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.

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

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

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.

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*

BACKGROUNDChinese medicine plays an important role in hepatoprotective treatment. This study was conducted to investigate the protective effects of emodin and astragalus polysaccharides (APS) in a rat model of chronic hepatic injury.

METHODSChronic hepatic injury was induced by hypodermic injection of an olive oil solution containing 40% carbon tetrachloride (CCl(4)) twice a week, in addition to a diet of 79.5% maizena, 20% fat, 0.5% cholesterol, and 10% alcohol in the drinking water ad libitum for 12 weeks. Meanwhile, the rats were exposed to different concentrations of emodin (40 mg x kg(-1) x d(-1)), APS (200 mg x kg(-1) x d(-1)), combination drug (emodin 40 mg x kg(-1) x d(-1) combined with APS 200 mg x kg(-1) x d(-1)) and colchicine (0.1 mg x kg(-1) x d(-1)) in parallel by oral gavage (once a day for 12 weeks). At the end of 12 weeks, blood serum and liver tissue were taken. Serum was collected to determine the levels of total bilirubin (TBIL), alanine transaminase (ALT), aspartate transaminose (AST), and albumin (ALB). Liver and spleen indexes were assayed, followed by the measurements of the liver associated enzyme superoxide dismutase (SOD) and malondialdehyde (MDA). Histopathological changes were studied using optical microscopy.

RESULTSSplenohepatomegalia was alleviated and serum levels of TBIL and ALT were reduced in the groups treated with emodin and APS when compared to the control group. In addition, the ALB level in the APS and combination groups was higher. Similarly, the SOD activity of liver homogenates was significantly higher in the groups treated with emodin and APS, while administration of the herbal derivatives prevented the elevation in MDA levels. Histological analysis showed that the APS and combination groups significantly ameliorated the hepatic injury.

CONCLUSIONSCo-administration of emodin and APS demonstrated a synergistic action in reducing ALT and restoring ALB in the serum from a rat model of chronic hepatic injury. Emodin and APS may ameliorate the CCl(4)-induced hepatic injury in rats by elevating antioxidant-enzyme activities and reducing lipid peroxidation.

Alanine Transaminase ; blood ; Animals ; Astragalus Plant ; chemistry ; Carbon Tetrachloride ; toxicity ; Chronic Disease ; Emodin ; pharmacology ; Liver ; drug effects ; pathology ; Male ; Malondialdehyde ; analysis ; Polysaccharides ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism

In this study, the HPAIV A/Tiger/Harbin/01/2002 (H5N1) used was originated from tigers and propagated in SPF embryonated hen eggs. TCID5, of the virus was 10(-7.36)/0. 05mL on MDCK cell. The cats were inoculated through bronchus route and then, the cats of dead and control were collected for histopathological and immunohistochemistry examination. Meanwhile, the emulsion supernatant fluid of organs and the pharyngeal swab samples of the dead cats were collected for RT-PCR, survived cats and the control cats were tested for the presence of HI antibody by standard method. The results indicated that the damage of lungs from the dead cats were most obvious, the wide range of red consolidation focus emerged on the lobus pulmonis, the fused focus of infection caused injury of lungs. Histology under the microscope revealed diffuse alveolar damage, confluence phlegmasia pathology, infiltration of lymphomonocytes, sackful of infiltration of macrophages and manipulus protein-like effusion in the alveolar. By immunohistochemistry, the positively stained virus particles were found on the epithelial cells of bronchus and alveolus, and also in the endochylema of lymphomonocytes. The specific electophoretic band of 464bp amplified by RT-PCR from samples of pharyngeal swabs, lungs, kidneys, hearts and brains was as same as the theory value. HI antibody titers of the survived cat were 1:32.
Animals ; Antibodies, Viral ; blood ; Cat Diseases ; pathology ; Cats ; Hemagglutination Inhibition Tests ; Immunohistochemistry ; Influenza A Virus, H5N1 Subtype ; immunology ; pathogenicity ; Orthomyxoviridae Infections ; pathology ; veterinary ; Reverse Transcriptase Polymerase Chain Reaction ; Tigers ; virology

OBJECTIVETo evaluate the forensic utility of Y-single nucleotide polymorphisms (SNPs) markers.

METHODSAllele-specific PCR, restriction enzyme digestion or direct PCR were performed to examine 10 different SNP loci on Y chromosome, namely M9, M15, M45, M89, M95, M122, M134, M145, M173 and P25 in 161 Chinese Han males.

RESULTSA total of 8 of the 10 SNPs are reported to be polymorphic in Chinese. The gene diversity for the loci showing polymorphism ranged from 0.988/0.012-0.752/0.248, with a power of discrimination 0.094-0.373. Loci M122 and M134 were the most polymorphic markers in Chinese Hans. Nine different haplogroups with frequencies from 1.2% to 51.6% were observed and 3 of the haplogroups-K*(x O2a, O3, P), O3*(x O3e) and O3e were found in 75.2% of Chinese Hans.

CONCLUSIONA comprehensive gene diversity data of Y chromosome and haplogroups were obtained in Sichuan Han population, which will be served as the base for using these Y-SNP markers in forensic medicine and individual identification in Sichuan Hans.

China ; Chromosomes, Human, Y ; genetics ; Female ; Haplotypes ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics