To establish an HPLC method for the determination of myricetrin, quercitrin, myricetin, quercetin, kaempferol, amentoflavone and hinokiflavone in Cacumen Platycladi and processed Cacumen Platycladi, and compare the difference before and after the processing. Methods:The method described in Chinese pharmacopoeia was used in the processing. The chromato-graphic column was Welch Ultimate LP-C18 (250 mm × 4. 6 mm,5 μm),the mobile phase was methanol and 0. 2% phosphoric acid with gradient elution, the detection wavelength was 330 nm, and the flow rate was 1 ml·min-1 . The column temperatuhe was 35℃, the injection volume was 20μl. Results:The linear relationship respectively of myricetrin, quercitrin, myricetin, quercetin, kaempfer-ol, amentoflavone and hinokiflavone was good(r≥0. 999 0). The average recovery was within the range of 97. 6%-101. 1% (RSD≤1. 14%, n=6). The content of myricetrin, quercitrin, amentoflavone and hinokiflavone in Cacumen Platycladi was reduced after the processing, and the content of quercetin and kaempferol was increased after the processing. Conclusion: The method is simple,and can be used in the determination of Cacumen Platycladi and its processing product. The results of the content show that the flavones in Cacumen Platycladi can be changed by the processing.

Objective To investigate the effect of propofol on the high-voltage-activated calcium currents [ICa(HVA)] in rat hippocampal neurons. Methods Hippocampal neurons were prepared from Wistar rats and cultured. ICa(HVA) was recorded using whole-cell patch clamp technique. Different concentrations of propofol were added to the culture. The effect of propofol on ICa(HVA) Was evaluated. Results ICa(HVA) was inhibited by propofol in 300 μmol/L reduced peak ICa(HVA) by (24±6)%, (33 ±5) %, (36±7)% and(38±3)% respectively with a mean IC50 of 3.8 μmol/L and Hill coefficient of 0.35. Vmax was shifted from (4.0± 2.0) mV to (3.8 ± 1.6) mV. The V1/2 of inactivation curve was shifted from (- 32 ± 5) mV to (- 35 ± 4) mV and the slope factor was 31 ± 5 and 35 ± 6 before and after administration respectively. Conclusion Propofol produces significant inhibition of calcium currents in the central neurons which may partly explain the action of propofol on central nervous system.

Objective:To investigate the expression and prognostic significance of CD19 in patients with Acute myelogenous leukemia with AML1-ETO positive. Methods: Clinical data of 66 patients AML with AML1-ETO positive who were newly diagnosed from Jan 2010 to Dec 2015 were collected. To retrospectively analyze the relationship between clinical characteristics and expression of CD19,so dose the prognosis. Results:The positive rate of CD19 expressing in AML with AML1-ETO positive was 50. 0%. There were no statistically significant differences in terms of age,gender,hemoglobin,platelet,percentage of bone marrow blasts,accompanied with chromosome ,gene mutations between patients with and without CD19 expression(P>0. 05). The white blood cell count(WBC) of the CD19 negative group was higher than CD19 positive group,while showed significant difference(P=0. 027). Although the relapse-free survival (RFS) of patients with CD19 expression was higher than those without,no significant difference was calculated (P=0. 105). Patients with CD19 expression had superior overall survival ( OS ) compared to those without CD19 expression ( P = 0. 030 ) . Multivariable analysis for OS identified CD19 positivity as an independent predictor associated with better prognosis. Conclusion: The expression of CD19 in AML with AML1-ETO positive may be an indicator associated with better prognosis.

