Objective To establish a radioresistant cell subline from a human A549 lung cancer cell line and investigate the mechanism of radioresistance. Methods Two proposals were applied for the non-small cell lung cancer A549 cells irradiated with X-rays:A group of A549 cell line was irradiated five times, the fractionated dose was 600 cGy, and the other group was exposed 15 times, the fractionated dose was 200 cGy. After the completion of irradiation, two monoclones were obtained from the survival of cells and named the subline A549-S1 and A549-S2. The radiosensitivity and cell cycle distribution of these two clones,together with its parental A549 cells were measured by clone formation assay and flow cytometry.The mRNA and protein levels of Notch1 in A549 cell line and the sublines were determined by RT-PCR and Western-blots. Results Compared with the parental A549 cells, A549-S1 cells showed significant resistance to radiation with D0, Dq and N values increased, and a broader initial shoulder as well as 1.38-fold increased value of SF2. The A549-S1 subline also showed higher percentage of cells in S phase and G2/M phase, but lower percentages in G0/G1 phase (P<0.05). The expression of Notch1 in A549-S1 was enhanced obviously than in A549 cells. But for A549-S2 the radioseasitivity was slightly increased compared with the parental cells with D0, Dq and N values decreased and a curve initial shoulder. The ratio of cells in S and G0/G1 phase ratio was lower than that in parental A549 cells, but that in G2/M phase ratio was higher significantly (P<0.05) .The expression of Notch1 had no marked change compared to A549 cell. Conclusions The radioresistance of the A549 cell subline is correlated with the irradiation program. The cell subline shows a different cell cycle distribution from their parental line. The cell cycle distribution has a close correlation with the expression of Notch1.

Body surface mapping is introduced in this paper. The research of body surface mapping involves two main aspects: data acquiring system and data processing methods. The recommended design utilizes the virtual instruments concept and high-speed open bus standard. The data processing methods involve qualitative and quantitative methods. Dynamic and static mapping techniques are introduced here as the qualitative methods while statistical methods, inverse calculation and principle component analysis are introduced as the quantitative methods.
Algorithms ; Automatic Data Processing ; methods ; Body Surface Potential Mapping ; methods ; Electrodes ; Myocardial Infarction ; diagnosis ; Signal Processing, Computer-Assisted

This article was aimed to study the optimum extraction process of paeonol. The extraction yield of paeonol was taken as investigation index. And the best extraction process was screened by orthogonal experimental design. The results showed that the optimum condition of extraction process was to soak coarse powder of Paeonia suffruti-cosa into 15-fold water for 0.5 h, and then the distillation lasted for 2.5 h. The distillate was collected and cooled to room temperature. The crystallization lasted for 24 h at 4℃, and then filtered and dried for 48 h at room tempera-ture. It was concluded that the selected technology was stable, reasonable and feasible. The extraction yield of paeonol is over 80%.

Objective To conduct morphological observation and gene identification of the strain of flagellate iso -lated from Cricetulus migratorius in the Xinjiang Research Center for Experimental Animals .Methods The ileocecal con-tents of C.migratorius were microscopically examined on direct smear with Wright-Giemsa staining , and the total RNA iso-lated from Xinjiang C.migratorius was extracted and 16S rRNA was amplified by PCR , and then sequenced .Furthermore the homology was compared and the phylogenetic tree was developed using MEGA 5.22 software.Results Morphological observation indicated that the isolated flagellate was Tritrichomonas muris.The 16S rRNA gene sequence of the Xinjiang C. migratorius isolate shared highly homology with that of other Tritrichomonas.Phylogenetic tree analysis indicated that the 16S rRNA gene of Xinjiang C.migratorius isolate was classified into a subgroup with T.muris 16S rRNA (U85966.1), but was relatively distant relative from other related tritrichomonas.Conclusions The flagellate isolated from Xinjiang C. migratorius is identified to be T.muris by both morphological observation and 16S rRNA gene analysis.

OBJECTIVE The objective of this study is to discuss when to dissect the lymph nodes behind the right recurrent laryngeal nerve (LN-prRLN) from the standpoint of the right cervical level Ⅵ-1 (superficial layer to the recurrent laryngeal nerve) lymph nodes in papillary thyroid carcinoma (PTC) patients.METHODS The clinical data of 306 bilateral or right PTC patients from the Hangzhou First People's Hospital who underwent dissection of level Ⅵ-1 lymph nodes and LN-prRLN between March 2014 and September 2015 were analyzed. We measured the number of level Ⅵ-1 metastatic lymph nodes and size of level Ⅵ-1 lymph nodes metastasis loci to predict the metastasis of LN-prRLN.RESULTS The number of level Ⅵ-1 metastatic lymph nodes and size of level Ⅵ-1 lymph nodes metastasis loci were risk factors of LN-prRLN metastasis(P<0.05). When the number of the level Ⅵ-1 metastatic lymph nodes was greater than 1.5, the AUC was 0.813 (the sensitivity was 78.43%, the specificity was 76.65%). The ROC showed that when the size of level Ⅵ-1 lymph nodes metastasis loci were more than 0.45 cm, the AUC was 0.726 (sensitivity was 90.20%, specificity was 48.90%).CONCLUSION In bilateral or right PTC patients with metastasis of level Ⅵ-1 lymph nodes, especially when the number of level Ⅵ-1 metastatic lymph nodes was greater than 2cm and the metastasis loci were more than 0.45 cm, we should dissect the LN-prRLN.

