This study aims to determine a reasonable clinical staging standard for patients with esophageal carcinoma who were receiving non-surgical treatment. The patients were staged on the basis of the (2004 and 2009 editions of clinical staging stan-dards. The prognosis of patients with different staging standards, as well as the effect of gross tumor volume-tumor (GTV-T) on clinical T stage and prognosis, was observed. Methods:Data on 219 patients with esophageal carcinoma who were receiving radical radiothera-py were retrospectively analyzed. Prior to radiotherapy, all patients underwent examinations, including esophageal barium meal and po-sitioning CT scan, for use in the radiation treatment planning system to outline the target range and to calculate the volume of GTV-T. All patients were staged with the use of the aforementioned clinical staging standards. Prognostic outcomes of the patients were ob-served. Results:For all patients, the one-, three-, and five-year overall survival rates were 70.8%, 35.6%, and 20.7%, respectively. The survival curve resolution of patients who were staged with the use of the 2009 edition of clinical staging standards was better than that of the patients who were staged with the use of the 2004 edition. Survival difference was significant (χ2=29.497, P<0.001). The clinical T stage positively correlated with GTV-T (r=0.615, P<0.001). GTV-T could thus affect prognosis at different T stages. Conclusion:Both esophageal carcinoma clinical staging standards could reflect the prognosis of patients undergoing radiotherapy, but the 2009 edi-tion appeared more accurate than the 2004 edition.

Transcriptions are regulated by transcription factors. Natural transcription factors usually consist of at least two functional domains: a DNA-binding domain and an effector domain. According to this, novel artificial transcription factors are designed to up or down regulate transcription and expression of a target gene. The Cys2-His2 zinc finger domain is a DNA-binding module that has been widely used as the DNA-binding domain in artificial transcription factors. Each zinc finger domain, which comprises about 30 amino acids that adopt a compact structure by chelating a zinc ion, typically functions by binding 3 base pairs of DNA sequence. Several zinc fingers linked together would bind proportionally longer DNA sequences. According to the "bipartite complementary" library strategy, a pair of zinc finger phage display libraries were constructed. After construction of the libraries, a 9bp sequence (5'-GCAGAGGCC-3') on the promoter of SV40 was chosen as a target for next step. After parallel selection, PCR amplification, desired fragments recovery, re-ligation, and additional rounds of selection, phage enzyme-linked ELISA experiments were performed to identify specific binding clones displaying the zinc fingers with predetermined sequence-specificity to our target sequence. Then two clones with strong ELISA signals were chosen to be tested for binding both to its full target site (5'-GCAGAGGCC-3') and to sites containing single transition mutations. The binding specificity of one of the two clones (clone 3) was shown to be fairly good. The three-finger DNA-binding domain targeted to SV40 promoter, that is, zinc finger sequences on clone 3, was fused to KOX1 suppression domain KRAB and cloned into pcDNA3.1 (+) (which expression product was artificial transcription factor). The zinc fingers (which expression product was the DNA-binding domain of artificial transcription factor) and KRAB domain only (which expression product was effector domain of artificial transcription factor) were also cloned separately into the same expression vector. All constructs contained an N-terminal nuclear localization signal. Every of the vectors (including pcDNA3.1 (+) without inserting sequences) were cotransfected with pGL3-Control and pRL-TK and the activity of luciferase was used to indicate the function of product from transfected expression vectors. Our artificial transcription factor was proved to repress the expression of reporter gene efficiently,while with only DNA-binding domain or effector domain the repression was not remarkable. By adding different effector domains and changing the DNA-binding domain, artificial transcription factor would have a wide range of potential applications.
Enzyme-Linked Immunosorbent Assay ; Genes, Synthetic ; genetics ; physiology ; Models, Theoretical ; Peptide Library ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Transcription Factors ; chemical synthesis ; chemistry ; metabolism ; Zinc Fingers ; genetics ; physiology

OBJECTIVEA total of 27 samples belonging to 5 cultivars of Fructus Aurantii (Citrus aurantium), i. e. cv. Xiucheng, cv. Xiangcheng, cv. Lecheng, cv. Jizicheng, and cv. Youzicheng, collected at Changfu and Huanggang, Zhangshu City, Jiangxi Province, were assayed to reveal the genetic relationship among the cultivars and the accordance between morphological and molecular markers.

