Role of glycogen synthase kinase 3ß in protective effect of propofol against hepatic ischemia-reperfusion injury.

BACKGROUND: It was previously reported that propofol, an intravenously administered hypnotic and anesthetic agent, protects organs from ischemia-reperfusion (I/R) injury. However, the underlying mechanisms are largely unknown. Glycogen synthase kinase 3ß (GSK-3ß) is known to play an important role in the oxidative stress-induced apoptosis. In this study, we investigated the role of GSK-3ß and mitochondrial permeability transition pore (MPTP) in the protective effects of propofol against hepatic I/R injury. MATERIALS AND METHODS: The left and median hepatic artery and the portal vein branches were blocked by no-damage artery clips to create the model of partial ischemia (70%), and liver lobes were subjected to warm ischemia for 30, 60, 90 min, respectively. Reperfusion of 120 min was then initiated by the removal of clamp. The MPTP opening was assessed by measuring mitochondrial large amplitude swelling and mitochondrial membrane potential. RESULTS: Pretreatment with propofol in conditions of hepatic I/R inhibits the apoptosis of hepatocytes as evidenced by decreased terminal deoxynucleotidyl transferase dUTP nick end labeling-positive cells. Importantly, propofol suppressed the mitochondrial GSK-3ß by promoting or preserving its phosphorylation at Ser9, thus restraining the opening of MPTP and preventing the mitochondrial swell and mitochondrial membrane potential collapse. CONCLUSIONS: Propofol protects liver from I/R injury by sustaining the mitochondrial function, which is possibly involved with the modulation of MPTP and GSK-3ß.

Testing the mutant selection window in rabbits infected with methicillin-resistant Staphylococcus aureus exposed to vancomycin.

OBJECTIVES: To test the mutant selection window (MSW) hypothesis with Staphylococcus aureus exposed to vancomycin in an animal model and to compare in vivo and in vitro exposures that restrict the enrichment of resistant mutants. METHODS: Local infection with S. aureus was established in rabbits, and the infected animals were treated with various doses of twice-daily vancomycin (half-life 6 h) for 3 consecutive days to provide antibiotic concentrations below the MIC, between the MIC and the mutant prevention concentration (MPC), and above the MPC. Changes in susceptibility and the numbers of surviving organisms were monitored daily on agar plates containing 2× and 4× MIC of vancomycin. RESULTS: S. aureus lost vancomycin susceptibility when drug concentrations at the site of infection fluctuated between the lower and upper boundaries of the MSW, defined in vitro as the MIC(99) and the MPC, respectively. Both boundaries were determined in vitro, before starting animal studies. The value at which resistant mutants are not enriched in vivo was estimated as an AUC(24)/MPC value of ∼15 h, where AUC(24) is the area under the drug concentration time curve in a 24 h interval. The estimated anti-mutant AUC/MIC ratio in vivo was ≥200 h. CONCLUSIONS: These findings support the MSW hypothesis and the anti-mutant AUC/MIC ratio estimated in vivo is consistent with that reported in in vitro studies. Keeping vancomycin concentrations above the MPC or AUC(24)/MPC >15 h is a straightforward way to restrict the acquisition of resistance.

[Effects of ischemic postconditioning on liver regeneration and energy metabolism in rats].

