Objective To use the fusion image of the end-inhalation holding (EIH) phase and endexhalation holding (EEH) phase to define the target volume of individual patient with liver cancer,and to evaluate the target geometry,feasibility,and clinical significance of the technology.Methods Eighteen patients with liver caucer who were treated in our hospital from 2012 to 2013 were enrolled as subjects.With the same posture and scan range,all patients underwent contrast-enhanced three-dimensional computed tomography (3DCT) scans in the phases of free breathing (FB),EIH,and EEH.Gross tumor volume (GTV),clinical target volume (CTV),and organ of risk (OAR) were delineated on the above images.CTVFB was defined as GTV on the FB phase image (GTVFB) plus a margin of 10 mm,while planning target volume (PTVFB) was defined as CTVFB plus a margin of 10 mm in the right-left and anterior-posterior directions and a margin of 20 mm in the superior-inferior direction.GTVEI and GTVEE were defined as GTV on the EIH and EEH images,respectively.Based on the EEH images,the registered EEH and EIH images were fused to form GTVEI+EI.CTVEI+EE was defined as GTVEI+EI plus a margin of 10 mm,while PTVEI+EE was defined as CTVEI+EE plus a margin of 5 mm in the right-left and anterior-posterior directions and a margin of 10 mm in the superior-inferior direction.The Pinnacle3 v8.0m treatment planning system was used to design two 3D conformal radiotherapy plans for each patient.The volume,degree of inclusion (DI),matching index (MI),and central displacement of CTVFB and CTVEI+EE,as well as PTVFB and PTVEI+EE,were compared between the two plans.Results In the 18 patients,the mean CTVFB was significantly smaller than the mean CTVEI+EE(149.00±87.54 cm3 vs.188.17± 125.72 cm3,P=0.014);there was no significant difference between the mean PTVFB and PTVEI+EE (276.68± 146.41 cm3 vs.253.66± 117.35 cm3,P=0.080).DI of CTVFB to CTVEI+EF,PTVFB to PTVEI+EE,CTVEI+EE to CTVFB,and PTVEI+EE to PTVFB were (99.83±0.09)％,(84.55±8.45) ％,(80.83± 12.31) ％,and (99.78±0.08) ％,respectively.MI of CTVEI+EE to CTVFB and PTVEI+EE to PTVFB were 0.83± 0.07 and 0.87± 0.03,respectively.The central displacements of CTVEI+EE from CTVFB in x,y,and z axes were 0.55± 1.07 cm,0.76±3.02 cm,and-0.26± 1.98 cm,respectively (P =0.432,0.971,0.587).Conclusions In the treatment of liver cancer,the target volume delineation and image fusion using 3DCT images in EIH and EEH phases may avoid target omission due to respiratory movement,making it possible to increase radiation dose to target volume and improve the efficacy of radiotherapy.

Objective To investigate the tissue remodeling and cell alignment of TDBM scaffolds seeded with rabbit tenocytes under the cycle dynamic tensile force or static tension-free culture in vitro. Methods TDBM were made by ourselves, and uniaxial cyclic tendon stretching device was designed and manufactured on our own. Primary tenocytes were isolated from the Achilles tendon of three-day-old New Zealand white rabbits and seeded into scaffolds, and were cultured collectively in DMEM in vitro. Samples were divided into two groups:dynamic tension-loaded group, and static tension-free group. Fresh natural tendons were used to be positive control. The experiment's time was six weeks. The scaffold-cell complexes were harvested at 3 and 7 days of culture for Inverted microscope and scanning electron micrograph (SEM) analysis. The morphological characters of the samples, including the general view, HE and Masson's dyeing, were observed at 2, 4 and 6 weeks. In addition, the gene expression of the I-type collagen and III-type collagen of the samples was detected by using Real time PCR at every week. Set fresh natural tendon as control. Results The inverted microscope and SEM showed that it was nice compatible condition between the tendon cells and TD-BM scaffold. In addition, the tendon of tension-loaded group revealed a structure of longitudinally aligned collagen fi bers and dense structure of collagen fibers arranged in orderly form a unique corrugated structure. Tenocytes layer located between the col-lagen fibers and aligned longitudinally along the force axis, with increased matrix deposition after the 3th week showed by RT-PCR. The cell/matrix ratio decreases. When cultured to 6 weeks, the tissue structure was very similar to that of fresh natural ten-don pattern. By contrast, HE and Masson's staining revealed the collagen fibro-tissue structure in tension-free groups with disorga-nized matrix structure and randomly distributed cells. Collagen fibers were gradually degraded with time. In tension-loaded group, Real-time PCR showed that gene expression of I-type collagen and III-type collagen gradually increase, but in tension-free group, the relative gene expression of I-type collagen and III-type collagen was highest at 3rd week, and from that time the relative expres-sion gradually decrease. Conclusion Under the dynamic stress, the TDBM scaffolds seeded with rabbit tenocytes can promote extra-cellular matrix biosynthesis and tendon structure remodeling, which is a promising method for tendon tissue engineering.

