Aim The aim of this work was to study the effect of diallyl disulfide (DADS) on human Leukemia cell line HL 60, and investgate the mechanisms of its antitumor effect. Methods HL 60 cells growth inhibition was measured by MTT assay. Cell apoptosis was inspected by flow cytometry,TUNEL assay and acridine orange fluorescent staining methods. The protein levels of Bcl 2, Bax were determined using immunohistochemical technique. Results MTT assay showed that DADS significantly inhibited the growth of HL 60 cells. After 24 hours of exposure to DADS, Partial cells presented characteristic morphological changes of apoptosis under the electron microscope, including cell shrinkage, nuclear condensation, and formation of apoptotic bodies. Some typical subdiploid peaks before G 0/G 1 phase were observed. Flow cytometry analysis revealed that the apoptosis rates were increased and the TUNEL assay showed the apoptosis index increased in occordance with increase of concentration of DADS. SP immunohistochemistry revealed that the Bax expression was increased while Bcl 2 expressed was decreased. Conclusion Diallyl disulfide could significantly induce apoptosis of human Leukemia cell HL 60, Apoptosis of tumor cells is closely associated with down regulation of the ratio of bcl 2/bax.

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

Objective: Cell cycle has recently become more appealing as a new target of anti-carcinogen-ic agent. Diallyl disulfide (DADS) inhibits growth and induces call cycle G_2/M arrest in human gastric cancer BGC823 cells. Cell division cycle protein 25C (Cdc25C) and CyclinB1 expression are involved in G_2/M arrest.However, mechanisms of G_2/M arrest are not yet fully understood. The aim of this study was to elucidate the mechanism of cell cycle G_2/M arrest in human gastric cancer BGC823 cells induced by DADS. Methods: The expression of chk1 and Chk2 mRNA associated with cell cycle arrest of BGC823 cells after the induction with DADS for 1 or 2 days was detected by RT-PCR. The protein expression of cycle-related proteins ATM-RAD3-related gene (ATR), checkpoint kinase1 (Chk1), checkpoint kinase 2 (Chk2), P-ATR, P-Chk1 and P-Chk2 was measured by Western blot. Interaction between Chk1/2 and Cdc25C was analyzed by immuno-precipitation. Results: After the cells were treated with 15 mg/L DADS for 1 or 2 days, the expression of Chk1 and Chk2 mRNA was not significantly different from that in untreated cells (P>0.05). Western blot analysis showed that the expression of total Chk1 and Chk2 treated with 15 mg/L DADS was not significantly different from that in untreated cells. But phospho-chk1 showed a significant increase after stimulation with 15 mg/L DADS for 2h to 12h and continued to increase gradually as time went on (P<0.05). Phospho-Chk2 showed a eak expression and a weaker expression after stimulation with DADS, but the changes were not statistically significant (P>0.05). Addition of 15 mg/L DADS to BGC823 cells for 15 rain to 120 min resulted in an increase in phospho-ATR expression, whereas no changes were found in ATR expression (P<0.05). The Chk1 Ab in-creasingly precipitated Cdc25C in BGC823 cells treated with DADS (P<0.05). In contrast, Chk2 Ab failed to change precipitation with Cdc25C by DADS (P>0.05). Conclusion: Activation of chk1 was involved in cell cy-cle G_2/M arrest in BGC823 cells treated with DADS. Cell cycle G_2/M arrest by DADS is associated with phos-phorylation of several cell cycle regulatory proteins including ATR and Chk1 which regulate expression of Cdc25C.

In the current study, nine flavonoids were isolated and purified from 75% ethanol extract of Selaginella uncinata (Desv.) Spring by column chromatographic techniques over macroporous resin, polyamide, silica gel, Sephadex LH-20 and pre-HPLC. On the basis of their physico-chemical properties and spectroscopic data analyses, these compounds were elucidated as cirsimarin (1), nepitrin (2), apigenin-6-C-α-L-arabinopyranosyl-8-C-β-D-glucopyranoside (3), apigenin-6-C-β-D-glucopyranosyl-8-C-α-L-arabinopyranoside (4), apigenin-7-O-β-D-glucopyranoside (5), 2,3-dihydroamentoflavone (6), 4'-O-methylamentoflavone (7), 2,3-dihydro-4'-O-methyl-amentoflavone (8), and 2,3,2",3"-tetrahydron-4'-O-methyl-robustaflavone (9). Compounds 1-5 belong to flavonoid glycosides and were isolated from the genus Selaginella for the first time.
Flavonoids ; analysis ; Selaginellaceae ; chemistry

