OBJECTIVE: To determine the content of netilmicin sulfate in serum by a high performance liquid chromatography-indirect photometric determination (HPLC-IPD) method. METHOD: The chromatographic system consisted of Soheisorb C18 column and mobile phase of a solution of methyl alcohol-a buffer of phosphoric acid (pH=2.0) (20∶80), that contained 0.5 mmolL-1 of nicotinamide and 0.3 mmolL-1 of sodium seventhalkyl-sulphonate. The detected wave length was 268 nm. The serum simple of 4 patients was determined. RESULTS: The mean recovery of was 93.41% and detection limits was 50 μgL-1. CONCLUSION: The method is constant, sensitive, and has a good concentration. It is good for determination of netilmicin sulfate concentration in serum.

Objective To investigate the natural infection status of murine norovirus ( MNV) in laboratory mice in Shanghai area and isolate MNV from mouse cecal feces .Methods To collect cecal contents and serum samples from 319 specific pathogen-free ( SPF) mice coming from different research institutions .Reverse transcription-polymerase chain reaction ( RT-PCR) and enzyme linked immunosorbent assay ( ELISA) were used to detect MNV infection in the mice , re-spectively.The positive stool samples were diluted and filtered through 0.22 μm membrane, inoculated into RAW 264.7 cells, and then identified by RT-PCR.Results There were 95 positive results in the 319 cecal samples by RT-PCR, and the positive rate was 29.78%.Among 180 serum samples which were tested by RT-PCR, 70 samples were positive by ELISA, and the positive rate was 38.89%.The infected RAW 264.7 cells showed cytopathic effect ( CPE) within 72 h. After 3 times of freezing and thawing , RT-PCR obtained a 187 bp band.Conclusions The results from the present study show that there is a high natural infection rate of MNV in laboratory mice in Shanghai area , and the strict breeding manage-ment must be strengthened .

AIM:To investigate the construction of oligonucleotide microarray specialized for pancreatic adenocarcinoma-associated genes and its preliminary application of detecting differential expressed genes in pancreatic cancer.METHODS:Pancreatic cancer related genes were purposely selected,and oligonucleotide microarray was prepared by spotting oligonucleotide probes on glass slides coated with APS-PDC.Labeled cDNA targets for hybridizations were synthesized by reverse transcription from total RNA in the presence of Cy5-dCTP and Cy3-dCTP,respectively.Hybridized microarray was scanned by Agilent laser scanner,and the aquired image was analyzed by Imagene3.0 software.The intensity ratios of Cy3 and Cy5 were calculated.To confirm the expression profiles of these genes,quantitative reverse transcription-PCR (QRT-PCR) was carried out for CDC25B and TUSC3 genes,and β-actin gene was taken as internal control.The product of PCR was quantitated by comparative Ct method.RESULTS:The signal of microarray hybridization was clear,and the images had a lower background and higher signal-noise ratio.In comparison with normal pancreas,twenty -four differentially expressed genes were identified which included seventeen up-regulated and seven down-regulated genes.The results of QRT-PCR demonstrated that the expressions of CDC25B and TUSC3 in pancreatic cancer were up-regulated and down-regulated respectively,which is consistent with microarray results.CONCLUSION:The oligonucleotide microarray specialized for pancreatic cancer is desirable for its specialty and sensitivity,which can simultaneously and parallelly detect multiple pancreatic cancer-associated genes.

Objective To study the principle of chemokine stromal-dirived factors-1(SDF-1)in acute rejection,we test the expression of CXCR4 which is the receptor of SDF-1 in acute rejection following liver transplantation. Methods needle-biopsy specimens after liver transplantation were divided into four groups according to Banff schema.We analyzed the mRNA level of CXCR4 in each group by RT-PCR.Results In non-rejection group and control group,all samples expressed CXCR4 in small and medium dose.In rejection group,high level expression of CXCR4 related to high degree of acute rejection were found.Conclusion The CXCR4 plays an important role in acute allograft rejection of human liver after transplantation.To restrain the expression of CXCR4 may be au effective method of anti-acute rejection.

Objective To determine the interference effect of H. hepaticus infection on the functional characteris-tics of dendritic cell ( DC) surface molecules and immune response in mice. Methods Male BALB/c mice were inocula-ted with H. hepaticus (ATCC 51450). Murine bone marrow-derived dendritic cells (DC) were isolated and co-cultured which were stimulated by GM-CSF and IL-4 at the fifth month after the last inoculation. Then the DCs were subjected to FACS analysis for surface markers (CD11c, CD40, CD80 and MHCII) detection. On this basis, virus suspension of New-castle disease virus( NDV) ZJ1 strain was inoculated into the mice. Serum was collected for detection of the NDV antibody titer in serum weekly to explore the difference of antibody titer between the two groups. Results The expression rates of CD40 and MHCII on the mouse DCs in experimental group were higher than that in the control group. The NDV antibody ti-ter of experimental group was slightly lower than that in the control group in the first week. During the 2nd to 5th weeks, the titer was higher than that in the control group, with a very significant difference. In the 6th week, the titer of both the two groups tended to fall. Conclusions H. hepaticus infection can promote bone marrow DC maturation in mice, stimulate the expression rates of MHC II and CD40, and enhance the NDV antibody levels.

