Objective To establish a method for determination of Icariin in Yishengling granules (Sucrose-free) by HPLC. Methods Using a column packed with C18 and mixture of acetonitrile-water (30∶70) as the mobile phase. The flow rate was 1.0 mL/min and the temperature was 30 ℃. The detection wavelength was at 270 nm. Result The linear range of Icariin was 4.1～20.5 ?g (r=1). The mean recovery was 102.90% and RSD was 1.72%. Conclusion The method is simple, reliable and rapid, and can be applied to the quality control of Yishenling granules (Sucrose-free).

Objective To evaluate the efficacy and feasibility of Flu/ivBu/Tl" conditioning regimen for the treatment of refractory or relapsed acute non-lymphocytic leukemia in patients receiving allogeneic hematopoietie stem cell transplantation. Methods Seven patients with refractory or relapsed acute non-lymphocytic leukemia received HLA identical peripheral blood hematopoietie stem cell transplantation (PBSCT) following Flu/ivBu/TY conditioning regimen, which consisted of fludarbine, busulfex and thiotepa. All patients received cyclos-porin A (CsA) and mycophenolet mofetil (MMF) for prophylaxis of graft - versus - host disease (GVHD). Results The Flu/IVBu/TT regimen was tolerated very well, without severe regimen related toxicity. In the 31-month median follow-up duration, 5 of 7 patients were a-live in disease-free situation. Conclusion The Flu/ivBu/TT conditioning regimen reduced transplantation-related toxicities and offered high long-term disease-free survival, and was tolerated very well. Allogeneie hematopoietie stem cell transplantation using Flu/ivBu/TT condition-ing regimen is a safe and effective option for the patients with refractory/relapsed acute non-lymphocytic leukemia.

Objective To evaluate the efficacy of Flu/CTX conditioning regimen for the treatment of severe aplastic anemia in pa- tients receiving allogeneic hematopoietic stem cell transplantation. Methods Nine patients with severe aplastic anemia received HLA identi- cal peripheral blood hematopoietic stem cell transplantation (PBSCT) using Flu/CTX conditioning regimen, which consisted of fludarbine [30 mg/(m2 d) for5 days (-7 to -3) ], CTX [50mg/(kg d) for4 days(-5 to-2)]. All patients received cyclosporin A (CsA) and mycophenolet mofetil (MMF) for prophylaxis of graft-versus-host disease(GVHD). Results The Fiu/CTX regimen was very well toler- ated, with no severe regimen related toxicity. In all patients, the median days of neutrephil exceeding 0. 5×109/L and platelet exceeding 20 ×109/L were 12 days (range 10-16 days) and 16 days (range 14-19 days), respectively. Complete chimerism was achieved in all pa- tients at one month after PBSCT. Two patients had acute GVHD and one had chronic GVHD. In the 39-month median follow-up duration, all patients were alive in disease-free situation. Conclusion The Flu/CTX conditioning regimen may reduce transplantation-related toxicities and can achieve full chimerism and high long-term disease-free survival. Allogeneic hematopoietic stem cell transplantation using intravenous Fiu/CTX conditioning regimen is a safe and effective treatment method for the patients with severe aplastic anemia.

Objective To investigate the effect of hosphocreatine therapy on the plasma cardiotrophin-1(CT-1) and N terminal probrain natriuretic (NT-proBNP) in elderly hypertensive patients with heart failure. Methods A total of 76 hy-pertensive patients with heart failure, aged 65 or over, were randomly divided into treatment group and control group (n=38 for each group). The control group received routine anti-heart failure treatment. The treatment group received conventional therapy plus creatine phosphate sodium for 2 weeks. The plasma levels of CT-1 and NT-proBNP were determined in two groups. The plasma CT-1 level was measured by a double antibody sandwich enzyme-linked immunosorbent assay (ELISA). The plasma level of NT-proBNP was tested by Rui Pu fluorescent dry quantitative analyzer. Results The plasma levels of CT-1 and NT-proBNP were significantly lower after treatment in two groups (P＜0.01). The plasma levels of CT-1 and NT-proBNP were significantly decreased in treatment group than those in control group (P＜0.05). The total effective rate was 89.47%in treatment group, which was significantly higher than that of control group (71.05%, P＜0.05). Symptoms of heart failure improved in one week (21 cases in treatment group/9 cases in control group) and in two weeks (13 cases in treatment group/18 cases in control group). Conclusion The conventional therapy plus creatine phosphate sodium can decrease the plasma CT-1 and NT-proBNP levels in elderly hypertensive patients with heart failure, which improves symptoms of heart failure in a shorter period of time.