Objective To investigate the effect of different injection rate of contrast medium on the quality of multislice spiral CT portography(SCTP). Methods Thirty patients with no prominent difference in the age and weight randomly divided into 3 groups (10 cases each group) underwent contrast-enhanced CT scan of abdomen. Non-ionic contrast medium (300 mgⅠ/ml, 1.5 ml/kg) was injected through antebrachium veins with power injector at the rates of 2.0 ml/s,3.0 ml/s and 4.0ml/s. Arterial phase acquisi-tion was made using the software of automated bolus triggering with a ROI placed on abdominal aorta when 100 HU was reached, and 7 seconds and 20 seconds later, portal phases were done respectively. The attenuations of portal vein(PV) and hepatic parenchy-ma of 3 groups were measured on source images. Then the oblique axial and coronal maximum intensity projection(MIP) maps were reconstructed at workstation and PVs were observed. Statistics analysis was made with software of SPSS 11.5. Results The attenu-ations of PV in 2.0ml/s, 3.0 ml/s and 4.0 ml/s groups were (150.80±21.16)HU, (170.90±17.26)HU and [181.90±22.88) HU respectively. There was obvious difference between 2.0ml/s and 4.0ml/s groups for atteuation of PV(P=0.017). The differences of CT attenuation of PV-hepatic parenchyma in three groups were (50.20±17.40) HU, (67.10±23.08) HU and (76.20±22.75) HU respectively. Prominent difference was also found between 2.0 ml/s and 4.0 ml/s groups(P=0.039). The grades of segment of PV displayed on SCTP maps were 4.20±1.14,4.90±0.99 and 5.50±0.53 in 2.0 ml/s,3.0 ml/s and 4.0 ml/s groups respectively, which were of obvious differences between them (P=0.013). Conclusion Injection rate of contrast medium dose influence the quality of SCTP of PV,SCTP quality is best with velocity of flow at 4.0 ml/s.

Objective To investigate the expression of adamalysin-12 (ADAM-12) and PCNA in human bladder carcinoma and to investigate their correlation with different grades and stages of bladder cancer.Methods Biopsies of 15 normal bladder and 43 bladder tumors were analyzed.Immunohistochemistry was conducted to detect the expression of ADAM12 and PCNA in the biopsies.Results Postive expression signals of ADAM12 were detected significantly higher in the bladder cancer biopsies than that in the normal ones (P = 0.010).Those with lower histological grade had a higher expression level of ADAM-12 compared to the higher histological grades (P ＜0.001 ).Positive expression signals of PCNA were detected significantly higher in the bladder cancer biopsies than that in the normal ones (P = 0.026).Those with lower histological grade had a higher expression level of PCNA (P =0.014).There was a positive correlation between the expression of ADAM-12 and PCNA in bladder cancer (r =0.997,P ＜ 0.001 ).Conclusions The overexpression of ADAM-12 and PCNA in the biopsies of bladder tumors shows that protein expression of ADAM-12 and PCNA correlated with tumor stage and grade.Furthermore,ADAM-12 may be a promising biomarker of bladder cancer in the clinical implication.

