Objective To establish a method for detecting K-rag gene point mutation in pancreatic adenocarcinoma based on the pyrosequencing and to compare its performance with that of Sanger sequencing.Methods Genomic DNA was extracted from formalin-fixed,paraffin-embedded pancreatic tissues including 49 pancreatic adenocarcinoma,10 normal pancreas,11 chronic pancreatitis,18 benign pancreatic tumor,7 insulin carcinoma,9 ampullary carcinoma,7 bile duct carcinoma and 7 duodenum papillary adenocarcinoma tissues.K-rag gene point mutations at codon 12 were analyzed by pyrosequencing and Sanger sequencing,respectively.Results No mutant K-ras gene Wag detected in normal pancreas,chronic pancreatitis,benign pancreatic tumor,insulin carcinoma,ampullary carcinoma,bile duct carcinoma and duodenum papillary adenocarcinoma tissues by either pyrosequencing or Sanger sequencing.K-rag gene point mutation was detection rate in pancreatic adenocarcinoma tissues was 71.4%(35/49)by pyrosequencing and 61.2%(30/49)by Sanger sequencing,respectively.Conclusions Pyrosequencing is more sensitive than Sanger sequencing and is also accurate,rapid and of high throushput in detecting mutant K-ras gene.It may serve as a practical method for fast batch analysis of clinical samples.

Abstract： Objective To evaluate the filtration effects of various nanofiltration systems on intravenous human immunog⁃ lobulin（IVIG）in order to screen the optimal nanofiltration system. Methods Various nanofilters were used for IVIG filtration to determine the best one and then various prefilters were selected to combine with the optimal nanofilter for IVIG filtration to determine the optimal nanofiltration system. Results The tangential flow（cross flow）nanofilter showed better filtering effect than dead end（direct current）nanofilter，and nanofilter C was the best one. The effect of deep filtration prefilter was better than that of absolute filtration prefilter，and prefilter Y1 in series with nanofilter C was the optimal nanofiltration system. Conclusion The optimal nanofiltration system was determined through the effect evaluation of various nanofiltration systems filtering for IVIG.

In recent years, research about sentinel lymph node mapping in endometrial cancer surgery has developed quickly. Several clinical studies indicate that sentinel lymph node mapping in early endometrial cancer can improve detectable rate and decrease the false-negative rate. However, the technology is still controversial. Therefore, further study is needed to assess the role of sentinel lymph node mapping in endometrial cancer. This review mainly focused on the current clinical practice of sentinel node mapping technique in endometrial cancer, and the existing problems and controversies.

OBJECTIVETo evaluate the effectiveness and safety of fluoroscopy-guided percutaneous intratumor injection of pingyangmycin lipiodol emulsion (PLE) in the management of recurrent sacrococcygeal chordomas.

METHODSSeven patients with recurrent sacrococcygeal chordomas presenting with severe local pain with visual analogue score (VAS)≥8 received treatment sessions of fluoroscopy-guided percutaneous intratumor injection of PLE. The patients were followed up every 3 months after the last session to assess their clinical responses and observe the changes in the tumor size measured by computed tomography. The changes in the VAS, tumor necrosis and pain relief as well as the adverse events were recorded.

RESULTSA total of 22 sessions of fluoroscopy-guided percutaneous intratumoral PLE injection was performed in these cases (3 or 4 sessions in each case). The total average pingyangmycin dose delivered was 48.0 mg and the average lipiodol dose was 40.0 ml in each case. Five patients showed low fever and vomiting 48 after the injection. During the follow-up (median time of 21.7 months, range 10-26 months), all the patients showed obviously reduced tumor size and VAS, and partial remission was achieved in 6 patients and stable disease (SD) in 1 patient. None of the patients had complications during the follow-up.

CONCLUSIONFluoroscopy-guided percutaneous intratumoral injection of PLE can be effective and safe and may serve as a alternative for treatment of recurrent sacrococcygeal chordomas.