OBJECTIVE: To explore the effects of ischemic postconditioning on the remnant liver regeneration under ischemic reperfusion after subtotal hepatectomy. METHODS: A total of 90 male SD rats weighting 230-280 g were randomly divided into 3 groups of simple subtotal hepatectomy (SH), ischemia reperfusion (IR) and ischemic postconditioning (IPO). The left and middle liver lobes were resected. And the remnant liver was treated as follows: blood flow was non-blocked in SH group, but blocked for 30 min in IR group, then reperfused. The IPO group was established by 30-30 s intermittent interruptions of blood flow thrice during the early phase of reperfusion. At 1, 6, 12, 24, 48 h respectively, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), hepatocyte growth factor (HGF) and the positive ratio of proliferating cell nuclear antigen (PCNA) were detected. Also the contents of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) were measured and the regenerated liver weight was calculated. RESULTS: The activities of ALT and AST in IPO group from 1 - 48 h were (138 ± 20), (340 ± 22), (770 ± 55), (389 ± 60), (113 ± 25) U/L and (247 ± 31), (509 ± 51), (972 ± 77), (955 ± 159), (222 ± 44) U/L respectively at each time point and they were significantly lower than those (294 ± 37), (560 ± 69), (1222 ± 162), (540 ± 40), (161 ± 15) U/L and (439 ± 34), (669 ± 94), (1347 ± 118), (1274 ± 223), (403 ± 68) U/L in IR group (P < 0.05); the regenerated liver weight (18.8 ± 3.2, 34.0 ± 3.5%) and positive cell ratio of PCNA (43.9 ± 2.7, 56.2 ± 4.0%) in IPO group at 24 and 48 h were respectively higher than those in IR group ((11.3 ± 2.9), (26.8 ± 2.5)% and (34.5 ± 2.8), (48.8 ± 1.8)%)(P < 0.05); The HGF levels ((261 ± 54) and (252 ± 103) pg/ml) at 24 and 48 h in IPO group were significantly higher than those ((99 ± 47) and (70 ± 19) pg/ml) in IR group (P < 0.05); the contents of ATP in hepatic tissue at 24 and 48 h in IPO group were (22.20 ± 3.6) and (12.87 ± 2.49) µg/g respectively. And they were significantly higher than those in IR group (P < 0.05), but lower than those in SH group (P < 0.05). CONCLUSION: Ischemic postconditioning improves the remnant liver regeneration under ischemic reperfusion after subtotal hepatectomy through its actions of stimulating HGF production and maintaining hepatic energy metabolism.

[The impact of ischemic postconditioning on the tumor necrosis factor-α/IL-6/signal transducers and activators of transcription-3 signal pathway of liver regeneration].

OBJECTIVE: To investigate the impact of the ischemic postconditioning on the tumor necrosis factor (TNF)-α/IL-6/signal transducers and activators of transcription (STAT)-3 signal pathway of liver regeneration. METHODS: Ninety healthy clean grade male Sprague-Dawley rats weighting 230 to 280 g were selected and assigned into three groups randomly: group I subtotal hepatectomy (SH), group II ischemia reperfusion (IR), group III ischemic postconditioning (IPO). The left and middle liver was resected, and the remnant liver was treated as followed: the blood flow was not blocked in SH group, but blocked 30 minutes in IR group, then reperfused; IPO groups received three cycles of 30 s-30 s intermittent interruptions of blood flow at onset of reperfusion. At 1, 6, 12, 24, 48 h after reperfusion, the serum TNF-α, IL-6 was detected and the mRNA of cyclinD1, Cdk4, STAT-3 was assayed by real-time PCR as well as the protein expression of cyclinD1 and Cdk4 by Western blot. RESULTS: Compared with SH group, the expression of IL-6 declined at each set time point in IR group (t = 5.076 to 8.334, P = 0.000), but the content of TNF-α increased in early stage (1 to 12 h) (t = 2.972 to 7.215, P = 0.000 - 0.014). The expression of STAT-3, cyclinD1 and Cdk4 mRNA and protein of cyclinD1 and Cdk4 at 24 and 48 h after reperfusion were lower in IR group than in SH group (t = 2.857 to 6.684, P = 0.000 to 0.017). However, there was a significant decrease in TNF-α from 1 to 12 h after reperfusion (t = 2.995 to 4.112, P = 0.002 to 0.017), but a significant increase in IL-6 in IPO group than in IR group (t = 2.458 to 3.543, P = 0.005 to 0.034). The expression of STAT-3, cyclinD1, Cdk4 mRNA and protein of cyclinD1 and Cdk4 at 24 and 48 h after reperfusion were all increased in IPO group in comparison with in IR group (t = 2.383 to 6.803, P = 0.000 to 0.038). CONCLUSIONS: The ischemic postconditioning could promote the remnant liver regeneration after subtotal hepatectomy with ischemia reperfusion injury. Its mechanism relates with the activation of the TNF-α/IL-6/STAT-3 signal pathway of and the cyclinD1-Cdk4 complex which enhances the proliferation of hepatocyte.

