Research progress on metabolism and pharmacological activities of several iridoid glycosides / 中国中药杂志

As a large category of natural products widely present in traditional Chinese medicine, iridoid glycosides have multiple pharmacological activities. Recent researches suggest that iridoid glycosides mainly exist in the forms of original form, aglycone and a series of their Ⅰ and Ⅱ metabolites under the biotransformation effect, and their metabolites have been proved to have multiple pharmacological activities. The research progress on metabolism and metabolite activities of several iridoid glycosides would be reviewed in this article, to provide a theoretical basis for the further development and utilization of iridoid compounds and their metabolites.

Impact of uncontrolled blood pressure on diagnostic accuracy of coronary flow reserve for detecting significant coronary stenosis in hypertensive patients / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Impaired coronary flow reserve (CFR) in patients with hypertension may be caused by epicardial coronary stenosis or microvascular dysfunction. Antihypertensive treatment has been shown to improve coronary microvascular dysfunction. The aim of this study was to evaluate the impact of uncontrolled blood pressure (BP) on diagnostic accuracy of CFR for detecting significant coronary stenosis.</p><p><b>METHODS</b>A total of 98 hypertensive patients scheduled for coronary angiography (CAG) due to chest pain were studied. Of them, 45 patients had uncontrolled BP (defined as the office BP ≥ 140/90 mmHg (1 mmHg = 0.133 kPa) in general hypertensive patients, or ≥ 130/80 mmHg in hypertensive individuals with diabetes mellitus), and the remaining 53 patients had well-controlled BP. CFR was measured in the left anterior descending coronary artery (LAD) during adenosine triphosphate-induced hyperemia by non-invasive transthoracic Doppler echocardiography (TTDE) within 48 hours prior to CAG. Significant LAD stenosis was defined as > 70% luminal narrowing. Diagnostic accuracy of CFR for detecting significant coronary stenosis was analyzed with a receiver operating characteristic analysis.</p><p><b>RESULTS</b>CFR was significantly lower in patients with uncontrolled BP than in those with well-controlled BP (2.1 ± 0.6 vs. 2.6 ± 0.9, P < 0.01). Multivariate linear regression analysis of the study showed that the value of CFR was independently associated with the angiographically determined degree of LAD stenosis (β = -0.445, P < 0.0001) and the presence of uncontrolled BP (β = -0.272, P = 0.014). With a receiver operating characteristic analysis, CFR < 2.2 was the optimal cut-off value for detecting LAD stenosis in all hypertensive patients (AUC 0.83, 95%CI 0.75 - 0.91) with a sensitivity of 75%, a specificity of 78%, and an accuracy of 77%. A significant reduction of diagnostic specificity was observed in patients with uncontrolled BP compared with those with well-controlled BP (67% vs. 93%, P = 0.031).</p><p><b>CONCLUSIONS</b>CFR measurement by TTDE is valuable in the diagnosis of significant coronary stenosis in hypertensive patients. However, the diagnostic specificity is reduced in patients with uncontrolled BP.</p>

Association between fractional flow reserve and quantitative coronary angiography parameters in intermediate coronary artery stenosis / 中华心血管病杂志

