Effect of Genkwa Flos on Transient Receptor Potential Vanilloid 1 of Thermosensitive Channel / 中国实验方剂学杂志

Objective:To study the effect of Genkwa Flos on the thermosensitive channel, transient receptor potential vanilloid 1 (TRPV1) by electrophysiological whole cell patch clamp technique and animal behavior experiment, in order to explore its mechanism. Method:The whole-cell patch clamp technique was used to measure transmembrane currents induced by 75%ethanol extract from different concentrations of Genkwa Flos in HEK293 cells that expressed human TRPV1.TRPV1 specific antagonist capsaicin was used to observe whether it could inhibit the transmembrane current induced by Genkwa Flos. Totally 30 C57BL/6 mice were taken for behavioral detection, and divided into blank group (6 mice), high-dose group (6 mice), medium-dose group (6 mice), low-dose group (6 mice) and ibuprofen positive control group (6 mice). Genkwa Flos treatment group was given low dose (0.195 g·kg-1·d-1), medium dose (0.39 g·kg-1·d-1) and high dose (0.78 g·kg-1·d-1) by gavage. One hour later, the changes of behavioral latency of cold pain and hot pain in mice were observed in cold plate at (0±2)℃ and hot plate at (55±1)℃. Result:Whole cell patch clamp experiment showed that 75%ethanol extract of Genkwa Flos in hTRPV1/HEK293 cells could activate TRPV1 channel to generate obvious transmembrane current, which was similar with that generated by the known TRPV1 agonist capsaicin in current density and current-voltage relationship. The dose-effect experiments showed that compared with extracellular fluid, 10 g·L-1 ethanol extract of Genkwa Flos could activate hTRPV1/HEK293 cells to induce significant transmembrane current (P-1 ethanol extract of Genkwa Flos was more than 3 g·L-1(P-1 ethanol extract of Genkwa Flos. In the experiment of cold plate and hot plate in mice, there was a dose-effect relationship between the latencies of cold pain behavior and hot pain behavior in mice prolonged by Genkwa Flos. In the experiment of cold plate, compared with the blank group, the cold pain behavior latency of mice in the medium-dose group was significantly prolonged (PPPPPConclusion:More than one TRPV1 agonist was included in 75%ethanol extract of Genkwa Flos. The warm, analgesic and anti-inflammatory effects of Genkwa Flos may be a series of effects after activation of TRPV1.

Advances in the study of New Delhi metallo-β-lactamase 1 / 药学学报

The New Delhi metallo-β-lactamase-1 (NDM-1) was first reported in 2010, detected in a Klebsiella pneumoniae isolate from a Swedish patient of Indian origin. It has recently attracted extensive attention for its biological activities to catalyze the hydrolysis of almost all of β-lactam antibiotics. The gene for NDM-1 can spread from one strain of bacteria to another by horizontal gene transfer. The most troubling aspect is that there are currently no clinically available inhibitors to block the metallo-β-lactamase action. Therefore, there is urgent need to develop new NDM-1 inhibitors, which can protect β-lactam antibiotics from the hydrolysis effect of NDM-1. In this review, the current research, drug-assistant mechanism and potential NDM-1 inhibitors are summarized.

Clinical Analysis of 4 AML Patients Aged Over 80 Years Treated with Chemotherapy Combined with Haploidentical Hematopoietic Stem Cell Infusion / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and safety of MA (mitoxantrone and cytarabine) regimen chemotherapy combined with granulocyte-colony stimulating factor (G-CSF)-mobilized family related HLA-haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) infusion for the treatment of acute myeloid leukemia (AML) patients aged over 80 years.</p><p><b>METHODS</b>Four elderly patients with AML were treated in Chinese Second Artillery General Hospital from August 2008 to September 2013. The proportion of male to female was 1 : 3 and the median age 83 (80-85) years. All patients received programmed infusions of G-PBHSC after MA regimen chemotherapy without graft-versus-host disease (GVHD) prophylaxis. After complete remission (CR), patients only received G-PBHSC infusion without chemotherapy.</p><p><b>RESULTS</b>Three cases achieved CR and their disease free survival (DFS) time was 18, 8, 6 months, respectively. 1 case did not reach remission after 2 cycles chemotherapy. The median overall survival (OS) time was 10 (3-20) months. No GVHD was observed in any of the patients during treatment. Concludsion: The combination of chemotherapy and programmed haploidentical G-PBHSC infusion is an alternative approach for AML patients aged over 80 years.</p>

