as blank control. These DCs above mentioned were cultured continuously for 72 hours. The T lymphocytes were co-cultured with DCs for another 72 hours to generate CTL. Then, the killing effects of them on K562 cells were determined by MTT assay. The results showed that all experimental groups pulsed by exosomes displayed stronger killing effect, compared with control group (p<0.05). DCexo and FTexo displayed stronger killing effect too, compared with Kexo (p<0.05). CPG ODN as an adjuvant could enhance the killing effect (p<0.05). It is concluded that the special killing effect on K562 cells can be induced by exosomes, CPG ODN as an adjuvant can enhance the killing effect. Exosome is hopeful as a remedial vaccine to be used for the leukemia therapy.
Adjuvants, Immunologic ; pharmacology ; Cancer Vaccines ; immunology ; Dendritic Cells ; immunology ; Exosomes ; immunology ; Humans ; K562 Cells ; Oligodeoxyribonucleotides ; immunology ; T-Lymphocytes, Cytotoxic ; immunology