One-pot hydrothermal synthesis of Si-doped carbon quantum dots with up-conversion fluorescence as fluorescent probes for dual-readout detection of berberine hydrochloride.

Here, the high fluorescent silicon-doped carbon quantum dots (Si-CQDs) were prepared by a facile and one-pot hydrothermal assay using 3-aminopropyltrimethoxysilane as the carbon and silicon source. The prepared Si-CQDs exhibit favorable water-soluble, high-temperature resistance, acid resistance, alkali resistance, high ionic strength resistance, high photostability, film-forming ability and solid-state fluorescence. Compared to other Si-CQDs that have been reported, the prepared Si-CQDs show unique up-conversion fluorescence. Furthermore, it is found that berberine hydrochloride (BH) can effectively quench the down- and up-conversion fluorescence of the Si-CQDs, making it can be used as a highly sensitive and specific probe for BH dual-mode sensing. Meanwhile, the linear range of down-conversion fluorescence detection for BH is 0.5-30.0 µmol/L with a limit of detection (LOD) of 50 nmol/L, and the linear range of up-conversion fluorescence assay for BH is 0-25.0 µmol/L. The mechanism of down-conversion fluorescence quenching by BH was investigated through a series of studies. The results show the quenching mechanism is the inner filter effect (IFE). Moreover, this proposed strategy has been well used to analyze BH in urine samples with satisfactory results.

[Randomized clinical case-control trial for the comparison of docetaxel plus thiotepa versus docetaxel plus capecitabine in patients with metastatic breast cancer].

OBJECTIVE: To evaluate the efficacy and safety of docetaxel plus thiotepa(TXT/TSPA) and docetaxel plus capecitabine(TXT/CAPE) in patients with metastatic breast cancer. METHODS: The patients were randomized to give intravenous TXT 35 mg/m2 on days 1 and 8 plus intravenous TSPA 60-65 mg/m(2) on day 1 every 3 weeks, or intravenous TXT 35 mg/m(2) on days 1 and 8 plus oral CAPE 1 000 mg/m(2) twice daily on days 1 to 14 every 3 weeks, at least 2 cycles applied. RESULTS: TXT/TSPA group (22 patients) and TXT/CAPE group (24 patients) had consistent baseline. Docetaxel thiotepa group (21 cases) and docetaxel combined with capecitabine group (22 cases) were evaluated for their clinical responses, which showed that 2 of the 21 (9.52%) from TXT/TSPA group and 6 of the 22 (27.27%) from TXT/CAPE group had achieved partial remission; 11 of the 21 (52.38%) from TXT/TSPA group versus 7 of the 22 (31.82%) from TXT/CAPE group for stable diseases; 8 of the 21 (38.10%) from TXT/TSPA group versus and 9 of the 22 (40.91%) from TXT/CAPE group for progressive diseases, respectively. The disease control rate was 61.90% (13/21) and 59.09% (13/22) for TXT/TSPA and TXT/CAPE groups, the median progression-free survival(PFS) was 7.9 months [95% confidence interval(CI) 0.77 to 15.03] from TXT/TSPA group versus 8.3 months (95% CI 4.01 to 11.79) from TXT/CAPE group. One year survival rate was 88.20% for TXT/TSPA versus 81.00% for TXT/CAPE group, respectively. P values all exceeded 0.05, and the two groups showed no difference. No chemotherapy-related deaths occurred. Myelosuppression was the major side effect. The adverse events of grades 3 to 4 respectively occurred in TXT/TSPA and TXT/CAPE groups:leucocytopenia was 45.45% vs. 26.09%; neutropenia 45.45% vs. 21.74%; thrombocytopenia 9.09% vs. 0%; hand-foot syndrome 0% vs. 13.04%. P values all exceeded 0.05, and the two groups showed no difference. CONCLUSION: Combination of docetaxel and thiotepa in the treatment of metastatic breast cancer has some curative effect and adverse reactions can be tolerated. It can be used as an economical and effective rescue plan.

