Objective　To investigate the significance of TGF-β1, TGFRl and TGFR2 in the pathogenesis and prognosis in patients with myelofibrosis. Methods　The expression of TGF-β1 and its receptors (TGFR1 and TGFR2 ) in bone marrow tissues and the level of TGF-β1 in the blood of 23 patients with myelofibrosis were detected by SABC immunocytochemistry and ELISA repectively. Results　Expression of TGF-β1 and TGFR 1 was significantly higher in primary and secondary myelofibrosis patients than that of the control. No significant difference of TGFR2 expression was found between the groups of myelofibrosis and the control (P>0.05). The level of TGF-β1 in the blood of the patients with myelofibrosis was significantly higher than that of the control (P<0.01) and more obvious in secondary cases while TGF-β1 decreased nearly to the normal level when patients were in clinical remission. Conclusion　TGF-β1 and it's receptors may be involved in the pathogenesis of myelofibrosis and might be of importance for the prognosis of the patients with myelofibrosis.

Akebia trifoliate has been reported to have many pharmacological activities and the roots, petioles and seeds are used to different symptoms. However, the structure and anatomy of its seeds was almost not reported until now. In the present study, we investigated the morphological characters of the fruit and seed, and the anatomical characters of the testa, micropyle, embryo and endosperm, which could provide evidences for the study on classification, identification and application of A. trifoliate. Our results showed that the testa of A. trifoliate consisted of an epidermic cell layer, the sclerenchyma cells layer, the parenchyma cells layer and an innermost pigment layer. At the micropylar region, the outermost epidermal cells were specialized the white caruncle-like structure and the testa included a lot of lignified tissues. Endosperm comprises two layer cells. Outermost yellowish-brown layer cells contains lots of fat droplets, and innermost white layer cells contains lots of aleurone grains and crystalloids.
Germination ; Magnoliopsida ; anatomy & histology ; growth & development ; Seeds ; anatomy & histology ; growth & development

BACKGROUND: Angiotensinogen (AGT) gene is the firstly discovered candidate gene for essential hypertension, both the T174M and M235T polymorphisms locate at the second exons of AGT gene, and there is existence of linkage disequilibrium. The polymorphism at A-6G and G-217A sites in promotor region plays an important role in regulating the gene expression, and the products of keep close correlation with the level of blood pressure. OBJECTIVE: To investigate the association between the polymorphism of AGT gene at A-6G, T174M and G-217A sites and the risk for the attack of essential hypertension in Chinese Han population, DESIGN: A cluster sampling and case-control analysis. SETTINGS: Department of Geriatrics and Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University; Southern Research Center of National Human genome; Department of Cardiology, Dongtai People's Hospital of Jiangsu Province. PARTICIPANTS: The experiment was carried out in the countryside of Dongtai county, Yancheng city, Jiangsu province. All the subjects were selected from the countryside of Dongtai county, Yancheng city, Jiangsu province. Totally 177 patients with essential hypertension who had never accepted any drug treatment, were taken as the essential hypertension group, and hypertension was diagnosed according to the diagnostic standard of hypertension set by WHO/ISH in 1999 (systolic blood pressure ≥ 140 mm Hg and/or diastolic blood pressure ≥ 90 mm Hg); Another 86 normal person were taken as the normal control group. ② Inclusive criteria: The enrolled subjects should be Han nationality; long-term local residents but not from other places; able to answer questions clearly; diagnosed by disease history, clinical symptoms, physical signs and assistant examinations; have complete data of investigation of uniform questionnaires by face-to-face interview (including demographic information, profession history, family history and life styles of smoking, drinking, drinking tea, etc.). ③ Exclusive criteria: The patients with secondary hypertension in the essential hypertension group, subjects having family hisory of hypertension in the normal control group, and those with chronic diseases of liver and kidney, and diabetes mellitus in both groups were excluded. METHODS: Peripheral venous blood samples (3 mL) were collected, and DNA was extracted from human peripheral blood with FlexiGene DNA Kit (250). The Primer3 software was applied to design primers, and the polymorphism sites in the primer sequence were excluded. After multiplex polymerase chain reaction (PCR), 3 μL products were selected to detected the amplified results by agarose gel electrophoresis. The successfully amplified PCR products were purified with the QIAquick PCR Purification Kit, and the purified products were fragmentized with Dnase Ⅰ . The fragmentized products of enzyme digestion were labeled with fluorescein by deoxynucleotide terminal transferase. Two allele specific probes and one mismatched probe were designed respectively for each single nucleotide polymorphism. The chips were prepared with the OmniGridTM 100 TLC samler, each probe was repeated for three times to form three matrix. The hyridization solution was degenerated at 95 ℃ for 10 minutes, and then immediately cut on ice. 10 μL hybridization solution was added onto the chip matrix, hybridized at 50 ℃ for 2 hours, then washed and dried. The chips were scanned with the GenePix 4000B laser confocal scanner (Figure 2),and the intensity of the fluorescent signal for each probe was extracted with GenePix Pro, and the allele score of each single nucleotide polymorphism was calculated to judge the genotype. MAIN OUTCOME MEASURES: ① Comparison of the frequencies of genotype distribution at each polymorphism site of AGT gene in both groups; ② Correlation analysis of the polymorphism of AGT gene at A-6G and T-174M sites with the risk for the attack of essential hypertension; ③ Effects of the polymorphism of AGT gene at A-6G, T-174M and G-217A sites on blood pressure.RESULTS: According to the intention-to-treat analysis,all the 263 subjects were involved in the analysis of results. ① At the A-6G site of AGT gene, the frequencies of AA, AG and GG genotypes (P=0.014) and A and G alleles (P=0.004, OR=0.44) had significant differences between the essential hypertension group and normal control group; At the T174M site, the frequencies of CC, CT and TT genotypes (P=0.031) and A and G alleles (P=0.014, OR=0.55) were significantly different; At the G-217A site, no obvious differences were found in the GG, AG and AA genotypes (P=0.722) and G and A alleles (P=0.403, OR=0.80). ② The risk of essential hypertension in the individuals carrying AA genotype of A-6G polymorphism and CC genotype of T174M polymorphism was reduced by 57% (95%CI= 0.23-0.82, P= 0.010) and 56% (95%CI= 0.25-0.79, P= 0.006) respectively. ③ There were no significant differences in the systolic blood pressure, diastolic blood pressure and mean arterial pressure among different genotypes at the A-6G, T174M sites and G-217A sites (F=0.100- 2.911, P ＞ 0.05). CONCLUSION: The AA genope at A-6G and the CC genotype at T174M site of AGT gene may reduce the risk for the attack of essential hypertension in Chinese Hun population, and no significant correlation was found between the genotype of G-217A polymorphism and the attack of essential hypertension.

