BACKGROUNDA nonsteroidal anti-inflammatory drug, indomethacin, has been shown to have anti-leukemic activity and induce leukemic cell apoptosis. This study was to elucidate the mechanism of indomethacin-induced K562 cell apoptosis.

METHODSK562 cells were grown in RPMI 1640 medium and treated with different doses of indomethacin (0 micromol/L, 100 micromol/L, 200 micromol/L, 400 micromol/L, 800 micromol/L) for 72 hours. The cells were harvested, and cell viability or apoptosis was analyzed using MTT assay and AO/EB stain, combining laser scanning confocal microscopy (LSCM) technique separately. For the localization and distribution of intracellular caspase-3 or caspase-8 protein, immunofluorescence assay was carried out. To reveal the activation of caspase-3 or caspase-8 in indomethacin-treated cells, Western blot detection was used. The change in intracellular free calcium was determined by Fluo-3/Am probe labeling combined with LSCM.

RESULTSIndomethacin could lead to K562 cell apoptosis and inhibit cell viability in a concentration-dependent manner. An increased expression of intracellular caspase-3 or caspase-8 was observed at higher doses of indomethacin (400 - 800 micromol/L). Western blot results showed upregulation and activation in both caspase-3 and caspase-8 protein. Under indomethacin intervention, the levels of intracellular free calcium showed a significant increase. Blocking the activity of cyclooxygenase did not abolish the effects of indomethacin on K562 cell apoptosis.

CONCLUSIONSActivation and upregulation of caspase-3 or caspase-8 protein were responsible for Indomethacin-induced K562 cell apoptosis. Variation of intracellular free calcium might switch on the apoptotic pathway and the proapoptotic effect of indomethacin might be cyclooxygenase-independent.

Apoptosis ; drug effects ; Calcium ; metabolism ; Caspase 3 ; Caspase 8 ; Caspases ; genetics ; metabolism ; Cyclooxygenase Inhibitors ; pharmacology ; Enzyme Activation ; Gene Expression Regulation, Enzymologic ; drug effects ; Humans ; Indomethacin ; pharmacology ; K562 Cells

Authors raised that staging based strategies and practice of integrative medicine (IM) by combining syndrome typing and disease identification, and choosing suitable measures in accordance with different persons and seasonal conditions after more than ten years' clinical practice and researches. Radical operation as prior (as evil eliminating) and strengthening vital qi in perioerative period are best strategy for promoting rapid rehabilitation of early stage prostate cancer patients. Strengthening body resistance to eliminate evil was used in treating advanced prostate cancer patients. For example, a comprehensive treatment program for hormone-dependent patients was combined with endocrinotherapy and Chinese herbs for synergisic efficacy-enhancing actions. In this way, these patients' quality of life (QOL) were improved and time to castration resistant prostate cancer (CRPC) was delayed, even some patients were clinically cured. There are lack of effective medicines and methods for CRPC patients. Greatly tonifying original qi is mainly used for improving their clinical symptoms and prolonging survivals. Practice has proved staging based strategies and practice of IM has favorable advantages in treating prostate cancer, especially showing prospect in prolonging survival and postponing progression of advanced prostate cancer patients. Besides, it also could provide beneficial considerations and inspiration for combination of syndrome typing and disease identification.
Disease Progression ; Humans ; Male ; Medicine, Chinese Traditional ; Neoplasm Staging ; Prostatic Neoplasms, Castration-Resistant ; diagnosis ; Quality of Life

Abstract: Objective　To investigate the clustered regularly interspaced short palindromic repeats (CRISPR) genotypes and regional distribution of Yersinia pestis strains in the natural plague foci of Hainan Tibetan Autonomous Prefecture of Qinghai Province (referred to as "Hainan prefecture") and provide a scientific basis for plague prevention and control in this area. Methods　A total of 36 representative Yersinia pestis strains, which were isolated from different host animals and insect vectors from 1954 to 2009 in Hainan Prefecture, were selected as experimental subjects. The DNAs were extracted using the traditional sodium dodecyl sulfate decomposition and phenol-chloroform method. Three pairs of CRISPR primers (YPa, Ypb, YPc) were used for PCR amplification, sequencing and analysis of the DNA of the tested strains, respectively, as a means to identify the CRISPR genotypes of Yersinia pestis in Hainan Prefecture. Results　A total of 17 spacers were observed among 36 strains of Yersinia pestis, including 9 of YPa, 5 of YPb and 3 of YPc. All strains were divided into 5 CRISPR gene clusters (Cb2, Cb4 ', Ca7, Ca7 ', Ca35 ') and 6 genotypes (G1, G9, G22, G22-A1 ', G26-A1 ', G26-A1 'A4 -). The G26-a1 ' was the main genotype, which was distributed in Gonghe, Guide and Xinghai County, and the G22 is the second type, which was distributed in Gonghe and Guide County. Conclusions　The genetic polymorphism of CRISPR loci of Yersinia pestis strains in Hainan was high, and the regional distribution characteristics of Yersinia pestis strains with different genotypes were significant.