2,2'-[(1E)-3-Phenyl-prop-2-ene-1,1-di-yl]bis-(3-hy-droxy-5,5-dimethyl-cyclo-hex-2-en-1-one).

In the title mol-ecule, C(25)H(30)O(4), the two cyclo-hexene rings adopt envelope conformations. The two hy-droxy groups are involved in the formation of intra-molecular O-H⋯O hydrogen bonds. In the crystal structure, weak inter-molecular C-H⋯O hydrogen bonds link mol-ecules related by translation along the axis a into chains.

Mobilization of Peripheral Blood Stem Cells Using Regimen Combining Docetaxel with Granulocyte Colony-stimulating Factor in Breast Cancer Patients.

OBJECTIVE: To evaluate the effectiveness and safety of the mobilization of peripheral blood hematopoietic stem cells by combining docetaxel with granulocyte colony-stimulating factor (G-CSF) in breast cancer patients. METHODS: A total of 57 breast cancer patients were treated with docetaxel 120 mg/m(2). When the white blood cell (WBC) count decreased to 1.0×10(9)/L, patients were given G-CSF 5 µg/kg daily by subcutaneous injection until the end of apheresis. Peripheral blood mononuclear cells (MNC) were isolated by Cobe Spectra Apheresis System. The percentage of CD34(+) cell was assayed by flow cytometry. RESULTS: At a median 6 of days (range 3-8) after the administration of docetaxel, the median WBC count decreased to 1.08×10(9)/L (range 0.20-2.31). The median duration of G-CSF mobilization was 3 days (range 2-7). The MNC collection was conducted 8-12 days (median 10 days) after docetaxel treatment. The median MNC was 5.35×10(8)/kg (range 0.59-14.07), the median CD34(+) cell count was 2.43×10(6)/kg (range 0.16-16.69). The CD34(+) cell count was higher than 1.00×10(6)/kg in 47 of 57 cases (82.46%) and higher than 2.00×10(6)/kg in 36 cases (63.16%). The CD34(+) cell count was higher than 2.00×10(6)/kg in 27 collections (23.68%). The MNC count and the CD34(+) cell count were correlated with the bottom of WBC after docetaxel chemotherapy (r=0.364, 0.502, P=0.005, 0.000). The CD34(+) cell count was correlated with the MNC count (r=0.597, P=0.000). The mobilization and apheresis were well tolerated in all patients. Mild perioral numbness and numbness of hand or feet were observed in 3 cases. No serious adverse events were reported. CONCLUSION: Mobilization of peripheral blood hematopoietic stem cell by combining docetaxel with G-CSF was effective and safety in breast cancer patients.

Multicenter clinical study for evaluation of efficacy and safety of transdermal fentanyl matrix patch in treatment of moderate to severe cancer pain in 474 chinese cancer patients.