Kidney injury and decreased chemosensitivity of tumor cells are obstacles with cisplatin (CDDP) chemotherapy. Down-regulation of the organic cation transporter 2 (OCT2) and multidrug resistance-associated protein 2 (MRP2) is a key means to alleviate CDDP-induced kidney injury and increase chemosensitivity. Astragaloside IV (AS IV) is obtained from the well-known traditional Chinese herb Astragalus membranaceus. This study explored the role of AS IV in preventing kidney injury and enhancing the antitumor effect of CDDP by suppressing OCT2 expression in kidney and MRP2 in tumors. This project was reviewed and approved by the Animal Ethics Committee of the First Hospital of Jilin University. The effects of AS IV on CDDP inhibition of tumor growth and promotion of apoptosis were assessed in Lewis lung tumor (LLC)-bearing mice by H&E and TUNEL staining. Kidney injury was assessed by serum biochemical parameters and H&E staining. We used Western blotting and immunohistochemistry assays to detect OCT2 and MRP2 expression in kidney and tumor. The concentration of CDDP in kidney and tumor was measured by HPLC-MS/MS. AS IV enhanced CDDP chemosensitivity by increasing tumor cell apoptosis and slowing tumor growth, and decreased kidney injury as evidenced by lower blood creatinine (Cr) and blood urea nitrogen (BUN). Co-administration of AS IV suppressed MRP2 overexpression induced by CDDP in tumor tissues and may be an important mechanism for enhancing CDDP chemosensitivity. Moreover, AS IV reduced CDDP-induced kidney injury in mice along with suppression of OCT2 expression in kidney. The concentration of CDDP was increased in tumor but decreased in kidney. In total, AS IV not only enhanced the antitumor effect of CDDP by suppressing MRP2 expression in tumor cells, but also decreased kidney injury induced by CDDP. The results provide new insight into the combined use of a chemotherapy drug and natural ingredients to treat cancer.

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P<0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.

OBJECTIVETo investigate the induction of antitumor immune responses and therapeutic effects of 10-hydroxycamptothecinc-treated (HCPT) DC-Hepa fusion vaccines by DC fused with hepal-6 cell from hepatoma.

METHODSThe fused cells were isolated by magnetic cell sorting and adherent culture. Cell apoptosis was detected by Rhodamine123/PI double-labeled assay, CTL activity by 4 h (51)Cr releasing assay. Protective and therapeutic effects of the fusion vaccine to the tumor-bearing mice was also observed.

RESULTSThe apoptosis rate was 29.7%+/-4.1% when DC-Hepa fusion vaccine was treated with 50 microg/ml HCPT for 24 h. After treatment with the HCPT-DC-Hepa fusion vaccine, the tumor grew obviously slowly, survival period of the mice was prolonged, induced more potent CTL cytotoxicity, and resisted against the rechallenge of Hepal-6 cells.

CONCLUSIONVaccination with HCPT-DC-Hepa fusion vaccine could elicit potent antitumor responses, which will provide a new approach to the DC-mediated therapeutic antitumor immunity.

Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Camptothecin ; analogs & derivatives ; pharmacology ; Cancer Vaccines ; administration & dosage ; genetics ; immunology ; Cell Fusion ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; transplantation ; Female ; Liver Neoplasms ; immunology ; pathology ; Mice ; Mice, Inbred C57BL ; Neoplasm Transplantation ; Rats ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Cells, Cultured

OBJECTIVETo investigate the value of intensity modulated radiation therapy (IMRT) for patient with gynecological malignancies after treatment of hysterectomy and chemotherapy/radiotherapy.

METHODSAll 32 patients with cervical or endometrial cancer after hysterectomy received full course IMRT after 1 to 3 cycles of chemotherapy (Karnofsky performance status(KPS) > or =70). Seventeen of these patients underwent postoperative preventive irradiation and the other 15 patients were pelvic wall recurrence and/or retroperitoneal lymph node metastasis, though postoperative radiotherapy and/or chemotherapy had been given after operation.

RESULTSThe median dose delivered to the PTV was 56.8 Gy for preventive irradiation, and 60.6 Gy for pelvic wall recurrence or retroperioneal lymph node metastasis irradiation. It was required that 90% of iso-dose curve could covere more than 99% of GTV. However, The mean dose irradiated to small intestine, bladder, rectum, kidney and spinal cord was 21.3 Gy, 37.8 Gy, 35.3 Gy, 8.5 Gy, 22.1 Gy, respectively. Fourteen patients presented grade I (11 patients) or II (3 patients) digestive tract side-effects, Five patients developed grade I or II bone marrow depression. Twelve patients had grade I skin reaction. The overall 1-year survival rate was 100%. The 2- and 3- year survival rate for preventive irradiation were both 100%, but which was 5/7 and 3/6 for the patients with pelvic wall recurrence or retroperioneal lymph node metastasis.

