Discussion on the Problems and Suggestions in Ethical Review of Clinical Scientific Research Projects in China / 中国医学伦理学

Objective: To standardize the capacity construction of medical ethics committee,improve the quali-ty of ethical review of clinical scientific research projects,and better protect the rights and interests of subjects.Methods: This paper reviewed the literature about standardization construction of ethical review of clinical scientif-ic research projects and analyzed them comprehensively.Results: Based on the problems of regulatory system,re-view standards,education training,ethic awareness and tracking review facing in the practice of ethical review,this paper put forward some reflections and suggestions for the ethical development trend and solving strategies of clinical scientific research projects in future.Conclusion: It should standardize the construction of ethical review system of clinical scientific research projects,strengthen education and training,promote the construction capacity of medical ethics committee of our country,and promote the scientificity and standardization of the ethical review of clinical scientific research projects.

Existing Problems and Solution of Informed Consent in Clinical Research / 中国医学伦理学

Objective:To standardize the present situation of informed consent in China' s clinical research projects, and to better play the role of protecting the rights and interests of subjects, in order to improve the ethicality of research. Methods:According to the actual work,this paper reviewed literatures about the existing problems and solutions in the informed consent process in clinical research to carry out comprehensive analysis and discussion. Results:Combining with the problems about researchers, subjects and ethical committees faced in informed conent process in clinical research, this paper put forward thinking and suggestions on standardizing the informed consent for future medical clinical research. Conclusion:We should standardize the informed consent form and carry out the training of the subjects and researchers about medical ethics knowledge, so as to improve the status of informed consent, reflect the scientificity and ethicality of clinical research project, and to contribute to the development and progress of clinical medicine.

Application of Enhanced Recovery After Surgery in Complicated Hepatic Alveolar Echinocoecosis Surgery / 昆明医科大学学报

Objective To observe the application of enhanced recovery after surgery (ERAS) in complicated hepatic alveolar echinocoecosis surgery. Me thods We selected 117 patients from 2013 to 2016 and divided them into 3 groups in chronological order: Group A (n =35), the traditional management group. All patients underwent perioperative management according to previous procedures. Group B (n=44), the transitional management group. The management in the transition period was improved as far as possible in accordance with the ERAS programme. Group C (n=38), the standard process group. All patients were treated in full accordance with standard process. The hemodynamics data, fluid volume and the perioperative complications were recorded in all groups. Re s ults Compared with group C, there was no significant difference in the general condition and the incidence of complications before operation, the anesthesia time, the operation time, blood loss, blood transfusion and the heart rate in group A and B. The incidence of hunger and thirst before operation, infusion volume, urine output, the incidence of postoperative complications and the average length of hospitalization were all higher than those in group C. The differences were statistically significant. Conclus ion The clinical path optimization of ERAS in liver hydatid surgery can significantly reduce the perioperative complications and reduce the average length of hospitalization.

Short-term efficacy comparison between Ivor-Lewis approach and McKeown approach in minimally invasive esophagectomy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare the perioperative complications between Ivor-Lewis approach and McKeown approach in minimally invasive esophagectomy and gastric tube reconstruction for the treatment of middle and lower thoracic esophageal cancer.</p><p><b>METHODS</b>Retrospective analysis of clinical data was performed on 288 patients with middle and lower thoracic esophageal cancer who underwent completely minimally invasive esophagectomy by one surgical team in Fujian Medical University Union Hospital from December 2010 to March 2014. Among the 288 patients, 103 patients underwent combined laparoscopic and thoracoscopic esophagectomy and intrathoracic esophagogastric anastomosis using a transoral anvil(Orvil)(Ivor-Lewis group, 2-incision) and 185 patients underwent combined laparoscopic and thoracoscopic esophagectomy and cervical anastomosis(McKeown group, 3-incision). Patients were stratified by surgical approach and perioperative outcomes were compared between the two groups.</p><p><b>RESULTS</b>There were no statistical differences between two groups in intra-operative blood loss, conversion to open, extubation time, time to resume oral intake, postoperative hospital stay, the median number of lymph nodes resected. The operation time of Ivor-Lewis group was significantly shorter than that of McKeown group [(283.4±32.0) min vs. (303.6±43.7) min, P=0.003). The hospital cost of Ivor-Lewis group was significantly higher than that of McKeown group [(76 492±18 553) yuan vs. (68 923±17 331) yuan, P<0.01]. There were no statistical differences between two groups in chylothorax, delayed gastric emptying, atrial fibrillation, postoperative bleeding, admission to ICU, short-term postoperative mortality (P>0.05). The total postoperative complication morbidity of Ivor-Lewis group was significantly lower than that of McKeown group(16.5% vs. 31.4%, P<0.01). Ivor-Lewis group had lower pulmonary complication(8.7% vs. 25.9%, P<0.01), anastomotic leakage(1.9% vs. 13.0%, P<0.01), anastomotic stricture (0% vs. 4.9%, P<0.05), recurrent laryngeal nerve injury(1.0% vs. 7.0%, P<0.05).</p><p><b>CONCLUSION</b>Ivor-Lewis approach is associated with less postoperative complications, but higher cost as compared to McKeown approach in the treatment of middle and lower thoracic esophageal cancer.</p>