<p><b>OBJECTIVE</b>To explore the relationship between quantitative coronary angiography (QCA) parameters and fractional flow reserve (FFR) for identifying ideal angiographic parameters predictive of myocardial ischemia.</p><p><b>METHODS</b>The study included 121 lesions with QCA and FFR data from 106 patients [mean age: (63 ± 10) years]. The lesions were grouped into FFR > 0.75 group and FFR ≤ 0.75 group. Assessed parameters by QCA included percentage diameter stenosis, minimum luminal diameter (MLD), percentage area stenosis, minimum luminal area (MLA), reference vessel diameter (RVD) and lesion length (LL). Correlation analysis was used to identify the relationship between QCA parameters and FFR value, and receiver operating characteristic (ROC) curve was used to determine parameters predictive of FFR ≤ 0.75.</p><p><b>RESULTS</b>LL was significantly higher [(14.8 ± 7.9) mm vs. (10.7 ± 5.4) mm, P = 0.024] while MLD [(1.47 ± 0.31) mm vs. (1.82 ± 0.51) mm, P = 0.028], RVD [(2.30 ± 0.50) mm vs. (2.81 ± 0.64) mm, P = 0.036], and MLA [(2.30 ± 1.50) mm(2) vs. (3.60 ± 2.30) mm(2), P = 0.038] were significantly lower in FFR ≤ 0.75 group than in FFR > 0.75 group. LL (r = -0.209, P = 0.040) was negatively correlated with FFR, and MLD (r = 0.414, P = 0.040), RVD (r = 0.303, P = 0.000) and MLA (r = 0.315, P = 0.002) were positively correlated with FFR. ROC analysis showed that MLD ≥ 1.6 mm was the best cut-off value to predict FFR > 0.75 with sensitivity 63%, specificity 82%, and positive predictive value 96%.</p><p><b>CONCLUSIONS</b>QCA derived anatomic parameters of intermediate coronary lesions correlate to FFR value in some extent. MLD ≥ 1.6 mm is the best cut-off value to predict FFR > 0.75 in patients with intermediate coronary lesions.</p>

Apoptosis-inducing effect of gambogic acid on K562 cells and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the apoptosis-inducing effect of gambogic acid (GA) on K562 cell line and its mechanism. The K562 cells were treated with GA at different concentrations and times, the inhibition rates were detected by MTT assay. Apoptosis induced by GA was assayed by Annexin-V/PI doubling staining. The influence of GA on cell cycle was studied by propidium iodide method. The mitochondrial membrane potential was measured by JC assay. The levels of caspase 3, caspase 8 and caspase 9 activated by fluorescein in living K562 cells were measured by caspGLOW(TM) fluorescein staining kit. The results showed that after incubation with GA, K562 cell proliferation was dramatically inhibited in concentration- and time-dependent manners. K562/A02 cells need higher GA concentration (> 2 microg/ml) to show antiproliferative effect, compared with that of K562 cells (> 0.5 microg/ml). Apoptosis could be induced by GA but the influence on cell cycle was not significant. GA could decrease the mitochondrial membrane potential and increase the activated caspase 3, caspase 8, caspase 9 positive cell levels by 2.19%, -1.95%, 34.01% in 24 hr and 60.4%, 71.3%, 77.7% in 48 hr respectively. It is concluded that the GA can significantly inhibit the proliferation of K562 cells without influence on cell cycles. The GA triggers K562 cell apoptosis through both intrinsic and extrinsic pathways.

Effects of platelet-derived membrane microparticles on the proliferation and apoptosis of human umbilical vein endothelial cells / 中国实验血液学杂志

This study was purposed to investigate the effects of platelet-derived membrane microparticles (PMP) on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVEC). Different concentrations of thrombin were adopted to activate the platelets so as to release PMPs. Flow cytometry (FCM) was adopted to evaluate the efficiencies of different concentrations of thrombin to release PMPs. By using the HUVEC cultivated in vitro as vector, the effects of PMPs on the proliferation and apoptosis of HUVEC were investigated by MTT and FCM. The results showed that the efficiencies releasing PMPs from platelets activated by 2.0, 1.5, 1.0, 0.5 U/ml thrombin were 28.7, 47.7, 50.1 and 43.9% respectively; PMPs induced proliferation of HUVEC in a dose dependent manner. At the concentration of 40 microg/ml PMPs, the proliferation rate of HUVEC was 1.8 +/- 0.3 times as much as blank control, the proliferation rate in group of vascular endothelial growth factor was 1.9 +/- 0.5 times of as much as blank control, there was no statistic difference (p > 0.05). The PMPs inhibited HUVEC apoptosis. Compared with the apoptosis rate of control (9.4 +/- 0.5)%, apoptosis rate in PMP group (40 microg/ml) was (3.9 +/- 0.4)% (p < 0.05). The addition of VEGF (10 microl/ml) did not successfully prevented apoptosis of HUVEC with apoptosis rate of (8.0 +/- 0.8)%. It is concluded that platelets activated by 1 U/ml thrombin gets the best efficiency of PMP release, which stimulates proliferation of HUVEC and inhibits its apoptosis.