Objective To investigate the clinic effect of the Salvia miltiorrhiza combined with dextran to prevent veno-occlusive disease after hematopoietic stem cell transplantation. Methods In the process of the pretreatment of the hematopoietic stem cell transplantation, patients were treated with salvia miltiorrhiza (20 ml/d), dextran(250 ml, twice a day) by venous transfusion and the drugs to protect the liver cell was used in the same time. When the count of platelet dropped to 30×109/L, salvia miltiorrhiza and dextranware stopped applying forever. Results Veno-occlusive disease and hemorrhage has not occurred during 85 times of the hematopoietic stem cell transplantation treated with salvia miltiorrhiza and dextran. Conclusion We conclude that the combined treatment with salvia miltiorrhiza and dextran is safe and effective to prevent veno-occlusive disease after hematopoietic stem cell transplantation.

OBJECTIVETo break through the restrictions of the evaluation model and the quantity of compounds by using the two-dimensional zebrafish model combined with chromatographic techniques, and establish a new method for the high-throughput screening of active anti-osteoporosis components.

METHODAccording to the research group-related studies and relevant foreign literatures, on the basis of the fact that the zebrafish osteoporosis model could efficiently evaluate the activity, the zebrafish metabolism model could efficiently enrich metabolites and the chromatographic techniques could efficiently separate and analyze components of traditional Chinese medicines, we proposed that the inherent combination of the three methods is expected to efficiently decode in vivo and in vitro efficacious anti-osteoporosis materials of traditional Chinese medicines.

RESULT AND CONCLUSIONThe method makes it simple and efficient in the enrichment, separation and analysis on components of traditional Chinese medicines, particularly micro-components and metabolites and the screening anti-osteoporosis activity, fully reflects that efficacious materials of traditional Chinese medicines contain original components and metabolites, with characteristic of "multi-components, multi-targets and integral effect", which provides new ideas and methods for the early and rapid discovery of active anti-osteoporosis components of traditional Chinese medicines.

Animals ; Chromatography ; methods ; Disease Models, Animal ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; trends ; Osteoporosis ; drug therapy ; physiopathology ; Phytotherapy ; methods ; trends ; Reproducibility of Results ; Zebrafish ; physiology

OBJECTIVETo explore the feasibility of membrane protein-based two-dimensional electrophoresis (2-DE) in the investigations of chondrocyte-related diseases and its complementarity with total protein-based 2-DE.

METHODSKnee cartilage samples were obtained to isolate the chondrocytes with type II collagenase/hyaluronidase digestion. The membrane proteins and total proteins were extracted and loaded separately onto PH3.0-10.0 non-linear gel strip for 2-DE analysis. The qualities of membrane protein-based 2-DE gels were evaluated, and the difference between the distribution profiles of the membrane protein spots and that of the total protein were observed and their complementarities were evaluated.

RESULTSMembrane protein-based 2-DE generated high-quality gel images, and on each gel 412.3±13.5 protein spots were identified. These spots were distributed in the region of isoelectric point pH 5.0-9.0. In the gel images generated by the total proteins, 564.3±5.9 protein spots were identified in each image, and the spots were distributed in the region of isoelectric point pH 3.0-7.0.