ObjectiveTo compare the therapeutic effects of percutaneous cervical discectomy (PCD group),percutaneous cervical disc nucleoplasty(PCN) and the association of them (PCDN) for the treatment of cervical intervertebral disk displacement and instability of cervical vertebral column.Methods From February 2003 to April 2011,171 consecutive patients with cervical disc herniation have presented at the authors' hospital and were retrospectively studied.The average age of patients was 47.8 years(ranged,21-74).Ninety-seven cases were treated with PCD,50 cases with PCN,and the other 24 cases with PCDN.Clinical result and the stability of cervical vertebral column after operation were evaluated and compared among the 3 groups.ResultsAll cases had been followed up for a median of 4.1 years.There was significant difference in the pre- and post-operation the Japanese Orthopaedic Association(JOA) scoring system on within 3groups (PCD:t=21.85,P＜0.05; PCN:t=14.50,P＜0.05; PCDN:t=8.56,P＜0.05).All cases had been successfully operated.There was no significant difference between groups among the 3 groups in terms of the clinical outcomes(The recovery rate of JOA standard evaluation,F=2.19,P=0.12).According to Odom criteria,the excellent and good rate are as follows:81.35％ in PCD,82.44％ in PCN,83.19％ in PCDN,respectively.There was no significant difference between groups among the 3 groups in terms of the clinical success rate (P＞0.05).There was no instability of cervical vertebral column cases in 3 groups after operation(P＞0.05),and no significant difference was found in terms of cervical vertebral column stability in pre- and post-operation in each group.ConclusionAll the three operations including PCN,PCD and PCDN are safe,minimally invasive spine surgery for the treatment of cervical intervertebral disk displacement; they achieve good clinical outcomes and there are no difference on the stability of cervical vertebral column between preoperation and postoperation.

Objective To study the growth inhibition,apoptosis induction effects of cinobufagin(CB)on human osteosarcoma(OS) cell line U2OS,MG63 and SaO2 in vitro and the underlying mechanism of action of cinobufagin in OS cells.Methods Cell viability was assessed by MTT assay.Cell-cycle status,apoptosis-inducing effects were evaluated by flow cytometry,fluorescent staining and DNA fragmentation assays.Inhibitors of apoptosis proteins (IAPs) and Bcl-2 family proteins including Bax,cleaved-PARP,xIAP,cIAP-1,survivin and p65 were tested by Western blot.Results MTT assay showed that CB could inhibited the growth of U2OS,MG63 and SaO2 cells in a dose-and time-dependent manner.The 48 h IC50 of CB on U2OS,MG63 and SaO2 cells were (104.83± 16.96) nmol/L,(47.07±7.5) nmol/L,and (136.72±10.08) nmol/L respectively.The induction of G2/M cell-cycle arrest was seen in the cells treated with CB.After cells were cultured for 12 h in the presence of 100 nmol/L CB,the percentages of cells in the G0/G1 phase were decreased,while G2/M phase were increased in U2OS,MG63 and SaOS2 cells,respectively.The results showed CB inhibited the proliferation of osteosarcoma cells through blocking the cell cycle in G2/M phase.Induction of apoptosis was confirmed by Hoechst 33258 and Annexin V/PI staining.After treating with 100 nmol/L CB for 48 h,the extents of apoptosis were 33.6％±6.4％,36.4％±7.8％ and 29.3％±5.1％,respectively.These results indicate that the anti-tumor activity of cinobufagin in osteosarcoma cells was due to a G2/M cell cycle arrest and apoptosis inducing effect.Western blot showed that CB could induce the apoptosis related family proteins Bax,cleaved-PARP up-regulation,xIAP,cIAP-1,survivin and p65 downregulation in OS cells.Conclusion CB can inhibit the cell viability and induce G2/M cell cycle arrest and apoptosis in U2OS,MG63 and SaO2 cells.The apoptosis-inducing effect of CB is confirmed by the regulation of apoptosis related proteins IAPs and Bcl-2 in vitro.