Test of activated plasma clotting time to assess efficacy of platelet transfusion / 中国实验血液学杂志

The study was aimed to investigate the value of activated plasma clotting time (APCT) for estimating the efficacy of platelet transfusion therapy. There were twenty patients with hematological diseases, who received transfusion of platelet, involved in the test. APCT was determined before and after transfusion of these patients, then APCT was contrasted with corresponding CCI and PPR. The results showed that 1 hour and 24 hour APCTs were shortened obviously. APCT before transfusion was (103.7 +/- 11.3) seconds, but the 1 hour and 24 hour APCTs were shortened to (60.0 +/- 9.7) seconds and (68.5 +/- 9.8) seconds respectively (P < 0.01). According to the judging criteria of CCI and PPR (CCI and PPR values at 1 and 24 hours after transfusion are < 7500, < 5000 and < 30%, < 20% respectively, the transfusion is invalid), two patients received invalid transfusion. Their 1 and 24 hour CCIs were 7415, 2966 and 6913, 4988 respectively. Their 1 and 24 hour PPRs were 28.0%, 11.2% and 25.2%, 14.1% respectively. One patient's PPR reached the standard of invalid transfusion, but his CCI showed a valid transfusion he received. Two patients' PPR reached the standard of invalid transfusion, but their 1 hour CCI reached the standard of valid transfusion, and their 24 hour CCI reached the standard of invalid transfusion. It is concluded that APCT reflects the variations of quantity and quality of platelet simultaneously, and can evaluate precisely the efficacy of platelet transfusion.

Effects of platelet-derived membrane microparticles on angiogenesis in chick chorioallantoic membranes / 中国实验血液学杂志

This study was purposed to investigate the angiogenesis effect of platelet-derived membrane microparticles (PMPs) in chick chorioallantoic membranes (CAM). Thrombin were adopted to activate the platelets and then PMPs were obtained. PMPs were isolated by high speed centrifugation. Flow cytometry (FCM) was adopted to evaluate the efficiency of thrombin to produce PMPs and BCA method was adopted to evaluate the content of PMPs. PMPs were put into CAM and the effects of PMPs on angiogenesis in CAM were observed. The results indicated that after incubation for 72 hours at the concentration of 80 microg/ml PMPs, the vessel nets in a 'spoked-wheel' pattern were shown around mixed fibrous filter membranes, number of vessel ramification was 112.5 +/- 11.31 and ratio of vessel area/CAM area was 6.19 +/- 1.29%, but there were not localized allantoic vessels developing in the control group, the number of vessel ramification and ratio of vessel area/CAM area in control group were 82.6 +/- 8.05 and 1.78 +/- 0.33 respectively, so there was significant difference between PMP and control groups. In above mentioned conditions, the number of vessel ramification and ratio of vessel area/CAM area in VEGF group were 128.4 +/- 10.02 and 7.44 +/- 1.36 respectively, so there was no difference between PMP and VEGF groups. It is concluded that PMPs show promotive effect on the formation of capillaries in chick chorioallantoic membranes.

Cold storage of rabbit platelet suspension by adding uridine diphosphate galactose / 中国实验血液学杂志

