Objective To construct a lentiviral vector of RNA interference (RNAi) of murine triggering receptor expressed on myeloid cells-1 (TREM-1) gene and to explore the effect of TREM-1 on the inflammatory response caused by Bacteroides fragilis. Methods Four target sequences were selected according to murine TREM-1 mRNA sequence, and then 4 pairs of double-strand DNA oligo according to these target sequences and one pair of negative control double-strand DNA oligo were designed and synthesized. These fragments were subcloned into pGCSIL-GFP/Lenti plasmid. After being identified by PCR and sequencing, these plasmids were cotransfected into 293T cells to package lentiviral particles. The lentiviral vector particles were transfected into Raw 264. 7 cells and TREM-1 expression in the transfected cells was assayed by real-time PCR and ELISA. Different concentrations of Bacteroides fragilis lipopolysaccaride (LPS) were administered in the Raw264. 7 cells, and the cells were stimulated with LPS for 12 h. TREM-1 expression was determined by real-time PCR and ELISA at the time points. Results PCR and sequencing confirmed that lentiviral vectors had the correct structure and could express high titer of virus. After being transfected into Raw264. 7 cells, TREM-1 expression was knocked down significantly by all of these lentiviral vectors at both protein and mRNA levels, and the pGCSIL-GFP/Lenti-1 had the most efficient interference. TREM-1 was upregulated in the presence of Bacteroides fragilis LPS, and this increase was partly abrogated in the TREM-1 siRNA-treated cell models of endotoxemia, depending on the sequence. Conclusion The lentivirus RNAi vector of TREM-1 was constructed successfully. The lentivirus RNAi vector of TREM-1 can inhibit the expression of TREM-1 in the murine endotoxemia model caused by Bacteroides fragilis LPS.

Palliative care focuses on improving the quality of life for patients and families facing life-threatening illness. Since its introduction in China,palliative care has developed rapidly with greater numbers of elderly adults and increasing chronic disease. However,palliative care education lags behind somehow,especially for undergraduate medical students. This article summarizes the history and status quo of palliative care education for undergraduate medical students in countries and regions including the United Kindom,the United States,Japan,and Taiwan Province,with an attempt to further promote the palliative care education for medical students in mainland China.

BACKGROUNDThe CKLF-like MARVEL transmembrane domain-containing family (CMTM) is a novel family of proteins linking chemokines and TM4SF. Different members exhibit diverse biological functions. In this study, the effect of intracellular CMTM2 on regulating human immunodeficiency virus type-1 (HIV-1) transcription was evaluated.

METHODSThe effects of CMTM2 on regulating full-length HIV-1 provirus and the HIV-1 long terminal repeat (LTR)-directed transcription were assessed by luciferase assay. Transcription factor assays, using the luciferase reporter plasmids of AP-1, CRE, and NF-κB were conducted to explore the signaling pathway(s) that may be regulated by CMTM2. The potential relationship between CMTM2 and the transcription factor AP-1 was further analyzed by Western blotting analyses to investigate the effect of CMTM2 on PMA-induced ERK1/2 phosphorylation.

RESULTSThe results from the current study revealed that CMTM2 acts as a negative regulator of HIV-1 transcription. CMTM2 exerted a suppressive action on both full-length HIV-1 provirus and HIV-1 LTR-directed transcription. Transcription factor assays showed that CMTM2 selectively inhibited basal AP-1 and CREB activity. Co-expression of HIV-1 Tat, a potent AP-1 and CREB activator, can not reverse CMTM2-mediated AP-1 and CREB inhibition, suggesting a potent and specific effect of CMTM2 on negatively regulating these two signaling pathways.

CONCLUSIONIntracellular CMTM2 can negatively regulate HIV-1 transcription, at least in part, by targeting the AP-1 and CREB pathways. Exploring the mechanisms further may lead to new ways to control HIV-1 replication.