OBJECTIVE: Although a new matrix formulation fentanyl has been used throughout the world for cancer pain management, few data about its efficacy and clinical outcomes associated with its use in Chinese patients have been obtained. This study aimed to assess the efficacy and safety of the new system in Chinese patients with moderate to severe cancer pain. METHODS: A total of 474 patients with moderate to severe cancer pain were enrolled in this study and were treated with the new transdermal fentanyl matrix patch (TDF) up to 2 weeks. All the patients were asked to record pain intensity, side effects, quality of life (QOL), adherence and global satisfaction. The initial dose of fentanyl was 25 µg/h titrated with opioid or according to National Comprehensive Cancer Network (NCCN) guidelines. Transdermal fentanyl was changed every three days. RESULTS: After 2 weeks. The mean pain intensity of the 459 evaluated patients decreased significantly from 5.63±1.26 to 2.03±1.46 (P<0.0001). The total remission rate was 91.29%, of which moderate remission rate 53.16%, obvious remission rate 25.49% and complete remission rate 12.64%. The rate of adverse events was 33.75%, 18.78% of which were moderate and 3.80% were severe. The most frequent adverse events were constipation and nausea. No fatal events were observed. The quality of life was remarkably improved after the treatment (P<0.0001). CONCLUSION: The new TDF is effective and safe in treating patients with moderate to severe cancer pain, and can significantly improve the quality of life.

The complete chloroplast genome sequence of rare and endangered Camellia pubipetala Y. Wan & S. Z. Huang (Theaceae) of South China.

The complete chloroplast genome sequence of rare and endangered Camellia pubipetala Y. Wan & S. Z. Huang (Theaceae) was mentioned in this research. By studying comparatively, we found that the C. pubipetala Y. Wan & S. Z. Huang chloroplast genome was 156,993 bp in length and composed of 86,590 bp LSC, 18,211 bp SSC, and two reverse repeating regions with 26,090 bp. The whole GC content was 37.33%. The genome encoded 116 functional genes, including 80 protein-coding genes, 32 tRNA genes, and 4 rRNA genes. In order to find the phylogenetic relationship of C. pubipetala Y. Wan & S. Z. Huang within Camellia genus, we reconstructed phylogenetic tree. The results indicate that C. pubipetala Y. Wan & S. Z. Huang was closely related to Camellia huana voucher and Camellia ptilosperma.

Vitamin D-vitamin D receptor system down-regulates expression of uncoupling proteins in brown adipocyte through interaction with Hairless protein.

Our previous study showed that feeding mice with vitamin D deficiency diet markedly alleviated high-fat-diet-induced overweight, hyperinsulinemia, and hepatic lipid accumulation. Moreover, vitamin D deficiency up-regulated the expression of uncoupling protein 3 (Ucp3) in white adipose tissue (WAT) and brown adipose tissue (BAT). The present study aimed to further investigate the effects of vitamin D and vitamin D receptor (Vdr) on Ucp1-3 (Ucps) expression in brown adipocyte and the mechanism involved in it. Rat primary brown adipocytes were separated and purified. The effects of the 1,25(OH)2D3 (1,25-dihydroxyvitamin D3; the hormonal form of vitamin D) and Vdr system on Ucps expression in brown adipocytes were investigated in basal condition and activated condition by isoproterenol (ISO) and triiodothyronine (T3). Ucps expression levels were significantly down-regulated by 1,25(OH)2D3 in the activated brown adipocyte. Vdr silencing reversed the down-regulation of Ucps by 1,25(OH)2D3, whereas Vdr overexpression strengthened the down-regulation effects. Hairless protein did express in brown adipocyte and was localized in cell nuclei. 1,25(OH)2D3 increased Hairless protein expression in the cell nuclei. Hairless (Hr) silencing notably elevated Ucps expression in activated condition induced by ISO and T3. Moreover, immunoprecipitation results revealed that Vdr could interact with Hairless, which might contribute to decreasing expression of Vdr target gene Ucps. These data suggest that vitamin D suppresses expression of Ucps in brown adipocyte in a Vdr-dependent manner and the corepressor Hairless protein probably plays a role in the down-regulation.

Efficacy of fiberoptic bronchoscopy and bronchoalveolar lavage in childhood-onset, complicated plastic bronchitis.

