Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.
Alkaloids ; biosynthesis ; genetics ; metabolism ; Atropa belladonna ; genetics ; metabolism ; Gene Expression Regulation, Plant ; genetics ; Hyoscyamine ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Tropanes ; metabolism

OBJECTIVETo investigate magnetic resonance imaging(MRI) manifestations of perianal abscess and to evaluate MRI diagnosis for perianal abscess.

METHODSFifty cases of perianal abscess between July 2007 and March 2009 were included in this study. MRI was performed using T1 weighted sequence in axial plane, T2 weighted sequence with fat saturation in axial, coronal and/or sagittal plane, and T1 weighted enhanced sequence with fat saturation in axial, coronal and/or sagittal plane. The location, size, shape, signal characteristics, and enhancement patterns of perianal abscess were analyzed.

RESULTSFifty cases with 51 perianal abscesses were identified. The abscess appeared slight hypo- or isointense signal on T1 weighted imaging, obviously hyperintense signal at cavity and isointense signal at wall on T2 weighted with fat saturation imaging, and marked enhancement at wall. The largest diameters ranged between 0.5 cm and 9.0 cm (mean 3.4±1.7 cm ) in abscess, between 0.2 cm and 8.0 cm(mean 2.7±1.7 cm) in cavity. The abscess was round, oblong and crescent in 23, 18 and 7 cases respectively, and was unilocular in 41 cases and multilocular in 10 cases. The abscess located between levator ani and intersphincteric groove in 23 cases, under intersphincteric groove in 3 cases, across levator ani and intersphincteric groove in one case, on levator ani in 7 cases, on intersphincteric groove in 16 cases and above levator ani in one case.

CONCLUSIONMRI is a noninvasive, convenient and highly accurate procedure for diagnosing perianal abscess, which can demonstrate the anatomical relationship between abscess and anal canal.

Abscess ; diagnosis ; pathology ; Adolescent ; Adult ; Aged ; Anus Diseases ; diagnosis ; pathology ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Vascular stent is an important medical appliance for angiocardiopathy. Its key deformation process is the expandable progress of stent in the vessel. The important deformation behaviour corresponds to two mechanics targets: deformation and stress. This paper is devoted to the research and development of vascular stent with proprietary intellectual property rights. The design of NiTinol self-expandable stent is optimized by means of finite element software. ANSYS is used to build the finite element simulation model of vascular stent; the molding material is NiTinol shape memory alloy. To cope with the factors that affect the structure of stent, the shape of grid and so on, the self-expanding process of Nitinol stent is simulated through computer. By making a comparison between two kinds of stents with similar grid structure, we present a new concept of "Optimized Grid" of stent.
Alloys ; chemistry ; Angioplasty, Balloon, Coronary ; instrumentation ; Computer Simulation ; Computer-Aided Design ; Humans ; Models, Biological ; Numerical Analysis, Computer-Assisted ; Prosthesis Design ; Stents

OBJECTIVETo study the disease spectrum and causes of death in hospitalized children in an upper first-class hospital in Hunan Province, China.

METHODSThe medical records of hospitalized children between 2010 and 2014 in this hospital were collected and analyzed retrospectively.

RESULTSFrom 2010 to 2014, the number of hospitalized children increased from 7 303 in 2010 to 10 902 in 2014, and the case fatality rate declined from 0.33% to 0.20% (P<0.05). The case fatality rate was highest (0.41%) in infants and lowest (0.11%) in newborns. As for the disease spectrum for these hospitalized children, in 2010, the top three diseases were leukemia, congenital heart disease, and pneumonia; in 2011, the top three diseases were tumor chemotherapy, congenital heart disease, and pneumonia; since 2012, tumor chemotherapy, epilepsy, and pneumonia had remained the top three diseases. The top three causes of death in hospitalized children were congenital malformation (39%, 39/99), tumor (13%, 13/99), and infectious diseases/parasitic diseases (8%, 8/99).

CONCLUSIONSThe number of hospitalized children is increasing, while the case fatality rate tends to decrease in this hospital. Tumor, epilepsy, pneumonia, and congenital heart disease are major diseases in hospitalized children, and congenital malformation is the primary cause of death.

Cause of Death ; Child ; Child, Hospitalized ; China ; Female ; Humans ; Male ; Time Factors

OBJECTIVETo detect the possible relationship between PARKIN gene and the Chinese pedigree with autosomal recessive early-onset Parkinson's disease(AREP).

METHODSClinical examination was carried out in 6 patients from 3 Chinese pedigrees with AREP and their 23 family members. PCR amplification of all exons of PARKIN gene was performed. The PCR products were analyzed by denaturing high-performance liquid chromatography(DHPLC) to screen for point mutation and polymorphism. And in the samples with abnormal DHPLC result, further sequencing was conducted to confirm the type of mutation and polymorphism.

RESULTSAll exons of PARKIN gene from the research subjects were successfully amplified. A heterozygous point mutation (Gly284Arg) in exon 7 was found in one pedigree. A polymorphism (Ser167Asn) in exon 4 was found in another pedigree. All the patients had the past history of exposure to environmental poison.