Clinical analysis of lymphoblastic lymphoma/leukemia treated with Hyper-CVAD/MA regimen chemotherapy combined with haploidentical hematopoietic stem cell infusion / 中国实验血液学杂志

This study was aimed to investigate the efficacy of Hyper-CVAD/MA regimen chemotherapy combined with haploidentical hematopoietic stem cell infusion for the treatment of lymphoblastic lymphoma/leukemia (LBL/ALL). Seven patients with LBL/ALL were treated in Second Artillery General Hospital from August 2009 to September 2012. All patients received programmed infusions of granulocyte-colony stimulating factor (G-CSF)-mobilized family related HLA-haploidentical donor peripheral blood hematopoietic stem cell (G-PBHSC) after each of cycle of Hyper-CVAD/MA regimen chemotherapy without graft-versus-host disease (GVHD) prophylaxis. A total of four cycles of therapy were planned. The interval between each cycle of treatment was 8 to 12 weeks. By April 2014, the median follow-up time was 41 (20-57) months. The results showed that the 7 patients totally received 30 cycles of treatment, and all patients achieved complete remission (CR). The patients were generally well-tolerated to therapy, and the most significant toxicities of grade 3 to 4 neutropenia and thrombocytopenia developed in nearly all of the patients after each course of the Hyper-CVAD/MA regimen. No GVHD was observed in any of the patients during treatment. Up to now, 5 patients were still alive, 2 patients were died of relapse. It is concluded that the combination of chemotherapy and programmed haploidentical G-PBHSC infusion is a promising approach to the treatment of LBL/ALL.

Prophylactic G-CSF mobilized donor lymphocytes infusion after non-myeloablative stem cell transplantation prevents relapse in patients with high-risk leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the anti-leukemia effects of prophylactic G-CSF mobilized donor lymphocytes infusion (pG-DLI) and its relationship with the incidence of graft-versus-host disease (GVHD) in high-risk leukemia patients with non-myeloablative stem cell transplantation (NST).</p><p><b>METHODS</b>12 patients with high-risk leukemia were analyzed, including Ph⁺ acute lymphocytic leukemia (n=1), acute leukemia (AL) with persistent non-complete remission (n=2), acute myeloid leukemia (AML) with central nervous system (CNS) relapse (n=3), hybrid AL (n=1), secondary AML evolving from myelodysplastic syndrome (MDS/AML) (n=2), chronic myeloid leukemia in accelerated phase (CML-AP) (n=1), CML in blastic phase (CML-BP) (n=2). All patients received non-myeloablative conditioning and pG-DLIs were administered 30-40 days post transplantation if no signs of GVHD were present. The percentage of donor cell chimera was analyzed by short tandem repeat-polymerase chain reaction (STR-PCR) just before and after pG-DLI. The incidence of leukemia relapse and GVHD were observed.</p><p><b>RESULTS</b>12 high-risk leukemia patients with a median age of 38 (range: 29-52) years received G-DLI at a median interval of 35 (32-40) days. The median numbers of infused mononuclear cells (MNCs), CD34⁺, and CD3⁺ cells/kg recipient body weight was 2.3×10⁸/kg, 1.7×10⁶/kg, and 0.6×10⁸/kg, respectively. 10 of 12 patients had full donor chimera before pG-DLIs and conversion from mixed to full donor chimera occurred in the other 2 patients shortly after pG-DLI. Grade II acute GVHD (aGVHD) was observed in only 2 patients and chronic GVHD (cGVHD) developed in 6 patients, including involvement of skin (n=3), skin and intestine (n=2), liver (n=1). 1 patient died of cGVHD. With a median follow-up of 40 (24-64) months, 7 patients are alive in remission, with 3-year actuarial overall survival (OS) and disease-free survival (DFS) rates of the same 58.3%.</p><p><b>CONCLUSION</b>Our findings indicate that pG-DLI after NST does not increase the risk of aGVHD, but could enhance the capacity graft-vs-leukemia and prevent relapse in high-risk leukemia patients.</p>

Role of G-CSF in the proliferation, differentiation and cell cycle distribution of mouse thymocytes after acute radiation / 中国实验血液学杂志

