OBJECTIVETo provide molecular evidences for its breeding by studying the genetic relationship among varieties of Rehmannia glutinosa.

METHODNineteen varieties were detected by Randomly Amplified Polymorphic DNA(RAPD) markers.

RESULTThe 20 selected primers produced 163 bands, among which 114(69.9%) were polymorphic. A DNA molecular dendrogram was established based on Hierarchical cluster analysis of 163 DNA bands amplified by 20 primers, which divided the 19 varieties into four groups: Group Beijing, Group 85-5, Group Guolimao and the other Group.

CONCLUSION8 varieties of Group Beijing have a close genetic relationship, and so have varieties of Group 85-5, which provides information for Rehmannia glutinosa's breeding.

DNA, Plant ; genetics ; Plant Leaves ; genetics ; Plants, Medicinal ; genetics ; Polymorphism, Genetic ; Random Amplified Polymorphic DNA Technique ; Rehmannia ; genetics

OBJECTIVETo evaluate the efficacy and safety of irinotecan combined with xeloda (CAPIRI regimen) in patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin.

METHODSTotally 38 patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin were enrolled. Patients received xeloda 1 000 mg/m2 orally twice daily on day 1 to 14 and intravenous irinotecan 100 mg/m2 on day 1 and 8 every 3 weeks.

RESULTSThe median age of 38 patients was 58.5 (27-77) years. CAPIRI regimen was used 11.0 (3.0-24.0) months after the diagnosis of metastatic colorectal cancer (CAPIRI regimen as second-line chemotherapy in 33 patients, third-line in 4 patients, and fourth-line in 1 patient). A total of 121 cycles of chemotherapy (median 3.0) were administered. Thirty-four patients were evaluable for response. The overall response rate and disease control rate were 5.9% and 61.8%, respectively. The median time to progression and overall survival were 4.5 months (95% CI, 3.4-5.6 months) and 11.0 months (95% CI, 10.2-11.8 months), respectively. All 38 patients were evaluable for safety. The most common adverse events were leukopenia (73.7%), neutropenia (65.8%), nausea and vomiting (60.5%), and diarrhea (28.9%). The occurrence rates of these grade 3-4 events were 10.5%, 13.2%, 10.5%, and 7.9%, respectively. All adverse events were tolerable.

CONCLUSIONCAPIRI regimen is effective and well-tolerated in Chinese patients with metastatic colorectal cancer after failure of chemotherapy with oxaliplatin.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Camptothecin ; administration & dosage ; analogs & derivatives ; Capecitabine ; Colorectal Neoplasms ; drug therapy ; Deoxycytidine ; administration & dosage ; analogs & derivatives ; Female ; Fluorouracil ; administration & dosage ; analogs & derivatives ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Organoplatinum Compounds ; therapeutic use ; Treatment Outcome

BACKGROUNDYes-associated protein (YAP) plays an important role in signal transduction and gene transcription regulation in normal cells, with elevated and over-expressed YAP levels observed in various malignant tumors. The aim of this study was to investigate the expression of YAP in non-small cell lung cancer (NSCLC), and to study the possible relationship of YAP expression with the occurrence and development of NSCLC.

METHODSYAP expression was assessed in 40 cases of NSCLC tumor tissues by immunohistochemistry, and their protein and mRNA levels were evaluated through Western blotting and reverse transcription-polymerase chain reaction (PCR), respectively. Normal lung tissues obtained from the same patient were used as control. Statistical analysis was performed to correlate the YAP expression to clinical pathological factors, such as tumor type, stage and grade.

RESULTSYAP-positive expression was found in 28 (70%) of the 40 cases of NSCLC, which included 10 cases of squamous cell carcinoma (25%), 17 cases of adenocarcinoma (42.5%) and 1 case of squamous adenocarcinoma (2.5%). In the 28 YAP-positive cases, 19 cases showed lymph node metastasis and were classified in TNM stage II + III (47.5%); the other nine cases showed no lymph node metastasis (22.5%) and were classified in the TNM stage I. There was no relationship between YAP expression and patients' age, gender or tumor histological grades. However, YAP showed significant over expression in late period of T stage (P = 0.012), TNM stage (P = 0.039), and lymph node metastasis (P = 0.013), respectively. Notably, YAP-positive expression was significantly higher in adenocarcinoma than that in squamous cell carcinoma (P = 0.041).