BACKGROUND: Plastic bronchitis (PB) is a rare, variable, and potentially fatal disease. This study aimed to assess the efficacy of fiberoptic bronchoscopy (FOB) and bronchoalveolar lavage (BAL) in treating children with PB. METHODS: In total, 15 children with PB, between 2012 and 2020, were enrolled in our study. Within 12 hours of admission, FOB and BAL were performed and reviewed under local anesthesia and sedation. Before and after FOB, clinical findings and chest imaging were evaluated. RESULTS: Regarding the onset of symptoms before FOB, all cases had prominent cough for 7.00 ± 4.55 days, and 14 had persistent high fever. In total, 13 cases had complete obstruction from bronchial casts, consistent with consolidated lesions; 2 had partial airway obstruction. Within 3 days, complete resolution was revealed in nine cases. Overall, six cases underwent repeated FOB (range, 2-3 times) for persistent atelectasis and airway obstruction. Except for two cases with type 2 PB, cast histology confirmed type 1 PB for all cases. Only eight children had minor intra- and post-procedure complications. Reverse transcription-polymerase chain reaction for Mycoplasma pneumoniae in sputum and BAL samples were positive in 13 cases. Next-generation sequencing of the BAL samples was positive for adenovirus and Human parainfluenza virus in one case, respectively. During 1 month to 7 years of follow-up, no patient developed PB recurrence, asthmatic attacks, or chronic cough. CONCLUSIONS: Early FOB and BAL were effective in alleviating clinical findings, atelectasis, and airway obstruction. Serial FOB could be performed in patients with recurrent symptoms.

Cloning and characterization of two chlorophyll A/B binding protein genes and analysis of their gene family in Camellia sinensis.

In this study, two chlorophyll A/B binding protein (CAB) genes (CsCP1 and CsCP2) in tea plant were cloned. The proteins encoded by these genes belong to the external or internal antenna proteins of PS II, respectively. They may be the targets of physiological regulation for tea leaf cell PS II because they all contain multiple functional domains and modifiable sites. The CAB gene family in the tea genome consists of 25 homologous genes. We measured the expression patterns of ten genes in the CsCP1 and CsCP2 subfamily under six different stresses. CsCP1 expression was inhibited in response to 6 kinds of stress; CsCP2 expression was slightly upregulated only after cold stress and ABA treatment. However, the expression levels of CSA016997 and CSA030476 were upregulated significantly in the six stresses. The results suggested that the 10 CAB genes may have different functions in tea leaves. Moreover, changes in the expression of the 10 genes under stress appear to be related to ABA- and MeJA-dependent signalling pathways, and their responses to MeJA treatment is faster than those to ABA. In addition, we introduced our experiences for cloning the genes in the context of complex genomes.

A molecular detection approach for a cotton aphid-parasitoid complex in northern China.

Aphid-parasitoid interactions have been widely used as a model system in research studies on the structure and functions of arthropod food web. Research on aphid-parasitoid food webs is hindered by their micromorphological characteristics and the high amount of labor associated with their development. Species-specific primers for cotton aphids and their parasitoids were designed and integrated into two multiplex PCRs and six singleplex PCRs, and all PCRs were optimized to achieve high specificity and sensitivity (100-10,000 DNA copies). One cotton aphid (Aphis gossypii) as well as three primary parasitoid and seven hyperparasitoid species or genera were detected using this molecular approach. This group comprises all the primary parasitoids and 97.2-99.6% of the hyperparasitoids reported in cotton fields in northern China. A tritrophic aphid-primary parasitoid-hyperparasitoid food web was then established. The described method constitutes an efficient tool for quantitatively describing the aphid-primary parasitoid-hyperparasitoid food webs and assessing the efficiency of the biological control of parasitoids in cotton fields in northern China.

[Treatment of malignant effusions with the injection of dendritic cells derived from autologous peripheral CD34+ stem cells].

