1.Expression and purification of spindlin 1,a novel cancer related protein,and preparation of its polyclonal antibody
Lin CHEN ; Quan ZENG ; Peng ZHANG ; Jingxue WANG ; Lipeng QIN ; Yang LYU ; Xue NAN ; Wen YUE ; Xuetao PEI
Chinese Journal of Pharmacology and Toxicology 2014;(3):321-328
OBJECTIVE Toprepareapolyclonalantibodyforspindlin1protein,anovelcancer related protein,and to provide the data for a better understanding of its functions and screening tu mor. METHODS Purifiedspindlin1proteinwasinjectedintorabbitstoproducethepolyclonalantiserumafter removing glutathione S-transferase (GST)from the fusion protein spindlin 1-GST that was expressed in Escherichia coli..The antiserum was purified through the Hitrap Protein A system,and the titer of spin-dlin 1 polyclonal antibody was detected by ELISA.The specificity of the polyclonal antibody was deter-minedbyWesternblottingandimmunohistochemistry.RESULTS Thetiterofspindlin1polyclonalanti-body was 1∶2000.Western blotting detection demonstrated that the spindlin 1 polyclonal antibody recog-nized myc-spindlin 1 reco mbinant fusion protein in HeLa cells transfected with pAdeasy-myc-spindlin 1 , which also corresponded with Myc.antibody.The HeLa cells were transfected with enhanced green fluo-rescence protein (EGFP)and spindlin 1 vector(pEGFP-C3-spindlin 1 ),which was confirmed by the in-dependent GFP fluorescence assay.The results of immunohistochemistry detection with the spindlin 1 polyclonal antibody suggested that spindlin 1 was mainly expressed in the nuclei of HeLa cells.More i m-portantly,in i mmunohistoche mical assays,the spindlin 1 antibody recognized nuclear spindlin 1 expres-sioninclinicalovariancancertissues.CONCLUSION Thespecificspindlin1polyclonalantibodyispre-pared,which may be used to detect cancer-related protein spindlin 1 in HeLa cells and ovarian cancer tissues.
2.The study on the clinical effect of modified four-step laparoscopic splenectomy and pericardial de-vascularization
Anding WU ; Lipeng WAN ; Yanqiong QIN
Journal of Clinical Surgery 2018;26(5):339-341
Objective To explore the clinical efficary of modified four-step laparoscopic splenec-tomy and pericardial devascularization.Methods The clinical data of 72 patients with portal hypertension who were admitted to our hospital were retrospectively analyzed.Patients were divided into modified four-step laparoscopic group(36 patients)and open group(36 patients). The clinical indicators(operation time,amount of intraoperative blood loss,hospital stay,total hospitalization expenses,and intraoperative bleeding rate)and the incidence of postoperative complications were compared among the two groups.Re-sults The intraoperative blood loss(755.16 ± 50.23),Intraoperative bleeding rate(13%)and hospital stay(14.77 ± 2.09)of open group was higher than the modified four-step laparoscopic group(462.02 ± 61.42,11.11%,9.45 ± 3.20,P<0.05).The median operation time(187.67 ± 32.10)and the cost of hospitalization(42368. 70)of open group was lower than the modified four-step laparoscopic group (265.79 ± 48.23,50143.46,P<0.05).There were no significant differences in perioperative complica-tion rate among the two groups(P>0.05).Conclusions The procedure of modified four-step laparo-scopic splenectomy and pericardial devascularization has safe,feasible and with good therapeutic effect.It can significantly reduce the blood flow and blood transfusion in patients with probability,shorten the hospi-talization,lower the incidence of postoperative complications to some extent.However,the cost of hospital-ization and the time of operation are still relatively high,and further improvement should be made on the clinical application.
3.Auto recognizing critical values from medical image examination reports by using regular expressions
Rongjie CAI ; Dapeng LI ; Fangyuan QIN ; Jingjing HUANG ; Xiaoyan YANG ; Lipeng MA
Journal of Practical Radiology 2018;34(3):444-446
Objective To introduce a method of automatically identifying critical values from medical image examination reports and prompt the physician to report it,to prevent the omission of the critical value reporting and improve the medical quality.Methods According to the requirement of critical value reporting system,regular expressions were made for each emergency situation of medical image examination,in order to form a critical value feature library.And an algorithm was designed to find critical value and prompt doctors automatically.Results According to this method,the critical value auto recognize software was developed and had been tested in Nanfang Hospital for 6 months.The software ran well.Conclusion Using regular expressions to define a criteria value feature library and design an algorithm of identifying criteria values,can recognize critical values and prompt physician automatically.