CONCLUSIONSOver-expression of YAP was associated with NSCLC, especially lung adenocarcinoma. The high YAP expression in late period of tumor stage and lymph node metastasis may indicate that YAP expression could be an early marker for NSCLC tumorigenesis.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adult ; Aged ; Blotting, Western ; Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Phosphoproteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo provide reference for breeding Scutellaria baicalensis from different germplasms,and to explore their genetic diversities and different strains.

METHODThe RAPD method was applied to the study on S. baicalensis from different germplasms by use of 14 random primers about 10 bp, and SAPD-analysis of 34 S. baicalensis from different germplasms was carried out by using the method of withingroups-linkage in SPSS 10.0.

RESULTS14 among 115 primers were selected to amplify about 165 segments of DNA. Among them, 132 segments of DNA, 80.0% of the total, can represent genetic of diversities of S. baicalensis. According to RAPD analysis, 34 germplasms of S. baicalensis were classified into A, B, C and D categories.

CONCLUSIONS. baicalensis from different germplasms shows abundant genetic diversities. Germplasms from Shandong like Mengyin 3, Menyin 2 and Pingyi have close distance (0.315) in genetic background, which can be chosen for breeding of cultivated. Though genetic characters are similar in the morphology, the geological distribution of S. baicalensis and its morphology have not certain correlation. The complex genetic background of S. baicalensis indicate that the work of the selective breeding and management for breeding of S. baicalensis have to be strengthened.

Breeding ; Cluster Analysis ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Ecosystem ; Genetic Variation ; Phylogeny ; Plant Leaves ; anatomy & histology ; genetics ; Plants, Medicinal ; anatomy & histology ; genetics ; Random Amplified Polymorphic DNA Technique ; methods ; Scutellaria baicalensis ; anatomy & histology ; genetics

OBJECTIVETo explore the mechanism of 'erythroblast island (EI)' formation in the bone marrow of patients with immun-related hemocytopenia (IRP).

METHODSThe category of BM-auto antibody (au Ab) in 48 patients with IRP was detected with FCM. The BM-au Ab in the 'EI' of these cases were explored with immuonhistofluorescence (IF). Clinical and laboratory characteristics of these cases were also analyzed retrospectively.

RESULTSIgG could be detected in the 'EI' on the BM smear of 14 cases (29.17%), BM-au Ab mainly deposited at the edge/membranes between macrophage and erythroblasts rather than cyto plasm. Positive reaction were seen in all the cases with GlycoAIgG. The red blood cell count [(1.8 ± 0.5) × 10(12)/L] and hemoglobin level [(59.6 ± 16.2)g/L] were significantly lower than that in the IF(-) group [(2.5 ± 0.9) × 10(12)/L and (83.4 ± 25.0) g/L] (P < 0.05). The percentage of reticulocyte [(2.0 ± 0.8)%], serum level of IBIL [(9.4 ± 4.7) µmol/L], percentage of erythroblats in sternum BM (0.441 ± 0.139) and response rate to therapy (85.7%) in IF(+) group were significantly higher than that in IF(-)group [(1.3 ± 1.0)%, (6.6 ± 6.7)µmol/L, 0.298 ± 0.082, 61.3%, respectively] (P < 0.05).

CONCLUSIONMacrophage was connected with erythroblasts through autologous IgG in the 'EI's of some patients with IRP. 'EI' were the places where macrophages devoured and destroyed erythroblasts rather than erythroid development and differentiation. The pathogenetic mechanism of IRP might be associated with macrophages phagocytosing and destroying BM hematopoietic cells.