OBJECTIVE: To evaluate the effect and side effects of CD34+ originated dendritic cells (DCs) infusion to treat malignant effusions. METHODS: Twenty-three patients with 26 malignant effusions received chemotherapy and granulocyte colony-stimulating factor (G-CSF) or G-CSF alone to mobilize peripheral blood hematopoietic stem cells. CD34+ stem cells were induced to differentiate into DCs by cytokines interleukin 4 (IL-4), granulocyte-macrophage colony stimulating factor (GM-CSF), and tumor necrosis factor-alpha (TNF-alpha). After being cultured for 10 to 14 days to stimulate proliferation and grow mature, DCs were refused into the body cavity by drainage tube weekly for 3 weeks. RESULTS: The response rate was 54% (14/26) and the benefit rate 81%. The median duration of response was 20 weeks (range:4-45 weeks). Karnofsky performance score (KPS) was improved in 15 patients. The response rate was 50% for the naive patients and 57% for the refractory patients. There were no severe side-effects. CONCLUSION: This pilot study demonstrates that DCs derived in vitro can exist viably after intropleural injection and can mediate biologic activity to malignant effusions. DCs immunotherapy is promising to treat malignant effusions with good tolerance.

(E)-2-(4-Fluoro-benzyl-idene)cyclo-octanone.

The title compound, C(15)H(17)FO, was prepared directly from the aldol condensation of cyclo-octa-none with 4-fluoro-benz-aldehyde, catalysed by Pd(Ni,Ce) in the presence of trimethyl-silyl chloride. The eight-membered ring adopts a boat-chair conformation.

[Structrue identification of biflavones and determination of Taxol from Taxus Madia].

OBJECTIVE: To study the flavonoids from Taxus Madia and determine the content of Taxol from different parts of Taxus Madia. METHODS: Various column chromatographic techniques were applied for isolation and purification. The structrues were elucidated by physico-chemical property and spectral evidence. The analysis of Taxol was carried out on C18 (150 mm x 4.6 mm i. d. 5 microm). The mobile phase was MeOH-H2O (3:1). The flow-rate was 0.8 ml/min. The detection wave-length was 228 nm. RESULTS: Three bifiavonyl compounds were isolated and identified as Sciadopitysin (Compound I), Ginkgetin (Compound II), 7,7",4'-Tri-O-Methylamentoflavone (Compound III). The contents of Taxol from branches and leaves, main root, fibrous root was 0.0308 mg/g, 0.02191 mg/g, 0.01983 mg/g respectively. CONCLUSION: Compound I-III are obtained from Taxus Madia for the first time. The contents of Taxol is the highest in the part of branches and leaves, and then is the main root, the lowest is the fibrous root.

[Role of molecular screening for common fusion genes in the diagnosis and classification of leukemia].

OBJECTIVE: To assess the value of common fusion genes analysis in the diagnosis and classification of leukemia by multiplex RT-PCR. METHODS: The multiplex RT-PCR, including 8 parallel PCR reactions, could screen 86 mRNA breakpoints or splice variants at the same time, which was important for the diagnosis and prognosis of leukemia. Bone marrow samples from 161 cases of leukemia and 8 cases of myelodysplastic syndrome (MDS) were involved in the study. The distribution of common fusion genes in leukemia was analyzed by the method mentioned above in combination with clinical and morphological features. RESULTS: Ten fusion genes were detected in 115 cases of leukemia, including AML1/ETO, PML/RAR alpha, PLZF/RAR alpha, dupMLL, MLL/AF6, MLL/AF10, CBFbeta/MYH11, BCR/ABL, Hox11, and EVI1 BCR/ABL was positive in all the 52 cases of chronic myeloid leukemia; PML/RAR alpha was found in 21 of 25 acute promyelocytic leukemia (APL), and PLZF/RAR alpha was detected in one case of APL. Sixteen cases of 17 AML1/ETO-positive acute leukemia (AL) belonged to FAB-M2 subtype, and one case was mixed leukemia. Three of 4 AL cases carrying CBFbeta/MYH11 were M4 subtype, and one was M5 subtype. MLL aberrations were found in 16 AL, in which all MLL/AF6 translocation existed in M5 subtype with classic monoblastic characters. Furthermore, BCR/ABL was detected in 5 acute lymphoblastic leukemia (ALL) cases. Fusion genes were also found in 2 MDS cases, of which AML1/ETO positive-MDS-RAEB progressed to AML rapidly. CONCLUSION: Screening of common fusion genes by multiplex RT-PCR is an important tool which could provide useful and reliable molecular genetic information for the diagnosis and treatment of leukemia.

