OBJECTIVETo observe the differentiation of bone mesenchymal stem cells (BMSCs) co-cultured with purified sinoatrial node cells (SNC) of neonate rats.

METHODSSNC from neonatal SD rat were cultured and purified with differential attachment method and labeled with BrdU. Rat BMSCs were isolated by a Percoll's gradient solution and cultured in DMEM. After 2 passages, these BMSCs were transfected with pEGFP-N1 by Lipofectamine and labeled with GFP. EGFP-BMSC were co-cultured with SNC in a rate of 1:5 for 1 week. EGFP-BMSC cultured in SNC culture medium served as controls. SNC marker hyperpolarization activated cyclic nucleotide gated cation channel 4 (HCN4) and connexin 45 (Cx45) expressions were determined by immunofluorescence staining.

RESULTPositive immunofluorescence staining against HCN4 and Cx45 were detected in EGFP-BMSC co-cultured with SNC but not in EGFP-BMSC cultured in SNC culture medium.

CONCLUSIONDirect cell-to-cell contact between BMSCs and SNC cells may induce BMSCs differentiation into sinus node-like cells.

Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; Cell Differentiation ; Cell Separation ; Cells, Cultured ; Coculture Techniques ; Mesenchymal Stromal Cells ; cytology ; Rats ; Rats, Sprague-Dawley ; Sinoatrial Node ; cytology

This study was aimed to explore the inhibitory effect of triptolide on proliferation and inducing apoptosis effect of K562/G01 cells and their possible mechanism. MTT assay was used to detect the effect of imatinib or triptolide alone and their combination on K562/G01 proliferation; the cell cycle, apoptosis rate, P-gp protein expression were detected by flow cytometry (FCM); the expression of P-gp was assessed by Western blot; the BCR/ABL gene expression was assayed by real time quantitative PCR. The results showed that triptolide could enhance the effect of imatinib on proliferation inhibition and apoptosis of K562/G01, arrested the cell cycle in G1 phase, down-regulated the expression of BCR/ABL gene and P-gp protein. It is concluded that triptolide induces K562/G01 cell proliferation inhibition and apoptosis, the mechanism may be related to cell cycle arrest, decrease of P-gp protein expression, inhibition of BCR/ABL gene expression.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Apoptosis ; drug effects ; Benzamides ; pharmacology ; Cell Cycle Checkpoints ; Cell Proliferation ; drug effects ; Diterpenes ; pharmacology ; Drug Resistance, Neoplasm ; Epoxy Compounds ; pharmacology ; Fusion Proteins, bcr-abl ; genetics ; Humans ; Imatinib Mesylate ; K562 Cells ; Phenanthrenes ; pharmacology ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology

Objective:To investigate the relationship between mitogen-activated protein kinase (MAPK) signaling pathway related signal molecules and the apoptosis of acute promyelocytic leukemia NB4 cells induced by puerariae radix flavones (PRF) and its significance.Methods:The cells were divided into control group [0.025% dimethyl sulfoxide (DMSO) to replace PRF] and 10, 30, 50 μg/ml PRF groups. The proliferation inhibition rate of NB4 cells exposed with PRF for 24, 48 and 72 hours was determined by methyl thiazolyl tetrazolium (MTT) method, and the nuclear morphology was determined by confocal laser scanning microscope after 48 hours. NB4 cells were divided into control group (adding 0.025% DMSO) and 10, 30 and 50 μg/ml PRF with or without 10 μmol/L c-Jun N-terminal kinase (JNK) inhibitor (SP600125) group, and the cells were treated for 48 hours and the changes in the expressions of MAPK pathway related proteins JNK, tumor necrosis factor α (TNF-α), extracellular signal-regulated kinase (ERK) and p38 MAPK were tested by Western blot.Results:10, 30 and 50 μg/ml PRF inhibited the proliferation of NB4 cells in 24, 48 and 72 hours, which was in time- and dose-dependent manners (all P < 0.05). The half-maximal inhibitory concentration (IC 50) at 24, 48 and 72 hours were (40.03±2.23) μg/ml, (22.92±1.72) μg/ml and (17.99±1.48) μg/ml, respectively. The confocal laser scanning microscope showed that NB4 cells displayed distinct apoptotic characteristics after PRF treatment. After co-cultivating NB4 cells with 10 μmol/L SP600125 and different concentrations of PRF for 48 hours, the expression of JNK1 in NB4 cells was suppressed ( P < 0.05), and the expressions of JNK2/3 and p38 MAPK decreased, but the differences were not statistically significant (both P > 0.05). The expressions of ERK1 and ERK2 gradually increased in the single-drug group, while the expression in the combined drug group decreased. The expression of TNF-α in the 50 μg/ml PRF+SP600125 group was down-regulated compared with the 50 μg/ml PRF single-drug group, while the expressions in the 10 and 30 μg/ml PRF+SP600125 groups were up-regulated compared with the 10 and 30 μg/ml PRF single-drug groups. Conclusion:10-50 μg/ml PRF may activate the MAPK signaling pathway through TNF-α. JNK, ERK1/2 and p38 MAPK interact with each other to activate pro-apoptotic related proteins and induce NB4 cells apoptosis.