Blood Cell Count ; Bone Marrow ; Bone Marrow Cells ; immunology ; Coombs Test ; Erythroblasts ; Humans

OBJECTIVETo investigate the expression of GLUT1, p63 and DNA-Pkcs in serous ovarian tumors and their significance.

METHODSGTUL1, p63 and DNA-Pkcs expression at protein level was detected by immunohistochemistry in patients with serous ovarian tumors. Chi-square analysis was used to assess if their expression is associated with clinicopathologic characteristics of the tumors.

RESULTSCells in the normal ovarian tissues were not stained with GTUL1 and p63 antiserum, but DNA-Pkcs was positively stained. The intensity of GTUT1 and p63 expression was stronger in malignant ovarian serous tumors compared with other subtypes (P < 0.01). There were significant differences of DNA-PKcs among normal ovaries (100.0%), benign (95.0%), borderline (90.0%) and malignant (60.0%) serious ovarian neoplasms (P < 0.01). The level of GLUT-1 expression was correlated with FIGO staging, intraperitoneal implantation, ascites and lymph node metastasis (P < 0.05). p63 expression was associated with clinicopathologic characteristics except ascites (P < 0.05). DNA-PKcs was only correlated with FIGO staging and lymph node metastasis (P < 0.05).

CONCLUSIONThe results suggest that the abnormal expression of GTUT1, p63 and DNA-Pkcs may perhaps participate in serous ovarian tumor occurrence and development and may be considered as a marker reflecting tumor malignant behavior.

Adolescent ; Adult ; Aged ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Cystadenoma, Serous ; metabolism ; pathology ; DNA-Activated Protein Kinase ; metabolism ; Epithelium ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glucose Transporter Type 1 ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Middle Aged ; Neoplasm Staging ; Nuclear Proteins ; metabolism ; Ovarian Neoplasms ; metabolism ; pathology ; Ovary ; cytology ; Trans-Activators ; metabolism ; Transcription Factors ; Tumor Suppressor Proteins ; metabolism ; Young Adult

OBJECTIVETo investigate the inductive factors of effect on Akebia trifoliata and establish culture method for A. trifoliata callus.

METHODTo study the possible effective factors of culture condition by comparing with different explantation, nutrient medium, pH, temperature, illumination, growth substance of plant and its ratio.

RESULTThe inductivity of leaves was the highest about 87.5%, followed with the stem section and leafstalk; The inductivity of nutrient medium such as MS, B5 callus was higher than the ones like H, SH and the White callus amended one; It was found that low-grade Phvalue benefits the growth of callus. The experiment result showed that different pH showed little difference in quality. The best condition of culture was 25 degrees C in temperature.

CONCLUSIONThe best culture condition for callus was the leaves as explantation. The A. trifoliata callus culture's best inductive condition was MS +2, 4-D 4.0 mg x L(-1) + NAA 1.0 mg x L(-1) + KT 1.0 mg x L(-1) (pH 5.8), cultural temperature was 25 degrees C, cultivation was dark.

Culture Media ; chemistry ; pharmacology ; Hydrogen-Ion Concentration ; Magnoliopsida ; drug effects ; growth & development ; Plant Leaves ; drug effects ; growth & development ; Plant Stems ; drug effects ; growth & development ; Plants, Medicinal ; drug effects ; growth & development ; Temperature ; Tissue Culture Techniques ; methods

OBJECTIVETo provide reference for breeding of cultivated ginseng by researching the genetic diversities among different strains.

METHODTo make up the systematic diagram of genetic relationship by calculating the proximity coefficient with SPSS 10.0 software and clustered by between-groups linkage method.

RESULTThe selected 18 random primers used in PCR amplification to produce 145 bands in 17 samples, among which 53 bands (36.5%) are polymorphic. The results of cluster analysis show that the genetic relationship between different strains of Da-maya is closer than that of Er-maya, which proves that it is feasible to differentiate the cultivated groups in molecular level on outside character.

CONCLUSIONRAPD is a favorable molecular marker to assist ginseng breeding.