This study was aimed to investigate the method to cold-store platelets with uridine diphosphate galactose (UDP-Gal). Rabbit heart blood was prepared for concentrated platelet suspension to which UDP-Gal was added, and then stored for ten days in 4 degrees C refrigerator. Thereafter, platelet count, mean platelet volume (MPV), platelet distributing width (PDW), platelet aggregation function, platelet activity to urge coagulation including PF3aT and APCT and apoptosis were determined. Meanwhile, survival time in vivo was tested after cold-stored rabbit platelets labeled with Cr51 were transfused into rabbits. The results showed that there was not significant difference for Plt count, MPV, PDW, PF3aT and APCT between UDP-Gal cold-stored platelet group and fresh platelet group (P > 0.05). On the contrary, platelet count decreased significantly, MPV, PDW jumped and PF3aT and APCT went down in cold control group as compared with fresh platelet group (P < 0.01). Apoptosis increased in UDP-Gal cold-stored platelet group as compared with fresh platelet group (P < 0.05), but was significantly lower than that in cold control group (P < 0.01). Although PagT (inducing reagent: C-PG) decreased, it could still be above 50% of fresh platelet. Survival time in rabbit in vivo was close between UDP-Gal cold-stored platelet group and fresh platelet group (P < 0.05). Survival rate in seventy-two hours after transfusion in the fresh platelet group, UDP-Gal cold-stored platelet group and cold control group was 57.5% +/- 7.2%, 50.3% +/- 6.3% and 0.1% +/- 0.5% respectively. It is concluded that the UDP-Gal can well protect cold-stored rabbit platelets and prolong the survival time of cold-stored platelets in vivo.

Killing activity in DC and CIK co-culture against hepatocarcinoma cells / 中国实验血液学杂志

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.

Platelet-derived microparticles stimulate proliferation of granulocyte-macrophage progenitor cells from umbilical cord blood / 中国实验血液学杂志

This study was aimed to investigate the effect of platelet-derived microparticles (PMP) on stimulating the proliferation of granulocyte-macrophage progenitors (CFU-GM) from umbilical cord blood. Different concentrations of thrombin were adopted to activate the platelets for releasing PMP. Flow cytometry was adopted to evaluate the efficiencies of different concentrations of thrombin to produce PMP. Umbilical cord blood mononuclear cells (MNC) were obtained from healthy donors and the MNC were isolated by Ficoll density gradient centrifugation. MNC were cultured in 2.7% methylcellulose containing different concentration of PMP and colonies were counted under an inverted microscope after 7 days. The result showed that the release rate of PMP activated by 2.0, 1.5, 1.0 and 0.5 U/ml thrombin were 28.7%, 47.7%, 50.1% and 43.9% respectively. The PMP enhanced colony formation in dose-dependent manner. The number of colonies per 2 x 10(5) MNCs in groups of PMP at different concentrations (10, 50 and 100 microg/ml) were 119.8 +/- 32.2,142.8 +/- 45.2 and 180.8 +/- 85.1 respectively. The number of colonies in the groups of PMP at 100 microg/ml and 50 microg/ml were statistically significant when compared with control group (103.0 +/- 24.8) (P < 0.05). The number of colonies per 2 x 10(5) MNC in the group of PMP (10 microg/ml) was 119.8 +/- 32.2 which was higher than that in control group, but there was no statistical significance between two groups. It is concluded that platelet activated with 1.0 U/ml thrombin can get the best release efficiency of PMP and PMP can enhance the proliferation of granulocyte-macrophage progenitor cells of umbilical cord blood.

Evaluation of the effects of glycosylation on in vivo survival of cold-storage human platelets by using rabbit model / 中国实验血液学杂志

To study the effects of glycosylation on survival of cold-storage human platelets by using rabbit model. (51)Cr-labeling platelets were used to detect the platelet storage survival. The human platelets (2.0 x 10(12)/L) treated with 5 g/L uridine diphosphate galactose (UDP-Gal) were stored in 4 degrees C refrigeratory up to 10 days. The survival of human platelets in rabbits whose reticuloendothelial system was inhibited by the administration of ethyl palmitate was monitored in blood drawn at various times after the platelet transfusion. The results showed that the survival rate of platelets was significantly increased in cold-storage human platelets by UDP-Gal treatment. The survival rates of platelets at 2 hours after transfusion into rabbits in groups of fresh platelets group, UDP-Gal + cold platelets group and cold platelets group were (68.9 +/- 8.5)%, (65.4 +/- 8.0)% and (5.0 +/- 2.6)%, respectively. Compared with cold platelets group, significant differences were seen among all groups (P < 0.01). UDP-Gal + cold platelets group had no significant differences compared with fresh platelets group (P > 0.05). It is concluded that UDG-Gal can provide the protective effect on cold-storage human platelets and prolong the survival time of refrigerated human platelets in rabbit model.