Adult ; Aged ; Bleomycin ; administration & dosage ; analogs & derivatives ; therapeutic use ; Chordoma ; drug therapy ; Emulsions ; administration & dosage ; therapeutic use ; Ethiodized Oil ; administration & dosage ; therapeutic use ; Female ; Humans ; Injections, Intralesional ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; drug therapy ; Sacrococcygeal Region ; pathology

Adolescent ; Adult ; Analgesics ; administration & dosage ; Anesthetics, Intravenous ; administration & dosage ; Flurbiprofen ; administration & dosage ; Humans ; Hypnotics and Sedatives ; administration & dosage ; Midazolam ; administration & dosage ; Middle Aged ; Molar, Third ; Pain Measurement ; Pain, Postoperative ; prevention & control ; Patient Satisfaction ; Preoperative Care ; Prospective Studies ; Tooth Extraction ; Young Adult

Coronary artery disease (CAD) is a multifactorial disease in which inflammation plays a central role.This study aimed to investigate the association of inflammatory markers such as the neutrophil to lymphocyte ratio (NLR),the Global Registry of Acute Coronary Events (GRACE) score with in-hospital mortality of elderly patients with acute myocardial infarction (AMI) in an attempt to explore the prognostic value of these indices for elderly AMI patients.One thousand consecutive CAD patients were divided into two groups based on age 60.The laboratory and clinical characteristics were assessed retrospectively by reviewing the medical records.The NLR and GRACE score were calculated.In the elderly (≥60 years),patients with non-ST-elevation myocardial infarction (NSTEMI) and ST-elevation myocardial infarction (STEMI) had significantly higher NLR than did those with unstable angina (UA) and stable angina pectoris (SAP) (P＜0.01).The NLR was considerably elevated in older AMI patients compared with their younger counterparts (＜60 years) (P＜0.05).In elderly AMI patients,the NLR was considerably higher in the high-risk group than in both the low-risk and medium-risk groups based on the GRACE score (P＜0.05 and P＜0.01,respectively),and the NLR was positively correlated with the GRACE score (r=0.322,P＜0.001).Either the NLR level or the GRACE score was significantly higher in the death group than in the surviving group (P＜0.05).By curve receiver operator characteristic curve (ROC) analysis,the optimal cut-off levels of 9.41 for NLR and 174 for GRACE score predicted in-hospital death [ROC area under the curve (AUC) 0.771 and 0.787,respectively,P＜0.001].It was concluded that an elevated NLR is a potential predictor of in-hospital mortality in elderly patients with AMI.

Objective To explore the treatment methods and surgical techniques of the third ventricle tumor combined with hydrocephalus under neuroendoscope. Methods The clinical data and imaging findings of 4 patients with third ventricle tumor combined with hydrocephalus, treated with surgery under neuroendoscopy, were retrospectively analyzed; and related literatures were reviewed to conclude the surgical experiences and skills. Results Three of the 4 patients were performed surgery only by neuroendoscopy, and 1 by neuroendoscopic auxiliary microscope for the tumor complete resection. The clinical symptoms improved obviously after the surgery, and no significant complications and no dead case were noted.We followed up the 4 patients for 3-18 months; MRI showed that the tumor did not relapse and the hydrocephalus got improvement. Conclusion Endoscopic navigation can help to directly reach the locations of third ventricle tumor and decrease the unnecessary damage, which enjoys its advantages in tumor resection,relieving obstructive hydrocephalus and rebuilding the cerebrospinal fluid circulation, indicating that surgery under neuroendoscope is a safe, effective and minimally invasive method.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Cross-Sectional Studies ; Homosexuality, Male ; psychology ; Humans ; Male ; Middle Aged ; Sexual Behavior ; Surveys and Questionnaires ; Young Adult

Adult ; Homosexuality, Male ; psychology ; Humans ; Male ; Students ; psychology ; Surveys and Questionnaires ; Unsafe Sex

Objective:To obtain a high specificity and high affinity anti-PcrV protein monoclonal antibody which can be used for the treatment of Pseudomonas aeruginosa infected.Methods: The PcrV gene was amplified by PCR using P.aeruginosa PAO1 genome DNA as the template.The expression vector(pET-28a-PcrV) was constructed and transformed into E.coli BL21(DE3).The re-combinant PcrV protein was expressed by IPTG induction and purified by Ni2+affinity chromatography.The specific binders of PcrV were screened by phage display.The genes encoding VH and VL were amplified respectively by PCR using the plasmid of positive clone as the template.Then the recombinant expression vectors were constructed and transfected into 293E cells.Monoclonal antibody were purified by the Protein A affinity resin from the culture supernatants.The affinity of antibody was detected by ELISA and the function of YG5 was verified in murine pneumonia model caused by P.aeruginosa.Results: Recombinant PcrV protein was expressed and purified.A full human monoclonal antibody(named as YG5) against PcrV was obtained by phage display.The results of ELISA showed that YG5 had a high affinity with EC50=61 ng/ml.Furthermore,it was found that YG5 could protect mice from infection caused by P.aeruginosa.Conclusion:Our findings present a novel human monoclonal antibody YG5 against PcrV,which inhibits the infection casued by P.aeruginosa and may be a potential drug for treatment of P.aeruginosa infection.