Chemokines ; physiology ; Cyclic AMP Response Element-Binding Protein ; antagonists & inhibitors ; HIV Long Terminal Repeat ; HIV-1 ; genetics ; Humans ; Intracellular Space ; metabolism ; Jurkat Cells ; MARVEL Domain-Containing Proteins ; Tetradecanoylphorbol Acetate ; pharmacology ; Transcription Factor AP-1 ; antagonists & inhibitors ; Transcription, Genetic ; U937 Cells

OBJECTIVETo study the effect of pregnancy-induced hypertension syndrome (PIH) on complications in very low birth weight (VLBW) preterm infants.

METHODSThe VLBW preterm infants were enrolled as research subjects, and according to the presence or absence of PIH in their mothers, they were divided into PIH group and non- PIH group. The incidence of major complications and length of hospital stay were compared between the two groups.

RESULTSThere were no significant differences between the two groups in gestational age, birth weight, sex, incidence rate of maternal diabetes, and use of antepartum hormone. The PIH group had a significantly higher rate of birth of small-for-gestational-age infants than the non-PIH group. The PIH group had a significantly lower incidence rate of bronchopulmonary dysplasia (BPD) than the non-PIH group, while there were no significant differences between the two groups in the incidence rates of apnea of prematurity, necrotizing enterocolitis, retinopathy of prematurity, and intraventricular hemorrhage-periventricular leukomalacia, and the length of hospital stay. There was no significant difference in the incidence rate of neonatal respiratory distress syndrome between the two groups, but the PIH group had a significantly lower proportion of infants who used pulmonary surfactant than the non-PIH group.

CONCLUSIONSPIH can alleviate respiratory complications and reduce the use of pulmonary surfactant and the incidence rate of BPD in preterm infants.

Bronchopulmonary Dysplasia ; epidemiology ; etiology ; Female ; Humans ; Hypertension, Pregnancy-Induced ; Incidence ; Infant, Premature ; Infant, Very Low Birth Weight ; Pregnancy ; Pulmonary Surfactants ; therapeutic use ; Respiratory Distress Syndrome, Newborn ; epidemiology

OBJECTIVE: To analyze the changes of DC subsets and the expression of CD80 and CD86 in peripheral blood of ITP patients and their correlation with dexamethasone efficacy. METHODS: Peripheral blood sample of 80 cases of ITP and 20 normal controls from June 2015 to June 2017 in our hospital were retrospectively analyzed. The specific distribution of DC subsets in the peripheral blood of all the subjects was detected by flow cytometry, and the expressions of CD80 and CD86 were detected by ELISA. RESULTS: The proportion of DC2 in DC subsets of ITP patients before treatment was significantly higher than that in normal control group (P<0.05). The proportion of DC2 in DC subset of ITP patients was still significantly higher than that of the control group (P<0.05). The level of CD80 expression on DC1 and DC2 in ITP patients before treatment was significantly higher than that in the normal control group (P<0.05), and the expression level of CD86 on DC2 was significantly higher than that of the normal control group (P<0.05). Both IL-2 and IFN- γ levels in the patients before the treatment were significantly higher than those in the normal control group (P<0.05), and the expression levels after treatment with dexamethasone decreased significantly. Before treatment, both IL-4 and IL-10 levels in ITP patients were significantly lower than those in the normal control group (P<0.05), and their expression levels after treatment with dexamethasone significantly increased (P<0.05). CONCLUSION The incidence of ITP patients closely relates with the level and dysfunction of DC subsets in peripheral blood and the expression levels of IL-2, IL-4, IL-10, IFN- γ, which significantly correlates with the efficacy of dexamethasone.
B7-1 Antigen ; Dendritic Cells ; Dexamethasone ; Humans ; Inosine Triphosphate ; Retrospective Studies

OBJECTIVETo investigate the effect of triggering receptor expressed on myeloid cells 1 (TREM-1) vshRNA vector on expression of inflammatory cytokines and survival rate in septic mice infected by Bacteroides fragilis.