CONCLUSIONIntensity modulated radiation therapy can provide a better dose distribution than traditional radiotherapy for both prevention and pelvic wall recurrence or retroperioneal lymph node metastasis. The toxicity is tolerable. The adjacent organs at risk can well be protected.

Adult ; Aged ; Combined Modality Therapy ; Diarrhea ; etiology ; Endometrial Neoplasms ; drug therapy ; radiotherapy ; surgery ; Female ; Follow-Up Studies ; Humans ; Hysterectomy ; methods ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Recurrence, Local ; Radiotherapy Dosage ; Radiotherapy, Intensity-Modulated ; adverse effects ; methods ; Survival Analysis ; Uterine Cervical Neoplasms ; drug therapy ; radiotherapy ; surgery

OBJECTIVETo establish a set of procedure for recovery and species identification of Legionella from the surface environmental water.

METHODSForty-four water samples were collected in eight parks of Guangzhou city from August to November in 2006. The bacteriologic examination was performed by cultivation on BCYEalpha plate, and 108 presumptive Legionella colonies were picked and their homogeneous relationship was analyzed by using an amplified fragment length polymorphism (AFLP) method. Species identification was carried out by latex agglutination test, biochemical characterization, analysis of cellular fatty acids composition, 16 S rRNA gene and mip gene sequencing.

RESULTSLegionella was recovered among 27 (61.36%) samples of all eight parks, and 31 different strains were identified from those 108 presumptive Legionella isolates by AFLP method, including 20 strains of L. pneumophila, five strains of L. feeleii, four strains of L. longbeachae, one strain of L. oakridgensis and one strain of L. sainthelensi, and L. pneumophila could be easily differentiated by phenotypic and biochemical characteristics, latex agglutination test or analysis of the cellular fatty acids composition . However, uncertain factors were existing in those phenotypic identification methods as compared to the sequence analysis.

CONCLUSIONThe taxonomic analysis of the Legionellae family should be dependent on the 16 S rRNA gene or mip gene.

Bacteriological Techniques ; DNA, Bacterial ; genetics ; Environmental Monitoring ; methods ; Legionella ; genetics ; isolation & purification ; RNA, Bacterial ; RNA, Ribosomal, 16S ; Water Microbiology

OBJECTIVETo determine the antibiotics resistance, aminoglycoside-modifying enzymes and homology of high-level gentamycin resistant enterococcus in clinical specimens.

METHODSThe high-level gentamicin resistant (HLGR) isolates were screened by the agar method and the resistance of 14 antimicrobial agents was determined by K-B method. The aminoglycoside-modifying enzyme genes were detected by polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) was used to analyze the homology of HLGR isolates.

RESULTSThe ratio of HLGR was 64.2% (68/106). Among the HLGR,there were no isolates resistant to linezolid, vancomycin and tecoplanin, and Enterococcus faecium was more resistant to beta-lactam antibiotics and quinolone than Enterococcus faecalis. The positive rate of aac(6')-Ie-aph(2')-Ia was 92.6% and 3 isolates had the resistance gene mostly similar to aph(2')-Id. And among 51 HLGR isolates from the hospitalized patients, PFGE grouped 17 E. faecalis isolates into 4 clusters (A-D), and 33 E. faecium isolates into 8 clusters (A-H) with A cluster as predominant.

CONCLUSIONHLGR has become the important antibiotic resistance bacteria which results in nosocomial infection; and aac(6')-Ie-aph(2')-Ia is the main aminoglycoside-modifying enzyme gene which causes HLGR.

Drug Resistance, Bacterial ; genetics ; Electrophoresis, Gel, Pulsed-Field ; Enterococcus ; drug effects ; genetics ; Enterococcus faecalis ; drug effects ; genetics ; Enterococcus faecium ; drug effects ; genetics ; Gentamicins ; pharmacology ; Humans ; Kanamycin Kinase ; genetics ; Microbial Sensitivity Tests