Functional role of protein kinase D1 in Aspergillus fumigatus-induced activation of nuclear factor-κB signal pathway and transcription / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the functional role of protein kinase D1 (PKD1) in the activation of nuclear factor-κB (NF-κB) signal pathway and NF-κB transcription mediated by Aspergillus fumigatus.</p><p><b>METHODS</b>A549 cells and HEK293 cells were transfected with green fluorescence protein (GFP) or GFP-PKD1 followed by treatment with 1×10(5) CFU/ml Aspergillus fumigatus conidia for different time lengths. The phosphorylation levels of PKD1, IκB and p65 (pS276) in the transfected cells were measured by Western blotting. A549 cells were transfected with GFP-PKD1 or siRNA-PKD1, and the phosphorylation of IκB and p65 (pS276) was examined. Finally, NF-κB-luc and renilla luciferase reporter pRL-SV40 were cotransfected into GFP- or GFP-PKD1-transfected A549 cells before exposure of the cells to Aspergillus fumigatus conidia for 24 h, and NF-κB transcriptional activity in the cells was determined using dual-luciferase reporter assay.</p><p><b>RESULTS</b>Overexpression of PKD1 significantly increased Aspergillus fumigatus conidia-stimulated phosphorylation of PKD1, IκB and p65 (pS276), whereas PKD1 knockdown by siRNA-PKD1 suppressed IκB and p65 (pS276) phosphorylation. Dual luciferase assay demonstrated that PKD1 overexpression markedly enhanced Aspergillus fumigatus-induced NF-κB transcription in A549 cells.</p><p><b>CONCLUSION</b>PKD1 may contribute to the activation of NF-κB signal pathway and NF-κB transcription induced by Aspergillus fumigatus.</p>

AXIN1-related CSRNP1 mRNA expression and its transcriptional regulation in TGF-β1-induced tumor cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate AXIN1-related CSRNP1 gene expression and the mechanism of its transcriptional regulation in TGF-β1-induced tumor cells.</p><p><b>METHODS</b>Human lung carcinoma A549 cells or human prostate cancer PC3 cells were treated with TGF-β1 at different doses (0, 20, 40, and 80 ng/ml) or at 20 ng/ml for 0, 8, 12, or 24 h, and the dose and time effect of TGF-β1 on CSRNP1 mRNA expression in the tumor cells were evaluated with real-time RT-PCR. A549 cells were also treated with TGF-β1 and cycloheximide to clarify whether CSRNP1 expression induced by TGF-β1 required de novo protein synthesis. A549 cells transfected with pcDNA3.1, flag-SMAD3, or flag-SMAD3-mu, after serum starvation, were treated with or without TGF-β1 (20 ng/mL) for 24 h, and the overexpression of wild-type SMAD3 and dominant negative SMAD3-mu mutant were confirmed by Western blotting. The effect of SMAD3 or SMAD3-mu overexpression on CSRNP1 mRNA expression was also measured by real-time RT-PCR.</p><p><b>RESULTS</b>In both A549 and PC3 cells, TGF-β1 dose- and time-dependently stimulated CSRNP1 expression, which required de novo protein synthesis in A549 cells. Overexpression of wild-type SMAD3 significantly increased the expression of CSRNP1 mRNA induced by TGF-β1, while overexpression of dominant negative SMAD3 mutant remarkably reduced CSRNP1 mRNA expression in response to TGF-β1 in A549 cells.</p><p><b>CONCLUSION</b>TGF-β1 may contribute to CSRNP1 expression through SMAD3 activation and downstream signaling in tumor cells.</p>

Mast cells and tumor angiogenesis / 国际肿瘤学杂志

Mast cells (MCs) are bone marrow-derived and tissue-homing immune cells which have important functions.Recruitment and activation in tumors are demonstrated to be mainly mediated by tumorderived stem cell factor (SCF) and its receptor c-kit on MCs.The numbers and distributions of MCs in tumors are closely related to the growth of tumors and angiogenesis.MCs can mediate angiogenesis by releasing a series of angiogenesis factors,so as to promote the progress of tumors.

