Objective To construct eukaryotic expression vector of antisense MBD1 gene fragment and to provide a tool for studying MBD1 gene function. Methods PCR primers were designed according to the coding sequence of MBD1 gene. Xba I and Kpn I recognition sequences and cutting sites were added to the 5' end of the sense and antisense primer respectively. The 342 bp specific PCR fragment was obtained from the cDNA of biliary tract carcinoma cell line QBC-939 using RT-PCR, the purified PCR fragment was then inserted reversely into the multiple cloning site of eukaryotic expression vector pcDNA3. 1 ( + ). The constructed recombinant plasmid was identified by PCR confirmation, Xba I and Kpn I double enzyme digestion and DNA sequencing. Results The 322 bp specific DNA band was obtained by PCR, Xba I and Kpn I double digestion produced a 327 bp and a 5. 4 kb DNA band which represent the inserted target gene fragment and the vector respectively. The sequencing result confirmed that the sequence of inserted fragment was correct. Conclusion The eukaryotic expression vector of antisense MBD1 gene fragment was constructed successfully by using gene cloning technique. It will be a useful tool for studying MBD1 gene function in DNA methylation and tumorigenesis.

Objective To study the reliability of Gleason score by prostate biopsy with prostatic cancer(PCa)in prediction of tumor location and analyse related influencing factors of positive surgical margins after radical prostatectomy.Methods The retrospective study recruited 72 patients with PCa who had been diagnosed by trans rectal ultrasound-guided prostate biopsy,and eventually treated with laparoscopic radical prostatectomy.The correlation of each index were analyzed by statistical software.Results In this study,Kappa test indicated a good agreement that Gleason score between prostate biopsy and radical prostatectomy.Biopsy positive more focused on four peripheral points near the bottom.Biopsy tumor location was not suitable for predicting tumor distribution.Biopsy Gleason score and biopsy positive percentage in biopsy specimen were independent predictors of positive surgical margins.F and L points had good correlation with the positive surgical margins.Conclusion The biopsy Gleason score is a good evidence to pathological grading in patients.These information will lead to positive surgical margins rise possibly:needle biopsy Gleason score exceed 7.25,positive biopsy percentage more than 45% and the location positive biopsy close to prostate apex.

According to the latest data from the National Cancer Center of China, prostate cancer (PCa) has become the highest incidence tumors of urinary since 2008, and its incidence and mortality has occupied great attention in the past decades. Therefore, the choice of safe and effective diagnostic method, to detect the occurrence of PCa, is necessary. The way of clinical diagnosis mainly includes digital rectal examination, serum PSA, transrectal ultrasound, MRI spectroscopy imaging, and prostate biopsy, ect. Prostate biopsy is the most reliable diagnosis method and agold standardfor the diagnosis of PCa. The systematic use of the prostate biopsy has greatly improved the diagnosis of PCa. However, there are several prostate biopsy scheme and scholars proposed a variety of programs on the choice of puncture point. It has not yet formed a standard method at this stage. In this study, we will review the development of prostate biopsy and the status recent research to explore application value of different method.

OBJECTIVETo introduce the framework of evidence-based practice with a case of castration-resistant prostate cancer (CRPC) as an example.

METHODSA clinical question was formulated according the clinical scenario. A systematic search was conducted for the published literature in the databases of PubMed, EMBASE, Cochrane Library, Clinical Trial Registries, and Web of Knowledge up to Dec 2014. The identified literature was reviewed for quality appraisal before the evidence was applied to clinical practice.

RESULTSThe treatment was effective and the patient achieved disease remission.

CONCLUSIONEvidence-based practice should be integrated with clinical scenario, current evidence, and patients' willingness, and follow a systematic framework.