Objective To investigate the effects of Yangxueqingnao Granule on the proliferation and differentiation of endogenous neural stem cells and their neuroprotective mechanism after brain ischemia. Methods Cerebral ischemia was induced in rats by temporary middle cerebral artery occlusion. Ischemic animals received Yangxue Qingnao Granule at high and low doses at days 3,7, 14,and 21 after MCAO. Neurological score and TTC staining were used to assess the neurological deficits and the volume of cerebral infarction, respectively. Immunohistochemistry was used to detect Brdu and Nestin expression in the subventricular zone(SVZ)and hippocampal dentate gyrus subgranular zone(SGZ)region. Results The rats in MCAO control group, low dose group and high dose group showed varying degrees of neurological def-icits and the volume of cerebral infarctions. The neurological defect scores and volume of cerebral infarction were signifi-cantly different at different time points in each group ( P<0.01). 2. The mean fluorescence intensity values of Brdu posi-tive cells or the Nestin positive cells in SVZ and SGZ regions were significantly different ( P< 0.01) at different time points after treatment for 3,7,14 or 21 days. The mean fluorescence intensity values of Brdu positive cells or the Nestin positive cells in SVZ and SGZ regions were significant different among low dose group, high dose group and MCAO con-trol group. The mean fluorescence intensity values of Brdu positive cells or the Nestin positive cells in SVZ and SGZ re-gions were significantly different among low dose group, High dose group and control group. High dose group significantly increased the mean fluorescence intensity values of Brdu positive cells or the Nestin positive cells in SVZ and SGZ re-gions. Conculsions Yangxue Qingnao Granule is able to promote the functional recovery and reduce the volume of cere-bral infarction in ischemic rats. Yangxue Qingnao Granule can enhance the proliferation and differentiation of endoge-nous neural stem cells in ischemic rats.

ObjectiveTo investigate the variation of peripheral blood CD34+ cells during the hematopoietic stem cell mobilization,and its influence on the collecting timing and results.Methods Twenty-seven cases of peripheral blood hematopoietic stem cell mobilization and collection from April 2010 to December 2011 were analyzed,including 13 autologous cases mobilized with chemotherapy combined with granulocyte colony-stimulating factor (G-CSF,10 μg· kg-1 · d-1) and 14 cases of healthy donors mobilized with only G-SCF (7.5 μg · kg- 1 · d- 1 ).The number of peripheral blood CD34+ cells was counted,and its correlation with the yield of mononuclear cells (MNCs) and CD34+cells was analyzed.ResultsMNCs (5.84 ± 1.48) × 108/kg and CD34+ cells (3.93 ± 2.16) × 106/kg were obtained in healthy donors,and (6.58 ± 3.72) × 108/kg MNCs and (3.98 ± 3.06) × 106/kg CD34+ cells were obtained in autologous cases,respectively.There was only 1 failure in autologous cases.The peak of peripheral blood CD34+ cells in autologous cases appeared at day 4 after the treatment of G-CSF,and in healthy donors the number of peripheral blood CD34+ cells at day 5 was still in ascendant phase.The CD34+ cells/kg in the collection products were positively correlated with the percentage and absolute value of peripheral blood CD34+ cells.The cases ratio of CD34+ cells≥2× 106/kg in the products of single collection was up to 76.2％ (16/21) in the cases with peripheral blood CD34+ cells absolute value greater than 20/μl.ConclusionThe number of peripheral blood CD34+ cells was an important monitoring indicator in hematopoietic stem cell mobilization and collection,CD34+ cell absolute value ≥20/μl could be used as collection threshold.

Objective To study the immune re-constitution after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in patients with hematological malignancies.Methods From June 2011 to May 2015,65 patients with hematological malignancies were analyzed retrospectively.Lymphocyte subsets were determined by flow cytometry (FCM),including total T lymphocytes (CD3+),helper T cells (CD3+ CD4+),cytotoxic T cells (CD3+ CD8+),CD4/CD8 ratio,nature killer (NK) cells (CD3-CD56+),NKT cells (CD3+ CD56+),B lymphocytes (CD19+),naive T cells (CD3+ HLA-DR+),static T cells (CD3+ HLA-DR-),and regulatory T cells (CD4+ CD25high Foxp3+) on the day 14,28 and 42,and on the month 2,3,6,9,12,15,18 and 24 after allo-HSCT.Results The percentage of CD3+ T cells located normal range after hematological recovery,and its absolute number recovered to normal range at + 15 months.The percentage of CD3+ CD4+ T cells recovered to normal range at + 24 months.However,the absolute number of CD3+ CD4+ T cells did not recover to normal range until 24 months after allo-HSCT.The percentage of CD3+ CD8+ T cells was higher than normal range at + 42 day,and its absolute number was greater than normal range at + 3 months.Hence,low CD4/CD8 ratio was observed for a long period.The re-constitution time points of the percentage and absolute number of CD3+ HLA-DR+ T cells were + 3 months and + 24 months respectively.The re-constitution time points of the percentage and absolute number of CD3 + HLA-DR-T cells were + 2 months and + 15 months respectively.The re-constitution time points of the percentage and absolute number of regulatory T cells were + 12 months and + 15 months respectively.The percentage of NKT cells located in normal range after hematological recovery,and its absolute number retumed to normal range at + 12 months.The re-constitution time points of the percentage and absolute number of B cells were + 9 months and + 18 months respectively.The percentage of NK cells located in normal range after hematological recovery,and its absolute number returned nearly to normal range at + 3 months.Conditioning regimen containing ATG,source of stem cells,CD34+ cell number,GVHD,and CMV reactivation were all associated with immune re-constitution after allo-HSCT.Conclusion Different immune cells showed different re-constitution models after allo-HSCT,and the percentage recovered faster than absolute number for a certain kind of immune cells.Studying immune re-constitution and its associated factors may offer beneficial information for insight into transplantation immunology and improve the management of allo-HSCT.