Protein kinase D3 is involved in negative regulation of MMP-7 in prostate cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the role of protein kinase D3 (PKD3) in the regulation of matrix metalloproteinases 7 (MMP-7) expression in prostate cancer cells.</p><p><b>METHODS</b>PC-3 cells were either stimulated with 100 nmol/L PMA to activate PKD3 kinase activity, or transiently transfected with PKD3 siRNA, and the relative expression level of MMP-7 mRNA were analyzed by real-time PCR using 2(-delta delta Ct) method. MMP-7 mRNA levels were also analyzed and quantified in HEK293 cells with over-expression of wild-type PKD3, PKD3 knockdown (using PKD3 siRNA), or over-expression of wild-type PKD3 followed by PKD3 knockdown.</p><p><b>RESULTS</b>MMP-7 mRNA expression in PC3 cells was significantly decreased after PMA-induced PKD3 kinase activation. In contrast, PKD3 knockdown by siRNA transfection markedly increased MMP-7 mRNA level (P<0.01). MMP-7 mRNA level in HEK293 cells was significantly decreased by PKD3 over-expression, whereas obviously increased by PKD3 knockdown. Down-regulation of MMP-7 mRNA level in HEK293 induced by PKD3 over-expression was rescued by PKD3 knockdown.</p><p><b>CONCLUSION</b>PKD3 may contribute to the malignant progression of prostate cancer cells through negative regulation of MMP-7 expression.</p>

PKD3 contributes to up-regulation of prostate-specific antigen in prostate cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the role of PKD3 in prostate-specific antigen (PSA) expression regulation in androgen-dependent prostate cancer cells and explore the mechanism.</p><p><b>METHODS</b>LNCaP cells containing low level of PKD3 were transfected with pEGFP-C2 or pEGFP-PKD3 plasmid followed by dihydrotestosterone (DHT) treatment, and PSA mRNA level was analyzed by RT-QPCR using 2(-delta delta Ct) method. Wild-type or kinase-dead PKD3 plasmids, human androgen receptor plasmid pSVAR0, pMMTV-luc of AR luciferase reporter and renilla luciferase reporter pRL-SV40 were cotransfected into HEK293 cells, and after treatment with DHT for 24 h, the cells were harvested and AR transcriptional activity were determined by dual-luciferase reporter assay. The subcellular localization of endogenous PKD3 and AR and their colocalization induced by DHT were observed by confocal microscopy.</p><p><b>RESULTS</b>PSA mRNA level triggered by DHT was significantly increased by overexpression of pEGFP-PKD3 in LNCaP cells compared with that in pEGFP-C2 control cells (P<0.001). AR transcription in response to DHT treatment was also significantly up-regulated by wild type PKD3 expression (P<0.001), but partially down-regulated by kinase-dead PKD3 mutant (P<0.01). Endogenous PKD3 and AR in LNCaP cells not only translocated from the cytoplasm to the nucleus, but also colocalized with each other after DHT stimulation.</p><p><b>CONCLUSION</b>Elevated AR transcriptional activity and enhanced expression of PSA induced by PKD3 in response to DHT treatment suggest that PKD3 contributes to the proliferation and malignant growth of androgen-dependent prostate cancer cells.</p>

Prokaryotic expression and bioactivity of human platelet-derived growth factor B chain mature peptide / 南方医科大学学报

<p><b>OBJECTIVE</b>To express human platelet-derived growth factor (hPDGF) B chain mature peptide gene in a prokaryotic expression system and detect the bioactivity of the expressed protein.</p><p><b>METHODS</b>hPDGF B chain mature peptide gene was amplified and expressed in E. coli, and the recombinant protein, rhPDGF-BB, was purified and renatured in GSSG/GSS system. The bioactivity of rhPDGF-BB in vitro was evaluated with SD rat osteoblasts.</p><p><b>RESULTS</b>The full-length PDGF-B mature peptide gene was obtained and verified, and successfully expressed in E. coli. Bioactivity detection results showed that the expressed rhPDGF-BB obviously promoted the proliferation and DNA replication of SD rat osteoblasts in vitro (P<0.01).</p><p><b>CONCLUSION</b>he PDGF-B chain mature peptide cDNA has been successfully cloned and the PDGF-B precursor highly expressed in E. coli, and renatured rhPDGF-BB displays high bioactivity as shown by MTT assay and flow cytometry. This success provides the basis for production of functional PDGF-BB and facilitates further studies of its role in fracture healing and trauma reconstruction.</p>

Phospholipase Cγ1 and NF-κB are required for cell-matrix adhesion of colorectal cancer cells / 第二军医大学学报

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Phospholipase Cγ1 and NF-κB are required for cell-matrix adhesion of colorectal cancer cells / 第二军医大学学报

