Cytomegalovirus ; Cytomegalovirus Infections ; prevention & control ; therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; T-Lymphocytes, Cytotoxic ; virology ; Transplantation, Homologous

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.

Objective To compare the prognosis of sentinel node-positive breast cancer patients forgoing axillary lymph node dissection.Methods A systematic literature search (Medline,Embase,Cochrane Library)ended in April 2014 was performed to identify all eligible articles.Two reviewers independently screened and extracted data.RevMan5 was used for statistical analysis.Results A total of 1026 abstracts were retrieved and 18 clinical controlled studies finally included,the total number of patients were 47 894,7389 had micrometastases in sentinel lymph node,35 217 had macrometastases in sentinel lymph node and 5288 had positive sentinel lymph node regardless of micrometastases or macrometastases.For patients with MIC,the 5-year axillary recurrence rate,5-year disease free survival and 5-year overall survival had no significant difference between patients who only received sentinel lymph node biopsy and patients who received further axillary lymph node dissection,(OR =1.78;95％ CI:0.72-4.39,P=0.21),(OR =0.76,95％CI:0.56-1.04,P=0.08),(OR=0.77,95％CI:0.43-1.40,P=0.39).For patients with MAC,the 5-year axillary recurrence rate had no significant difference between patients who only received sentinel lymph node biopsy and patients who received further axillary lymph node dissection,(OR =1.21;95％ CI:O.76-1.91,P =0.42).For patients with positive sentinel lymph node regardless of micrometastases or macrometastases,the 5-year axillary recurrence rate and 5-year overall survival had no significant difference between patients who only received sentinel lymph node biopsy and patients who received further axillary lymph node dissection,(OR =1.29;95％ CI:0.92-1.80,P =0.14),(OR =0.96,95％ CI:0.64-1.45,P =0.84).Conclusions Among patients with limited positive SLN of breast cancer,patients forgoing ALND compared with ALND did not have obvious affect on long-term survival.

Objective: To study the impact of interferon-inducible protein 16 (IFI16) inhibition on apoptosis of human aortic adventitial fibroblasts (HAAFs). Methods: Our research included 3 groups: ① IFI16-siRNA group, specific small interference RNAs (siRNAs) of IFI16 were transfected into HAAFs in vitro to make IFI16 gene silence, ②Con-siRNA group, non-specific siRNAs were transfected into HAAFs as negative control and ③Untreated HAAFs group, blank control. HAAFs cell cycle and apoptosis rate were examined by flow cytometry, IFI16 mRNA expression was measured by real time qRT-PCR, protein expressions of IFI16, p53, p21, Bax and Bcl-2 were detected by Western blot analysis. Results: Compared with Con-siRNA group and Untreated HAAFs group, IFI16-siRNA group showed decreased apoptosis rate of HAAFs (3.33±0.41) % vs (7.42±1.51) % and (6.49±1.10) %, P<0.05, reduced ratio of G0/G1 phase cells (56.64 ± 4.77 ) % vs (69.67±3.54) % and (68.29±4.14) %, P<0.05, while increased ratio of S phase cells (25.23±5.19)% vs (13.76±2.07) % and (14.04±3.00) %, P<0.05. Meanwhile, IFI16-siRNA group presented down-regulated IFI16 mRNA and protein expressions, decreased protein levels of p53, p21, Bax and increased protein level of Bcl-2, all P<0.05. Conclusion: Inhibited IFI16 expression could decrease HAAFs apoptosis, promote cell cycle transition from G1 to S phase which might be related to the suppression of p53/p21 signaling pathway and regulation of Bax/Bcl-2 expression.