CONCLUSIONWhen acting together with risky environmental factors, the heterozygous mutation Gly284Arg in PARKIN gene may cause AREP. The polymorphism Ser167Asn in PARKIN gene increases the risk of developing Parkinson's disease and may cause AREP when acting together with hydrargyrism.

Age of Onset ; Aged ; China ; epidemiology ; Chromatography, High Pressure Liquid ; Exons ; genetics ; Female ; Genes, Recessive ; Humans ; Male ; Middle Aged ; Parkinson Disease ; epidemiology ; genetics ; Pedigree ; Point Mutation ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Ubiquitin-Protein Ligases ; genetics

Osteosarcoma is the most common primary malignant bone cancer in children and adolescents. Emerging evidence has suggested that the capability of a tumor to grow is driven by a small subset of cells within a tumor, termed cancer stem cells (CSCs). Although several methods have been explored to identify or enrich CSCs in osteosarcoma, these methods sometimes seem impractical, and chemotherapy enrichment for CSCs in osteosarcoma is rarely investigated. In the present study, we found that short exposure to chemotherapy could change the morphology of osteosarcoma cells and increase sarcosphere formation in vitro, as well as increase tumor formation in vivo. Furthermore, methotrexate (MTX)-resistant U2OS/MTX300 osteosarcoma cells were larger in size and grew much more tightly than parental U2OS cells. More importantly, U2OS/MTX300 cells possessed a higher potential to generate sarcospheres in serum-free conditions compared to parental U2OS cells. Also, U2OS/MTX300 cells exhibited the side population (SP) phenotype and expressed CSC surface markers CD117 and Stro-1. Notably, U2OS/MTX300 cells showed a substantially higher tumorigenicity in nude mice relative to U2OS cells. Therefore, we conclude that chemotherapy enrichment is a feasible and practical way to enrich osteosarcoma stem cells.
Animals ; Antigens, Surface ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Neoplastic Stem Cells ; drug effects ; pathology ; Osteosarcoma ; metabolism ; pathology ; Phenotype ; Proto-Oncogene Proteins c-kit ; metabolism

OBJECTIVETo study the growth inhibition and apoptosis induction effects of bufalin on human osteosarcoma cell lines in vitro.

METHODSU-2OS and U-2OS/methotrexate (MTX) 300-resistant cell lines were treated with bufalin. Cell viability was assessed by MTT assay. Cell-cycle status, apoptosis-inducing effects, and the expression of apoptosis-related proteins were evaluated by flow cytometry, fluorescent staining, DNA fragmentation assay, and Western blotting.

RESULTSBufalin inhibited cell growth in both U-2OS and U-2OS/MTX300 cells. The IC(50) values of bufalin for U-2OS and U-2OS/MTX300 cells were (8.49 ± 2.1) ng/ml and (10.19 ± 1.7) ng/ml, respectively. The induction of G(2)/M cell-cycle arrest was also seen in the bufalin-treated cells. The bufalin-induced apoptosis was confirmed by increased expression of tumor suppressor protein p53, bax and decreased expression of bcl-2.

CONCLUSIONBufalin inhibits the growth of and induces apoptosis in both MTX-sensitive and MTX-resistant human osteosarcoma U-2OS cell lines. The apoptosis-inducing effect of bufalin is not influenced by the presence of high levels of DHFR.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Bone Neoplasms ; metabolism ; pathology ; Bufanolides ; pharmacology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; Humans ; Methotrexate ; pharmacology ; Osteosarcoma ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tetrahydrofolate Dehydrogenase ; metabolism ; Tumor Suppressor Protein p53 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVETo evaluate the application of multi-detector row CT (MDCT) and CT angiography (CTA) for detecting early signs of acute bowel ischemia (ABI) in experimental porcine models.

METHODSTwelve pigs were assigned to four groups with 3 in each group. The digital subtraction angiography of superior mesenteric artery (SMA) and the embolization of branches of SMA with gelatin sponge and blood clot were performed by percutaneous transfemoral artery puncture and catheterization. MDCT pre- and post-contrast scanning in the arterial, venous and delay phase and CTA with three-dimensional reconstruction were carried out at pre-operation, 3 h, 6 h, 9 h, and 12 h after occlusion. The normal mesenteric vascular anatomy, arterial occlusion, mesentery and bowel changes, and dynamic change were evaluated.

RESULTSABI changes were identified pathologically in all the 12 experimental pigs, and the severity of ischemia increased over time after embolization. CTA showed all 57 embolized branches of SMA and 29 of 34 unoccluded arterial branches with 5 false-positive vessel occlusions. The sensitivity and specificity of CTA were 100% and 85.3%, respectively. Thin-slab maximum intensity projection (TSMIP) revealed the disappearance of distal comb-like vessel branches and brush-like vasa recta, which were clearly delineated in the normal bowel segments. Using this criterion, TSMIP correctly defined 23 of 24 ischemic bowel segments and all the 12 normal bowel segments with a sensitivity of 95.8% and a specificity of 100%.