OBJECTIVETo examine whether the polymorphisms of endothelial nitric oxide synthase (eNOS) gene are associated with the susceptibility to high altitude pulmonary edema (HAPE) in Chinese railway construction workers at Qinghai-Tibet where the altitude is over 4 500 m above sea level.

METHODSA case-control study was conducted including 149 HAPE patients in the construction workers and 160 healthy controls randomly recruited from their co-workers, matching the patients in ethnicity, age, sex, lifestyle, and working conditions. Three polymorphisms of eNOS gene, T-786C in promoter, 894G/T in exon 7, and 27bp variable number tandem repeat (VNTR) in intron 4, were genotyped using polymerase chain reaction (PCR) and confirmed with DNA sequencing.

RESULTSThe frequencies of 894T allele and heterozygous G/T of the 894G/T variant were significantly higher in HAPE patients group than in the control group (P=0.0028 and P=0.0047, respectively). However, the frequencies of the T-786C in promoter and the 27bp VNTR in intron 4 were not significantly different between the two groups. Haplotypic analysis revealed that the frequencies of two haplotypes (H3,T-T-b, b indicates 5 repeats of 27 bp VNTR; H6, C-G-a, a indicates 4 repeats of 27 bp VNTR) were significantly higher in HAPE patients (both Pü0.0001). On the contrary, the frequencies of H1 (T-G-b) and H2 (T-G-a) were lower in HAPE patients than in healthy controls (both Pü0.001).

CONCLUSIONSTwo haplotypes (T-T-b and C-G-a) may be strongly associated with susceptibility to HAPE. Compared with the individual alleles of eNOS gene, the interaction of multiple genetic markers within a haplotype may be a major determinant for the susceptibility to HAPE.

Adolescent ; Adult ; Altitude ; Base Sequence ; Case-Control Studies ; DNA Primers ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Nitric Oxide ; blood ; Nitric Oxide Synthase Type III ; genetics ; Occupational Diseases ; enzymology ; genetics ; Polymorphism, Genetic ; Pulmonary Edema ; enzymology ; genetics ; Tibet ; Young Adult

OBJECTIVETo evaluate the results of surgical procedures for pulmonary embolism.

METHODSFifty-four patients of pulmonary embolism received surgical treatment from October 1994 to June 2007, of which 9 were acute pulmonary embolism underwent pulmonary embolectomy and 45 patients were chronic thromboembolic pulmonary hypertension (CTEPH) underwent pulmonary thromboendarterectomy.

RESULTSThe mortality rate was 44.4% in acute pulmonary embolism group and 13.3% in CTEPH group (P < 0. 05). Thirteen patients had residual pulmonary hypertension and 23 patients had severe pulmonary reperfusion injury postoperatively. The pulmonary artery systolic pressure changed from (89.4 +/- 36.3) mm Hg (1 mm Hg =0.133 kPa) preoperative to (55.6 +/- 22.4) mm Hg postoperative. The pulmonary vascular resistance changed from (89. 7 +/- 56.7) kPa L(-1) S(-1) preoperative to (38.9 +/- 31.1) kPa L(-1) S(-1) postoperative. The arterial partial pressure of oxygen changed from (52. 3 +/- 6.7 ) mm Hg preoperative to (87.6 +/- 6.5) mm Hg postoperative. The arterial oxygen saturation changed from (88.9 +/- 4.5)% preoperative to (95.3 +/- 2.8 )% postoperative (P < 0.05). With the follow-up of (41.8 +/- 36.4) months, there were 4 patients died. According to NYHA, there were 28 patients for class I , 10 patients for class II and 2 patients for class III. According to Kaplan-Meier survival curve, the 3-year, 4-year, 5-year and 8-year survival rate were (97.1 +/- 2.8 )%, (94.0 +/- 4.1)%, (90.8 +/- 5.2)% and (85.0 +/- 7.3)% respectively. Linear rate of bleeding and thromboembolic related to anticoagulation were 0. 63% patient-years and 0. 62% patient-years respectively.

