Objective To summarize the experience and the insufficiency of Nanjing Drum Tower Hospital on patent management,and find out the problemsand countermeasures for the management.Methods The authorized patents during 2003-2014 of Nanjing Drum Tower Hospital were analyzed statistically,and the date was overall analyzed combined withthe number,type,content,maintenance and transfer of patent and inventors' educational background,title and subject;and results are further analyzed;Results Patent grant increased significantly more than the invention patent in recent years.In terms of patent types,most patents are utility model patents,and conversion rate of patent outcome is very low.Conclusions Suggestions are proposed to improve hospital patent management:strengthening the guidance of the management of patent policy;promoting patent work;enhancing the professional quality of patent managers;optimizing the patent management processe andstrengthening the patent examination and data analysis;strengthening strategic cooperation among intellectual property management department,patent agency and biological pharmaceutical enterprises.

OBJECTIVE To study and find reasonable methods of nursing care of incision through analyzing the risk factors in liver transplant patients.METHODS A total of 416 liver transplantation operations in our hospital from Sep 2004 to May 2006 were evaluated.The risk factors that resulted in the incision infection were considered.RESULTS The incision infection rate was decreased from 13.9% to 4.3% after improvement.CONCLUSIONS Nursing care is of great significance in preventing the incision infection in liver transplant patients.

An analytical method was developed for the simultaneous extraction and determination of eighteen fluoroquinolones (FQs), tetracyclines (TCs) and sulfonamides (SAs) antibiotics from soils using solid phaseextraction and liquid chromatography tandem mass spectrometry.Soil sample was firstly extracted by phosphate buffer at pH 3 in combination with 50% of organic modifier acetonitrile, then purified and concentrated by SAX and HLB column.Qualitative and quantitative analysis were carried out for the analyte under the MRM mode after the chromatography separation on Kromasil C_(18)(250 mm x4.6 mm, 5 μm) column.The range of recoveries (in percent) for FQs, TCs, SAs, in the soil matrix was 67.20%-88.98%, 62.23%-85.36%, 55.76%-97.37% with 1.1%-17.2% of relative standard deviation respectively in two different concentra tions.The limits of quantification (LOQ, S/N = 3) were 3.36-8.88 jig/kg, 0.56-0.91 μg/kg and 0.07-1.85 μg/kg for FQs, TCs and SAs, respectively.This method was successfully used to detect 18 anti biotics in 6 soil samples with different land types in Tianjin.Results showed some of the antibiotics in the arable soil were detected, with concentrations of 1.72-119.57 μg/kg.

The expression of N-myc down-regulated gene 1 (NDRG1) has previously been reported to be involved in the proliferation, differentiation, invasion and metastasis of cancer cells, but its role in cervical cancer is still unclear. This study aimed to investigate the expression of NDRG1gene in human cervical cancer and its effect on aggressive tumor behaviors. The NDRG1 expression in cervical tissues and cells was detected by RT-PCR. Specific expression plasmid pEGFP-N1-NDRG1-GFP was used to enhance the expression of NDRG1 in human cervical cancer cell lines. The mRNA and protein level of NDRG1 was assessed by RT-PCR and Western blotting, respectively. Its effects on cell proliferation, migration, invasion, cell cycle and apoptosis were detected by MTT, transwell migration assay and flow cytometry (FCM), respectively. The results showed that the expression of NDRG1 in cervical cancer tissues and cells was significantly lower than in normal cervical tissues (P<0.001). After transfection with pEGFP-N1-NDRG1-GFP, the mRNA and protein expression of NDRG1 was up-regulated in Siha cells, which suppressed cell proliferation (P<0.001), induced cell cycle arrest (P<0.05), reduced invasion and migration of Siha cells (P<0.05), but caused no cell apoptosis. Moreover, vascular endothelial growth factor (VEGF), a tumor-induced angiogenesis factor, was markedly reduced and E-cadherin, a cell adhesion molecule, was increased in the cells transfected with pEGFP-N1-NDRG1-GFP. It was concluded that up-regulated NDRG1 may play a role in the suppression of malignant cell growth, invasion and metastasis of human cervical cancer.