Seven guaiane-type sesquiterpenoids, a new compound 6-formyl-5-isopropyl-3-hydroxymethyl-7-methyl-1H-indene (1), a new natural product 5-isopropyl-3, 7-dimethyl-1H-indene-1-one (2), along with five known compounds: guaiazulene (3), 4-formyl-7-isopropyl-10-methylazulene (4), sesquiterpene ketolactone (5), alismoxide (6) and guaia-1 (5), 6-diene (7), were isolated from gorgonian Muriceides collaris collected in South China Sea. Their structures were elucidated on the basis of extensive spectroscopic analysis [MS, IR, 1H NMR, 13C NMR (DEPT), HMQC, HMBC, NOESY] and by comparison of the spectral data with those of the literatures.
Animals ; Anthozoa ; chemistry ; Azulenes ; China ; Sesquiterpenes ; chemistry ; isolation & purification ; Sesquiterpenes, Guaiane

Objective To study the clinicopathological and imaging features,key points of diagnosis and prognosis of endolymphatic sac tumor.Methods Seven cases of endolymphatic sac tumor were studied with routine histopathological,immunohisochemical staining and CT,MRI scanning.Patients' prognosis and life quality were aesessed by follow-up.Results The clinical symptoms of patients were progressive sensorineural deafness,tinnitus,vertigo and affection of cranial nerves.CT revealed reticular of moth-eaten bony destruction of the petrosal region,and MRI revealed a region of high-signal intensity void of vascular flowing signals.The histopathologic examination of the tumor presented a papillary-cystic pattern.The tumor cells were round or cubaid in shapen,with nuclei in similar shape and size.There were cyst-like structares filled mith pinkish gelatinous.Tumor cells were roundor cuboid in shape,and some of them contained clear cytofrlasm.There was materials,which were stained strongly with PAS.Hemorrhage,cholesterol crystal and foreign body giant cells were found in the tumor stroma with abundant small blood vessels and extravasation of blood,cells with intracellular hematin,and giant cells collagen.There was also proliferation of fibrous connective tissues.Necrosis and bone destruction were also found in some cases.Immunohistochemical results showed that CK and NSE were positive in all the tumors,and CD56 and S-100 were positive in part of cases,GFAP,EMA,CgA,TG and Syn were negative in all the specimens.One patient had received radiotherapy after partial resection.Relapse occurred only in one patient two years after the first surgery,and no metastasis or relapse occurred in the rest of patients.Conclusions Endolymphatic sac tumor is a rare neoplasm.The diagnosis relies mainly on histopathological and immunohisochemical findings.The patients often have good prognosis.

Objective:To determine the contents of paraceltamol and caffeine in New compouad Folium Isatidis Tablets simultaneously by HPLC. Methods:The determination was carried out with C 18 chemical bonded silica gel as a solid phase, methanol-water (25∶75) as a mobile phase and UV deterction wavelength at 215 nm. Results: The average recovenies of the added sample were 99.6%( RSD=0.67, n=5) for paracetamol and 99.3%(RSD=0.58, n=5) for caffine, There was a good linear relationship between the concentration and absorption area value in the rang of 1.6?g～6.4?g for paractamol or 0.16?g～0.64?g for caffeine.Conclusions: The method is simple, quick and accurate.