Objective To explore the long-term outcomes and complications of patients with hematological malignancies (HM) after haploidentical donor transplantation (HDT) or siblingidentical donor transplantation (SDT).Methods From June,2011 to July,2016,89 patients with HM receiving allo-HSCT were retrospectively analyzed,including 57 patients undergoing HDT and 32 cases undergoing SDT.Results The median time to achieve neutrophil engraftment was 2 days shorter after HDT than SDT,whereas that of platelet engraftment was 3 days longer after HDT than SDT.The cumulative incidence for 3 to 4 grade acute graft-versus-host disease (GVHD) was not obviously different between HDT and SDT (8.77％ versus 12.5％ respectively;x2 =0.313,P =0.576).The cumulative incidence for chronic GVHD was not significantly different between HDT and SDT (45.6％ versus 37.5％;~ =0.551,P =0.458).Cytomegalovirus (CMV) reactivity was significantly higher in patients after HDT (77.19％) than those after SDT (21.88％) (x2 =25.633,P＜0.001).The occurrence of hemorrhagic cystitis was also obviously higher in patients after HDT (26.32％)than those after SDT (3.85％) (x2 =5.340,P =0.021).The 1-,2-,and 3-year relapse-free survival rate of patients receiving HDT and SDT was 63.9％,55.4％,44.3％ and 71.2％,58.3％,51.8％,respectively (P =0.541).The 1-,2-,and 3-year overall survival rate of patients receiving HDT and SDT was 75.3％,65.3％,52.3％ and 76.9％,62.9％,62.9％,respectively (P =0.777).Conclusion Considering similar incidence of severe GVHD and long-term outcomes,haploidentical donors should be recommended as a potential alternative donor source when an identical donor is lacking for patients with HM.

Objective To analyze the effect of haploidentical hematopoietic stem cell transplantation (HID-HSCT) on high-risk acute lymphoblastic leukemia (ALL),and to explore the influence of minimal residual disease (MRD) before transplant on the outcomes.Methods A retrospective analysis was performed on 39 high risk ALL patients receiving HID-HSCT in our hospital from Jan.2013 to Jan.2018.The clinical features,stem cell engraftment,complications,survival and recurrence were compared between patients with pretransplant MRD + and MRD-.Results All the 39 patients presented with successful engraftment.The overall survival (OS) was 54.67％;the disease free survival (DFS) was 40.96％;the incidence rate of acute graft versus host disease (aGVHD) was 53.8％,including 23.1％ Ⅱ-Ⅳ degree aGVHD and 2.6％ Ⅲ-Ⅳ degree aGVHD.There was no significant difference in stem cell engraftment,GVHD,cytomegalovirus infection and hemorrhagic cystitis between MRD + and MRD-patients.DFS and OS in MRD + patients were significantly lower than those in MRD-patients;the cumulative RR rate increased significantly,and there was no significant difference in cumulative TRM.Conclusion HID-HSCT was an effective method to treat high-risk ALL,but MRD + patients had high recurrence rate and poor prognosis.Strategy adjustment should be considered to reduce tumor residual and the transplantation strategy should be optimized for these kinds of high risk patients,so as to improve long-term outcomes.