Objective To examine the effects of Shenqi Bufei Tang decoction on the expression of histone deacetylase-2 ( HDAC2) and nuclear factor-κB p65 ( NF-κB p65) in the airway smooth muscle tissues of COPD rats with lung-qi deficiency syndrome. Methods A total of 40 male rats were randomly divided into normal control group,model control group,Shenqi Bufei Tang decoction group,and aminophylline group.The COPD rat model with lung-qi deficiency syndrome was established by intra-tracheal instillation of lipopolysaccharide (LPS) and passive smoking for 28 days.Pathological changes of lung tissues were ob-served under the light microscope and the thickness of the small airway wall and airway smooth muscle ( ASM) layer analyzed by the image analysis.Immunohistochemistry,real-time PCR and Western blotting were used to detect the expressions of NF-κB p65 and HDAC2 in ASM. Results The thickness of the airway wall and ASM,and the expression levels of NF-κB p65 mRNA and protein were significantly increased in the model control group when compared with those in the normal control group ( P<0.05) . They were significantly reduced and the expression levels of HDAC2 mRNA and protein were markedly increased in Shenqi Bufei Tang decoction group and aminophylline group,which were compared with those in the model group (P<0.05).There were no sig-nificant differences in these indices between Shenqi Bufei Tang decoction group and aminophylline group (P>0.05). Conclu-sion Shenqi Bufei Tang decoction can inhibit the proliferation of ASM in COPD rats with lung-qi deficiency syndrome,which may be associated with the increased expression of HDAC2 and decreased expression of NF-κB p65.

Objective To investigate the expression of microRNA 27a (miR-27a) and relationship with drug resistance in human ovarian cancer A2780/Taxol cells.Methods A stem-loop-mediated real-time PCR was used to detect miR-27a expression in A2780 and A2780/Taxol cells.The cells were transfected with the mimics or inhibitors of miR-27a or negative control RNA ( NC) by lipofectamine 2000.The expressions of MDR1 gene,P-glycoprotein (P-gp) and homeodomain-interacting protein kinase 2 (HIPK2) protein levels were measured by real-time PCR and western blot respectively.Methyl thiazolyl tetrazolium (MTT) assay was used to analyze drug sensitivity.Apoptosis analysis was measured by fluorescence activated cell sorter ( FACS).Results (1) miR-27a was an average of 2.2-fold higher expression level in A2780/Taxol cells than that in A2780 cells,with a significant difference between the two groups (P <0.05).(2) A2780/Taxol cells transfection with inhibitors of miR-27a showed that the levels of MDR1 mRNA was decreased by 39%,P-gp protein level[(26 ±5)%]decreased than that in the NC group[(43 ±7)%],HIPK2 protein level[(30 ±6)%]increased than that in the NC group[(19 ±4)%],the 50% inhibitionconcentration (0.5 (μmol/L) was less than that in the NC group (6.8 μmol/L),apoptosis rate[(32.5 ± 3.6) %]was higher than that in the NC group[(5.6 ±2.1) %],and there were significant differences between two groups (all P < 0.05 ).( 3 ) Transfection of A2780 cells with mimics of miR-27a led to increase MDR1 mRNA expression by 121% as compared with one transfection with NC (P<0.05).Conclusion The expression of miR-27a is upregulated in A2780/Taxol cells,which may regulate MDR1 and P-gp expression by targeting HIPK2.

AIM and METHODS: Electron cytochemical methods were used to study the changes of calcium and reactive oxygen species in rat kidney during ischemia and reperfusion period.RESULTS:By the end of 1h ischemia, intra-cellular calcium increased. There were no H 2O 2 generation at this time. In the early reperfusion period, large amount of H 2O 2 generated. At this time, there were no evident changes of intra-cellular calcium compare with 1h ischemia group. In the later reperfusion period, less H 2O 2 generated. Intra-cellular calcium increased continuously.CONCLUSION:Calcium and reactive oxygen species all participated in ischemia-reperfusion injury, but the time they participated and their effects were different.

OBJECTIVETo investigate the anti-metastasis effect of emodin on the pancreatic cancer in vitro and in vivo.

METHODHuman pancreatic cancer cell line SW1990 was treated with different concentrations of emodin (10, 20, 40 micromol x L(-1)) for 2 h, the effects of emodin on the migration and invasion of SW1990 cells were examined by using wound assay and matrigel counting. Western blot was used to detect the protein expression of NF-kappaB and MMP-9 in SW1990 cells after various concentrations of emodin (10, 20, 40 micromol x L(-1)) treatment for 48 h. Metastatic model simulating human pancreatic cancer was established by orthotropic implantation of histologically intact human tumor tissue into pancreatic wall of nude mice, and then divided into three groups: control group, low-dose emodin group (L-EMO) and high-dose emodin group (H-EMO). Eight weeks after implantation, the presences of metastasis were evaluated respectively after the mice were sacrificed. Immunohistochemistry was used to detect the positive expression of CD34, NF-kappaB and MMP-9 in the tumors.