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Phospholipase Cγ1 and NF-κB are required for cell-matrix adhesion of colorectal cancer cells / 第二军医大学学报

Objective: To investigate the function and mechanism of phospholipase Cγ1 (PLCγ1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-κB) were used to study the effect of PLCγ1 and NF-κB on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLCγ1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLCγ1 or NF-κB resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P<0.05), but had no marked effect on SW480 cells. Western blot analysis showed that epidermal growth factor (EGF) stimulated the phosphorylation of PLCγ1 in LoVo. The results of EMSA indicated that inhibition of PLCγ1 signaling pathway also down-regulated the activity of NF-κB while EGF reversed the function. Conclusion:These data suggest that PLCγ1 plays a pivotal role in the EGF-induced cell-matrix adhesion of highly metastatic colorectal cancer cells and that NF-κB is also functional in this signaling pathway.

Molecular cloning and expression of human PDGF-B chain mature peptide gene / 中华外科杂志

<p><b>OBJECTIVE</b>To acquire sufficient PDGF-BB protein and provide the basis for the further studies of its role on the fracture healing and trauma reconstruction and its clinical applications.</p><p><b>METHODS</b>Constructed the prokaryotic expression vector pQE-PDGF-B with the gene rearrangement technique, and the monomeric form of recombinant PDGF-B expressed in E. coli M15.</p><p><b>RESULTS</b>PDGF-B mature peptide gene was inserted into prokaryotic expression vector pQE30, which was confirmed by PCR, enzyme digestion and sequencing identification; the expressed products of pQE-PDGF-B in E. coli showed a single protein on SDS-PAGE, and their expression level was about 15% of the total bacterial protein. The molecular weight of the purified PDGF-B protein was about 15 KDs on SDS-PAGE.</p><p><b>CONCLUSIONS</b>The construction of recombinant plasmid and preparation of the monomeric protein of PDGF-B provides a solid foundation for further studying the function of PDGF-BB and producing biologically PDGF-BB protein.</p>

CONSTRUCTION AND EXPRESSION OF HUMAN Axud1 GENE WITH HA EPITOPE TAG IN LUNG ADENOCARCINOMA SPC-A1 CELL LINE / 解放军医学杂志

To construct the eukaryotic vector that expresses the fusion protein of Axud1 and influenza virus hemagglutin HA epitope tag, the total RNA was isolated from the peripheral blood lymphocytes, and reverse transcription reaction was used to amplify the full length of human Axud1 cDNA. PCR product of Axud1 was then amplified using specific primers containing HA epitope sequence, and inserted into eukaryotic expression plasmid pcDNA3.1(+)digested with BamH Ⅰand Xba Ⅰ. The recombinant plasmid was identified by polymerase chain reaction, restriction endonuclease mapping and sequencing, and then transfected into human lung adenocarcinoma SPC-A1 cell lines.The fusion HA-Axud1 protein expression in anti-G418 clones was verified by Western blot. This study might be instrumental in further study of the function of Axud1 protein in tumor cells.

Mathematical model of the hydrolization of PIP2 with the stimulation of EGF / 医疗卫生装备

To investigate dynamic characteristics of the hydrolization lipid phosphatidylinositol(4,5)-bisphosphate(PIP 2 )with the stimulation of epidermal growth factor(EGF),the mathematical model to simulate the metabolizability of PIP 2 is established based on Law of Mass Action and Law of the Quasi-steady-State Approximation.Differential equations of concentration relations between PIP 2 and EGF receptor are formulated,and the effect of the parameters on the changing trend of PIP 2 is analyzed.This mathematical model describes biology characteristics of metabolized PIP 2 and dependency relationships of concentration between key signal producuts.

Phospholipase C?1 and NF-?B are required for cell-matrix adhesion of colorectal cancer cells / 第二军医大学学报

Objective: To investigate the function and mechanism of phospholipase C?1 (PLC?1) in cell-matrix adhesion in colorectal cancer. Methods: Highly metastatic colorectal cancer cell line LoVo and lowly metastatic colorectal cancer cell line SW480 were subjected to cell-matrix adhesion assay. U73122 (a specific inhibitor of PLC) and pyrrolidine dithiocarbamate (PDTC) (an inhibitor of NF-?B) were used to study the effect of PLC?1 and NF-?B on cell-matrix adhesion. Furthermore, Western blot and gel electrophoresis mobility shift assay (EMSA) were performed to detect the mechanism of PLC?1 in colorectal cancer cell adhesion to matrix. Results: Inhibition of PLC?1 or NF-?B resulted in reduction of cell-matrix adhesion in a dose-dependent manner in LoVo cells(P