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Four compounds were isolated from the lateral roots of Aconitum carmichaeli by Al2O3,silica gel and preparative HPLC.Their structures were identified as:(13R,15S,19S)-13,15,19-triol-hetisan (1),fuziline (2),neoline (3) and beiwutine (4),by methods of NMR,MS spectroscopy as well as single crystal X-ray crystallography.Among them,compound 1 was considered as a new diterpenoid alkaloid.
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@#This study aimed at evaluating the antioxidant effects of Guanfu base A(GFA)on acetylcholine(Ach)/CaCl2(CaCl2 10 mg/mL, Ach 66 μg/mL)-induced atrial fibrillation(AF)in rats. SD rats were rando-mized into normal group, model group, GFA treatment groups(6 mg/kg, 12 mg/kg), Amiodarone(Ami)treatment group(50 mg/kg)and Lovastatin(Lov)treatment group(10 mg/kg). The AF durations were measured by electrocardiogram(ECG). The effective refractory periods(AERP)were measured in the left atrial appendage. Oxidative stress-related gene and protein expression was evaluated by RT-PCR and Western blot. The activity of antioxidant enzymes was measured by enzymatic assay. Results indicated that, in comparison with that in the vehicle-treated AF rats, treatment with GFA(6 mg/kg, 12 mg/kg, po), significantly shortened the AF duration and prolonged the AERP in rats. In addition, treatment with GFA reduced the levels of plasma and myocardium malondialdehyde, increased the activity of plasma superoxide dismutase in a dose-dependent manner. Moreover, treatment with GFA mitigated AF-up-regulated p22phox, p47phox, gp91phox, and p67phox NADPH oxidase expression, and AF-increased ratios of membrane to cytosolic Rac-1 in the atrium. It also significantly prevented AF-down-regulated atrial connexin40 expression in rats. Data suggested that GFA(6 mg/kg, 12 mg/kg)has potent anti-oxidant activity and inhibits oxidative-stress-related AF in rats.
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OBJECTIVE To compare two different procedures of blood donation in volunteer donors,which lead to different discard rates of blood,different donation reaction rates and the satisfaction of the donor agency,so as to seek the better procedure bringing less discard of blood and more convenience for the military donor agency and blood center. METHODS In group A,3 667 donors blood was collected before the tests and retests for transfusion transmitted diseases(TTD) were done.While in group B,4 185 donors were taken blood samples for pre-donation test.The blood collection was performed 4 hours later. RESULTS In group A,3 652 units of blood were collected,of which 69 units were discarded on account of positive results in test and retest.Meanwhile,in group B 3 718 units of blood were collected from the donors who passed the pre-donation test for TTD.As a result,34 units of blood were discarded because of the positive results in retest.The discard rates of blood were 1.89% and 0.91% while the donation reaction rates were 2.22% and 3.98%,respectively.in two procedures.The discard rates of blood in group A were higher than those in group B.But the donation reaction rate in group B was higher than that in group A. CONCLUSIONS The discard rate of blood in the procedure collecting before test is higher than that in the procedure testing before collection.But the donation reaction rate is low and the waiting period for donation is short in the former procedure,which is suitable for low TTD infections population of military agencies.
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Objective To compare the specificity, sensitivity, titers, and rapidity of four methods including papain technique, anti-globulin technique, polybrene test, and micro-column gel test for determination of IgG red blood cell antibodies. Methods Twelve kinds of IgG red blood cell antibodies such as anti-D, anti-E, anti-C, anti-c, anti-e, anti-Jk a, anti-Jk b, anti-Fy a, anti-Fy b, anti-k, anti-S, and anti-s were checked by the four methods. Results Seven kinds of IgG red blood cell antibodies including anti-D, anti-E, anti-C, anti-c, anti-e, anti-Jk a, and anti-Jk b were detected using papain technique (7/12). All of the 12 kinds of IgG antibodies were detected by anti-globulin technique (12/12). Eleven kinds of IgG red blood cell antibodies except anti-k were examined with polybrene test (11/12) and all the antibodies were also determinated by micro-column gel test (12/12). The titers of the antibodies determination suggested that papain technique was less sensitive than other three methods, while the micro-column gel test was more sensitive than other three methods in examination of all the antibodies. The lasting time of four techniques were: papain technique 45 min, anti-human globulin technique 60 min, polybrene test 5 min, and micro-column gel test 30 min. Conclusion Papain technique has some limitation in determination of IgG antibodies and anti-globulin technique is complicated because of long period incubation and multiple wash of red blood cells. Polybrene test is the most simple and convenient technique for determination of IgG antibodies. Micro-column gel test is the most sensitive method in determination of IgG antibodies.