OBJECTIVE: To explore the application of quantitative temperature testing (QTT) in forensic identification and clinical diagnosis of neurogenic erectile dysfunction (NED). METHODS: TSA-II-NeuroSensory Analyzer was used to measure the thresholds of four kinds of sensory, including cold, cold pain, heat, heat pain, in 22 normal and 35 NED patients at dorsal glans (DG), left thigh interior (LTI) and left thenar (LT). To calculate the relative thresholds of the sensory mentioned above between DG and LTI (DG/LTI), and between DG and LT (DG/LT). Then to analyze those thresholds and the relative thresholds. RESULTS: NED group showed significant higher threshold than the normal group in DG-heat, DG-heat pain, LTI-heat, LTI-heat pain, DG/LTI-heat, DG/LT-heat, DG/LT-heat pain (P < 0.05). CONCLUSION The threshold of QTT at dorsal glans could be used as an accessory indicator in forensic medicine and clinical diagnosis of NED.
Adult ; Case-Control Studies ; Erectile Dysfunction/physiopathology* ; Hand/physiology* ; Humans ; Male ; Nervous System Diseases/physiopathology* ; Neurologic Examination/methods* ; Pain Threshold ; Penis/physiopathology* ; Sensory Thresholds ; Temperature ; Thermosensing

Objective To investigate the efficacy, safety and tolerance of bosentan, a dual endothelin receptor antagonist, in Chinese patients with idiopathic pulmonary arterial hypertension (IPAH).Methods Totally 79 IPAH patients (hemodynamic criteria confirmed by right heart catheterization) were included in this open-label, prospective multicenter study. Patients received 62. 5 mg of bosentan twice daily for the first 4 weeks, and then up-titrated to 125 mg twice daily for another 12 weeks. The primary end point was the change in exercise capacity showed by six-minute walk distance (6MWD) from baseline to 16 weeks. Secondary end points included the change in World Health Organization (WHO) functional class,Borg dyspnoea scale and systolic pulmonary artery pressure measured by echocardiography. Results The 6MWD increased from (343. 7 ± 93.7) meters at baseline to (397.5 ± 104. 4) meters after 16 weeks ( P ＜0. 01 ), WHO functional class and Borg dyspnoea scale were also significantly improved after 16 weeks therapy compared to baseline levels (all P ＜0. 01 ). Furthermore, the systolic pulmonary artery pressure was significantly decreased from (97.8±25.2) mm Hg (1 mm Hg=0. 133 kPa) to (92.8 ±29.5) mm Hg (P ＜0. 05) after 16 weeks bosentan treatment. There was no patient withdrawal from this study for safety consideration. Conclusion Bosentan therapy is well tolerated and can improve the exercise capacity and WHO functional class in Chinese IPAH patients.

Objective To evaluate the influence and significance of preoperative design of surgical approach on stem cell transplantation via stereotactic surgery. Methods Six patients with stroke in the basal ganglia region were selected. The transplantation target and transcranial approach point were designed by magnetic resonance examination before stem cell transplantation via stereotacfic surgery to guarantee that the line connecting the transplantation target and transcranial approach point could avoid the important functional areas, the ventricular system and the softening focus. Postoperative magnetic resonance examination was performed to observe whether the practical target and surgical approach coincided with the preoperative design or not. Results The practical transplantation target was coincided with the designed transplantation target, distributed around the softening focus without implanted cells in the softening focus. Surgical approach was coincided with the preoperative design and it successfully avoided the important brain functional area, ventricular system and softening focus.Conelnsion The preoperative design of surgical approach can not only ensure the cells being exactly transplanted into the reservation target and guarantee the curative effect, but also promise the surgical approach successfully avoiding the important brain functional area, ventricular system and softening focus and reduce the operative injury.

OBJECTIVETo find out the vector ability and function of Nosopsyllus wualis leizhouensis in the transmitting plague.

METHODSIn T: 19 degrees C +/- 1 degrees C, RH: 85% +/- 5%, data regarding the vector ability as cluster spreading, single flea spreading, single flea transmitting plague to single animal, formative bacterial embolus and infection fleas life-span through experiments was gathered.

RESULTSThe rate of infection on fleas was 94.64%, with 100% transmission rate of colony to spread, and 30% from single flea spreading to single animal. In the experiment of single flea transmission, all of the 388 rattus loseas were bitten by the fleas with bacterial, but only 9 animals were characteristically infected with the transmission potential, vector efficiency, survival potential of embolus, vector index as 0.360, 0.257, 0.868 and 0.223 respectively. The mean survive days of infected flea feed with blood were 17.58 (1 - 58), and the mean survive days of hunger infected flea were 7.25 (1 - 16). Formative bacterial embolus days were 8.80 (2 - 16) and the rate of embolus flea was 78.12%.

CONCLUSIONNosopsyllus wualis leizhouensis could serve as vector and important in the mode of plague transmittion.

Animals ; Female ; Insect Vectors ; microbiology ; Male ; Plague ; transmission ; Rats ; Siphonaptera ; microbiology

OBJECTIVETo investigate the role of CD4+ CD25+ regulatory T (Treg) cells in patients with ITP.