METHODS(1) TREM-1 vshRNA vector was constructed. Bacteroides fragilis (2.5 x 10(9) CFU/mL, 0.5 mL) was intraperitoneally injected in each mouse, and septic model was reproduced after 12 hours. (2) One hundred and fifteen mice were divided into healthy control group (n = 3, HC), sepsis group (n = 28, S), TREM-1 vshRNA group (n = 28, T), TREM-1 vshRNA hd group (n = 28, Th), GFP group (n = 28, G) according to random number table. Mice in S, T, Th, G groups were firstly injected with isotonic saline, TREM-1 vshRNA 2 x 10(8) TU, TREM-1 vshRNA 1 x 10(8) TU, GFP siRNA through tail vein, and then sepsis was induced after 1 hour. Mice in HC group were injected with equal volume of isotonic saline through tail vein. Three mice in each group were sacrificed after 12 hours for determination of plasma level of TNF-alpha, IL-1 beta and IL-6, and level of TREM-1mRNA and its protein in hepatic tissue. The survival rate of other mice in each group was monitored for 72 hours. (3) In 125 mice sepsis was reproduced, among them 100 mice were injected with TREM-1 vshRNA 2 x 10(8) TU after 1, 2, 4, 6 hours through tail vein (25 mice at each time point), other 25 mice were injected with equal volume of isotonic saline as control. The survival rate of mice in each group was recorded 72 hours after injection.

RESULTS(1) Compared with those in S group, the plasma level of TNF-alpha, IL-1 beta and IL-6 lowered in T and Th groups (P < 0.05), especially in T group, while those in G group showed no obvious difference (P > 0.05). (2) Compared with those in G group, the level of TREM-1mRNA and its protein in hepatic tissue in T and Th groups decreased (P < 0.01), especially in T group. (3) The survival rate of mice in S and G group was 16%, which was obviously lower than that in T and Th groups (76%, 44%, respectively, P < 0.05 or P < 0.01). (4) The survival rate of mice at 1, 2, 4, 6 hours after injection was 72%, 56%, 40%, 16%, respectively, while all that except at 6 hour after injection were higher significantly than that of control (P < 0.05 or P < 0.01).

CONCLUSIONSThe intervention with TREM-1 vshRNA can effectively decrease hepatic level of TREM-1 in septic mice induced by Bacteroides fragilis, inhibit inflammatory response, and improve the survival rate.

Animals ; Bacteroides fragilis ; Disease Models, Animal ; Genetic Vectors ; Lentivirus ; Male ; Mice ; Mice, Inbred BALB C ; RNA, Messenger ; metabolism ; Receptors, Immunologic ; genetics ; Sepsis ; metabolism ; microbiology ; therapy ; Virosomes

Microbeam X-ray fluorescence (micro-XRF) spectrometry has been raised as an analytical technique of microbeam during the recent years. With its advantages of high sensitivity, small sample requirement, high testing accuracy and non-destruction, the technique is widely utilized in forensic science. This review bases on recent researches at home and abroad, describes its applications including identification of gunshot residue, visualization of fingerprints, discrimination of drug source, production process, and other material evidences of analysis in crime scene. Thanks to the advances in technology, intelligent and portable micro-XRF equipment has appeared to be applied. It is believed that it may be more popular and frequent in administration of forensic science in the near future.
Bone and Bones/chemistry* ; Crime ; Dental Enamel/chemistry* ; Dermatoglyphics ; Drowning/diagnosis* ; Forensic Medicine/methods* ; Humans ; Limit of Detection ; Spectrometry, X-Ray Emission/methods* ; Zinc/analysis*

