Objective To study the characteristics of mesenchymal stem cells(MSCs)derived from chronic myelogenous leukemia(CML)patients' bone marrow and examine their hematopoiesis support in vitro.Methods MSCs from CML were obtained and cultured.Immunophenotype and in vitro differentiation capacities were investigated.Moreover,the Ph chromosome and bcr/abl gene of CML derived MSCs were detected.The expression of cytokines was detected by RT-PCR,and hema- topoiesis support of MSCs in vitro was detected by long-term bone marrow culture and the methylcel- lulose progenitor assay.Results CML derived MSCs showed a typical fibroblast like morphology. They were positive for CD29,CD44 and CD105,while negative for CD14,CD31,CD45,CD34 and HLA-DR.Under suitable conditions,CML derived MSCs could differentiate into osteoblasts and adi- pocytes.Further,CML derived MSCs showed a normal choromosome,and did not express bcr/abl gene.At last,they expressed hematopoiesis cytokines and possessed ability of hematopoiesis support in vitro.Conclusion There are MSCs with multidirectional differentiation potential in CML bone mar- row and they possess the ability of hematopoiesis support.

According to the transcriptome dataset of Panax notoginseng, the key geranylgeranyl pyrophosphate synthase gene (GGPPS) in terpenoid backbone biosynthesis was selected to be cloned. Using specific primer pairs combining with RACE (rapid amplification of cDNA ends) technique, the full-length cDNA sequence with 1 203 bp, which containing a 1 035 bp open reading frame, was cloned and named as PnGGPPS. The corresponding full-length DNA sequence contained 2 370 bp, consisted of 1 intron and 2 exons. The deduced protein PnGGPPS contained 344 amino acids and shared more than 73% identity with GGPPS from Ricinus communis and Salvia miltiorrhiza. PnGGPPS also had specific Aspartic acid enrichment regions and other conserved domains, which belonged to the Isoprenoid-Biosyn-C1 superfamily. The quantitative real-time PCR showed that PnGGPPS expressed in different tissues of 1, 2, 3 years old root, stem, leaf and 3 years old flower, and the expression level in 3 years old leaf was significant higher than that in other organs, which suggested that it might not only be involved in the regulation of the growth and development, but also be associated with the biosynthesis of chlorophyll and carotenoids, the development of chloroplast, the shade habit and the quality formation of P. notoginseng.
Cloning, Molecular ; Computational Biology ; Geranylgeranyl-Diphosphate Geranylgeranyltransferase ; genetics ; Panax notoginseng ; genetics ; Real-Time Polymerase Chain Reaction

Mesenchymal stem cells (MSCs) are promising candidates to develop new cell-based therapeutic strategies and bone marrow represents the main source of MSCs for experimental and clinical application currently. How to obtain enough MSCs is the biggest challenge faced by the experimental and clinical application. The difference of morphology, growth pattern, immunophenotype and multilineage differentiation of MSCs derived from human umbilical cord vein and their multiple differentiation capacity were investigated. The results showed that MSCs derived from human umbilical cord vein and normal adult bone marrow were similar in biological characteristics. They not only grew in fibroblast-like cells mode but had great expansion potency and multilineage differentiation ability in vitro. The MSCs derived from human umbilical cord vein may be an excellent alternative source for experimental and clinical application.

Objective To explore the clinical features of transbronchial tuberculous mediastinal lymph-adenitis and value of bronchoscopic interventional therapy. Methods The clinical data of 50 patients who had been diagnosed as tuberculous mediastinal lymphadenitis and had received bronchoscopic interventional therapy in our hospital during the period from January 2008 to January 2013 were retrospectively analyzed. The bronchoscopic change , improvement in symptoms , and time to sputum smear and culture conversion were used to assess the therapeutic effect. The patients were followed up for six months. Results The mean age of the patients was (35 ± 15) years and the male to female ratio was 1:1.2. The lesions occurred mostly at the right middle lobe in 24% (12/50) of the patients. The total effectiveness rate was up to 98% (49/50) after chemotherapy and bronchoscopic interventional therapy. The major complication associated with interventional therapy was hemoptysis (8%, 4/50). After follow-up of 6 months , 49 patients with active lesions were stable , with smooth bronchial mucosa and no obvious obstruction by granulation and caseous necrosis tissues. Conclusions The relavent clinical symptoms of transbronchial tuberculous mediastinal lymphadenitis is mainly caused by tuberculosis inflammation which destroys and blocks the airway. The fiber bronchoscopic therapy with forceps clip and drug infusion has a definite effect and fewer complications.