Objective To investigate the risk factors of posttraumatic hydrocephalus (PTH) in patients with moderate to severe traumatic brain injury (TBI).Methods Aretrospective study was conducted for 183 patients with moderate to severe TBI (125 males,58 females;6-91 years of age,mean 48.23 years).According the presence of PTH,the patients were allocated into PTH group (n =34) and non-PTH group (n =149).Risk factors of PTh were assessed by univariate and logistic regression analysis,including gender,age,injury types,injury severity,intraventricular hemorrhage,subarachnoid hemorrhage,midline shift,subdural effusion,therapeutic strategies and skull defect.Association between the boundaries of skull defect and PTH was determined.Results Between-group differences were not significant regarding age,gender,injury types and intraventricular hemorrhage (P ＞ 0.05),but differed significantly in injury severity,subarachnoid hemorrhage,midline shift,subdural effusion,craniectomy and skull defect (P ＜ 0.05).Further Logistic regression analysis confirmed subarachnoid hemorrhage (OR =6.169),interhemispheric subdural effusion (OR =31.743),and unilateral (OR =17.602) and bilateral (OR =30.567) skull defects were risk factors of PTH.Of the patients with unilateral skull defect following decompressive craniectomy,the inferior limit ≤ 10 mm from the zygomatic arch also played a role in the development of PTH (OR =5.500,P ＜ 0.05).Conclusions Subarachnoid hemorrhage,interhemispheric subdural effusion and skull defect are risk factors of PTH.Unilateral skull defects with the inferior limit too close to the zygomatic arch can predispose to the development of PTH.

0. 05). However, the serum CYFRA21-1 level was related to the histologic classification (P

Objective:To study the change of microRNA during the early stage of high phosphorus in-duced vascular smooth muscle cell (VSMC)calcification and its related mechanism.Methods:The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol /L)for 7 d.The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content,real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules.Based on the cell calcification model,micro-RNA microarray array was applied to screen the profiles of microRNA expression in VSMC following high Pi stimulation for different periods (0,3 and 12 h).The array data were analyzed by TAMtool to explore the activated signaling pathway.Results:The calcium content of A7r5 cells induced by high Pi was in-creased 9.6 times high as cells without Pi treatment (P <0.05 ).VSMC contractile phenotype genes (SM-αactin,SM22)were down-regulated (P <0.05 ),while calcification-related genes (BMP2, MSX2,Runx2)were up-regulated (P <0.05)in VSMC stimulated by high Pi.The calcium nodules were obviously formed in cells after 7 d high Pi treatment.In microarray experiment,680 individual mi-croRNAs were detected in high Pi-treated VSMCs at different time points (0,3 and 12 h).Among these genes,miR-183,miR-664 and miR-9 * were increased whereas miR-542-5P,let-7f and miR-29a were decreased in time-dependent manners.Twenty-six kinds of signaling pathways,including cell apoptosis, differentiation and proliferation,were significantly activated.All these activated pathways were associated with calcification.Conclusion:This study implies that microRNA changed in high Pi-induced VSMCs may involve in the process of calcification.

Objective The aim of this study was to evaluate the individual and combined diagnostic value of five tumor markers in the elderly patients with pleural effusion. Methods Serum and pleural levels of cytokeratin fragment19(CYFRA21 1), neuron specific enalase(NSE), carbohydrate antigen15 3(CA15 3), carbohydrate antigen19 9(CA19 9) and carbohydrate antigen 125(CA125) were assayed in 32 elderly patients with malignant pleural effusions resulting from advanced lung cancer and in 30 elderly patients with benign pleural effusions by ELISA. Results Serum levels of CYFRA21 1, NSE, CA15 3, CA19 9 and CA125 in patients with malignant pleural effusions were (12 84?6 48) ?g/L, (22 07?11 25) ?g/L, (65 74?30 26) kU/L, (56 32?25 67)kU/L and (71 86?31 45) kU/L, respectively and were significantly higher than those patients with benign pleural effusions (P

Objective To study the signal transduction pathway for myocyte contraction induced by Substance K (SK) in rats. Methods The fluorescent Ca 2+ indicator Fluo 3AM was used to quantitate the calcium signal directly in the primary cultured myocardial cells. Changes of free Ca 2+ concentration in cardiac myocytes were detected by flow cytometry. The phospholipase C (PLC) inhibitor, neomycin and IP3 receptor antagonist, heparin, were used to block signal system of phosphates Ca 2+ in order to investigate whether they took part in the SK induced changes of [Ca 2+ ]i in myocytes. Results SK(1.78?10 -5 mol/L) elevated [Ca 2+ ]i in myocytes. The effects could be blocked by neomycin and heparin. Conclusion SK can elevate [Ca 2+ ]i in myocyte, which may be mediated by a signal system of phosphates Ca 2+ .