CONCLUSIONSMDCT and CTA reliably define normal and occluded mesenteric vessels in the pig. It can easily detect ischemic bowel segment by identified early changes of ischemia. The early direct ischemic signs are occluded vessels, the disappearance of distal comb-like branches or brush-like vasa recta, and poor bowel enhancement. The early indirect sign is bowel dilatation with fluid collection.

Angiography ; methods ; Animals ; Female ; Intestinal Diseases ; diagnostic imaging ; etiology ; Ischemia ; diagnostic imaging ; etiology ; Mesenteric Arteries ; diagnostic imaging ; Mesenteric Vascular Occlusion ; complications ; diagnostic imaging ; Mesentery ; blood supply ; Swine ; Tomography, X-Ray Computed ; methods

OBJECTIVEThis study was undertaken to observe the change in the local level of angiotensin II (Ang II) and the expression of its corresponding receptors AT1 and AT2 during wound healing, and explore the possible role of Ang II in wound healing .

METHODSA model of full-thickness cutaneous wound was developed on the back of C57/BL6 mice. Specimens were taken from the wound of each mouse on the day 0, 1, 3, 5, 7, 9, 11, 13 and 15 after wounding. The change in the generation of Ang II in wounded tissue during the healing process was detected with ELISA. The proliferation and the apoptosis of cells were detected by bromodeoxyuridine (Brdu) and terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method in wounded skin during the healing process, respectively. The cellular localization and the mRNA level change of Ang II receptors in wounded tissue during healing were detected with immunostaining and RT-PCR.

RESULTSAng II produced in wounded skin was increased in the first 7 days to reach the peak, and then gradually decreased during wound healing. BrdU labeling index was increased gradually in the first 7 days to reach the peak, and then gradually decreased during wound healing. The number of TUNEL-positive cells was increased slowly in the first 7 days after wounding. The increase in the number of TUNEL-positive cells was more markedly after epithelization of the wound. In normal mice, AT1 and AT2 receptor were found positively expressed in the whole epidermal layer, while positive expression was only found in the endothelial cells of the capillary vessels within the dermal layer, and positive expression was also found in appendages of the skin, i. e. hair follicle, sweat gland and sebaceous gland respectively. Positive staining signal of both AT1 and AT2 receptors were increased in the first 7 days to reach the peak, then gradually decreased. Expression of AT2R was increased again following the epithelization of wound. The result of RT-PCR showed that the expression of both AT1 and AT2 receptors was detectable, and AT1 receptor was increased in the first 7 days to the peak, and then gradually decreased during wound healing, while AT2 receptor expression reached its peak value on day 7, then gradually decreased, and increased again following the epithelization of wound.

CONCLUSIONSThese results indicate that Ang II participate in wound repair and related to remolding in the late stage of wound healing through the change in production of angiotensin II and expression of AT1 and AT2 receptors. AT1 receptor might be closely associated with cell proliferation, while AT2 receptor might play a role in cell apoptosis and remolding during wound healing.

Angiotensin II ; genetics ; metabolism ; Animals ; Apoptosis ; Cell Proliferation ; Male ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; genetics ; Receptors, Angiotensin ; genetics ; metabolism ; Skin ; injuries ; metabolism ; pathology ; Wound Healing ; physiology

OBJECTIVETo investigate the correlation between the proteasome subunit PSMA7 expression in colorectal cancer and its role in liver metastasis.

METHODSTo identify the PSMA7 protein expression in 62 primary site colorectal cancers, 34 lymph node metastatic sites and 13 liver metastatic sites by immunohistochemistry and clarify the correlation of its expression with the clinicopathological parameters.

RESULTSHigh expression of PSMA7 was detected in 38.7% (24/62) of primary site colorectal cancer, 52.9% (18/34) of lymph node metastatic sites and 100% (13/13) liver metastatic sites but not in the normal colorectal tissue. High expression of PSMA7 was significantly correlated with liver metastasis (P = 0.028). The survival rate was significantly lower in patients with high expression of PSMA7 than in those with low expression of PSMA7 (P = 0.0008). As well, in multivariate analysis, PSMA7 expression demonstrated to be an independent prognostic factor (P = 0.004, relative risk 5.057; 95% confidence interval, 1.682-15.201).

CONCLUSIONPSMA7 may play an important role in the colorectal cancer progression. Evaluation of PSMA7 expression in primary colorectal cancer at the time of surgery might be a valuable test in defining patients with a high risk of developing liver metastasis.

Adult ; Aged ; Colonic Neoplasms ; enzymology ; pathology ; surgery ; Female ; Follow-Up Studies ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; enzymology ; secondary ; Lymphatic Metastasis ; Male ; Middle Aged ; Multivariate Analysis ; Neoplasm Staging ; Proteasome Endopeptidase Complex ; metabolism ; Rectal Neoplasms ; enzymology ; pathology ; surgery ; Survival Rate ; Young Adult