Aged ; Drug Resistance, Bacterial ; Humans ; Intensive Care Units ; Male ; Microbial Sensitivity Tests ; Pneumoconiosis ; microbiology ; Sputum ; microbiology

BACKGROUNDKeratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.

METHODSA K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer. RPMI-1640 supplemented with 10% fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44(+)/CD24(-) as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog, N-cadherin, and E-cadherin) was analyzed with real-time PCR.

RESULTSMuch higher percentage of CSCs was achieved with K-SFM: 17.3% for MCF-7 cells, 17.4% for SKBR-3, and 20.0% for primary breast cancer culture. Less than 1% CSC was achieved using RPMI-1640 supplemented with 10% FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4, ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.

CONCLUSIONK-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; Cadherins ; genetics ; Cell Culture Techniques ; methods ; Cell Line, Tumor ; Culture Media, Serum-Free ; Female ; Homeodomain Proteins ; genetics ; Humans ; Keratinocytes ; cytology ; Nanog Homeobox Protein ; Neoplasm Proteins ; genetics ; Neoplastic Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; genetics ; Real-Time Polymerase Chain Reaction

OBJECTIVETo introduce a new procedure of laparoscopic adrenal surgery.

METHODSThirty patients with adrenal disease underwent surgery by a 70 degrees lateral decubital trans-abdominal laparoscopic approach, which wa compared with 30 degrees decubital laparoscopic operation in 26 patients.

RESULTSThe procedure is superior to open laparotomy. There was significant difference in average operative time (77.2 min vs. 215.7 min), blood lost volume (27.0 ml vs. 94.5 ml), and postoperative complications (13.3% vs. 46.0%). Postoperative feeding time and hospitalization time were shortened markedly.

CONCLUSIONSThis modified technique is safe, straightforward, with a good anatomic view during operation. It could be widely adopted for the treatment of benign adrenal disease, especially pheochromocytoma.

Adrenal Gland Diseases ; surgery ; Adrenalectomy ; methods ; Female ; Humans ; Intraoperative Complications ; prevention & control ; Laparoscopy ; methods ; Male ; Postoperative Complications ; prevention & control ; Posture ; Treatment Outcome

OBJECTIVETo evaluate the efficacy and safety of thrombopoietin (TPO) on platelet engraftment in hematological malignancies patients after allogeneic haematopoietic stem cell transplantation (allo-HSCT).

METHODSOne hundred and twenty patients were enrolled in a multicenter, open-label, randomized, controlled clinical trial, and were randomized into 4 treatment groups following allo-HSCT. Group A was the control arm without TPO, while group B, C and D were trial arms with received 300 U×kg(-1)×d(-1) of TPO starting from day +1, +4 and +7, respectively. A total of 89 cases were evaluated, of which 22 cases in group A, 23 in group B, 20 in group C and 24 in group D. Efficacy evaluation (the time of platelet engraftment, the number of platelet transfusion) and safety evaluation \[adverse events, routine blood tests, liver and renal function, coagulation function and occurrence of graft-versus-host disease (GVHD)\] were observed.

RESULTSThe median platelet engraftment time in experimental groups (groups B, C and D) were on day (13.17 ± 2.89), day (12.15 ± 2.08), day (12.33 ± 1.76), respectively, and that in control group was on day (14.82 ± 5.05). There was statistically significant difference between two groups (P = 0.029), There were no statistically significant difference in the average amount of platelet transfusion, platelet engraftment time, and platelet nadir value among the 3 experimental groups. No significant adverse events were observed in experimental groups.

CONCLUSIONSTPO administration following allo-HSCT for patients with hematologic malignancies appears to shorten platelet engraftment time. TPO given starting from day +7 is effective and safe.

Adolescent ; Adult ; Blood Platelets ; Child ; Female ; Hematologic Neoplasms ; surgery ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Platelet Transfusion ; methods ; Thrombopoietin ; therapeutic use ; Transplantation, Homologous ; Young Adult

AIMTo investigate the protective effect of lumbrokinase against myocardial ischemia and to further explore its underlying mechanisms.

METHODSThe effect of lumbrokinase on myocardial ischemia was observed by a model of acute myocardial infarction due to permanent ligation of the left anterior descending coronary artery in rats. Patch-clamp technique and laser scanning confocal microscopy were utilized to study the action of lumbrokinase on L-type calcium current (ICa-L) and intracellular calcium concentration ([Ca2+]i).

RESULTSLumbrokinase decreased the infarct size of myocardium in a dose-dependent manner. The inhibitory rate of lumbrokinase at the dose of 20, 40 and 80 mg x kg(-1) was 7.7%, 34.6% and 46.2%, respectively. The electrophysiological studies displayed that, at + 10 mV, the ICa-L was markedly reduced from (-14.42 +/- 1.53) pA/pF to (-11.33 +/- 1.40) pA/pF (decreased by 21.4%, P <0.01) and (-9.92 +/- 1.31) pA/pF (decreased by 36.5%, P <0.01) by lumbrokinase (10 and 50 micromol x L(-1)), respectively. Confocal experiments showed that 10 micromol x L(-1) lumbrokinase showed no obvious effects on [Ca2+]i at resting states (P > 0.05). However, the increase of [Ca2+]i induced by 60 mmol x L(-1) KCl was distinctly limited by 10 micromol x L(-1) lumbrokinase (P <0.01). Within 240 s, the no obvious peak value of fluorescent intensity (FI) was shown.

CONCLUSIONLumbrokinase showed protective action against myocardial infarction in rats. The possible mechanisms of anti-ischemia could be attributed to decreasing ICa-L and [Ca2+] of ventricular myocytes in rats.