METHODCultivar identification was based on morphology and cultivar relationship was based on Inter-simple sequence repeats (ISSR).

RESULTTwenty out of 40 ISSR primers screened generated 392 loci across all 27 samples with 315 informative loci. The UPGMA dendrogram showed that samples within cv. Xiucheng and cv. Xiangcheng from Changfu were closely related. However, samples of cv. Lecheng, cv. Jizicheng and cv. Youzicheng from Huanggang, or cv. Xiucheng and cv. Xiangcheng from both Changfu and Huanggang did not exhibited close relationships within each cultivars.

CONCLUSIONBased on morphology the same cultivar grown in different plantations, or even within a single plantation sometimes do not show close genetic relationship, indicating diverse origin of the cultivars. Synonyms or homonyms are believed to common phenomenon in Fructus Aurantii production. To solve the problem ISSR markers can serve a kind of molecular markers which are preferable to partition genetic variations within and between cultivars and to establish genetic relationships among them.

Citrus ; anatomy & histology ; classification ; genetics ; DNA Primers ; DNA, Plant ; genetics ; Fruit ; anatomy & histology ; genetics ; Genetic Markers ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; anatomy & histology ; classification ; genetics ; Repetitive Sequences, Nucleic Acid

OBJECTIVETo summarize the clinical features of cytopenia with bone marrow hypoplasia in 100 children and to investigate an effective treatment regimen for myelodysplastic syndrome (MDS) in children.

METHODSA retrospective analysis was performed on the clinical data of 100 children non-randomly selected from Japan and China who were diagnosed with cytopenia with bone marrow hypoplasia between 2006 and 2011. The data of patients from China were subjected to prognostic analysis.

RESULTSThere was no significant difference in the proportion of MDS cases and acquired aplastic anemia (AA) cases between the Japanese and Chinese children. Of the 100 patients, there were 29 cases of acquired AA, 58 cases of refractory cytopenia of childhood (RCC) and 13 cases of refractory cytopenia with multilineage dysplasia (RCMD). There were significant differences in reticulocyte absolute value in peripheral blood and degree of bone marrow proliferation among the three patient groups (P<0.05). The patients from China were followed up for 16-70 months (median, 41 months). After being treated with cyclosporine (CsA) combined with stanozolol, the patients with AA had response rates of 25% and 75%, the patients with RCC had response rates of 47.1% and 82.4%, and the patients with RCMD had response rates of 60% and 60% respectively at 3 and 6 months after treatment.

CONCLUSIONSThere are significant differences in reticulocyte absolute value in peripheral blood and degree of bone marrow proliferation among patients with RCC, RCMD and acquired AA. CsA combined with stanozolol has a good therapeutic efficacy in the treatment of acquired AA and hypoplastic MDS in children, but studies of more cases and a longer follow-up duration are needed.

Adolescent ; Anemia, Aplastic ; blood ; drug therapy ; pathology ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Cyclosporine ; therapeutic use ; Female ; Follow-Up Studies ; Hematopoietic Stem Cell Transplantation ; Humans ; Infant ; Male ; Myelodysplastic Syndromes ; blood ; drug therapy ; pathology ; Pancytopenia ; blood ; drug therapy ; pathology

Objective To analyze the spatio-temporal process on 2009 influenza A (HlNl) pandemic in Changsha and the influencing factors during the diffusion process. Methods Data were from the following 5 sources, influenza A (HlNl) pandemic gathered in 2009, Geographic Information System (GIS) of Changsha, the broad range of theorems and techniques of hot spot analysis, spatio-temporal process analysis and Spearman correlation analysis. Results Hot spot areas appeared to be more in the economically developed areas, such as cities and townships. The cluster of spatial-temporal distribution of influenza A (HlNl) pandemic was most likely appearing in Liuyang city (RR=22.70,P<0.01). The secondary cluster would include districts as Yuelu (RR=6A9,P< 0.01) , Yuhua (RR=81.63, P<0.01). Xingsha township appeared as the center in the Changsha county (RR=2.90, P<0.01) while townships as Yutangping (RR=19.31, P<0.01) , Chengjiao (RR=73.14,P<0.01) and Longtian appeared as the center in the west of Ningxiang county (RR= 14.43,P<0.01) and Wushan as the center in the Wangcheng county (RR= 13.84,P<0.01). As time went on, the epidemic moved towards the eastern and more developed regions. Regarding factor analysis, population, the amount of students, geographic relationship and business activities etc. appeared to be the key elements influencing the transmission of influenza A (H1N1) pandemic. At the beginning of the epidemic, population density served as the main factor (r=0.477, P<0.05) but during the initial and fast growing stages, it was replaced by the size of students to serve as the important indicator (r=0.831, P<0.01; r=0.518, P<0.01). However, during the peak of the epidemics, the business activities played an important role (r=-0.676, P<0.01). Conclusion Groups under high risk and districts with high incidence rates were shifting, along with the temporal process of influenza A(H1N1) pandemic, suggesting that the protection measures need to be adjusted, according to the significance of influencing factors at different stages.

