OBJECTIVETo explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.

METHODAcute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.

RESULTThe purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.

CONCLUSIONASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.

Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Neoplastic Stem Cells ; cytology ; drug effects ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects

OBJECTIVETo investigate the diagnostic value of serum CEACAM1 in patients with pancreatic cancer.

METHODSFifty patients with pancreatic cancer and 50 with chronic pancreatitis were examine for serum levels of CEACAM1 by enzyme-linked immunosorbent assay (ELISA). The cut-off values and area under curve (AUC) of CEACAM1 was obtained by receiver operating characteristic (ROC) curve. The diagnostic efficiency of the tumor markers for pancreatic cancer was assessed by the fourfold table.

RESULTSThe serum level and positivity rate of CEACAM1 in pancreatic cancer patients were higher than those in chronic pancreatitis patients (P<0.05). Based on the ROC curve, the cut-off values and AUC of CEACAM1 were 13.835 ng/ml and 0.780, respectively (P<0.05). In pancreatic cancer patients, the diagnostic sensitivities of the tumor markers decreased in the order of CEACAM1 < CA242 < CA19-9 (P<0.05), and the specificity in the order of CA242 < CA19-9 < CEACAM1 (P<0.05).

CONCLUSIONCEACAM1 shows a higher diagnostic sensitivity than CA19-9 and CA242 for pancreatic cancer, but due to its low specificity this marker alone is not sufficient for diagnostic purposes.

Aged ; Aged, 80 and over ; Antigens, CD ; blood ; Biomarkers, Tumor ; blood ; Cell Adhesion Molecules ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; ROC Curve

BACKGROUNDVon Hippel-Lindau disease (VHL), a heritable autosomal dominant disease characterized by neoplasia in multiple organ systems, has rarely been reported in Asia. We genetically investigated a unique Chinese family with VHL disease and performed an analysis of the VHL protein stability.

METHODSGenomic deoxyribonucleic acid (DNA) extracted from peripheral blood was amplified by polymerase chain reaction (PCR) to three exons of the VHL gene in 9 members of the Chinese family with VHL disease. PCR products were directly sequenced. We estimated the effects of VHL gene mutation on the stability of pVHL, which is indicated by the free energy difference between the wild-type and the mutant protein (ΔΔG).

RESULTSThe Chinese family was classified as VHL type 1. Three family members, including two patients and a carrier, had a T to G heterozygotic missense mutation at nucleotide 515 of the VHL gene exon 1. This missense mutation resulted in the transition from leucine to arginine in amino acid 101 of the VHL protein. There was low stability of the VHL protein (the ΔΔG was 12.71 kcal/mol) caused by this missense mutation.

CONCLUSIONSWe first reported a family with this VHL gene mutation in Asia. This missense mutation is predicted to significantly reduce the stability of the VHL protein and contribute to the development of the renal cell carcinoma (RCC) phenotype displayed by this family. The genetic characterization and protein stability analysis of families with VHL disease are important for early diagnosis and prevention of the disease being passed on to their offspring.

Adult ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Protein Stability ; Von Hippel-Lindau Tumor Suppressor Protein ; chemistry ; genetics ; von Hippel-Lindau Disease ; genetics

Objective:To observe serum levels of high sensitive C reactive protein(hsCRP),interleukin(IL)-6, tumor necrosis factor(TNF)-α and hepatocyte growth factor(HGF)in patients with ischemic cerebrovascular dis-ease(ICVD),and analyze their correlation with blood lipids and anticardiolipin antibody(ACA)levels.Methods:A total of 45 ICVD patients treated in our hospital from Mar 2015 to Aug 2016 were regarded as ICVD group.An-other 45 non-ICVD patients treated in our hospital during the same period were regarded as non-ICVD control group.Serum levels of hsCRP,IL-6,TNF-α,HGF,blood lipids and ACA were observed and compared between two groups,and correlation among serum levels of hsCRP,IL-6,TNF-α and HGF,blood lipids and ACA levels were analyzed in ICVD patients.Results:Compared with non-ICVD control group,there were significant rise in se-rum levels of hsCRP[(4.69 ± 1.31)mg/L vs.(8.87 ± 1.56)mg/L],IL-6[(12.17 ± 4.33)mg/L vs.(34.26 ± 5.15)mg/L],TNF-α[(28.45 ± 2.18)pg/ml vs.(48.35 ± 3.15)pg/ml],HGF[(502.34 ± 15.36)pg/ml vs. (876.25 ± 18.15)pg/ml],ACA[(4.11 ± 0.65)IU/L vs.(7.89 ± 1.02)IU/L],total cholesterol[(4.68 ± 1.12) mmol/L vs.(5.57 ± 1.21)mmol/L],low density lipoprotein cholesterol[(2.62 ± 0.49)mmol/L vs.(3.24 ± 0.87) mmol/L]and triglyceride[(1.42 ± 0.31)mmol/L vs.(1.84 ± 0.37)mmol/L],and significant reduction in serum level of high density lipoprotein cholesterol[(1.28 ± 0.25)mmol/L vs.(1.02 ± 0.22)mmol/L]in ICVD group,P=0.001 all.Pearson correlation analysis indicated that serum levels of hsCRP,IL-6,TNF-α and HGF were signifi-cant positively correlated with serum levels of ACA,TC,LDL-C and TG(r=0.468~0.632,P<0.05 or <0.01), and significant inversely correlated with HDL-C level(r= -0.571~ -0.511,P<0.05 or < 0.01)in ICVD pa-tients.Conclusion:Serum levels of hsCRP,IL-6,TNF-α and HGF significantly rise in ICVD patients,and they are closely correlated with levels of blood lipids and ACA,which can serve as important indexes monitoring ICVD.