4.NRSF Plays a Regulatory Role in Human Insulin Gene Transcription
Qingbin LIU ; Yanhua LI ; Yinxiang YANG ; Hongfeng YUAN ; Rui ZHANG ; Lipeng QIN ; Yang L ; Sainan ZHANG ; Dijin SUN ; Lin CHEN ; Cixian BAI ; Xue NAN ; Yunfang WANG ; Xuetao PEI
Progress in Biochemistry and Biophysics 2006;0(09):-
The transcriptional repressor RE1 silencer transcription factor(NRSF/REST) is an important factor that restricts some neuronal traits in neurons.Since these traits are also present in pancreatic islet cells,NRSF-regulated genes involved in islet function are searched.A NRSE-like motif was analysed in human insulin promoter.The role of NRSE was evaluated by generating a model of insulin-secreting cells that firmly express NRSF.The presence of NRSF led to a decrease in activity of human insulin promoter by stable or transient transfection with human insulin-promoter luciferase.The predicted NRSE-like motif also confers NRSF-dependent transcriptional repression in the context of a surrogate gene promoter.Specific binding activity of NRSF/REST to the NRSE-like motif was confirmed by EMSA.Moreover,the binding activity is competed by consensus NRSE sequence.These data showed that human insulin promoter is regulated by the transcriptional repressor NRSF/REST via the NRSE-like motif.
5. Condition assessment and treatment strategy selection for patients with renal cell carcinoma bone metastasis
Xiyi WEI ; Zebin YOU ; Guangyao LI ; Xinglin CHEN ; Xiaohan REN ; Lipeng YU ; Yimin MA ; Juming LI ; Chao QIN ; Zengjun WANG
Chinese Journal of Urology 2019;40(11):833-837
Objective:
To investigate the assessment and treatment strategy of patients with renal cell carcinoma.
Methods:
The clinical data of 43 patients with renal cell carcinoma and bone metastases admitted to the First Affiliated Hospital of Nanjing Medical University from January 2006 to December 2018 were retrospectively analyzed. The follow-up time was 6 years, with an average age of 55.4 years (21-87 years). There were 29 males, 14 females, 22 cases of limb bone metastasis, 12 cases of spinal metastasis, 9 cases of multiple bone metastasis, 21 cases of Fuhrman grade 1 and 2, 19 cases of T1, and 20 cases of N0. All patients were confirmed by postoperative pathological examination or imaging data suggesting that bone metastasis are from renal cell carcinoma. Forty-three patients underwent primary renal surgery, and molecular targeted therapy was used after the operation. The treatment process was smooth, no obvious discomfort, and postoperative pathology showed clear cell carcinoma.22 patients with limb bones metastasis and 12 patients with spinal metastasis included in the study all met the indications for secondary surgery after the disease assessment. After communicating with the patient, 13 patients with limbs metastasis and 6 patients with spinal metastasis received local treatment, including complete resection of the extremities and spinal fixation, the remaining 15 patients and 9 patients with multiple bone metastasis were treated conservatively. There were 19 patients in the local treatment group, 13 patients with limbs bone metastasis, 6 patients with spinal bone metastasis, the average age was 54.9 years, the average diameter of the primary tumor was 4.7 cm. There were 24 patients in the conservative treatment group, 9 patients with limbs metastasis, 6 patients with spinal metastases and 9 cases with multiple bone metastasis, with an average age of 56 years and a primary tumor diameter of 5.6 cm. Limb metastatic lesions were evaluated according to the patient's general condition, bone pain, fracture risk, and bone metastasis. Spinal lesions were evaluated according to Tokuhashi score, Harrington score, Tomita score, vertebral stability assessment, and molecular targeted therapy. Aminokinase inhibitors, conservative treatment with local radiotherapy and bisphosphonate treatment.
Results:
During the follow-up period, the 1-year overall survival rate of the local treatment group was 100.0%, the 2-year overall survival rate was 89.4%, and the 5-year overall survival rate was 73.7%. The 1-year overall survival rate of the conservative treatment group was 87.5%, and the 2-year overall survival rate was 62.5%. The 5-year overall survival rate was 16.7%. The 2-year and 5-year survival rates of the local treatment group were statistically different (