Objective The aim of this study is to detect the mRNA and protein expression levels of ERCC1 and XPF genes among different age groups of healthy Chinese Han individuals,and to analyze the correlation between the mRNA and protein expression levels andthe age of individuals in order to find new molecular markers for forensic age estimation.Methods Peripheral blood samples were obtained from 150 unrelated healthy Chinese Han individuals.The plasma was centrifuged from the whole blood by gradient centrifugation,and the totalRNA was extractedwithTrizol fromperipheral blood mononuclear cells(PBMCs).Real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) was used to quantitatively analyze the mRNA relative expression levels of ERCC1and XPF in PBMCs.Enzyme linked immunosorbent assay (ELISA) was used to quantitatively analyze the protein expression levels of ERCC1and XPF in plasma.Results There were no significant differences in the mRNA relative expression levels of ERCC1 and XPF in PBMCs between males and females(P＞0.05).Significant differences were found in the mRNA relative expression levels of ERCC1 and XPF between different age groups (P＜0.05).Regression analysis showed thatthe mRNA relative expression levels of ERCC1 and XPF were both negatively correlated with age.The correlation coefficients(r) were-0.578 and-0.844,respectively.When the age was used as independent variable(x) and the mRNA expression relative level as dependent variable (y),the fitting curveswere Y=3.3E-5X2-0.0261X+1.9175 (R2=0.3244,P＜0.01),Y=0.0003X2-0.0459X+2.0439 R2=0.729,P＜0.01),respectively.There were no significant differences inthe protein expression levels of ERCC1 and XPF in plasma between different age groups or genders (P＞0.05).Conclusion The mRNA relative expression levels of ERCC1 and XPF in PBMCsdeclined with the increase of age,however,the protein expression levels in plasma were unrelated to age.ERCC 1 and XPF genes can be used asnew molecular markers for forensic age estimation,so as to providetheoretical basis for establishing the mathematical model of ERCC1/XPF genesin concern ofindividual ages.

BACKGROUND:In the perspective of traditional Chinese medicine, few studies have focused on the compound preparations though there are many investigations. The present study was undertaken to investigate the effect of Zhenwu Tang Granule on chronic pressure-overloaded left ventricular hypertrophy in rats. METHODS:The study was performed at the laboratory of Guangzhou Institute of Respiratory Disease. Male SD rats were divided randomly into 3 groups:sham operation group (n=8), operation group (n=15) and traditional Chinese medicine (TCM) group (n=15).The model of myocardial hypertrophy was made by gradually constricting the abdominal aorta. Sixteen weeks later, cardiac ultrasonography was performed in all groups in order to ascertain post-operational left ventricular (LV) hypertrophy. And Zhenwu Tang Granule was added at a dose of 12 g/kg in the mixed feedstuff for 8 weeks in the TCM group. In the 24th week, weight, structure as well as function of the heart in each group were measured by high-frequency ultrasonography, and Masson's staining was performed on the cardiac muscles. Meanwhile, total collagen volume fraction (CVF-T) and non-coronary vessel collagen volume fraction (CVF-NV) were analyzed. RESULTS:There was an increase in the weight of the heart in the operation group, with the left ventricule dominated (P<0.05). The heart was enlarged, with diastolic interventricular septal distance (IVSd) and left ventricular posterior wal distance (LVPWd) dominated (P<0.01).There was a significant decrease in the cardiac function (P<0.05). The weight (P<0.01) and volume of the heart decreased in the TCM group compared with the operation group, with IVSd and systolic left ventricular posterior wal dominated (P<0.01). And the cardiac function was improved (P<0.05). Significant interstitial and col agen hyperplasia was shown in the operation group based on pathological analysis, and various improvements were proved in the TCM group, i.e. there was a significant decrease in CVF-T and CVF-NV (P<0.01) compared with the operation group; but no difference (P>0.05) was found when compared with the pseudo-operation group. CONCLUSION:Zhenwu Tang Granule could reduce the weight and volume of the heart, improve the cardiac function, inhibit hyperplasia of collagen, and reverse myocardial hypertrophy in rats with pressure-overloaded left ventricular hypertrophy.

