Objective To construct the lentiviral overexpression vectors of mouse neuromedin B(NMB) and neuromedin B receptor(NMBR) genes and express them in RAW264.7 cells, so as to lay a foundation for further study on the effects of mouse NMB and NMBR gene overexpression on the proliferation and apoptosis of RAW264.7 cells. Methods The coding sequences(CDSs) of mouse NMB and NMBR genes were amplified by RT-PCR and inserted into vector pCD513B-1 to construct recombinant plasmids pCD513B-1-NMB and pCD513B-1-NMBR. Mouse NMB and NMBR gene overexpression lentiviruses were obtained through packaging by HEK-293T cells, and the virus titers were detected by double dilution method. After infection with lentivirus for 48 h, RAW264.7 cells were detected for the expression of NMB and NMBR mRNA by qPCR using the uninfected cells as control. Results The recombinant plasmids were constructed correctly as identified by double enzyme digestion. The virus titers of mouse NMB and NMBR gene overexpression lentiviruses were 6 × 106and 8 ×106TU/mL, respectively. The mRNA expression levels of mouse NMB and NMBR genes in RAW264.7 cells transfected with two lentiviruses were significantly higher than those in the control group(t = 24. 158 and 14. 958, respectively, each P <0. 01). Conclusion Mouse NMB and NMBR gene overexpression vectors were successfully constructed, which can significantly increase the expression of NMB and NMBR genes in RAW264.7 cells.

Surgical treatment is the only cure treatment for patients with inferior vena cava tumor thrombus in hepatic segment and upper hepatic segment.The accurate diagnosis of tumor thrombus is very important.In preoperative imaging examination,the abdominal enhanced CT scan and the inferior vena cava MRI scan were the best methods for the diagnosis and evaluation of the tumor thrombus in hepatic segment and upper hepatic segment.Compared with the tumor thrombus below the liver,the tumor thrombus in hepatic segment or above hepatic segment extend widely,and the operation are more difficult.For simple inferior vena cava tumor thrombus (the top of the thrombus has reached the level of hepatic vein),Retroperitoneal approach combined with transperitoneal approach should be used.Open surgery is the standard procedure for other tumor thrombus in hepatic segment and upper hepatic segment.In addition to exposure of inferior vena cava below the hepatic vein,the liver and the first hepatic hilum should be exposed.For tumor thrombus in the atrium,after the longitudinal incision of diaphragm,we use Milking technology to squeeze thrombus into inferior vena cava.Then we use catheterization technology to remove thrombus.For difficult atrial tumor thrombus,an extracorporeal circulation should be performed.The median incision in the chest should be performed to open the chest and open the pericardium and remove the tumor thrombus.Patients with tumor thrombus in hepatic segment or upper hepatic segment should be diagnosed as early as possible and they need actively treated by operation.

OBJECTIVE: To investigate the protective effect and probable mechanism of 20-Hydroxyecdysone on SH-SY5Y cells injured by MPP+. METHODS: MPP+ induced SH-SY5Y cells injury model was established. 20-Hydroxyecdysone was added into the culture to test its protective effect. The morphological changes of cells were observed. Cell viability was detected by method of MTT. The contents of LDH, MDA and activity of SOD were inspected by kits. Apoptosis rate of cells was detected by flow cytometry. RESULTS: Compared with MPP+ group, 20-Hydroxyecdysone group (50, 100, 150 μmol·L-1) alleviated the damage of SH-SY5Y cells induced by MPP+, significantly improved cell survival rate, reduced the release of LDH, increased the activity of SOD, decreased the contents of MDA and reduced the apoptosis of cells. CONCLUSION: 20-Hydroxyecdysone has a significant protective effect on SH-SY5Y cells injured by MPP+. The probable mechanism may be anti-oxidation and anti-apoptosis.

Objective To determine the effect of ABCC4 gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3.Methods PANC1 and BxPC-3 pancreatic cancer cells were transfected with a lentivirus expressing an ABCCA short hairpin RNA (shRNA).ABCC4 mRNA and protein expression of transfected cells was determined by RT-PCR and Western blot,colony formation ability was measured by colony assay,and cell cycle change was investigated by the flow cytometric analysis.Results Lentivirus expressing an ABCC4 short hairpin RNA (shRNA) was successfully established.After transfection with shRNA lentivirus,ABCC4 mRNA and protein expression were significantly inhibited (0.28 ±0.01 vs 1.00 ±0.03,0.22 ±0.02 vs 1.00 ±0.03,P ＜0.05).Colony formation ability was significantly decreased (4 vs 65,P ＜0.05),and cell cycle was significantly blocked at G1 phase [(54.98 ±1.78) ％ vs (42.93 ± 0.88) ％,(68.55 ± 0.75) ％ vs (54.76 ± 0.29) ％].Conclusions ABCC4 gene silencing can significantly inhibit cell proliferation of human pancreatic cancer cell lines PANC1 and BxPC-3,and block the cells at G1 phase.

Objective To investigate the level change and correlation of serum IL-18 and soluble Fas/Fas ligand(sFas/sFasL) in patients with chronic obstructive pulmonary disease (COPD) to evaluate their roles in the pathogenesis of COPD. Methods The levels of serum IL-18 and sFas/sFasL in 36 patients with COPD of acute aggravation stage and 20 healthy control subjects were detected by ELISA. Results The levels of serum sFasL and IL-18 in patients with COPD were significantly lower than those in healthy control subjects, while there was not significant differene in serum sFas level between the two groups. Conclusion The serum sFasL and IL-18 level decreass in patients with COPD of acute aggravation stage indicated that cell apoptosis level and immune function reduced in the patients.