AIMTo optimize the method of investigating structural integration between proteins and study the integration between arrhythmia related proteins in molecular level.

METHODSImmunostaining the normal ventricular myocytes was used to observe the distribution of connexin 43 and muscarinic acetylcholine receptor (mAChR). The five mAChR subtypes were precipitated using immunoprecipitation. Then, SDS-PAGE and Western blotting with the anti-connexin 43 antibody were performed to observe whether they were structurally integrated. Further, different concentrations of detergent were used to observe whether this relationship could be broken.

RESULTSThe five subtypes of mAChR existed in the cardiac myocyte of the rat, and all the five mAChR subtypes combined with connexin 43. In the normal rat ventricular myocyte membrane, connexin 43 and M3 receptor are co-located. When adding certain concentration of detergent to the membrane protein, the integration between M3 receptor and connexin 43 was broken, and the phosphorylated form of connexin 43 integrated with M3 receptor.

CONCLUSIONThe results indicated that the structural integration between mAChR and phosphorylation of connexin 43 existed in rat ventricular myocardium, and this integration could be broken by certain concentration of detergent.

Animals ; Cell Membrane ; metabolism ; Connexin 43 ; metabolism ; Heart Ventricles ; Immunoprecipitation ; Male ; Microscopy, Confocal ; Myocytes, Cardiac ; metabolism ; Phosphorylation ; drug effects ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; metabolism ; Receptors, Muscarinic ; metabolism ; Sodium Dodecyl Sulfate ; pharmacology

This study is to explore whether the protective effect of resveratrol on ischemia-reperfusion injury is correlated with the structural and functional association between M3 receptor (M3 subtype of muscarinic acetylcholine receptor) and Cx43 (connexin 43 gap junction proteins). Immunoprecipitation, immunoblotting and immunofluorescence were applied to investigate whether resveratrol has an effect on structural and functional association between M3 and Cx43. The effect of resveratrol on electrocardiogram Lead II ex vivo in rats, SOD (superoxide dismutase) activity and MDA (malondialdehyde) content was also observed in order to evaluate the protective effect of resveratrol on ischemia-reperfusion injury. Resveratrol could restore the structural and functional association between M3 receptor and Cx43 gap junction proteins that was partially destroyed under ischemia-reperfusion injury. The phosphorylation and spatial distribution disturbances in Cx43 expression caused by ischemia-reperfusion injury were also restored. Also, the QRS duration, SOD activity and MDA content were restored. Resveratrol could restore the structural and functional association between M3 receptor and Cx43 gap junction proteins.
Animals ; Connexin 43 ; metabolism ; Electrocardiography ; Heart ; drug effects ; physiopathology ; In Vitro Techniques ; Male ; Malondialdehyde ; metabolism ; Myocardial Reperfusion Injury ; metabolism ; physiopathology ; Myocardium ; metabolism ; Phosphorylation ; drug effects ; Protective Agents ; pharmacology ; Random Allocation ; Rats ; Rats, Wistar ; Receptor, Muscarinic M3 ; metabolism ; Stilbenes ; pharmacology ; Superoxide Dismutase ; metabolism

OBJECTIVETo study the surgical management of metastatic disease in the conjunctive area between the neck and thorax and its efficacy.