Objective To investigate the inhibitory effects of norcantharidin (NCTD),paclitaxel (PTX) and their combination on androgenic independent prostate cancer cells.Methods The experiment was divided into 3 groups:NCTD group,PTX group and combination group (NCTD + PTX).NCTD group and PTX group:In the exponential growth of the prostate cancer cell line DU145,different concentrations (12.5,25.0,50.0,100.0,200.0,400.0,800.0 μmol · L-1)of NCTD and PTX were added.Combination group:NCTD and PTX according to the ratio of 1:1 concentration,repeat first group operation.The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution 20 μL was added after 48 h.The culture supematant liquid was extracted after train 4 h,DMSO 200 μL was added in order to dissolve.The recovery rate of DU145 and the inhibition rate of cell growth were measured by centrifugation.half inhibitory concentration was determined by MTT.Whether the two drugs have synergistic effect was investigated by simulating the joint index.Results The inhibition rate of the above 7 concentration of NCTD group were 0.12％,0.14％,0.25％,0.30％,0.58％,0.70％,0.74％.That of the above 7 concentration of NCTD group were 0.12％,0.14％,0.25％,0.30％,0.58％,0.70％,0.74％.The inhibition rate of combined group was higher than that of NCTD group and PTX group.The difference was statistically significant (all P ＜ 0.05).With the increase of the dosage of two drugs,the simulated joint index (CI) value was higher and lower than 1.The lowest of CI was 0.07 ± 0.01,with a maximum of 0.59 ± 0.20.Compared with the lowest concentration,the difference was statistically significant (P ＜ 0.05).The results showed that when NCTD and PTX were combined,there was a synergistic effect between the two groups.Conclusion Both NCTD and PTX have inhibitory effects on androgenic independent prostate cancer cells.The higher of the drug contents,the stronger of the inhibitory effect.The combined use of NCTD and PTX has stronger inhibitory effect on cancer cells.

Objective To investigate the protective effects of polydatin on performance of sepsis injury of renal glomerular vascular endothelial and the activity of adenosine monophosphate (AMP)-activated protein kinase(AMPK) and mammalian target protein (rmTOR)in the process.Methods The sepsis model was established by injection of lipopolysaccharide through the tail vein of rats.The rats were divided into four groups:sham operated group (implementation of anesthesia,intubation),model group (anesthesia,intubation and sepsis),control group (anesthesia,intubation and application of saline resuscitation),experimental group (anesthesia,intubation and sepsis,saline combined with tiger polydatin 10 kU · kg-1).The activity of AMPK and rmTOR in each groups were detected by ELISA.Results After administration for 6 h,the heart rate (beat per minute) in experimental group,control group,model group were 88.21 ± 17,102.36 ± 12.83,118.04 ± 16.51.The urea nitrogen(BUN) in the three groups were (49.77 ±9.67),(77.92 ± 12.01),(122.19 ± 12.87) mg · dL-1.Compared with the control and model groups,all of them in experimental group were statistically significant (all P ＜ 0.05).The protein activeties of AMPK and mTOR in experimental group,model group were respectively 3.06 ± 1.34,0.77 ± 0.21;2.48 ± 1.12,3.95 ± 1.02.Compared with model groups,all of them in experimental group were statistically significant (all P ＜ 0.05).Conclusion Polydatin had protective effect on septic renal damage glomerular vascular endothelial,which might be correlated with its inhibition on mTOR activity through AMPK pathway.

OBJECTIVETo investigate the clinical application value of 8.5/11.5 F transurethral seminal vesiculoscopy in the diagnosis and treatment of refractory hematospermia.

METHODSWe retrospectively analyzed 78 cases of refractory hematospermia diagnosed and treated by 8.5/11.5 F transurethral seminal vesiculoscopy from June 2012 to June 2014. The patients underwent serum PSA examination, transrectal ultrasonography, seminal vesicle ultrasonography, and pelvis CT or MRI before surgery, and all received transurethral seminal vesiculoscopy under the 8.5/11.5 F rigid ureteroscope.