[Objective]Examine the expression of microsomal prostaglandin E synthase-1(mPGES-1)and nuclear transcription factor kappa B p65(NF-κB p65)in diffuse large B cell lymphoma(DLBCL),assessing their correlation with clinical variables,prognosis and potential clinical valve.[Methods]The immunohistochemistry was uesd to investigate the expression of mPGES-1 and NF-κB p65 in 83 DLBCL patiens'tissues.The relationship between these two proteins and the clinical variables and prognosis of these patients was evaluated.[Results]The high expression of mPGES-1 and NF-κB p65 were observed in 65.1%(54/83)and 73.5%(61/83)cases of DLBCL,respectively.The expression level of NF-κB p65 was positively correlated with mPGES-1 expression(P<0.05).The expression of these two proteins was found to be significantly associated with B cell lymphoma/leukemia-2 protein(BCL-2),higher expression of Ki67,higher lactate dehydrogenase (LDH),more extranodal lesions,advanced Ann Arbor stage and higher international prognostic index(IPI)score(P<0.05). In addition,NF-κB p65 was related with multiple myeloma oncogene 1(MUM1),pathological type(P<0.05). Both mPGES-1 and NF-κB p65 overexpression was correlated with worse overall survival(OS)while NF-κB p65 was an in-dependent prognostic factor for OS of DLBCL(P<0.05).[Conclusions]mPGES-1 and NF-κB p65 were highly expressed in DLBCL and closely linked with each other. The overexpression of mPGES-1 and NF-κB p65 was correlated with tumor progression and unfavorable prognosis in patients with DLBCL.

Objective To evaluate the feasibility and safety of endoscopic retrograde cholangiopancreatography (ERCP) after Roux-en-Y reconstruction. Methods 22 cases underwent ERCP after Roux-en-Y reconstruction from January 2015 to January 2017 were collected, the operating time, success rate of endoscopy and treatment, related complications were analyzed. Results ERCP was performed in 22 cases about Roux-en-Y reconstruction of digestive tract, the mean insertion and cannulation time was 74.1 and 22.5 minutes; the overall success rate was 81.8% (18/22) and 77.2% (17/22), and no major complications occurred. Conclusions ERCP can be used as a safe and effective method for the diagnosis and treatment on the Roux-en-Y reconstruction of digestive tract.

OBJECTIVETo explore the effects of chloride channels on the regulation of platelet cytoplasmic free calcium concentration ([Ca2+]i) and platelet aggregation (PAG).

METHODSFreshly separated platelets were activated by thrombin. Chloride channel blockers DIDS or NFA and calcium channel blockers SK&F96365 or nifedipine were added to study the effects on platelet [Ca2+]i and PAG by a single reagent or the combination of reagents and find out the interactions among DIDS, NFA, SK&F96365 and nifedipine.

RESULTSBoth DIDS and NFA could inhibit the thrombin (1 U/ml) induced PAG in a dose-dependent manner, whereas had little effect on resting [Ca2+]i. As compared with the control group, DIDS, SK&F96365 and Nifedipine could significantly reduce the PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05). The combination of DIDS and SK&F96365 had greater effects in reducing the PAG, Ca2+ release and Ca2+ influx than either reagent alone (P < 0.05). The combination of DIDS and nifedipine also had greater effect than each alone in reducing Ca2+ release (P < 0.05). The combination of NFA and SK&F96365 weakened each other's effect on Ca2+ release (P < 0.05), while NFA and nifedipine weakened each other's effects on PAG, Ca2+ release and Ca2+ influx in thrombin activated platelet (P < 0.05).

CONCLUSIONDIDS and NFA have no effect on the resting [Ca2+]i and the leak calcium influx of platelet. DIDS can inhibit the Ca2+ release, Ca2+ influx and PAG of platelet induced by thrombin, while NFA can only inhibit the Ca2+ release. The chloride channel and calcium channel blockers have interactions in affecting resting [Ca2+]i and PAG of platelet. The opening of chloride channel can influence the cellular calcium movement of platelet.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid ; pharmacology ; Adult ; Blood Platelets ; cytology ; drug effects ; metabolism ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; physiology ; Cytoplasm ; drug effects ; metabolism ; Drug Interactions ; Humans ; Imidazoles ; pharmacology ; Nifedipine ; pharmacology ; Niflumic Acid ; pharmacology ; Platelet Aggregation ; drug effects ; Thrombin ; pharmacology