This study was aimed to investigate the incidence of JAK2V617F mutation in BCR-ABL negative patients with myeloproliferative disorders (MPD) and its relation with clinical characteristics of MPD. The sensitive and specific test for JAK2V617F mutation was established for improving diagnosis level in Gansu province. 47 BCR/ABL negative MPD patients and 12 healthy people were enrolled in this study. Allele specific polymerase chain reaction (AS-PCR) was used to amplify the exon 12 of JAK2 gene which harbours V617F mutation. The PCR products were identified by DNA sequencing. And its relation with clinical characteristics of MPD was analyzed also. The results indicated that the incidence of JAK2V617F positive mutation in 47 patients with BCR-ABL negative MPD was 74.5 % (35/47), including 83.9 %(26/31) in patients with polycythemia vera (PV), 60 % (9/15) in patients with essential thrombocythemia (ET), only in one patient with idiopathic myelofibrosis (IMF). In PV group, the patients with JAK2V617F positive mutation had higher counts of WBC and Plt than patients with JAK2V617F negative mutation. In ET group, the patients with JAK2V617F positive mutation had higher WBC count and Hb level than those in the patients with JAK2V617F negative mutation with tendency of suffering from complications such as hepatosplenomegaly, haemorrhage and thrombosis. It is concluded that JAK2V617F mutation is more frequent in BCR-ABL negative patients with MPD, the AS-PCR method is sensitive and specific for detection of the mutation and may successfully use in clinical examination.
Adult ; Aged ; Case-Control Studies ; Female ; Humans ; Janus Kinase 2 ; genetics ; Leukocyte Count ; Male ; Middle Aged ; Mutation ; Myeloproliferative Disorders ; diagnosis ; genetics ; Polycythemia Vera ; diagnosis ; genetics ; Primary Myelofibrosis ; diagnosis ; genetics

Objective:To translate the Incontinence-Associated Dermatitis Intervention Tool (IADIT) into Chinese and to test the reliability and validity.Methods:The IADIT was translated into Chinese after getting the approval of original author, professor Junkin. We selected 15 experts with a high academic in incontinence nursing and rich clinical experience for two rounds of consultation to determine the Chinese version and its validity. At the time, the reliability of the Chinese version of IADIT was tested at a ClassⅢ Grade A hospital.Results:The item-content validity index ( I- CVI) of the Chinese version of IADIT ranged from 0.80 to 1.00, and the scale-content validity index ( S- CVI) of the tool was 0.98. The reliability of assessors was r=0.928 ( P<0.01) , and the test-retest reliability was 0.996 ( P<0.01) . Conclusions:The Chinese version of the IADIT has a good reliability and validity which could guide nurses to carry out assessment and intervention for incontinence patients.

OBJECTIVETo investigate the potential effects of arsenic trioxide (As(2)O(3)) combined with 8-(4-chlorophenylthio) adenosine 3', 5'-cyclic monophosphate (8-CPT-cAMP) on the retinoic acid (RA)-resistant acute promyelocytic leukemia (APL) cells.

METHODSThe RA resistant APL cell lines NB4-R1 and NB4-R2 were used as in vitro models. The effect of As(2)O(3) and/or 8-CPT-cAMP was evaluated according to cellular morphology, cell surface antigen and nitroblue-tetrazolium (NBT) assay. Meanwhile, immunofluorescence analysis and Western blot assay were used to detect the degradation of PML-RAR alpha fusion protein and the change of several key cell cycle regulatory proteins in these cells before and after the treatment.