ObjectiveTo study the technique and therapeutic effect of percutaneous cannulate screw fixation to treat the traumatic diastasis of the pubic symphysis.MethodsFrom February 2003 to December 2010,46 patients (27 men and 19 women,mean age of 34.6 years) with unstable pelvic fractures and diastasis of the pubic symphysis were admitted.All the patients were treated by closed reduction and percutaneous cannulate screw fixation of the symphysis pubis.Then to complete the fixation of posterior ring,by percutaneous screw inserted through the sacroiliac joint or posterior iliac wing.According to Tile's classification,the patients were classified into type B1.1 in 4 cases,B1.2 in 7,B2 in 2,B3 in 2,C1.1 in 7,C1.2 in 7,C1.3 in 10,C2 in 5 and C3 in 2.All patients were performed posterior ring fixation except 4 cases of type B1.1.Anteroposterior,inlet and outlet X-ray radiographs and CT scans of the pelvis were taken preoperatively to evaluate the stability and deformities,and after surgery the plain radiographs and CT scans were taken to evaluate the reduction and the location of screws.ResultsThe average operative time was 45 min (range,15-65 min),and the average intraoperative blood loss was 25 ml (range,15-50 ml).Each patient was inserted one screw in the symphysis pubis.Thirty-five patients underwent postoperative pelvic CT scan and 3screws of them penetrated the wall of symphysis pubis.The average follow-up period was 23.5 months (range,5-48 months).In the follow-up period,no infection and displacement were found.The last follow-up result:according to Matta and Tornetta's standard,the result was excellent in 43 cases and good in 3.Among all patients,31 cases (67.39％) had returned to their original works,6 were to give up their original works as complicating injuries; 28 cases had no pain,13 cases had slight pain of the symphysis pubis while to make an effort,5 cases had pain of sacroiliac joint.There were no iatrogenic injury of nerve.ConclusionClosed reduction and percutaneous cannulate screw fixation of the traumatic diastasis of the pubic symphysis is a safe,feasible and easy manipulated method.The operative injuries is small and the clinical outcome is satisfactory.

Objective To investigate the methods of X-ray diagnosis of various displacement of unstable pelvic fracture in three-dimensional space and its instructive significance in closed reduction.Methods A normal adult pelvic specimen was selected and fixed in a wood-frame at supine position after soft tissue rejection and ligament preservation.With the breakage at the unilateral anterior-posterior ring,models of hemipelvic rotation in the transverse and sagittal planes and hemipelvic vertically upward displacement were induced.Anteroposterior radiographs of the pelvic specimen were made and picture archiving and communication system (PACS) was used to measure width of iliac wing,suprainferior diameter of hemipelvis,vertical displacement of iliac crest,acetabulum roof,pubic tubercle and sciatic tuber and area of obturator foramen.Methods of X-ray diagnosis of various displacements of hemipelvis were concluded and applied in treatment of 43 patients with unstable pelvic fractures.Operation time and intraoperative blood loss were recorded.Postoperative images were evaluated by Matta standard.Results Hemipelvic rotation in transverse plane included eversion and inversion.Width of iliac crest was enlarged and area of obturator foramen was shrunk while extroversion ; on the contrary,an opposite result was observed while inversion.Sagittal rotation included pronation and supination.Pubic tubercle had obvious downward shift,iliac crest presented no change or slight upward shift,suprainferior diameter of hemipelvis was lengthened,iliac roof and sciatic tuber remained their position unchanged and area of obturator foramen was shrunk while pronation; on the contrary,pubic tubercle had obvious upward shift,iliac crest presented no change or slight downward shift,suprainferior diameter of hemipelvis was shortened,iliac roof and sciatic tuber remained their position unchanged and area of obturator foramen was enlarged while supination; iliac crest,acetabular roof,pubic tubercle and sciatic tuber presented equidistant upward shift while the hemipelvis displaced upward vertically.Average operation time was 55 minutes (range,15-85 minutes) and intraoperative blood loss was 26 ml (range,10-50 ml).According to Matta standard,pelvic radiography evaluation at postoperative 3 days was excellent in 31 cases and good in 12 cases,with excellent and good rate of 100％.Conclusion Pelvis X-ray films are able to diagnose various three-dimensional displacement of unstable pelvic fractures and guide closed reduction timely and rapidly to achieve satisfactory result.

Biotechnology supremacy is a newly-advanced power theory. It is a superior dominance of military biotechnological application based on the microcosm of life structure within a certain period of time. The advancement of biotechnology supremacy and modern biotechnology has created the concept of bio-micro-frontier, which involves information and defense resources of all living ultra-micro-organisms with national and regional characteristics. Being feasible both in theory and practice, the implementation of bio-micro-frontier system is strategically important. This article explores the implementation of bio-micro-frontier in terms of strategy and tactics, which will add a unique dimension to future military transformation and active defense.