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Objective To study the preservative indices of stored platelet and verify the effect of a new platelet incubator made in China. Methods 12 bags of fresh platelet rich plasma (PRP) were collected. Each bag was equally divided into 2.One bag was stored for 5 days in XHZ-IA platelet incubator which was made in China, and the other bag stored in FORMA-3606 platelet incubator made in USA. Some preservative indices of stored platelet e.g. aggregation rate, recovery rate after response to hypotonic shock, CD62p expression rate etc. were tested daily. Results There were no significant differences in these preservative indices between platelets stored in two platelet incubatorshas. Conclusion XHZ-IA platelet incubator has the same effect on PRP preservation as FORMA-3606 platelet incubator.
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Objective To explore the changes in plasma fibronectin level in patients with SARS in the acute stage, and the feasibility of the use of plasma fibronectin in acute stage of SARS. Methods Plasma fibronectin contents were assayed with ELISA in patients with SARS in acute stage and 24 healthy donors. Results The plasma fibronectin level of control group was 476.2?221.0mg/L, while that of four SARS patients was 42.2?23.0 mg/L(P
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To study the formation and change of platelet leucocyte aggregation (PLA) in red blood cell concentrations(CRCs) stored at 4℃ and the preventive effectiveness of filtration, 8 units of CRCs were each divided into two equal parts in two integrative bags and two paired groups were formed: control group and filtration group. The following items of control group and filtration group were detected weekly. Platelet and leukocyte counts, concentration and percentage of platelet lympocyte (P lym), platelet monocyte (P mon), platelet neutrophil (P neu) aggregatoins, and concentration of total PLA. The results showed that rate and concentration of PLA in the control group were significanly higher than those in the circulating blood. The rates of P lym and P mon in CRCs stored at 4 ℃ were two to five times of those before storage, and the rate of P neu in CRCs stored at 4 ℃ was ten to twenty times of that before storage. PLA in the CRCs separated from plasma rose to a high level at the first day of preparation. The concentration of PLA in CRCs reached the highest level at one week after stored at 4 ℃ and then decreased, but a high concentration was retained all the while. P neu is the major part (about 80%～90%) of whole PLA in CRCs. PLA was not detectable by flow cytometry during the whole storage course. The results suggested that high concentrated PLA was detected in CRCs stored at 4 ℃, which could be prevented by leucocyte depleting filtration.
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Objective To analyze the curative effect of blood transfusion on 100 patients undergoing liver transplantation,to efficiently enhance hemostasis and coagulation function of the patients,to lower the incidence of the operative complications,and to put forward the guideline of blood transfusion to the patients undergoing liver transplantation.Methods The volumes of red blood cells concentrates(RCCs),fresh frozen plasma(FFP),cryopreserved platelets,and cryoprecipitate transfused to the patients undergoing liver transplantation were measured before,during and after the operation respectively.The volumes of blood components transfused to the 50 patients in the former stage and 50 patients in the later stage of liver transplantation in our hospital were also measured respectively.Results The mean volume of the blood components transfused to each patient was 3 334ml.The ratios of the transfused blood volumes to 100 patients were 4.3%,53.4% and 42.3% before,during and after operation respectively.The rate of blood components transfusion was 100% and the ratio of transfused blood volumes to the patients between RCCs and FFP was 1.45:1.The mean volumes of the transfused blood and the operative complications of 50 patients in the former stage of liver transplantation of our hospital were significantly higher than those of the 50 patients in the later stage.Conclusion It is very vital in the scientific,safe and reasonable blood transfusion for the success of liver transplantation,which significantly lowers the incidence of operative complications,such as massive haemorrhage during and after operation,and thrombosis at inosculation of liver artery and portal vein.
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Objective To study the technology of loading trehalose into cytoplasm of human platelets before lyophilization,and to determine the optimal experimental conditions of pretreatment technology.Methods The curves of loading efficiency,the internal trehalose concentration versus incubated time,temperature,and external trehalose concentration were drawn up to screen the optimal loading parameters.Results The loading efficiency was linear to the incubated time(2 hours later) and incubated temperature(range from 30℃ to 40℃),respectively.The loading efficiency almost reached 60 percent when the platelets were incubated at 37℃ for 4h.The intracellular trehalose concentration was increased with the raise of the external trehalose concentration(
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Objective To prolong the shelf life of RBCs stored at non-4℃, and improve the ability of blood supplying during wartime. Methods 12 bags of packed RBC were collected from 12 volunteers presenting blood, respectively, and each was averagely divided into 5 parts(groups): combination group, new solution group, ammonia group, control group and 4℃ group. The fist 4 groups were stored at room temperature, and were tested for ATP concentration daily. 4℃ group was stored at 4℃, and RBC ATP concentration was measured at the end of its shelf life, which served as a critical ATP concentration. When RBC ATP concentration of the fist 4 groups decreased to the level of critical ATP, the time of preservation was their shelf life. 6 bags of packed RBC were added the same solution as the combination group, and each was averagely divided into 4 parts(groups)after stored at room temperature for 12 days, which were washed for 0, 5,10 and 15min before centrifugation, respectively. The residual ammonia and the recovery ratio of RBC were measured after washing. Results The shelf life of combination group,new solution group,ammonia group and control group was 12,6,6 and 5 days, respectively. The residual ammonia could be effectively cleaned up after either washing 4 times with 5 min balance time or 3 times with 10 or 15 min balance time. The recovery ratio of RBC was 88.5% and 83.1% after washing 3 and 4 times, respectively. Conclusion The combination of new solution and ammonia was a good additive solution for RBC preservation at non-4℃, and the residual ammonia can be cleaned up using normal saline.