OBJECTIVETo investigate value of X-ray, CT and MRI for the diagnosis of lumbar spinal stenosis.

METHODSThe data of 130 patients with clinical diagnosis and typical imaging signs of lumbar spinal stenosis were analyzed. The present study included 83 males and 47 females with an average age of 43.5 years (range from 27 to 75 years). CT examination was performed in all patients, routine X-ray examination in 23 patients and routine MRI in 57 patients.

RESULTSThe lumbar spinal stenosis showed at L(3,4) plane in 25 patients, L(4,5) in 48 patients and L5S1 in 57 patients. CT showed hyperostosis of lumbar posterior marginal, vertebral lamina, inferior articular process in 46 patients, hypertrophy of superior and inferior articular processsus in 7 patients, calcification or ossification of ligamentum flavum in 13 patients, vertebral body spondylolisthesis in 5 patients, lateral recess stenosis in 24 patients, and intervertebral foramen stenosis in 35 patients. MRI showed intervertebral disk hernia with disc associated with ligamentum flavum hypertrophy in 23 patients, ligamentum flavum symmetrical hypertrophy in 18 patients,extensive multi-segmental ligamentum flavum hypertrophy in 9 patients,and local ligamentum flavum hypertrophy in 7 patients.

CONCLUSIONThe main cause of secondary lumbar spinal stenosis is degeneration. Traditional X-ray examination has great limitations in diagnosis of lumbar spinal stenosis. CT and MRI have advantages of multi-directional imaging and the high resolution. CT can show well ligament calcification and ossification and other bone change which are showed not well on MRI, so CT is recommended to lumbar spinal stenosis.

Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Spinal Stenosis ; diagnosis ; diagnostic imaging ; Tomography, X-Ray Computed

This study was aimed to investigate the effect of GATA-2 over-expression on function of mouse fetal liver hematopoietic stem cells. GATA-2 was introduced into mouse fetal liver cells via retrovirus mediated transduction with GFP as a detecting marker. Flow cytometry, colony-forming assay and cell cycle assay were used to detect the biologic changes of these retrovirus infected mouse fetal liver hematopoietic stem cells. The results showed that GATA-2 over-expression increased the Lin(-)Sca1(+)C-Kit(+) (LSK) population dramatically. Cell cycle of LSK cells didn't show abnormal, while colony forming ability decreased significantly. These data indicated that GATA-2 over-expression inhibited definitive differentiation of mouse fetal liver hematopoietic stem cells. It is concluded that over-expression of GATA-2 can significantly raise the LSK cell proportion in mouse fetal liver and inhibit the differentiation capability, the underlying mechanisms may be related to up-regulation of Hes-1, which may lead to the blocking of cell differentiation at the stem/progenitor cell stage.
Animals ; Cell Differentiation ; Cells, Cultured ; Female ; GATA2 Transcription Factor ; genetics ; Hematopoietic Stem Cells ; cytology ; Liver ; cytology ; Male ; Mice ; Mice, Inbred C57BL