Cluster Analysis ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Genetic Variation ; Panax ; classification ; genetics ; growth & development ; Phylogeny ; Plants, Medicinal ; genetics ; growth & development ; Random Amplified Polymorphic DNA Technique ; methods

Objective:To conduct a glucagon like peptide-1(GLP-1)controllability model rat by chemical genetics, and observe the impact of GLP-1 neuron excitability on appetite.Methods:Fifteen rats were evenly divided into Green fluorescent protein(GFP)group, HM3D group, and HM4D group. Various combinations of adeno-associated virus(rAAV)were injected into the nucleus tractus solitarius(NTS). rAAV-GLP-1-cre and rAAV-GFP-dio were administered in rats of GFP group. The rats of HM3D group were injected with rAAV-GLP-1-cre and rAAV-HM3D-mCherry-dio while rAAV-GLP-1-cre and rAAV-HM4D-mCherry-dio were injected in rats of HM4D group . The optimal dose of clozapine N-oxide(CNO)was selected based on feeding behavior and body weight changes of rats after intraperitoneal injection of different doses of CNO. The controllability of GLP-1 neurons was confirmed by comparing with intraperitoneal injection of saline. The number of activated GLP-1 neurons in the NTS area and the expression of POMC neurons in the hypothalamus were detected 30 minutes after CNO injection.Results:GLP-1 neurons in the NTS area of rats were successfully labeled. The rat of HM3D group revealed a decrease in food intake( P=0.021)while the rat of HM4D group showed an increase( P=0.002), when given 1 mg/kg of CNO, no changes at the dose of 0.5 mg/kg and 3.0 mg/kg. Immunofluorescence showed that the activity of GLP-1 neurons in NTS of GFP group was lower than that of HM3D group( P=0.022), and higher compared with that of the HM4D group( P=0.049). The expression of GLP-1 neurons in NTS and POMC neurons in the hypothalamus of the HM3D group after intraperitoneal injection of CNO was also higher than that in the HM4D group( P=0.003). Conclusion:Using chemical genetics technology, GLP-1 controllability model rat could be successfully established via injecting varying combinations of rAAV into the NTS area of rat. Injection of 1 mg/kg CNO can effectively activate or inhibit the neuron to regulate appetite.

Construction of recombinant adenovirus, which contain human microdystrophin, and then transfection into mesenchymal cells( MSCs) of mdx mice were done, and genetically-corrected isogenic MSCs were acquired; the MSCs transplantation into the mdx mice was then done to treat the Duchenne muscular dystrophy( DMD). Microdystrophin cDNA was obtained from recombinant plasmid pBSK-MICRO digested with restrictive endonuclease Not I ; the production was inserted directionally into pShuttle-CMV. The plasmid of pShuttle-CMV-MICRO was digested by Pme I , the fragment containing microdystrophin was reclaimed and transfected into E. coli BJ5183 with plasmid pAdeasy-1. After screening by selected media, the extracted plasmid of positive bacteria was transfected into HEK293 cells with liposome and was identified by observing the CPE of cells and by the PCR method. Finally, MSCs of mdx mice were infected with the culture media containing recombinant adenovirus, and the expression of microdystrophin was detected by RT-PCR and immunocytochemistry. Recombinant adenovirus including microdystrophin was constructed successfully and the titer of recombinant adenovirus was about 5.58 x 10(12) vp/mL. The recombinant adenovirus could infect MSC of mdx mice and microdystrophin could be expressed in the MSC of mdx mice. Recombinant adenovirus including microdystrophin was constructed successfully, and the microdystrophin was expressed in the MSC of mdx mice. This lays the foundation for the further study of microdystrophin as a target gene to correct the dystrophin-defected MSC for stem cell transplantation to cure DMD.
Adenoviridae ; genetics ; Animals ; Cells, Cultured ; Dystrophin ; genetics ; metabolism ; Gene Expression ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Immunohistochemistry ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred mdx ; Muscular Dystrophy, Duchenne ; genetics ; pathology ; therapy ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transduction, Genetic