RESULTEmodin suppressed the migration and invasion of SW1990 cells in a dose-dependent manner. Western bolt assay indicated that emodin down-regulated the expression of NF-kappaB and MMP-9 proteins in SW1990 cells. The incidences of metastasis were decreased significantly in L-EMO group and H-EMO group as compared with that in control group. The percentage of CD34, NF-kappaB and MMP-9-positive cells in the tumors were significantly reduced by the administration of emodin.

CONCLUSIONEmodin exerts anti-metastatic activity in pancreatic cancer both in vitro and in vivo, which may be related to down-regulation of NF-kappaB and MMP-9.

Angiogenesis Inhibitors ; pharmacology ; Animals ; Cell Line, Tumor ; Emodin ; pharmacology ; Female ; Humans ; Matrix Metalloproteinase 9 ; analysis ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred BALB C ; NF-kappa B ; analysis ; antagonists & inhibitors ; Neoplasm Metastasis ; prevention & control ; Pancreatic Neoplasms ; blood supply ; drug therapy ; pathology

To character a novel chimera(1b/2a) hepatitis C virus cell culture (HCVcc) system carrying envelope E1E2 coding gene from Hebei strain of China, chimera HCVcc (cHCVcc) was developed from Huh7.5-CD81 cells after transfection with in vitro transcribed full-length 1b/2a chimera RNA, which carrying envelope E1E2 coding gene from Hebei strain of China. Then the replication, expression and infectious titer of serial passage HCVcc were assessed by Real Time RT-PCR, indirect immunofluorescence assay (IFA) and Western blotting (WB). In addition, chimeric envelope gene from HCVcc was sequenced after serial passage. We found that the number of HCV positive focus increased gradually in cell post-transfection with chimera HCVcc (1b/2a) RNA and reach a peak platform (80% to 90%) at 41 days post-transfection; the expression of HCV protein was also confirmed by WAB during serial passage. At meantime, HCV RNA copy number in the supernatant peaked at 10(4)-10(7) copies/mL and the highest infectious titer of this 1b/2a cHCVcc reinfection were tested as 10(4) ffu/mL. Sequence analysis indicated 6 of adaptive amino acid substitutes occur among chimeric envelope E1E2 during serial passages. We con:luded that a novel 1b/2a chimera HCVcc carrying envelope E1E2 coding gene from Hebei strain of China was developed and its infectious titer increased after serial passage of HCVcc. This novel cHCVcc will be an effective tool for further evaluation of anti-virus drugs and immune effects against the major genotype from Chinese.
Cell Line ; China ; Hepacivirus ; genetics ; growth & development ; metabolism ; Hepatitis C ; virology ; Humans ; Serial Passage ; Viral Envelope Proteins ; genetics ; metabolism

Objective To observe the long-term prognosis and analyze the predictive factors of esophageal cancer patients treated with three-dimensional radiotherapy.Methods A total of 373 patients with esophageal squamous carcinoma who received three-dimensional radiotherapy were retrospectively enrolled in this study.Among these,231 cases received three dimensional conformal radiotherapy (3D-CRT) and the other 142 received intensity modulated radiotherapy (IMRT);202 cases received radiotherapy alone,and the other 171 received chemoradiotherapy;249 cases received involved-field irradiation(IFI),and the other 124 received elective nodal irradiation(ENI);60 cases received a total radiation dose of 50-60 Gy,and the other 313 received 60-70 Gy.Kaplan-Meier method was used to calculate the overall survival (OS) and progression-free survival (PFS).The Logrank single factor analysis and Cox multivariate analysis were used to evaluate predictive factors of PFS and OS.Results The 1-,3-,5-year OS and PFS were 69.4％,33.7％,22.9％ and 63.8％,32.8％,22.4％,respectively.The median OS and PFS were 22.7 months (95％ CI 18.6-25.4 months) and 19.2 months (95％ CI 16.7-21.3 months) respectively.Univariate analysis showed that age,gender,tumor location,three-dimensional technology (3D-CRT vs.IMRT),chemotherapy,prophylactic irradiation to lymphatic drainage area and irradiation dose did not influence OS and PFS (P ＞ 0.05).T-stage,N-stage,TNM-stage and gross tumor volume (GTV) were significantly correlated to OS and PFS (x2 =5.836-14.526,P ＜ 0.05).The multivariate analysis showed that N-stage and GTV were independent predictive factors of OS and PFS (x2 =5.345-12.216,P ＜0.05).The OS and PFS of patients with two fields of lymph node metastases were worse than those with only one lymph node field metastasis (x2 =4.467,4.169,P ＜ 0.05).Conclusions The long-term efficacy for esophageal cancer patients could be significantly improved through 3D-CRT technology.N-stage and tumor volume were independent prognostic factors of OS and DFS.The number of lymph node metastasis field is significantly related to prognosis.