Objective:To analyze and evaluate the teaching effect of the Problem-Based Learning and intelligent simulation training in emergency medicine.Methods:Based on Problem-Based Learning,129 residents of our hospital accepted the cardiopulmonary resuscitation training and test by intelligent simulation training.And their feedback questionnaires were analyzed.Results:After training,their abilities to deal with the cardiac arrest have made a great progress(P

Health self-management group adopts the mode of chronic disease self-management and offers a series of courses for community residents in order to help them carry out health self-management. For healthy people, the program aims at disease prevention by means of improving health literacy, acquiring health knowledge and skills, and developing a healthy lifestyle.While for patients, the program aims to prevent or slow the progression of diseases by means of helping them deal with their condition, achieving disease self-management. By nearly ten years of exploration and practice in Shanghai communities, with the program a working system had been established which was dominated by patriotic health campaign committee office at the levels of city and district, and technologically supported by centers for disease control and prevention with support and cooperation of the town government.The program was implemented by community health service centers and neighborhood ( village ) committee.A large-scale and whole-society promotion had been really achieved in community residents with health self-management. By the end of 2015, health self-management groups had covered all of the city's streets ( town ) and neighborhood ( village ) committee, 26 thousand groups set up and nearly 440 thousand community residents participated in group activities.

OBJECTIVETo investigate the incidence and clinicopathologic significance of MSI and LOH on 3P in breast carcinoma and its precancerous lesions, intraductal papillary adenoma and ductal carcinoma in situ.

METHODS41 paired sporadic invasive breast carcinomas, 13 archival precancerous lesion specimens of the breast and 14 couples of benign hyperplasia were collected. Twelve microsatellites on chromosomes 2p, 3p, 5q, 6q, 16q, 17q, eleven markers on chromosome 3p were amplified for MSI and LOH, respectively, by polymerase chain reaction ( PCR ) with designed primers and detecting after polyacrylamide gel electrophoresis. In addition, the expression of protein of hMSH2, hMLHI, FHIT, ER, and PR were detected by immunohistochemistry.

RESULTSMSI was observed, at least two microsatellite markers, in 15 out of 41 (36. 6%) of the carcinomas, almost all belonging to poorly or intermediately differentiated carcinoma. Instability was shown in 9 of the 13 cases of precancerous lesions, but only 2 among them had more than 2 MSI sites. There was no MSI in benign hyperplasia. MSI was targeted predominately at D3S1766, D2S2739 in both carcinomas and precancerous lesions. Of the 11 loci examined, D3S1295, D3S1029 and D3S1038 were identified as the locus with most frequent LOH which were all correlated significantly with some clinicopathological parameters such as histological type, lymph node metastasis in breast cancer, while D3S1295 and D3S1029 were the most frequent markers in precancerous lesions. LOH of D3S1295 had significant correlation with negative expression of FHIT. Positive expression of hMLH1 and hMSH2 protein was detected in breast carcinomas in scattered distribution and their positive rate was 45% and 40% , respectively. In precancerous lesions, hMLH1 and hMSH2 protein showed diffuse expression and their positive rate was 61. 54% and 76. 92% , respectively, significantly lower than that in the control tissues.

CONCLUSIONDefective expression of MMR genes is closely associated with the development of breast cancer. Genomic instability might play a role in the early stage during multi-step mammary carcinogenesis. MSI indicates poor histological differentiation in breast carcinoma. D3S1766 and D2S2739 might be the sensitive sites to detect MSI in breast carcinoma and precancerous lesions. The smallest common LOH deletion regions seem likely to be situated between 3p14 and 3p25, indicating the existence of breast tumor related genes in those regions and some of them might affect tumor development.

Acid Anhydride Hydrolases ; metabolism ; Adaptor Proteins, Signal Transducing ; metabolism ; Adenoma ; genetics ; metabolism ; pathology ; Adult ; Aged ; Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; genetics ; metabolism ; pathology ; Chromosome Deletion ; Chromosomes, Human, Pair 3 ; genetics ; DNA Mismatch Repair ; Female ; Humans ; Hyperplasia ; Immunohistochemistry ; Loss of Heterozygosity ; Lymphatic Metastasis ; Microsatellite Instability ; Middle Aged ; MutL Protein Homolog 1 ; MutL Proteins ; Neoplasm Proteins ; metabolism ; Neoplasm Staging ; Nuclear Proteins ; metabolism ; Polymerase Chain Reaction ; Precancerous Conditions ; genetics ; metabolism ; pathology ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