This study was purposed to investigate the effect of G-CSF on the proliferation, differentiation, and cell cycle distribution of thymocytes in sublethally irradiated mice. Female BALB/c mice were exposed to 6.0 Gy γ-ray irradiation and then randomly divided into control and G-CSF treatment group. In the treatment group rhG-CSF 100 µg/(kg·d) was given subcutaneously for 14 continuous days and to make sure the first injection was given within 1 hour after irradiation. Cell cycle distribution and apoptosis of thymocytes were detected within 72 hours after irradiation. Subpopulations of CD4(-)CD8(-) cells and sequential changes in the distribution of CD4(+)CD8(+), CD8(+)CD4(-), CD8(-)CD4(+) cells were detected by a three-color flow cytometry during a four-weeks period after irradiation. The results showed that in G-CSF treatment group marked increase of cells in G(0)/G(1) phase (G-CSF vs control: 82.0 ± 5.0% vs 75.9 ± 2.8%) (p < 0.05) and a decrease of cells in S phase (G-CSF vs control: 10.2 ± 4.8% vs 15.7 ± 2.3%) (p < 0.05)could be observed as early as 6 hours after irradiation, but G-CSF seems have no evident effects on the cells in G(2)/M phase. G-CSF could also protect thymocytes against apoptosis. 6 and 12 hours after irradiation the apoptosis rates of thymic cells in G-CSF treatment group were 11.5 ± 2.4% and 15.5 ± 3.3% respectively, while in the control group the apoptosis rates were 16.5 ± 2.2% and 22.6 ± 0.7% respectively. Comparison between the two group demonstrated significant difference (p < 0.05). CD4(-)CD8(-) double negative thymocytes (DN)can be defined as DN1-4 according to their maturation. G-CSF treatment resulted in a significant increase in DN1 thymocytes and promoted their proliferation and differentiation to a more mature DN3 and DN4 stage. G-CSF could enhance the recovery of CD4(+)CD8(+) thymocytes and mitigate their relapse during reconstitution. The percentage of CD4(+)CD8(+) thymocytes in the G-CSF treatment group 28 days after irradiation was significantly higher than that of the control group (71.0 ± 6.3% vs 25.5 ± 6.3%) (p < 0.05). It is concluded that G-CSF has a positive effects on the thymic cell cycle distribution, proliferation and differentiation, which may contribute to the reconstitution of central immune system after acute irradiation.

Effect of G-CSF on murine thymocyte emigration and cell cycle alteration after a sublethal dose of irradiation / 中国实验血液学杂志

This study was aimed to investigate the effect of recombinant human granulocyte colony-stimulating factor(G-CSF) on murine thymocyte emigration and cell cycle alteration after a sublethal dose of gamma-irradiation. Female BALB/c mice were given 6.0 Gy γ-ray total body irradiation and then randomly divided into G-CSF and control groups. Mice in the G-CSF group were injected recombinant human G-CSF 100 µg/(kg·d) subcutaneously once daily for 14 consecutive days and mice in the control group were given the same volume of phosphate buffered solution. Thymocyte cycle alteration and the proportion of apoptosis cells were detected by flow cytometry within 72 hours after irradiation. Real-time PCR was used for detection and quantitation of murine T cell receptor rearrangement excision circles (sjTREC) of the thymic cells at 30 and 60 day after the irradiation. The results showed that at 6 hour after irradiation G-CSF could significantly increase the thymic cells in G(0)/G(1) phase, G-CSF vs control: (82.0 ± 5.0)% vs (75.9 ± 2.8)% (p < 0.05), and decrease the thymic cells in S phase, G-CSF vs control: (10.2 ± 4.8)% vs (15.7 ± 2.3)% (p < 0.05), but G-CSF seemed have no evident effects on the percentage of thymic cells in G(2)/M phase. G-CSF could also protect thymocytes from apoptosis at 6 hour and 12 hour after irradiation the percentages of apoptosis cells in G-CSF group were (11.5 ± 2.4)% and (15.5 ± 3.3)%, respectively, which were significantly lower than that of the control group (16.5 ± 2.2)% and (22.6 ± 0.7)%, respectively (p < 0.05). The sjTREC copy amount was conspicuously higher in G-CSF group than that in the control at 30 day after irradiation (p < 0.01), but the preponderance disappeared 60 days later. It is concluded that G-CSF has a positive effect on the thymic cell cycle alteration to protect thymocytes from apoptosis and enhance the recent thymocyte emigration, which may contribute to the central immune reconstitution after irradiation.