Objective: To develop a health management system for outpatient follow-up of kidney transplant patients. Methods: Access 2010 database sotfware was used to establish the health management system for kidney transplantation patients in Windows XP operating system. Database management and post-operation follow-up of the kidney transplantation patients were realized through 6 function modules including data input, data query, data printing, questionnaire survey, data export, and follow-up management. Results: The system worked stably and reliably, and the data input was easy and fast. The query, the counting and printing were convenient. Conclusion: Health management system for patients after kidney transplantation not only reduces the work pressure of the follow-up staff, but also improves the effciency of outpatient follow-up.

ObjectiveTo study the effect of behavior training on the expression of NR2B around the infarcted focus and in the cortex of temporal lobe in rats with bilateral hippocampal infarction.Methods54 SD rats were randomized into pre-training, 7 d, 14 d, 21 d after training groups, pre-immobilization, 7 d, 14 d, 21 d immobilization groups and control group. Behavior training and immobilization were performed on the 3rd day after the infarction. Immunohistochemistry was used to detect the expression of NR2B around the infarcted focus and in the cortex of temporal lobe at different points of time.ResultsThe expression of NR2B was abundant in the normal hippocampus and cortex of temporal lobe of rats. The expression of NR2B decreased after infarction and increased after behavior training. However, the expression of NR2B had little increased in immobilization groups, and showed significant difference compared with that in behavior training groups (P<0.01).ConclusionBehavior training can accelerate the expression of NR2B around the infarcted focus and in the cortex of temporal lobe in rats with bilateral hippocampal infarction.

The success of solid organ transplantation in the treatment of end-stage organ failure has fuelled a growing demand for transplantable organs worldwide that has far outstripped the supply from brain dead heart-beating donors. In Singapore, this has resulted in long waiting lists of patients for transplantable organs, especially kidneys. The Human Organ Transplant Act, introduced in 1987, is an opt-out scheme that presumes consent to removal of certain organs for transplantation upon death. Despite this legislation, the number of deceased organ donors in Singapore, at 7 to 9 per million population per year, remains low compared to many other developed countries. In this paper, we reviewed the clinical challenges and ethical dilemmas encountered in managing and identifying potential donors in the neurological intensive care unit (ICU) of a major general hospital in Singapore. The large variance in donor actualisation rates among local restructured hospitals, at 0% to 56.6% (median 8.8%), suggests that considerable room still exists for improvement. To address this, local hospitals need to review their processes and adopt changes and best practices that will ensure earlier identification of potential donors, avoid undue delays in diagnosing brain death, and provide optimal care of multi-organ donors to reduce donor loss from medical failures.
Brain Death ; Donor Selection ; Humans ; Intensive Care Units ; Presumed Consent ; ethics ; Referral and Consultation ; Singapore ; Terminally Ill ; Tissue and Organ Procurement ; organization & administration ; Waiting Lists

BACKGROUND: Loss of nipple projection is a common problem following nipple reconstruction. The aim of this study was to demonstrate that the use of a tightly rolled dermal graft is effective in the long-term maintenance of nipple projection. METHODS: Nipple reconstruction was performed using the C-V flap technique. A dermal graft was harvested from the dog-ear portion of previous scars. The graft was rolled tightly into a compact cylinder and used to augment the nipple reconstruction. Postoperatively, stacked Allevyn dressing was used for protecting the nipple from compression for a minimum of two months. Nipple projection was measured at the time of surgery and at 12 months postoperatively. RESULTS: Forty nipple reconstructions were performed using this technique. There were 19 transverse rectus abdominis musculocutaneous (TRAM) flaps, 10 latissimus dorsi (LD) flaps, and 11 tissue-expanded breast mounds. At one year, the mean projection was 0.80 cm (range, 0.62-1.22 cm). The twelve-month average maintenance of nipple projection was 70.2% for the TRAM flap group, 76.3% for the LD flap group, and 61.8% for the tissue-expanded group. In two patients with previous irradiation of the reconstructed breasts, relatively poor maintenance of nipple projection was noted (45.7%). No complications were noted, and all of the donor sites healed well primarily. CONCLUSIONS: Our results demonstrated that the use of a C-V flap with a tightly rolled dermal graft for nipple reconstruction improves the long-term maintenance of nipple projection. Its advantages include reproducibility, technical simplicity, cost-effectiveness, and minimal donor site morbidity.
Bandages ; Breast ; Cicatrix ; Dermis ; Female ; Humans ; Mammaplasty ; Nipples* ; Rectus Abdominis ; Superficial Back Muscles ; Tissue Donors ; Transplants*