This study was aimed to observe the biological characteristics of cryopreserved bone marrow mesenchymal stem cell (MSC) and to examine their abilities to support in vitro hematopoiesis. Bone marrow MSC were cryopreserved in -196 degrees C liquid nitrogen for 4 weeks (short term) and 9-15 months (medium term) with IMDM containing 10% DMSO, 40% fetal calf serum as cryoprotectant. The viability, proliferation, immunophenotype, in vitro differentiation and ability of supporting hematopoiesis of thawed MSC were investigated and compared with these of pre-cryopreserved MSC. The results showed that the cell viability were (93 +/- 2.51)% and (90 +/- 3.75)% for MSC cryopreserved as long as 4 weeks or 9-15 months respectively. However, there were no changes detected, as compared with pre-cryopreserved MSC in immunophenotype, abilities of proliferation and supporting colony forming of CFU-GM, CFU-E and CFU-GEMM. It is concluded that bone marrow-derived MSC can be stored in liquid nitrogen for short-term (4 weeks) or medium-term (9-15 months) without changes of abilities of proliferation, differentiation and hematopoiesis support.
Blood Preservation ; methods ; Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Colony-Forming Units Assay ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Dimethyl Sulfoxide ; pharmacology ; Hematopoiesis ; physiology ; Humans ; Mesenchymal Stromal Cells ; cytology ; physiology

OBJECTIVETo study the effect of p38 MAPK activity on isolated rabbit liver during the period of cold preservation and reperfusion.

METHODSBased on the cold preservation-reperfusion model of isolated rabbit livers, according to the concentration of SB202190 in the preservation solution which was a specific p38 MAPK inhibitor, the isolated livers were divided into 4 groups, six in each A, B, C and D. Liver tissue samples and blood samples were harvested at different time points: before and end of cold preservation, reperfusion for 5, 10, 15, 30, 60, and 120 minutes. The activity levels of p38 MAPK were detected by both western blot and immunoprecipitation. The function markers of isolated livers were detected with automatic biochemistry analyzer, and the levels of total bile after reperfusion were measured.

RESULTSIn normal rabbit liver tissues, p38 MAPK had low activity. During the cold preservation period, the activity of p38 MAPK elevated slightly. But during the reperfusion period, the activity of p38 MAPK changed markedly which elevated rapidly at the early stage and reached its peak value at 10 minutes, then decreased gradually to the normal level (7.6 +/-0.9) at 120 minutes. SB202190 could inhibit the activity in a dose-dependent manner. The peak values of p38 MAPK activity in group B,C and D were 42.5 +/-2.4, 10.1+/-1.4, and 7.6 +/-0.6 respectively, while 78.6 +/-6.1 in group A. During the reperfusion period, the levels of serum ALT, AST and ALP were higher in group A than those in any other group, alike the p38 MAPK activity, especially at 15 and 30 minutes. On the other hand, the total bile secretion volume was (10.2 +/-2.9) ml/100 g, (13.9 +/-1.3) ml/100 g, (15.6 +/-1.2) ml/100 g, and (16.0 +/-1.3) ml/100 g in group A, B, C and D respectively.

CONCLUSIONSDuring the cold preservation- reperfusion period, p38 MAPK specific inhibitor SB202190 can lower the p38 MAPK activity, and ameliorate the isolated liver injury. Activation of p38 pathway is one of the important mechanisms to cause ischemia-reperfusion injury of isolated liver.

Animals ; Enzyme Inhibitors ; pharmacology ; Female ; Hepatocytes ; cytology ; physiology ; Imidazoles ; pharmacology ; In Vitro Techniques ; Liver ; blood supply ; cytology ; Male ; Protein Kinase C ; metabolism ; Pyridines ; pharmacology ; Rabbits ; Reperfusion Injury ; enzymology ; physiopathology ; Signal Transduction ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism

Objective To establish a method for qualitative and quantitative analysis of Zhendian Kaiqiao Granules. Methods Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba in the preparation were identified by TLC. Gentiopicrin and paeoniflorin were determined by HPLC. The analysis was performed on a Phenomenex C18 column (250 mm × 4.6 mm, 5 μm), with the mobile phase of methyl alcohol-water (12:88). The flow rate was 1.0 mL/min and the column temperature was maintained at room temperature; the detection wavelengths were set at 273 nm (gentiopicrin) and 230 nm (paeoniflorin). Results Gentiane Radix et Rhizoma, Scuteliariae Radix, Rhei Radix et Rhizoma, Radices Paeoniae Alba could be detected by TLC. Gentiopicrin showed a good linear relationship at a range of 2.396–11.980 μg, r=0.999 6. The average recovery was 99.72%, and RSD was 2.70%. Paeoniflorin showed a good linear relationship at a range of 2.728–13.640 μg, r=0.999 0. The average recovery was 98.74%, and RSD was 2.42%. Conclusion The established method is simple, reliable and reproductive.The method can be used for control quality of Zhendian Kaiqiao Granules.