OBJECTIVE:To investigate the effects of different doses of tanshinone ⅡAon hemodynamics in patients underwent non-extracorporeal circulation coronary artery bypass grafting(NECCABG). METHODS:A total of 66 patients underwent selective NECCABG during Nov. 2016-Apr. 2017 were selected and divided into control group(33 cases)and observation group(33 cases) according to random number tablet. Both groups were given Tanshinone ⅡAsodium sulfonate injection with loading dose of 0.4 μg/kg for 10 min at the beginning of surgery. Control group was given Tanshinone ⅡAsodium sulfonate injection 0.2 μg/(kg·h) continuously till the end of surgery;observation group was given 0.4 μg/(kg·h)continuously till the end of surgery. HR,MAP, central venous pressure(CVP),mean pulmonary arterial pressure(MPAP),pulmonary capillary wedge pressure(PCWP),cardiac index(CI),systemic vascular resistance index(SVRI),pulmonary circulation resistance index(PVRI)and the occurrence of ADR were observed in 2 groups before anesthesia induction(T0),5 min after tracheal intubation(T1),at the moment of sternum cut(T2),at the end of anastomosis of anterior descending branch(T3),at the end of anastomosis of right coronary artery(T4),at the end of operation(T5). RESULTS:At T0,there was no statistical significance in HR,MAP,CVP,MPAP,PCWP, CI,SVRI or PVRI between 2 groups(P>0.05). At T1-T5, HR,MAP,CVP,MPAP,PCWP,SVRI and PVRI of control group were significantly higher than at T0,with statistical significance(P<0.05). There was no statistical significance in CI of control group among at T1-T5and with at T0;there was no statistical significance in HR,MAP,CVP,MPAP,PCWP,SVRI or PVRI of observation group between at T1-T5with at T0 (P>0.05). At T1-T5,CI of observation group was significantly higher than at T0and than control group;HR,MAP,CVP, MPAP,PCWP,SVRI and PVRI were significantly lower than control group,with statistical significance(P<0.05). No obvious ADR was found in 2 groups during medication. CONCLUSIONS:Tanshinone ⅡA0.4 μg/(kg·h)is helpful to keep hemodynamics stable and improve heart function in patients with NECCABG with good safety.

Objective:To explore the incidence rates, clinical features, risk factors and its impacts on survival of central nervous system complications (CNSC) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:From June 2011 to October 2018, 237 consecutive patients undergoing allo-HSCT were retrospectively analyzed.Results:The incidence of CNSC was 10.5%(25/237) and the median time 82(-4 - 810) days post-transplantation. The most common instances of CNSC were drug-associated encephalopathy (n=6), CNS infection (n=5), unexplained convulsions (n=4), metabolic encephalopathy (n=3), immune-related encephalopathy (n=3), primary central relapse (n=3) and cerebrovasculopathy (n=1). The most common clinical symptom was epileptic seizure (n=11). CsA-related encephalopathy was manifested mainly as posterior reversible encephalopathy syndrome on brain MRI. Metabolic encephalopathy is mostly demyelination. Most hippocampal lesions were caused by immune-related encephalopathy or CNS infection. Analysis of risk factors indicated that umbilical cord blood transplantation, HLA incompatible transplantation and delayed platelet implantation were high risk factors for post-transplantation occurrence of CNSC. Survival analysis suggested that non-relapse mortality rate (42.9%, 9/21) in group with CNSC of malignant hemoblastosis was higher than that in group without CNSC (15.3%, 27/176) and inter-group difference was statistically significant ( χ2=9.511, P=0.005). The 1/3-year OS rates in group with CNSC were lower than those in group without CNSC (56.6% vs 77.8%; 37.1% vs 65.7%). And the difference was statistically significant ( P=0.022). Conclusions:With a complex etiology, CNSC is one of serious complications after allo-HSCT and it significantly reduces the overall survival rate of patients. Umbilical cord blood transplantation, HLA incompatible transplantation and delayed platelet implantation are high-risk groups for CNSC.