Animals ; Calcium ; metabolism ; Calcium Channels, L-Type ; metabolism ; Cardiotonic Agents ; pharmacology ; Endopeptidases ; pharmacology ; Female ; Heart Ventricles ; Male ; Myocardial Infarction ; metabolism ; pathology ; Myocardium ; pathology ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar

Background Keratinocyte serum-free medium (K-SFM) is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells (CSCs) using K-SFM.Methods A K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer.RPMI-1640 supplemented with 10％ fetal calf serum (FCS) was used as a control. CSCs were identified with flow cytometry using CD44+/CD24-as molecular markers. The expression of a variety of CSC markers (Oct-4, ABCG2, Nanog,N-cadherin, and E-cadherin) was analyzed with real-time PCR.Results Much higher percentage of CSCs was achieved with K-SFM: 17.3％ for MCF-7 cells, 17.4％ for SKBR-3, and 20.0％ for primary breast cancer culture. Less than 1％ CSC was achieved using RPMI-1640 supplemented with 10％ FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4,ABCG2, Nanog and N-cadherin, and lower level of E-cadherin.Conclusion K-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

OBJECTIVETo investigate the effect of phenyl-hexyl isothiocyanate (PHI) on acetylation and methylation of histone in acute lymphoblastic leukemia cell line Molt4.

METHODSThe inhibition of cell proliferation was observed by MTT method and clone suppression test. Apoptosis and cell cycle arrest were measured by flow cytometry. The alterations in histone acetyltransferase and acetylation and methylation of histones were detected by Western blot.

RESULTSPHI could up-regulate the expression of acetyltransferase (P300/CBP), markedly induced the accumulation of acetylated histone H3, H4 and methylated histone H3 lysine 4 (H3K4), and inhibited methylation on lysine 9 of H3 (H3K9). The epigenetic regulation resulted in cell cycle arrest at G0/G1 phase, and induction of apoptosis.

CONCLUSIONSPHI can modulate both histone methylation and acetylation. It may serve as a histone deacetylase inhibitor, and might be a potential novel anti-leukemia agent.

Acetylation ; drug effects ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Epigenesis, Genetic ; Histone Deacetylases ; metabolism ; Histones ; metabolism ; Humans ; Isothiocyanates ; pharmacology ; Methylation ; drug effects

OBJECTIVETo construct a multiple myeloma (MM)-specific APE1siRNA expression vector, and detect the specific knock-down effect of the siRNA on expression of APE1 protein.

METHODSAPE1siRNA cDNA sequence was designed, synthesized and inserted into pSilencer 2.0-U6 linear expression vector. pSilencer APE1siRNA was digested by enzyme EcoRI and BamHI, then linear vector and IgP fragments were conjugated by T4 DNA ligase. pSilencer IgP-APE1siRNA and pSilencer IE-IgP-APE1siRNA were digested by enzyme EcoRI or XhoI. Linear vector and IE or Kappa fragments were conjugated by T4 DNA ligase. Then a MM specific pSilencer K-IE-IgP-APE1siRNA was cloned. The recombinant products were identified by DNA sequencing and enzyme digestions at each step. pSilencer K-IE-IgP-APE1siRNA plasmid was transfected to KM3, HOS, MDA-231 cells by liposome. APE1 gene silence induced by RNAi was analysed by Western blot.

RESULTSAPE1 protein in KM3 cells could be knocked down effectively and specifically by pSilencer K-IE-IgP-APE1siRNA vector. After 2 days, the level of APE1 protein in KM3 cells transfected with siRNA was 0.118 +/- 0.047, while that transfected with plasmid only was 0.988 +/- 0.029. The efficiency of gene silence was 90%.

CONCLUSIONA MM specific APE1siRNA expression vector was successfully constructed.

Base Sequence ; Cell Line, Tumor ; Cloning, Molecular ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; genetics ; Genetic Vectors ; genetics ; Humans ; Molecular Sequence Data ; Multiple Myeloma ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection

The distribution information of Lomatogonium rotatum. was collected by interview investigation and field survey, and 55 related environmental factors were collected, the habitat suitability study was conducted based on geographic information system (GIS) and maximum entropy model. The AUCs of ROC curve were both above 0.99, indicating that the predictive results with the maximum model were highly precise. The results showed that 13 major environmental factors have obvious influence on ecology suitability distributions of L. rotatum, including month average temperature of February et al., the suitable distribution areas are mainly concentrated in the east-central of Inner Mongolia, including Hexigten banner, Duolun county, Zhenglan banner et al., The zoning results basically coincide with the genuine producing areas, and further afford new suitable distribution areas, which can provide reference for L. rotatum's wild nursery and the siting of introduction and cultivation.
China ; Ecosystem ; Environment ; Gentianaceae ; growth & development ; Geographic Information Systems ; Rain ; Temperature

Objective To assess the laparoscopic technique combined with open surgical technique in pyeloplasty.Methods Overall,45 patients with ureteropelvic junction obstruction underwent laparo- scopic dissection of the renal pelvis and upper ureter transperitoneally,and pyeloplasty was performed through a expanded trocar-incision(extension of 1-2 cm)as open surgery was performed.Results The opera- tion was successful in all 45 patients.The mean operative time was 58 min(range,40-85 min),and the mean blood loss was 22 ml(range,15-30 ml).No complication was observed during and after operation. Follow-up for 3-36 months was available in 34 patients.Intravenous urography(IVU)showed no obstruc- tion of the anastomotic stoma,and B-ultrasound indicated relief of hydronephrosis.Conclusions Laparo- scopic approach combined with open surgery in pyeloplasty is an effective way to treat ureteropelvic junction obstruction.This technique can simplify the operative manipulation and shorten the operative time without more trauma to the patients.It is worth general application in clinical practice.