CONCLUSIONMembrane protein-based 2-DE of the chondrocytes can generate high-quality gel images, and the isoelectric distribution of the protein spots is complementary to that of total protein, which provides valuable information for chondrocyte-related diseases.

Adult ; Chondrocytes ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Isoelectric Point ; Male ; Membrane Proteins ; isolation & purification ; Middle Aged

Objective To investigate the association of angiotensinogen(AGT)gene A-6G、T174M and G-217A polymorphisms with the risk of essential hypertension(EH)in the elderly of Han nationality.Methods Genotypes of AGT gene A-6G,T174M and G-217A polymorphisms in 177 aged EH patients and 86 sex and age-matched controls were analyzed with gene chip technology.Results The A-6G and T174M polymorphisms of AGT gene were significantly associated with EH.The numbers of the three genotypes of A-6G were 113,58 and 6 in the patient group and 70,15 and 1 in the control group(P= 0.014)and those of T174M were 94,77 and 6,60,25 and 1(P=0.031),respectively.G-217A polymorphism was not related to EH.Individuals carrying A-6G AA and T174M CC genotypes showed 57% and 56% lower risk of EH(OR=0.43;95%CI=0.23-0.82 and OR=0.44;95%CI=0.25-0.79, respectively).Conclusions The A-6G AA and the T174M CC genotype may be related with decreased risk of EH and G-217A polymorphism may have little role in the etiology of EH in Han nationality.

Objective From May 2009-January 2010,a total of 3768 biosamples were tested for influenza A ( H1N1 ) infection at Zhengzhou center for disease control and prevention,China.1452 cases were laboratory confirmed H1N1 infection and 2316 were considered suspected victims.To evaluate the current protocol of influenza A (H1N1) based on the epidemic situations of Zhengzhou,relationships among features were explored and whether additional clinical characteristics should be part of H1N1 diagnosis protocols were determined.Methods Both clinical and epidemiologic findings as well as statistical analyses were described in this article.Test for independence between features related to the disease diagnosis has been proposed.Furthermore,logistic regression was carried out to measure the association among features and latent class analysis was performed to identify additional crucial features in laboratory confirmed H1N1 by building various latent models with different combinatorial features.Results The mean generation time for H1N1 was estimated as 3.59 ± 1.41 days (range =2.01 -7.26).The estimated infection rate was 0.258 ±0.088 3,and reproduction number was 1.94 (95％ CI =1.12 - 3.18 ).Our results revealed that the six features,including molecular detections using three separate primer/probe sets,gender,age and temperature,are all associated with clinical diagnosis of H1N1,and that three separate primer/probe sets for laboratory confirmed H1N1,age and temperature are associated with each other.Conclusion Additional clinical features applied into the H1N1 diagnosis with current three primers/probe sets can increase the diagnostic efficiency.

Adult ; Carcinoma, Renal Cell ; pathology ; surgery ; Cerebellar Neoplasms ; pathology ; surgery ; Cystadenoma, Papillary ; pathology ; surgery ; Diseases in Twins ; pathology ; surgery ; Epididymis ; pathology ; surgery ; Genital Neoplasms, Male ; pathology ; surgery ; Hemangioblastoma ; pathology ; surgery ; Humans ; Kidney Neoplasms ; pathology ; surgery ; Male ; von Hippel-Lindau Disease ; pathology ; surgery

OBJECTIVETo detect the transcriptional level of a novel gene F10 associated with the pathogenesis of hydatidiform mole in human cell lines and screen the cell lines with low F10 expression to construct a stable eukaryotic expression system for F10 gene.

METHODSThe expression level of F10 mRNA was detected with fluorescent quantitative PCR in A549, 16HBE, Bel7402, HIC, HepG2, 293, PC and MGC cell lines. A549 cell line was transfected with plasmid pRc-CMV2-F10 via electroporation to allow stable F10 expression, and the positive cell clones were selected by G418. The insertion and expression of F10 gene in the A549 cells was analyzed using fluorescent quantitative PCR.

RESULTSF10 mRNA was expressed differentially in these cells lines, and the Bel7402 cells, PC and MGC cells showed the highest F10 mRNA expression, followed by HepG2 and HIC cells and further by 293 cells, and 16HBE and A594 cells had the lowest expression. After transfection, A594 cells showed genomic integration of F10 gene and high expression level of F10 mRNA.

CONCLUSIONThe pulmonary carcinoma cell line A549 with stable expression of F10 gene has been established, which may facilitate further study of the biological functions of F10 gene.

Cell Line, Tumor ; Eukaryotic Cells ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Genes, Neoplasm ; genetics ; Humans ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Trophoblastic Neoplasms ; genetics ; pathology ; Uterine Neoplasms ; genetics ; pathology