METHODSFourteen cases with metastatic node disease in the area between neck and thorax were collected and analysed. Eleven tumors were from the thyroid cancer, and the other three were from the hypopharyngeal cancer, esophagual cancer and malignant pheochromocytoma, respectively. The clavicle was displaced or resected, and the upper half of the manubrium might also be resected when necessary. The recurrent laryngeal nerve and phrenic nerve were exposed and protected. The metastatic disease was completely removed with the internal jugular and/or the brachiocephalic vein resected or spared, depending on the disease condition.

RESULTSIn 10 cases with metastases from the thyroid, no local recurrence was found within the follow-up period from 2 to 5 years. In contrast, no patient with metastatic disease from hypopharyngeal or esophageal cancer survived more than 11 months. No serious complications were found in this group.

CONCLUSIONSThe surgical treatment of node metastases in the conjunctive area between neck and thorax from the well-developed thyroid cancer has promising effect and is comparatively safe.

Adenocarcinoma, Follicular ; surgery ; Adult ; Aged ; Carcinoma, Papillary ; surgery ; Esophageal Neoplasms ; surgery ; Female ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Lymph Node Excision ; Lymphatic Metastasis ; Male ; Neck Dissection ; Thoracotomy ; Thyroid Neoplasms ; surgery ; Thyroidectomy

OBJECTIVETo evaluate the efficacy and safety of postoperative adjuvant chemotherapy with imatinib in gastrointestinal stromal tumor(GIST) patients who had high risk of recurrence.

METHODSA prospective, open-label, multi-center trial conducted in sixteen teaching hospitals in China was carried out. The criteria of the enrolled patients included age more than 18 years old, CD117 positive GIST, tumor size more than 5 cm, pathological mitosis counts more than 5/50 HPF, and treatment beginning within 4 weeks after complete resection and with imatinib (400 mg, once a day) for at least 12 months. The 1, 3 year recurrence rates, disease free survival, overall survival rate and quality of life were evaluated.

RESULTSFrom Aug. 16th 2004 to Sep. 13th 2005, there were totally 74 patients screened and 57 patients (34 men, 23 women) enrolled in the imatinib treatment group. The primary tumors were located in the stomach in 50.9%, the small intestine in 38.6% and the colorectum in 10.5% of the cases. All the patients received radical resection. Until the cut-off date of interim analysis, there was no evidence of tumor relapse or metastasis in all patients and no death was reported either. Among the 57 enrolled patients with intention to treat(ITT), twelve patients finished the protocol (per protocol, PP). The disease free survival was (268.3 +/-120.2) d in ITT analysis, and (396.7+/-38.2) d in the PP analysis. The incidence of adverse effect was 44.4% . The score in quality of life showed no statistically significant difference between the baseline visit and the follow-up visits.

CONCLUSIONImatinib is a promising postoperative adjuvant chemotherapy in GISTs patients with high risk of recurrence, and the adverse effects are receivable.

Adult ; Aged ; Aged, 80 and over ; Benzamides ; Chemotherapy, Adjuvant ; Female ; Gastrointestinal Stromal Tumors ; drug therapy ; Humans ; Imatinib Mesylate ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Piperazines ; therapeutic use ; Postoperative Period ; Prospective Studies ; Pyrimidines ; therapeutic use ; Young Adult

Objective To elucidate the prevalence and the mutation pattern of N348I that related to the resistance seen in the AIDS patients, in China. Methods Partial pol gene of HIV-1 comprising of full protease (PR) and reverse transeriptase (RT) was obtained from plasma samples of therapy-failure individuals (n=614) and therapy-naive individuals (n=619) by using reverse transcription polymerase chain reaction(RT-PCR). 1233 sequences were then submitted to the HIV-1 drug resistance database of the Stanford University to analyze the prevalence and the emergence pattern of N348I. Results The prevalence of N348I was 6.5％ in the therapy-failure patients and 0.8％ in the naive individuals, respectively. The prevalence of N348I was more popular among those patients whose ART regimens containing zidovudine (AZT or ZDV) than those without AZT in regimens( 14.1％ vs. 4.7％, x2=10.21, P＜0.01 ). N3481 always emerged, and combined with others mutations among patients of ART, whose frequencies were: 85.0％ in combination with thymidine analog mutations (TAMs) and 52.5％ with M184V/I, respectively. Conclusion N348I was somehow prevalent in the therapy-failure patients when using the first-line antiretroviral drugs,and it emerged as unique patterns. This study laid the ground in improving the techaology of drug resistance genotypes detection and providing theoretical basis to study the mechanism of resistance and the law of molecular evolution.

The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.
Endophytes/genetics* ; High-Throughput Nucleotide Sequencing ; Plants, Medicinal ; Research Design