Study on the mechanism of protective effect of oral L-arginine on intestine after scald injury in rats / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the mechanism of protective effect of oral L-arginine (L-Arg) on the intestine after scald injury in rats.</p><p><b>METHODS</b>Sixty-six Sprague-Dawley (SD) rats were randomly divided into three groups: i.e. normal control (N, n = 6, without treatment), oral L-arginine group (A, n = 30, with 1 ml 70 g/L of L-Arg per os 2 times a day from 2 post scald hour (PSH)) on with normal enteral feeding and group B (n = 30, with oral feeding of cold boiled water after scald). The changes in the content of superoxide dismutase (SOD), malondialdehyde (MDA), nitric oxide (NO), endothelin (ET), ET/NO ratio in the intestine and the level of plasma endotoxin (LPS) in portal vein were assessed at 6, 12, 24, 48, 72 PSH. Ileum tissue samples were harvested for pathological examination.</p><p><b>RESULTS</b>The ET content in the intestinal tissue in A group at 6, 12 and 24 PSH (0.80 +/- 0.26 ng/g, 0.75 +/- 0.30 ng/g, 0.63 +/- 0.22 ng/g) was obviously lower than that in B group (1.26 +/- 0.38 ng/g, 1.34 +/- 0.37 ng/g, 0.97 +/- 0.19 ng/g, P < 0.05), but the NO contents in the intestine in A group at the same time points were significantly higher than that in B group (P < 0.01). The ET/NO ratio and the level of plasma endotoxin in A group were significantly lower than those in B group at each time point (P < 0.05 or 0.01). Pathological examination showed that the intestinal mucosal injury in the A group was obviously milder than that in the B group.</p><p><b>CONCLUSION</b>Oral L-arginine was shown to have the effects to ameliorate ischemia reperfusion injury of the intestine and to protect the barrier function of the intestinal mucosa. This might be related to an increase in the NO level in intestinal mucosa resulting in maintenance of a stable ET/NO ratio.</p>

Effect of dendritic cells co-cultured with cytokine induced killer cells on cytotoxicity against drug resistant K562 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of dendritic cells (DC) co-cultured with cytokine induced killer (CIK) cells on cytotoxicity against K562 and K562 drug-resistant (K562/ADM) cells.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (MNC) isolated from healthy adult donors were induced to obtain CIK cells and DC respectively and then these two kinds of cells were co-cultured. The cytotoxicity of the co-cultured cells against K562 and K562/ADM cells was measured with MTT assay.</p><p><b>RESULTS</b>The cytotoxicity CIK cells alone to K562 and K562/ADM cells was (20.0 +/- 1.2)% - (61.1 +/- 2.2)% and (17.5 +/- 2.1)% - (45.2 +/- 3.3)% respectively at low effector to target ratios (2.5 - 20.0). This effect was significantly enhanced by co-culturing with DCs being (25.2 +/- 2.3)% - (70.9 +/- 4.1)% and (22.4 +/- 2.7)% - (62.3 +/- 5.0)%.</p><p><b>CONCLUSION</b>CIK cells showed high cytotoxicity against K562 and K562/ADM cells and the activity could be enhanced by co-culturing with DC.</p>

Effect of S-2-(3-aminopropylamino) ethyl phosphorothioic acid on apoptosis and proliferation inhibition of HL-60 cell line / 中国实验血液学杂志

