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.

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.

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 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 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 determ ine the norm al reference values of 33 elem ents, Ag, Al, As, Au, B , B a, Be, Ca, Cd, Co, Cr, Cs, Cu, Fe, G a, H g, Li, Mg, Mn, Mo, N i, Pb, Rb, Sb, Se, Sr, Th, Ti, Tl, U , V , Zn and Zr, in the blood and urine sam ples from the general population in Sanm en County of Zhejiang province, a typical coastal area of eastern China. Methods The 33 elem ents in 272 blood and 300 urine sam ples w ere determ ined by inductively coupled plasm a-m ass spectrom etry (ICP-MS ). The norm ality test of data w as conducted using SPSS 17.0 Statistics.The data w as com pared w ith other reports. Results The norm al reference values of the 33 elem ents in the blood and urine sam ples from the general population in Sanm en County w ere obtained, w hich of som e elem ents w ere found to be sim ilar w ith other reports, such as Co, Cu, Mn and Sr, w hile As, Cd, H g and Pb w ere generally found to be higher than those previously reported. There w as a w ide variation betw een the reports from different countries in blood B a. Conclusion The norm al reference values of the 33 elem ents in the blood and urine sam ples from the general population in Sanm en County are established, and successfully applied to tw o poisoning cases.

BACKGROUND:Metabonomics has been proved to analyze and observe the pathological process of rat myocardial infarction and the underlying mechanism. OBJECTIVE:To further analyze the metabolomic pathways of bioinformatics in rat models of myocardial infarction. METHODS:The experimental studies about rat myocardial infarction were retrieved from CNKI, WanFang, CqVip, PubMed and Embase databases. The metabolic products described in the literatures were col ected and summarized. Signaling pathways were analyzed using KEGG database molecular function annotation, the enzymes, translocators and their properties were analyzed by HMDB database. Metabolites pathway were visualized with MetPA. RESULTS AND CONSLUSION:A total of 26 metabolic products were identified in the included literatures and mainly participated in 29 metabolic pathways. Through topology analysis, 5 of the 10 metabolic pathways were selected and regarded as the metabolic pathways of myocardial infarction in rats, including aminoacyl-tRNA biosynthesis;glycine, serine and threonine metabolism;valine, leucine and isoleucine biosynthesis;biosynthesis of unsaturated fatty acids;phenylalanine, tyrosine and tryptophan biosynthesis. In conclusion, the bioinformatics analysis of metabolites in rats with myocardial infarction show that myocardial infarction is related to the metabolism and metabolic pathways of carbohydrates, proteins, fat and RNA.

Objective To prepare celastrol ethosomes and to observe the permeability characteristics of the ethosomes which act as the transdermal delivery carriers of celastrol in vitro. Methods Celastrol ethosomes were prepared by ethanol injection method, and then the encapsulation efficiency, particle size, polydispersity index ( PDI) and zeta potential of the ethosomes were analyzed. TP2A intelligent percutaneous penetration instrument was used to compare the skin penetration properties of celastrol ethosomes, celastrol solution and the mixture of blank ethosomes with celastrol solution. Results The prepared celastrol ethosomes were spherical, and the average particle size was (401.3 ± 5.5) nm, PDI was 0.21± 0.02, steady zeta potential was (-2.75 ± 0.1) mV, and average encapsulation efficiency was ( 80.6 ± 0.7) %. The amount of accumulative penetration of celastrol ethosomes at 48 h was 76.86 μg·cm -2 and the permeation rate was 1.640 9 μg·cm -2·h -1, which were significantly higher than the celastrol solution and the mixture of blank ethosomes with celastrol solution. Conclusion The prepared ethosomes have high encapsulation efficiency, uniform particle size and stable quality, and are beneficial to the transdermal absorption of celastrol.

Objective To investigate and determine the dietary intake level and food source of elements in Kashin-Beck disease(KBD) and non-endemic KBD areas in parts of Aba Autonomous Prefecture , Sichuan Province. Methods Three KBD villages were selected from each county including the counties of Songpan , Jiuzhaigou and Ruoergai in Aba Autonomous Prefecture with multi stage random sampling method in 2010 , and 122 KBD patients who were diagnosed at county-level or higher-level hospitals were selected from villages. One non-KBD village was selected randomly from each county and 43 controls without KBD or other bone and joint disease were selected in proportion to the number of investigated KBD patients. Dietary intake of each object was recorded 24 hours before the survey using a 24 hour dietary survey method and intake of each kind of food(cooked food) was recorded. Three day dietary intake of each surveyed object was recorded in continuous and the average value was calculated. The local resident representative food samples were collected and 20 kinds of elements in these samples were determined with inductively coupled plasma mass spectrometry ( ICP-MS ) and inductively coupled plasma atomic emission spectrometry(ICP-AES), and daily elements intake was calculated and compared with the relevant intake standards. Results Daily intakes of vegetables and meat were 126.72, 28.25 g, respectively, in patients with KBD. More than 3 times of brick tea was drank by KBD patients than non-KBD people (P<0.05). Per capita daily intakes of Ca, Fe, Mg, K, Na, Zn, I, Cu and Co were less than recommended nutrient intake or allowable daily intake in both groups. More than 80% patients with KBD overtook more Al and Pb than tolerable upper intake levels. Conclusion There is a single dietary structure and a serious shortage of a variety of essential elements intake in parts of Aba Autonomous Prefecture, and the dietary intake level of elements is different between two groups.