CONCLUSIONSThe operational mortality of acute pulmonary embolism is significantly higher than CTEPH, and the mid-long term survival rate is agreeable and the complication rate related to anticoagulation is relatively low.

Adolescent ; Adult ; Aged ; Embolectomy ; methods ; Endarterectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Pulmonary Artery ; surgery ; Pulmonary Embolism ; pathology ; surgery ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo compare the relative merits between chordal shortening and artificial chordae to repair the anterior leaflet prolapses (ALP).

METHODSThe clinic data of 50 cases underwent repair of ALP from March 1990 to March 2008 were analyzed retrospectively. There were 29 male and 21 female patients with a mean age of (42.6 +/- 11.3) years old. There were 23 patients in chordal shortening group and 27 patients in artificial chordae group.

RESULTSThere were 3 operative deaths in chordal shortening group (13.0%), and 1 death in artificial chordae group (3.7%, P = 0.199). With a mean follow-up of (5.8 +/- 4.8) years and a total follow-up of 278 patient-years, there were 3 late deaths respectively in each group. According the Kaplan-Meier survival curve, the actuarial survival rate at 5-8 years was 70.0% +/- 18.2% for chordal shortening group and 86.8% +/- 9.2% for artificial chordae group (chi(2) = 8.17, P = 0.046). There were 5 reoperations, of which 4 in chordal shortening group and 1 in artificial chordae group. According to the Kaplan-Meier freedom from reoperation curve, the freedom from reoperation at 5 years was 83.3% +/- 15.2% for chordal shortening group and 100% for artificial chordae group (chi(2) = 12.06, P = 0.007). The COX proportional hazard regressions revealed that chordal-shortening technique was the independent risk predictor for the late cardiac event after ALP surgical repair.

CONCLUSIONArtificial chordae techniques has a relative superiority to chordal shortening for repair of mitral valve ALP.

Adolescent ; Adult ; Aged ; Child ; Chordae Tendineae ; surgery ; Female ; Follow-Up Studies ; Heart Valve Prosthesis Implantation ; Humans ; Male ; Middle Aged ; Mitral Valve Prolapse ; surgery ; Retrospective Studies ; Treatment Outcome

OBJECTIVETo explore the way of promoting the efficacy of surgical treatment for ventricular septal rupture (VSR) after acute myocardium infarction in terms of perioperative and long term survival.

METHODSThe clinic data of 37 VSR cases underwent surgical treatment from October 1994 to October 2007 were analyzed retrospectively. There were 24 male and 13 female, and the age was (63.4 +/- 7.6) years old. The whole group was divided into the VSR repair plus revascularization group (group A, 26 cases) and simple VSR repair group (group B, 11 cases).

RESULTSThere were 4 operative deaths in group A (15.4%), 7 deaths in group B (63.6%), P = 0.006. With the follow-up of (34.0 +/- 29.8) months ranged from 2 to 103 months of the 26 operational survivors, there were 5 late deaths, of which 2 deaths in group A and 3 deaths in group B. According to the Kaplan-Meier survival curve, the actuarial survival rate at 6 to 8 year was (64.3 +/- 21.0)% for group A and the actuarial survival rate at 4 year was (25.0 +/- 21.7)% for group B, P = 0.011. Of the 21 mid-long term survivors, 17 cases were in NYHA class I to II and 4 cases in NYHA class III to IV. There were 4 cases suffered from VSR recurrence. According to Logistic regression, the risk factors for the early death were not adoptive of revascularization, cardiogenic shock and emergency surgical procedure, while the risk factors for late death were not adoptive of revascularization and low cardiac output after the procedures.

CONCLUSIONSVSR repair plus revascularization could improve the perioperative and mid-long term survival for the surgical treatment of VSR. The appropriate timing and procedures of the surgical operation are very important to promote perioperative survival and to prevent VSR recurrence.

Aged ; Female ; Follow-Up Studies ; Heart Rupture, Post-Infarction ; surgery ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; complications ; Retrospective Studies ; Treatment Outcome ; Ventricular Septal Rupture ; etiology ; surgery

OBJECTIVETo compare the efficacy of conservative or pulmonary thromboendarterectomy (PTE) therapy for chronic thromboembolic pulmonary hypertension (CTEPH) patients according to a new clinical classification scheme.

METHODSThis retrospective study analyzed 63 cases of CTEPH admitted to our hospital from February 1995 to October 2007 and 45 cases were treated surgically (Group A) and 18 cases received conservative therapy (Group B). Results were analyzed using Fisher exact test and t test according to San Diego medical center quartering classification scheme and Anzhen Hospital modified bifurcate classification scheme.