RESULTSOperations were all successfully accomplished, which revealed abnormal opening of the ejaculatory duct in 5 cases, mucosal inflammatory hyperemia in the prostatic utricle and seminal vesicle in 78, dark red mucilage substance in the seminal vesicle in 34, seminal vesicle stones in 19, small polyp in the seminal vesicle in 2, and ejaculatory duct or seminal vesicle cyst in 4. All the patients received symptomatic treatment during the surgery. After surgery, hematouria was found in 13 cases, which disappeared within 2 weeks, pelvic hematoma in 1 case, which was cured by conservative treatment within 3 months, and epididymitis in 2 cases, which was controlled by anti-infection treatment. Hematospermia recurred in 3 cases during the 1-year postoperative follow-up.

CONCLUSION8.5/11.5 F transurethral seminal vesiculoscopy, with its advantages of easy operation, wide field of vision, large channel for operation, and few complications, deserves general clinical application in the diagnosis and treatment of refractory hematospermia.

Calculi ; Ejaculatory Ducts ; Endoscopy ; methods ; Epididymitis ; etiology ; Hemospermia ; diagnosis ; therapy ; Humans ; Magnetic Resonance Imaging ; Male ; Postoperative Period ; Recurrence ; Retrospective Studies ; Seminal Vesicles ; Tomography, X-Ray Computed ; Urethra

OBJECTIVETo study the feasibility of regenerating a whole menisci using poly-(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) scaffolds loaded with meniscal cells in rabbits undergoing total meniscectomy, and to explore its protective effect on cartilage degeneration.

METHODSA solvent casting and particulate leaching technique was employed to fabricate biodegradable PHBV scaffolds into a meniscal shape. The proliferated meniscal cells were seeded onto the polymer scaffolds, transplanted into rabbit knee joints whose lateral menisci had been removed. Eight to 18 weeks after transplantation, the rege- nerated neomenisci were evaluated by gross and histological observations. Cartilage degeneration was assessed by Mankin score.

RESULTSEighteen weeks after transplantation, the implants formed neomenisci. Hematoxylin and eosin (HE) staining of the neomenisci sections revealed regeneration of fibrocartilage. Type I collagen in the neomenisci was also proved similar to normal meniscal tissue by immunohistochemical analysis and Sirius scarlet trinitrophenol staining. Articular cartilage degeneration was observed 8 weeks after implantation. It was less severe as compared with that in total meniscectomy controls and no further degeneration was observed at 18 weeks. At that time, the regenerated neomenisci strongly resembled normal meniscal fibrocartilage in gross and histological appearance, and its mechanical property was also close to that of normal meniscus.

CONCLUSIONSThe present study demonstrates the feasibility of tissue-engineering a whole meniscal structure in total meniscectomy rabbit models using biodegradable PHBV scaffolds together with cultured allogeneic meniscal cells. Cartilage degeneration is decreased. But long-term in vivo investigations on the histological structure and cartilage degeneration of the neomenisci regenerated by this method are still necessary to determine the clinical potential of this tissue engineering avenue.

Animals ; Cartilage, Articular ; Cells, Cultured ; Knee Joint ; Menisci, Tibial ; Polymers ; Regeneration ; Tissue Engineering

OBJECTIVETo investigate the effects of estrogen receptor alpha (ERa) and insulin-like growth factor 1 (IGF1) on the proliferation of prostatic smooth muscle cells (PSMCs) in vitro.

METHODSThe ERalpha shRNA expression frame was subcloned to the pGSadeno adenovirus vector by homologous recombination technology to construct the pGSaaeno-ERalpha vector. After the mouse PSMCs were transfected in vitro by pGSaaeno-ERalpha, the mRNA and protein expression levels of ERalpha were detected by RT-PCR and Western blot respectively. The expression of IGF1 in the ERa-reduced cells was determined by Western blot 6 hours after treatment with 17beta-estradiol (E2) at 10(-8) mol/L. The post-transfection activity of estrogen or exogenous IGF1 in the proliferation of PSMCs was evaluated by MTT chlormetric analysis.

RESULTSAfter treatment with E2, the proliferation of PSMCs and the expression of the IGF1 gene were significantly increased in the normal control group (P <0.05), but not obviously changed in the ERalpha-siRNA group (P> 0.05). And exogenous IGF1 failed to induce the proliferation of the ERalpha-reduced PSMCs.

CONCLUSIONE2 induces the expression of IGF1 via ERalpha, and IGFl, with the interaction of ERalpha, promotes the proliferation of PSMCs.

Animals ; Cell Proliferation ; Cells, Cultured ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mice ; Myocytes, Smooth Muscle ; cytology ; Prostate ; cytology ; RNA, Messenger ; genetics