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0.05).⑵ With the increase of titers of anti-A(B) antibodies of IgG type, both the incidence and severity of HDN elevated (P
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Objective To explore the factors affecting the collection of allogenic peripheral blood stem cells in donors. Methods Using COBE-spectra blood cell separator to collect peripheral blood stem cells in 21 allogenic donors. The impact of the factors including the number of white blood cell(WBC) and nucleated cell(NC), and mono-nucleated cell(MNC) ratio on the collecting rate of peripheral blood stem cells was analyzed before and after collection. The output of CD 34 +stem cells stimulated by different doses of G-CSF was also analyzed. Results When the mean count of peripheral WBC and MMC in donors was (41?7.7)?10 9/L and 12?2.9% respectively, the count of collected NC was 2.7?0.8?10 8/L, and the number of the collected CD 34 + stem cells was also higher. Statistical analysis showed that the number of NC was significantly related to the collecting rate of CD 34 stem cells(P
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Objective To study the effect of leukoreduction transfusion on improving medical quality. Methods The cases in our hospital which had RBC transfusion records in the past 3 years were collected and divided into two groups: leukoreduction group(LR) and non-leukoreduction group(NLR). Through statistical analysis, the items related to leukoreduction were found out and the differences in these items between the two groups were compared. Results Operation complication, non-operation complication, infectious rate, transfusion reactions and mortality rate in LR group were significantly lower than those in NLR group. The transfusion of single components-RBC had less clinical side-effect than the transfusion of complex components including RBC. Conclusions Leukoreduction transfusion can effectively improve clinical therapy and promote the medical quality.
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Objective To study the injurous effect of leukocytes on vital organs of hemorrhagic shock dog and role of leukocyte depletion filter in reducing the injury.Methods A canine model of hemorrhagic shock was established.12 male dogs were divided into blank group,whole blood transfusion group and leukocyte depletion group. Same amount of 0 9% physiological saline,whole blood of the canine itself and filtrated whole blood (i.e. lack of leukocytes) were back transfused respective correspondingly, changes in parameters such as white blood cells counting, LDH, MDA, GOT and the pathological changes in heart, lung, liver and kidney were detected among different treatment groups.Results Higher level of WBC,LDH, AST and MDA were observed in the group with whole blood transfused than in the group of leukocyte-depletion transfused. The pathological changes in liver and kidney indicated more serious injury happened in whole blood group than in the leukocyte-depletion group.Conclusion The Leukocyte depletion filter in itself can definitely alleviate the damages in the canine vital organs when reperfusion is inevitable as a result of hemorrhagic shock.
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Objective To study the effect of oxigenated leukodepletion filter(OLF), in order to evaluate its potential value in battlefield trauma application.Methods 12 bags of whole blood were filtered by OLFs, while the other 12 by normal filters,some indexes including ,partial pressure of oxygen(PO 2),O 2 saturation of oxygen(SO 2),HbO 2 WBC,RBC counter and volume of RBC supernatant were measured before and after leukocyte filtration.Results PO 2,SO 2 and HbO 2 increased 57%,51% and 53% respectively after filtration by OLFs, the residual leukocyte count decreased to (3 37?0 54)?10 7 each bag,and the leukocyte clearance rate was (97 52?0 74)%.Conclusions OLF can improve the effect of blood transfusion and reduce some acute transfusion adverse reactions.Thereby,application of the OLF has binefits significantly to heal the wounded patients in battlefield trauma.