Objective To study the relationship between gene polymorphism of apolipoprotein E (apolipoproteinE,ApoE)of peripheral blood and Mycoplasma pneumoniae infection in children.Methods Collected 236 cases serum of inpatient and outpatient screening in children with Mycoplasma pneumoniae infection and healthy children between March 2011 and March 2014 in the First Affiliated Hospital of Xi’an Medical University and Xi’an Children’s Hospital,at the age of 3~8 years old,divided into two groups:110 cases of control group and 126 cases of Mycoplasma pneumoniae infection in chil-dren.Used multiple allele-specific PCR (multi-AS PCR)to detect gene polymorphism of ApoE in each group.Results ApoE gene was polymorphic and 6 genotypes:3 homozygous (ε2/2,ε3/3,ε4/4)and 3 heterozygote (ε3/2,ε3/4,ε4/2).Theε3/2 had four bands,ε3/3,ε3/4 and 4/2 had three bands,ε2/2 andε4/4 had two bands.ε3/3 of ApoE genotype distribution in two groups was the most common,control group was 66.7%,infection group was 46.4%.Allele frequencies ofε3 and genotype frequencies ofε3/3 inMycoplasmapneumoniae infection of children were lower than those in control group (P<0.05).But allele frequencies ofε4 and genotype frequency ofε4/4 in Mycoplasma pneumoniae infection of children were increased, which were compared with those in control group (P<0.05).Conclusion There were an association between ApoE gene polymorphism and the incidence of Mycoplasma pneumoniae infection in children.Allelesε3 seems to be a protective factor and allelesε4 may contribute to the development of Mycoplasma pneumoniae infection of children.

Objective To investigate the expression of miR-29s in the glioma stem cells,and explore how the members of miR-29s affect the bio-logical behaviors of glioma stem cells. Methods Eight patient specimens were used to culture glioma stem cells. Real-time PCR was adopted to test the expression of miR-29s. CCK-8 analysis was performed to test the proliferation ,Transwell was used to test cell migration and invasion ,and flow-cytometry analysis was carried out to test apoptosis. Results miR-29a,miR-29b and miR-29c were decreased in glioma stem cells. Over-ex-pression of miR-29s could inhibit the proliferation,cell migration and invasion,but promote apoptosis of glioma stem cells. Conclusion miR-29s acts as a cancer suppressor gene in the glioma stem cells ,and miR-29a plays the dominant functional role in the family.

BACKGROUND:With more and more elderly patients suffering lumbar degenerative disease undergoing internal fixation, infection after spinal internal fixation is a common complication in orthopedic surgeries, but its treatment strategy remains controversial.OBJECTIVE: To explore the curative efficacy of incision infection after internal fixation in the elderly with lumbar degenerative disease.METHODS: 197 patients with lumbar degenerative disease undergoing internal fixation and followed up for more than 2 years admitted in Department of Orthopedics, Beijing Shijitan Hospital Affiliated to Capital Medical University from January 2012 to January 2015, were analyzed retrospectively. The follow-up time was 2-4.9 years. There were 97 cases of lumbar stenosis, 29 cases of lumbar disc herniation, 33 cases of lumbar spondylolisthesis, 17 cases of degenerative scoliosis and 21 cases of lumbar compression fractures.RESULTS AND CONCLUSION: (1) Eleven patients experienced incision infection, including ten acute, and one delayed infection. (2) Among acute infected cases, three were superficial infection in three cases and seven had deep infection,who characterized as exudation (10/10), local pain (8/10) and fever (9/10). Acute infected cases received bacterial culture, drug sensitive test, antibiotic therapy, and debridement of the infected wound, and leaving all internal fixators in situ in all but one case. (3) For delayed infection, one patient had local pain, incision exudation, and intermittent fever,and then the internal fixators were removed. (4) Pseudarthrosis was not founded during 2-year follow-up in all patients.(5) These results suggest that for the elderly patients suffering lumbar degenerative disease with infection after internal fixation, intravenous antibiotics, debridement plus drainage are recommended, but without internal fixator removal, and repetitive debridement and drainage is a rational choice if necessary.

Objective To analyze the expression of urine exosomal miR-375 in prostate tumors and investigate its clinical utility.Methods A total of 45 patients with PCa,24 with benign prostate hyperplasia (BPH) and 24 healthy individuals were enrolled into this study.Exosomes were isolated from the urine of PCa,BPH and healthy individuals and the total RNA was extracted from the exosomes.The exosomal miR-375 expression was assessed by quantitative real-time PCR and analyzed with the comparative quantification cycle method (2-△△CT).We performed comprehensive biostatistical analyses to explore the clinical value of miR-375 in prostate cancer.Results The urine exosomal miR-375 expression was significantly downregulated in the patients with PCa compared with BPH and the healthy controls (P ＜ 0.01).No statistically significant difference of the urine exosomal miR-375 expression levels between the patients with BPH and healthy individuals was observed (P ＞ 0.05).The urine exosomal miR-375 expression level was also found to be associated with clinical stage and bone metastasis status of the patients with PCa (P ＜0.05),and with the increase of Clinical stage.The expression level of miR-375 decreased.No significant relationship was detected between miR-375 level and the patient's age,gleason score and serum prostate-specific antigen level (P ＞ 0.05).Receiver operator characteristic analyses demonstrated that the urine exosomal miR-375 expression could better differentiate PCa from BPH patients:AUC 0.715 (95％ CI:0.589-0.842) vs PSA AUC 0.632 (95％ CI:0.492-0.771) (P＜0.01).Conclusion The urine exosomal miR-375 could serve as a non-invasive biomarker for the diagnosis of PCa.