To study the effects of S-2-(3-aminopropylamino) ethyl phosphorothioic acid (WR-2721, amifostine) on proliferation inhibition and apoptosis of HL-60 human leukemia cell line, the cell apoptosis rate of HL-60 was determined by annexin V/PI double staining method. Cell proliferation and chemotherapy sensitivity were analyzed with XTT assay, and the changes of cell cycle were observed through flow cytometry. The results showed that WR-2721 could significantly inhibit HL-60 cell proliferation. After treatment (30 min, 37 degrees C) with WR-2721, the sensitivity of HL-60 cells to VP16 was enhanced, and the IC(50) descended from 52.5 micro g/ml to 40.5 microg/ml. After 72 hours treatment of HL-60 cells with WR-2721, the early apoptotic cells (annexin V-FITC positive/PI negative) were increased from (5.5 +/- 1.9)% to (48.5 +/- 8.4)% (P < 0.001), late apoptotic cells (annexin V-FITC positive/PI positive) were increased from (1.2 +/- 0.5)% to (39.0 +/- 4.0)% (P < 0.001), and HL-60 cells were arrested in G(2)-M phase. In conclusion, WR-2721 treatment can enhance HL-60 cell chemotherapy sensitivity to VP16, inhibit proliferation, induce apoptosis and accumulation of cells in G(2)-M phase.

Protective effects of amifostine on hematopoietic stem/progenitor cells against chemotherapeutic damage / 中国实验血液学杂志

The aim was to study the protective effects of amifostine (AMF) on normal hematopoietic stem/progenitor cells against the chemotherapeutic damage from etoposide (VP-16). The cord blood mononuclear cells (CBMNC), fresh and frozen peripheral blood stem cells (PBSC), and HL-60 cells were divided into AMF, AMF + VP-16, VP-16 and control groups, each group cell viability was determined by using trypan blue exclusion test, the CFU-GM culture was used to count cells, the apoptosis was detected by flow cytometry. The results showed that in CBMNC, fresh and frozen PBSC samples, cell viability and the number of CFU-GM in AMF + VP-16 group were all significantly higher than those in VP-16 group (P < 0.05); the CFU-GM incidence in AMF + VP-16 group was higher than that in VP-16 group, and the GFU-GM life in AMF + VP-16 group was also longer than that of latter, in CBMNC samples, the number of CFU-GM in AMF groups was higher than that in control group, but there was no statistical significance between the two groups (P > 0.05), in HL-60 cell apoptotic rate in AMF + VP-16 group was little higher than that in VP-16 group, but no statistical significance between these two groups (P > 0.05). It is concluded that AMF can significantly protect normal hematopoietic stem/progenitor cells against the damage from VP-16. Moreover, AMF does not affect cytotoxity of VP-16 on HL-60 cells, and can not stimulate the growth and differentiation of cord hematopoietic stem/progenitor cells directly.

Effect of Autologous or Allogeneic Anti-CD3 Monoclonal Antibody Activated Killer Cells on Normal Hematopoietic Cells / 中国实验血液学杂志

In order to investigate the effect of autologous and allogeneic anti-CD3 McAb activated killer cells (CD3AK) on normal hematopoietic cells, the immobilized anti-CD3 McAb and low concentration IL-2 were used to activate CD3AK. Flow cytometry was used to assay the phenotype change of CD3AK to analyze the proportional change of CD34(+) cells in normal bone marrow mononuclear cells (BMMNC) cocultured with autologous or allogeneic CD3AK. The effect of CD3AK on normal hematopoietic progenitor cells was also assayed by methylcellulose clonogenic culture of CFU-GM. It was found that 3 - 5 micro g/ml immobilized anti-CD3 McAb and 100 U/ml IL-2 could activate CD3AK effectively. There were 99.51% CD3(+) cells in CD3AK groups. When BMMNCs from healthy volunteers were cocultured with allogeneic CD3AK for six hours, the percentage of CD34(+) cells was decreased 32.37%. CD3AK had no significant influence on autologous BMMNC. Allogeneic and autologous CD3AK were cultured with BMMNC from healthy volunteers for six hours, and then CFU-GM was evaluated. Allogeneic CD3AK inhibited 20.44% CFU-GM formation, but autologous CD3AK had no inhibition on CFU-GM. It is concluded that CD3AK has no inhibition to autologous normal hematopoietic progenitor cells after cocultured with them from these results, while allogeneic CD3AK inhibits the normal hematopoietic progenitor cells significantly.