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Objective To establish a simple and feasible method of preparing young erythrocytes and test some indexes of evaluating the mean cell age.Methods We collected 34 bags of whole blood from healthy blood donors,400ml/each bag,then eliminated WBCs by leukoreduction filter.After twice centrifuging and manual speration,we finally obtained young erythrocytes from the upper section of the red cell layer,the remains were old erythrocytes.These different age erythrocytes were separated by blood bags.Some indexes as pyruvate kinase,reticulocyte count,mean corpuscular volume(MCV),mean corpuscular hemoglobin concentraion(MCHC),erythrocytes deformation index EI were tested on the non-separation erythrocytes,young erythrocytes and old erythrocytes,simultaneouosly,the detecting hemoglobin recovery rate.Results There was significant difference of all indexes among the three kinds of erythrocyte products.The PK activity ratio of young erythrocytes' to non-preparating erythrocytes was 1 175?0 13,showed the mean cell age was about 40 days.The hemoglobin recovery rate was(48 37?3 82)%.Conclusions The method is effective for preparing young erythrocytes,PK,REI,MCV,MCHC and EI can be used to evaluate the young erythrocytes' mean cell age.
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0.01).After incubation with trehalose for 4h,the expression of CD62p increased,and it decreased when platelets activation inhibitor PGE-1 and adenosine were added to the incubation buffer.The expression of PAC-1 showed no difference among the three groups. Conclusion The preprocessing before platelets lyophilization has no significant effects on the function and activity of platelets when the platelets activation inhibitor is added.
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Objective To establish the technique of loading trehalose into human platelets including temperature, time and concentration of trehalose loading. Methods Intracellular trehalose concentration versus loading time curves at 37℃ and 16℃, mean plateletee volume(MPV) versus loading time curves, and loading concentration curves were investigated and compared. Then loading time and temperature for high loading efficiency of trehalose into human platelets were ascertained. Intracellular trehalose versus trehalose loading concentration curves were made, and loading efficiency was caculated. Results The trehalose loading efficiency of 4 hours at 37℃ was significantly higher than that at 16℃ (11.6% vs 5.6%). MPV at 16℃ increased 43.2% compared with at 37℃, but had no changes along with the change of loading time and concentration. When loading at 37℃, MPV change was positively related with loading time and concentration, and loading time and loading concentration had a cooperative effect. MPV had no changes in 1～4hours loading when trehalose loading concentration was under 50mM. MPV increased with the increase of loading time and concentration when trehalose loading concentration was above 50mM. When trehalose loading concentration was 45mM, loading efficiency at 37℃ reached 13.86% 4 hours aftr loading. Conclusion The basic parameter of trehalose loading technique of human platelets was loading temperature at 37℃, loading time for 4 hours, and loading concentration of 45mM.
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Objective To study the changes of CPDA whole blood's shelf life after stored at non-4℃. Methods 200 ml whole blood was collected from each of 10 donors and anticoagulated by CPDA. Then 50 ml whole blood from each one was marked as control group, the rest was marked as test group. The blood of the control group was stored at 4℃ and RBC ATP concentration at the end of its shelf life served as critical ATP. The blood of the test group was stored at 10℃ for 24 hours, then it was transferred to 4℃ refrigeratory and continued to preserve. RBC ATP concentration were tested daily to ensure it eligible. When RBC ATP concentration decreased to the level of critical ATP, the time of preservation at 4℃ was blood's succeeding shelf life. Using the same method, the succeeding shelf life of CPDA whole blood after stored at different temperature for 48 to 72 hours was measured. Results The succeeding shelf life of CPDA whole blood did not obviously change after stored at non-4℃ for 24 hours, while it shortened dramaticly after stored at non-4℃ for more than 48 hours. The higher the temperature, the shorter the succeeding shelf life. Conclusion The results suggested that CPDA whole blood's succeeding shelf life should be adjusted after stored at non-4℃ for 24 to 72 hours to ensure the blood quality.
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Objective To evaluate the biological activity of anti-Rh(D) antibodies in vitro. Methods The titres of plasma Anti-Rh (D) antibodies in the RH(-) patients uncorrectly infused RH(+) blood were detected by various methods. The anti-D antibodies were evaluated for their capacity to sensitize erythrocytes, trigger monocyte and K-cell mediated antibody-dependent cellular cytotoxicity (ADCC), mediate binding to monocyte and lymphocyte Fc gamma R, and stimulate phagocytosis by monocytes. The synergistic effect of anti-D antibodies in promoting hemolysis was detected by complement fixation test (CFT). Results The titre of plasma anti-D antibodies measured by micro-column gel indirect anti-globulin technique(MGIAT) was the highest compared with the other detection methods. When red cells were sensitized with anti-D antibodies, the binding and phagocytosis of red cells by monocytes and the lysis of red cells by monocytes or lymphocytes were great, and the hemolysis of red cells by alexin was great too. Conclusion Anti-Rh (D) antibodies had great biological activity in vitro, and MGIAT had the highest sensitivity for detecting anti-Rh(D) antibodies.