RESULTSThere were 6 operational deaths in Group A and 2 deaths during hospital stay in Group B. During follow-ups (mean 3.6 +/- 2.5 years), there were 4 deaths in Group A and 9 deaths in Group B. the totality survival rate is significantly higher in Group A than that in Group B (P < 0.05). For patients with San Diego Type I CTEPH, survival rate was significantly higher in Group A compared with Group B (P = 0.009) and was similar for patients with type II and III and IV CTEPH between the two groups (P = 0.338, 0.455, 0.800). Survival rate was significantly higher in Group A than that in Group B for patients with Anzhen central type CTEPH (P = 0.009), but was similar between the two groups for patients with Anzhen peripheral type CTEPH (P = 0.125). The Kaplan-Meier survival curve 5 years survival rate in the Group A was (91.7 +/- 8.0)% for Anzhen central type and (76.0 +/- 8.5)% for Anzhen peripheral type (P = 0.04), and the 5 years Kaplan-Meier survival rate in the Group B was (42.9 +/- 18.7)% for Anzhen central type and (56.2 +/- 10.8)% for Anzhen peripheral type (P = 0.851).

CONCLUSIONAnzhen Hospital modified bifurcate classification scheme is a simple and effective classification to predict the prognosis and choose treatment method of CTEPH.

Adult ; Chronic Disease ; Female ; Humans ; Hypertension, Pulmonary ; mortality ; surgery ; therapy ; Male ; Middle Aged ; Prognosis ; Pulmonary Embolism ; surgery ; therapy ; Retrospective Studies ; Survival Rate ; Treatment Outcome

Osteosarcoma is a rare primary malignancy of bone that is prone to early metastasis. Resection surgery and chemotherapeutic regimens are current standard treatments for osteosarcoma. However, the long-term survival rate of patients with osteosarcoma is low due to a high risk of metastasis. Hence, a new approach is urgently needed to improve the treatment of osteosarcoma. Compared with chemotherapy, natural active constituents isolated from herbs exhibit less adverse effects and better anti-tumor effects. This study aimed to summarize the anticancer effects of constituents of herbs on the progression and metastasis of osteosarcoma cells. It showed that many constituents of herbs inhibited osteosarcoma by targeting proliferation, matrix metalloproteinases, integrin and cadherin, and angiogenesis. The findings might be beneficial for the development of new drugs and treatment strategies.

Objective:To study the effect of combined decoction and single decoction of Houpu Wenzhongtang on rats with deficiency-cold of spleen and stomach from the perspective of metabonomics, to find out the relevant potential biomarkers and metabolic pathways, and to explore the similarities and differences between the combined decoction and single decoction, so as to provide reference for the feasibility analysis of replacing traditional decoction with single dispensing granule of this formula. Method:SD rats were randomly divided into normal group, model group, Houpu Wenzhongtang combined decoction group and simgle decoction group. Rats in the normal group were given distilled water by intragastric administration, rats in the other three groups were given cold vinegar at 4 ℃ in the morning and refined lard in the afternoon for 10 days (the dosage of 10 mL·kg^-1). After the model was successfully established, rats in the combined decoction group and the single decoction group were given corresponding decoction with dosage of 1.8 g·kg^-1 (according to the amount of crude drugs), once a day for 7 days. Ultra-high liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) technique was used to analyze the small molecular endogenous metabolites in urine. Orthogonal partial least squares-discriminant analysis (OPLS-DA) and principal component analysis (PCA) were used to compare the changes of differential metabolites among the normal group, model group, Houpu Wenzhongtang combined decoction group and single decoction group, and the differential metabolites were introduced into Kyoto Encyclopedia of Genes and Genomes (KEGG) for metabolic pathway analysis. Result:Compared with the model group, the Houpu Wenzhongtang combined decoction group and single decoction group jointly regulated 13 potential biomarkers, including phosphatidylcholine(PC), lysophosphatidic acid(LysoPA) and cholic acid, etc. They played a role in treating deficiency-cold of spleen and stomach by influencing metabolic pathways such as glycerophospholipid metabolism, glycerolipid metabolism, phosphatidylinositol signaling system and so on. The combined decoction and single decoction of Houpu Wenzhongtang could obviously restore the body weight, motilin and gastrin contents of rats with deficiency-cold of spleen and stomach to normal levels. Conclusion:According to biochemical indexes, there is no obvious difference between combined decoction and single decoction of Houpu Wenzhongtang, but according to metabonomics, the combined decoction may be slightly better than the single decoction. The research shows that it is feasible to replace traditional decoction with single dispensing granule of Houpu Wenzhongtang in clinical application.