METHODSFlow cytometry was used to examine the number of CD4+ CD25+, CD4+ CD25 high, CD4+ Foxp3+ and CD4+ CD25+ Foxp3+ T cells. The level of Foxp3 mRNA expression was analyzed by realtime quantitative reverse transcriptase polymerase chain reaction (RT-PCR) . CD4+ CD25 high T cells was cocultured with CD4+ CD25 - T cells from patients or controls for assessing the regulatory properties of CD4+ CD25 Treg cells.

RESULTSThe proportion of CD4+ CD25+ T cells in the peripheral blood of patients with ITP [(15.64 +/- 5.82) %] was significantly higher than that in normal control group [(9.30 +/- 3.95)%] (P <0.01). There was no significant difference in the percentages of CD4+ CD25 high T cells between ITP patients and controls [(1.53 +/- 0.66)% versus (1.36 +/- 0.55)% (P = 0.317)]. But the number of CD4 Foxp 3+ T cells and CD4 + CD25+ Foxp3+ T cells in patients were significantly lower than that in control group (P <0.01). The expression of Foxp3 mRNA reduced (P < 0.01) and the suppressive activity of CD4+ CD25 high T cells is lower than that of healthy controls (P <0.01).

CONCLUSIONIn patients with ITP, both the number and immuno-regulative function of CD4+ CD25+ Treg cells are reduced.

Adolescent ; Adult ; Aged ; Female ; Forkhead Transcription Factors ; metabolism ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; metabolism ; RNA, Messenger ; metabolism ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo investigate the relationship between angiotensin converting enzyme (ACE) gene and endothelial dysfunction.

METHODSOne hundred and ten type 2 diabetic patients without angiopathy were selected randomly, and PCR technique was used to determine their ACE genotypes. High resolution ultrasonography was performed to measure the changes in brachial artery diameter at rest, after reactive hyperemia (with increased flow producing an endothelium-dependent dilation) and after sublingual glyceryltrinitrate (GNT, an endothelium-independent dilator). Meanwhile, 50 healthy individuals were selected randomly as controls.

RESULTSIn type 2 diabetes mellitus and control groups, the percentages for flow-mediated arterial dilation in patients with DD genotypes were 3.38% and 3.67% respectively, which were significantly lower than those in patients with II genotypes (4.12% and 4.68% respectively, P<0.05). The baseline blood vessel size, baseline blood flow and GNT induced dilation in both groups showed no significant differences among ACE genotypes (P>0.05). By multiple stepwise regression analysis, reduced flow-mediated arterial dilation was associated with age, baseline vessel size, low density lipoprotein cholesterol(LDL-C), Lp(a), D allele, fasting blood glucose (FBG), postparandial blood glucose (PPBG), HbA1c, duration of diabetes in type 2 diabetic patients (P<0.0005).

CONCLUSIONACE DD genotype is related to endothelium-dependent arterial dilation in the early stage of type 2 diabetes mellitus and in healthy individuals.

Adult ; Aged ; Brachial Artery ; physiopathology ; Diabetes Mellitus, Type 2 ; enzymology ; genetics ; physiopathology ; Diabetic Angiopathies ; genetics ; physiopathology ; Endothelium, Vascular ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymerase Chain Reaction

OBJECTIVETo study the effect of superparamgnetic iron oxides (ferumoxides) on the survival and proliferation of neural stem cells (NSCs) and determine the optimal ferumoxides concentration for labeling.

METHODSBone marrow stromal cells (BMSCs) were obtained from rat femoral marrow and cultured in vitro to induce their differentiation into NSCs. Ferumoxides labeling of the NSCs was performed with different final concentrations of ferumoxides, and the labeling efficiency and viability of the labeled NSCs were evaluated by Prussian blue staining, MTT assay, flow cytometry and transmission electron microscope.

RESULTSThe NSCs could be effectively labeled with ferumoxides with a labeling efficiency of around 90%. Prussian blue staining showed numerous fine granules with blue staining in the cytoplasm of the labeled NSCs, and the intensity of the blue staining was in positive correlation with the ferumoxide concentration for labeling. Transmission electron microscopy of the labeled NSCs revealed the presence of numerous vesicles spreading in the cytoplasm and filled with electron-dense magnetic iron particles. The ferumoxides vesicles increased with the labeling concentration of ferumoxides, and at the final concentration exceeding 25 microg/ml, ferumoxides vesicles in the NSCs gave rise to conglomeration which hampered observation of the cellular ultrastructure by transmission electron microscope. The results of flow cytometry and MTT assay demonstrated that the cell viability, proliferation, differentiation and apoptosis of the labeled cells were affected by ferumoxides at the concentration above 25 microg/ml, but such effects could be minimal at lower concentrations.

CONCLUSIONFerumoxides might be feasible for in vitro labeling of the NSCs with the optimal concentration of 25 microg/ml.

Animals ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Dextrans ; Ferrosoferric Oxide ; Iron ; pharmacology ; Magnetite Nanoparticles ; Male ; Microscopy, Electron, Transmission ; Neurons ; cytology ; ultrastructure ; Oxides ; pharmacology ; Rats ; Stem Cells ; cytology ; ultrastructure