Expression of NOV and BNIP3 gene in mouse myelomonocytic leukemia and its significance / 中国实验血液学杂志

This study was aimed to investigate the expression level of NOV and BNIP3 mRNA in mice myelomonocytic leukemia (AML-M(4)) and its significance. The mice were inoculated intravenously with myelomonocytic leukemia cells of WEHI-3, and divided randomly into chemotherapy group and control (untreated) group. Bone marrow samples were then collected from both groups at different times. The NOV and BNIP3 mRNA expression were detected by TaqMan quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR), and the relationship between these expression levels and clinical significance in leukemia incidence and progression were analyzed with β-actin as the housekeeping gene. The results showed that the mean values of NOV and BNIP3 increased gradually from 2 weeks after inoculation and achieved highest level at death in control group. Expression level of NOV increased from 1.85E-05 before inoculation to 3.57E-02 at death (p < 0.05), and BNIP3 from 3.44E-03 to 3.48E-02. While 2 gene expression in the chemotherapy group decreased quickly to 2.51E-05 and 1.58E-03 (p < 0.05) respectively after chemotherapy, which were close to the level before inoculation (p > 0.05). The 2 gene expressions again rose at relapse, and difference of expression level between 2 group at death were no statistically significant (p > 0.05). It is concluded that the expression of NOV and BNIP3 in leukemia AML-M(4) is significantly higher than that in normal controls, of which high level expression is an important factor in the development of leukemia. Close relation between the therapeutic effect and expression level of these two genes suggests the great value in prognostic evaluation and MRD detection.

Studies on chromosome numbers of Salvia miltiorrhiza, S. flava and S. evansiana / 中国中药杂志

<p><b>OBJECTIVE</b>To study the numbers of chromosome in Chinese herb Salvia miltiorrhiza from 7 provinces in China, and S. flava as well as S. evansiana from Yunnan province in China.</p><p><b>METHOD</b>The young root was treated with the mixture of ice and water for 24 h, fixed with Carony's fixative for 6-12 h. After differentiating for 10-12 min with 1 mol x L(-1) hydrochloric acid at 60 'C and staining with carbol fuchsin,the section was observed under microscope.</p><p><b>RESULT</b>Chromosome numbers of S. miltiorrhiza and S.flava were 2n = 2x = 16. The numbers of S. evansiana were 2n = 4x = 32. The basic numbers of the chromosomes were x = 8. And tetraploids were observed in S. miltiorrhiza from Sichuan provices and Shandong provices.</p><p><b>CONCLUSION</b>The basic number of the chromosomes are x = 8. The chromosome numbers of S. miltiorrhiza, S.flava and S. evansiana are 16,16 and 32 respectively. As the chromosomes are the small or micro-small ones, it is difficult to use them for karyotype.</p>

Significance and Assaying of Serum Interleukin-1?,Interleukin-6,Interleukin-8 and Tumor Necrosis Factor-? in Highly Pathogenic Avian Influenza Viral Pneumonia in Mice / 实用儿科临床杂志

Objective To study the interrelation between highly pathogenic avian influenza viral pneumonia(HPAIVP) and cellular factors interleukin-1?(IL-1?),interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor-?(TNF-?) in HPAIVP.Me-thods Sixty Kunming mice were divided into 2 groups randomly:experiment group and control group,each group consisted of 30 mice.The highly pathogenic avian influenza virus was used to establish HPAIDP models.The expressions of serum IL-1?,IL-6,IL-8 and TNF-? in experiment group of different moment and control group were detected by enzyme linked immunosorbent assay(ELISA).Result The levels of serum IL-1?,IL-6 and TNF-? in experiment group were significantly higher than those in control group(Pa

Studies on mechanism of antiasthmatic and anti-inflammatory actions of CDCA / 中国中药杂志

<p><b>OBJECTIVE</b>To observe antiasthmatic and anti-inflammatory actions mechanisms of CDCA.</p><p><b>METHOD</b>The content of NO was determined by method of nitroreductase chromatometry in serum and trachea tissue. The content of cAMP was analysed by method of competitive protein-binding assay. The content of PGE2 was determined by method of ultraviolet spectrophotometry.</p><p><b>RESULT</b>CDCA significantly decreased the content of NO of serum and trachea tissue in mice. CDCA increased greatly the content of cAMP of trachea tissue in rats. CDCA significantly decreased the content of PGE2 of trachea and lung tissue in mice.</p><p><b>CONCLUSION</b>Mechanisms of antiasthmatic and anti-inflammatory actions of CDCA are related to increasing the content of cAMP in trachea tissue and decreasing the constituent of NO and PGE2 in body.</p>

Change of Interleukin-2 in Nasopharyngeal Secretion of Children with Acute Respiratory Syncytial Virus Bronchitis / 实用儿科临床杂志

0.05).Conclusion There is no association between IL-2 levels in NS and RSV bronchitis.The IL-2 levels show a heterogenous behavior.