RESULTSLow dose of As(2)O(3) (0.25 micromol/L) synergized with 8-CPT-cAMP (200 micromol/L) in inducing differentiation of NB4-R1 and NB4-R2 cells, while neither of these two drugs alone could induce differentiation of these cells. In addition, 8-CPT-cAMP was able to inhibit the cell growth by modulating the expression of some important cell cycle regulators and to facilitate the As(2)O(3)-mediated degradation of PML-RAR alpha fusion protein.

CONCLUSIONSAs(2)O(3) combined with 8-CPT-cAMP could induce differentiation of RA-resistant APL cells.

Antineoplastic Agents ; pharmacology ; Arsenicals ; pharmacology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cyclic AMP ; analogs & derivatives ; pharmacology ; Dose-Response Relationship, Drug ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; Leukemia, Promyelocytic, Acute ; pathology ; Oxides ; pharmacology ; Thionucleotides ; pharmacology ; Tretinoin ; pharmacology

In order to investigate the role of calcium pathway in myeloid differentiation, the expression level of genes related to calcium pathway in all-trans retinoic acid (ATRA)-induced NB4 cell differentiation was detected by cDNA microarray, some of which were further confirmed by quantitative real time RT-PCR. At the same time, the expressions of these genes in NB4-R1 cells treated with ATRA and 8-CPT-cAM P alone or in combination, and in differentiation of primary cells from ATRA-induced newly diagnosed APL patients were detected by real time RT-PCR. The results showed that during differentiation of ATRA-induced NB4 cells, the expressions of genes related to calcium concentration had changed, the expression of downstream effectors in calcium pathway was up-regulated and confirmed by real time RT-PCR assay. The expression of genes related to calcium concentration did not change significantly when NB4-R1 cells were treated by ATRA or 8-CPT-cAMP alone, but expression changes of those genes were similar to the changes in ATRA-induced NB4 cell differentiation when NB4-R1 cells were treated by ATRA combined with 8-CPT-cAMP. In addition, the expression changes of those genes in ATRA-induced primary cells of patients with APL were also similar to changes in ATRA-induced NB4 cell differentiation. It is concluded that calcium pathway may be involved in ATRA-induced differentiation in APL cell.
Calcium ; metabolism ; Cell Differentiation ; drug effects ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia, Promyelocytic, Acute ; genetics ; metabolism ; Signal Transduction ; Tretinoin ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo analyze the periodicity of pandemic influenza A (H1N1) in Changsha in year 2009 and its correlation with sensitive climatic factors.

METHODSThe information of 5439 cases of influenza A (H1N1) and synchronous meteorological data during the period between May 22th and December 31st in year 2009 (223 days in total) in Changsha city were collected. The classification and regression tree (CART) was employed to screen the sensitive climatic factors on influenza A (H1N1); meanwhile, cross wavelet transform and wavelet coherence analysis were applied to assess and compare the periodicity of the pandemic disease and its association with the time-lag phase features of the sensitive climatic factors.

RESULTSThe results of CART indicated that the daily minimum temperature and daily absolute humidity were the sensitive climatic factors for the popularity of influenza A (H1N1) in Changsha. The peak of the incidence of influenza A (H1N1) was in the period between October and December (Median (M) = 44.00 cases per day), simultaneously the daily minimum temperature (M = 13°C) and daily absolute humidity (M = 6.69 g/m(3)) were relatively low. The results of wavelet analysis demonstrated that a period of 16 days was found in the epidemic threshold in Changsha, while the daily minimum temperature and daily absolute humidity were the relatively sensitive climatic factors. The number of daily reported patients was statistically relevant to the daily minimum temperature and daily absolute humidity. The frequency domain was mostly in the period of (16 ± 2) days. In the initial stage of the disease (from August 9th and September 8th), a 6-day lag was found between the incidence and the daily minimum temperature. In the peak period of the disease, the daily minimum temperature and daily absolute humidity were negatively relevant to the incidence of the disease.

CONCLUSIONIn the pandemic period, the incidence of influenza A (H1N1) showed periodic features; and the sensitive climatic factors did have a "driving effect" on the incidence of influenza A (H1N1).

China ; epidemiology ; Climate ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; epidemiology ; virology ; Regression Analysis ; Risk Factors ; Seasons ; Temperature