Bioethics, one of the focus subjects in 21 century,attracts the attention in the worldwide.Originated from western developed countries,it has been branded with distinct western civilization,which is disadvantageous to the development of the subject.Some suggestions are put forward as following: 1.Communication between the East and the West should be promoted in order to form a solid multi-cultural base for bioethics. For instance, in America ethics bears the imprint of religious conventionality,while in China politics is infiltrated into traditional ethical ideas. Both of them are not perfect. So they cannot form the base of bioethics. 2.Research on the "base line" of the bioethics should be emphasized in order to provide freer academic atmosphere. Transplanting the western bioethical ideas to the developing countries will never work. As for the bioethical"base line",different countries should establish feasible ethical codes according to their own actual situations. 3.Scientists should be encouraged to adopt various technical ways to explore the problems in the field of bioethics.Such methods as tissue-engineering,antologous stem-cell replanting and cell diffrenttation inducing may bring about earlier breakthrough than fetus stem-cell.Equal attention,encouragement and support should be given to bioethics.

Objective To establish a calcification mode in vitro of human vascular smooth muscle cells (HVSMCs) induced by β-GP. Methods Primary HVSMCs were obtained from human embryo by plant method and confirmed by stain with α-sm-actin antibody. The cells after 4-6 passages were divided into two groups.The control group was incubated with normal DMEM medium while the calcification group was incubated with the medium containing 10 mmol/L β-glycerophosphate for 10 days.Calcification was confirmed by alizarin red S stain and alkaline phosphatase(ALP) assays. Results The primary cells observed by S-P stain were positive and the cells after being stained were pale yellow. After being induced with β-GP, the cells of calcification group began concentric growth and formed vesicles. Alizarin red S staining showed that the reaction of calcifying nodules was red,ALP activity was higher than that of controls at various time points(4 d,6 d,8 d and 10 d,P＜0.01 ).Conclusion The HVSMCs could be induced into calcification in vitro by β-GP, and this model contribates to further studies of vascular diseases.

Adult ; Female ; Humans ; Hyperlipoproteinemia Type III ; pathology ; Male ; Middle Aged ; Nephrosis, Lipoid ; pathology

Humans ; Infant ; Infant, Low Birth Weight ; Infant, Newborn ; Male ; Tyrosinemias ; diagnosis ; therapy

Objective: To observe the clinical effects of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) in the treatment of shoulder and arm pain. Methods: A total of 90 patients with shoulder and arm pain were randomly divided into an observation group and a control group, 45 cases in each group. The treatment of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) was adopted in the observation group, routine acupuncture was used in the control group. The two groups were treated once every day, with 5 treatments as one course, and a 2-day rest between two courses. After 3 courses, pain was assessed by visual analog scale (VAS), and the clinical effects were compared between the two groups. Results: After the treatment, VAS scores were significantly changed in both groups (both P<0.01). The VAS score was lower in the observation group than that in the control group, with a statistical difference between the two groups (P<0.05). The total effective rate was 100% in the observation group, versus 91.1% in the control group, the difference was statistically significant (P<0.05). Conclusion: The therapeutic effect of Shu-acupuncture method in Nei Jing (Classic of Internal Medicine) is better than that of routine acupuncture in treating shoulder and arm pain.

Objective To compare the effectiveness of hyperbaric oxygen (HBO) therapy and normal pressure oxygen inhalation for lung infection in patients with severe brain injury and tracheotomy.Methods Sixty lung infection patients with severe brain injury and tracheotomy were stratified by age and Glasgow coma scale (GCS) score into an HBO group (30) and a conventional therapy group (30).Both groups were given treatments to reduce intracranial hypertension and protect the brain,surgery,antibiotics,and symptomatic and support therapy.In the conventional group the patients received oxygen therapy at normal oxygen pressure by tracheal cannula.In the HBO group,the patients received HBO therapy through self-made oxygen devices by the authors.The body temperature,cough and expectoration,respiratory sounds and white blood cell (WBC) count were compared between the groups.Results There were statistically significant differences in body temperature,cough and expectoration,respiratory sounds and the normalized time for the count and classification of WBC between the groups.Conclusion HBO had good treatment effectiveness for excretion and expectorant in lung infection patients with severe brain injury and tracheotomy.