[Comparison of therapeutic effects of low-dose versus high-dose interferon alpha-2b treatment on chronic myelocytic leukemia: a prospective randomized study].

OBJECTIVE: To compare the therapeutic effects of low-dose and high-dose interferon alpha-2b (IFN) treatment on chronic myelocytic leukemia (CML). METHODS: A real-time quantitative reverse transcriptase PCR (RQ-PCR) method was established to detect the fusion gene bcr-abl expression, thereby studying the reduction of leukemic cells. Thirty newly diagnosed CML patients, 21 males and 9 females, aged 14 - 69, were treated with hydroxyurea to keep the white blood cell count less than 20 x 10(9)/L, and then randomized into 2 groups: high-dose IFN group receiving IFN alpha-2b 5MIU 6 times per week for 3 - 6 months and low-dose IFN group receiving IFN alpha-2b 3MIU every other day for 3 - 6 months. Bone marrow was collected every month to Real-time PCR was used to detect the expression of bcr-abl mRNA. Mononuclear cells were isolated and RNA was extracted to detect the expression of fusion gene bcr-abl and a control gene GAPDH. The results were reported as the number of bcr-abl copies/GAPDH copy. RESULTS: The established real-time quantitative PCR method could detect the bcr-abl molecules as low as 50 copies. The intra-assay coefficient of variation (CV) was less than 5% and the inter-assay CV was 5.13%. The median bcr-abl fusion gene expression level of 30 CML patients before IFN therapy was 0.098 (range: 0.010 - 5.799). The bcr-abl expression level decreased by 19.37% and 24.86% in the low-dose and high-dose IFN groups respectively after 3 months' therapy. No significant difference was observed between the two groups (P = 0.398). Relatively more side effects were observed in the high-dose IFN group than in low-dose group. CONCLUSION: RQ-PCR is a reliable method to monitor CML therapy by analyzing fusion gene bcr-abl expression. There is a difference in bcr-abl fusion gene expression levels among the newly diagnosed patients, and low-dose IFN is as effective as high-dose IFN in reducing bcr-abl expression but with less side effects.

Use of Monte Carlo simulation to evaluate the development of vancomycin resistance in meticillin-resistant Staphylococcus aureus.

Monte Carlo simulations were performed for various vancomycin dosage regimens to evaluate the potential for development of vancomycin resistance in meticillin-resistant Staphylococcus aureus (MRSA). When the target of free AUC(24)/MIC≥200 was considered (where AUC(24) is the area under the drug concentration-time curve in a 24-h interval and MIC is the minimum inhibitory concentration), a standard dose regimen (1000 mg every 12 h) yielded unacceptable simulated outcomes in patients with normal renal function; in particular, the probability of target attainment (PTA) was only 30.5% at an MIC of 1mg/L. For the same dosage regimens and the mutant prevention concentration (MPC)-based pharmacokinetic target (total AUC(24)/MPC>15), the cumulative fraction of response exceeded 80% for all renal function strata; low values of PTA (<80%) were obtained only for isolates with MPCs of ≥22.4 mg/L, which consisted of all 21 strains of heterogeneous vancomycin-intermediate S. aureus (hVISA) and a handful of non-hVISA strains with MICs of 2mg/L (32%; 16/50). Based on the current status of vancomycin resistance, we conclude that total AUC(24)/MPC>15, derived from in vivo experiments, is more suitable to predict the development of vancomycin resistance. In clinical practice, individualised vancomycin therapy should be considered to minimise selection of resistance mutations.