OBJECTIVETo examine the changes of expressions of orexin A, orexin receptor-1 (OX1R), prepro-orexin (Prepro-OX) mRNA, OX1R mRNA and ob-R of hypothalamus in rats with chronic renal failure (CRF).

METHODSSixty-two male Wister rats weighing 200-250 g were divided into three groups, including group 1 (normal, n = 5), group 2 (sham-operated, n = 25) and group 3 (CRF, n = 32). Hypothalamus orexin A was assayed by radioimmunoassay. Serum leptin was assayed by enzyme linked immunosorbent assay. The expression of Prepro-OX mRNA and OX1R mRNA of hypothalamus were measured by reverse transcription polymerase chain reaction, and expression of orexin A, OX1R and ob-R by immunohistochemistry. Automatic biochemical analyzer was used to measure the serum creatinine.

RESULTSHypothalamus orexin A levels were negatively correlated (r = -0.63, P < 0.001) with serum leptin levels in the rats. The expression of hypothalamus Prepro-OX mRNA in CRF rats was significantly lower than that of sham-operation at week 12 (P < 0.01). Hypothalamus Prepro-OX mRNA levels were negatively correlated (r = -0.81, P < 0.001) with the levels of serum leptin and serum creatinine (r = -0.68, P < 0.05) in the rats at week 12. The expression of hypothalamus OX1R mRNA in CRF rats was lower than that of sham-operation at week 12 (P > 0.05). Specific immunoreactivity for orexin A was present in perikeryon of the hypothalamus neuron. Specific OX1R-like immunoreactivity was observed in some nerve fibres. Specific immunoreactivity for ob-R was present in membranes of the hypothalamus neuron. Hypothalamus neurons of orexin A-like specific immunoreactivity in CRF rats were significantly fewer than those in shamoperated rats at week 8. Hypothalamus neurons of OX1R-like specific immunoreactivity in CRF rats were similar to those in sham-operated rat at week 8. Hypothalamus neurons of ob-R-like specific immunoreactivity in CRF rats were significantly more than those in sham-operated rats at week 8.

CONCLUSIONSThe lower hypothalamus orexin A levels may be induced by high serum leptin level in CRF rats. The lower expression of hypothalamus Prepro-OX mRNA in CRF rats may be one of the main causes inducing lower hypothalamus orexin A. The expression of OX1R in hypothalamus neurons is somewhat reduced and the expression of ob-R in hypothalamus neurons is somewhat raised in CRF rats. These remain to be studied further.

Animals ; Carrier Proteins ; genetics ; metabolism ; Hypothalamus ; metabolism ; Intracellular Signaling Peptides and Proteins ; Kidney Failure, Chronic ; metabolism ; Leptin ; genetics ; metabolism ; Male ; Neuropeptides ; genetics ; metabolism ; Neurotransmitter Agents ; genetics ; metabolism ; Orexin Receptors ; Orexins ; Protein Precursors ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Receptors, Cell Surface ; genetics ; metabolism ; Receptors, G-Protein-Coupled ; Receptors, Leptin ; Receptors, Neuropeptide ; genetics ; metabolism

Objective To explore knowledge, attitude, practice (KAP) and related determinants on nutrition among caregivers of those rural stranded children under 7 years of age in China and to provide evidence for setting up relevant health education program. Methods 1691 caregivers of the stranded children randomly selected were surveyed by a standard questionnaire. Logistic regression models were used to screen the determinants on KAP regarding nutrition. Results Rates on awareness, positive attitude and approprite behavior were lower in caregivers of children whose parents both left (47.8%, 55.4%, 41.8%, respectively) the countryside, when compared with those only one parent was away from home (59.9%, 59.5%, 38.0%, respectively). Data from multivariable logistic regression models showed that caregivers' KAP on nutrition was related to age, educational background, average family income, and willingness on the job as well as the age of the child. Conclusion Improving caregivers' KAP on nutrition and setting up appropriate health education program were in urgent need.