Improving oral absorption of BCS Ⅱ drugs by increasing solubility: frequently overlooked permeability / 药学学报

BCS Ⅱ drugs are characterized by low solubility and high permeability. Improving their solubility is considered an important approach to improve its oral absorption. Recent strategies to increase the solubility of poorly-soluble drugs may unexpectedly result in greatly depressed permeability, ultimately leading to failure in improving oral absorption. Based on the mathematics of membrane permeability coefficient of a drug, the membrane/aqueous partition coefficient is dependent on the drug's solubility in the gastrointestinal milieu, suggesting a unique interplay between the solubility and permeability of the drug, and treating the one irrespectively of the other may be insufficient. When we focus on the increase of drug solubility and overlook the efficacy of drug permeability, the positive effect of increased solubility to drug oral absorption might be traded off by depressed permeability. To provide rational formulary designs, by optimizing excipients and evaluation, this review summarizes solubility- permeability interplay for different types of solubilizing techniques, such as cyclodextrin, surfactants-based vehicle, cosolvent, amorphous solid dispersions, other infectors such as P-gp transporters and new techniques for simultaneous evaluation of drug solubility and permeability.

An in vitro method for preliminarily screening competing agents to enhance drug permeation in cyclodextrin inclusion complexes / 药学学报

Cyclodextrin can increase the solubility of poorly soluble drugs, but also decrease the permeability of poorly soluble drugs in inclusion complexes simultaneously, which partially or completely counteracts the contribution of improvement in solubility to the oral absorption of poorly soluble drugs. If a competing agent is added to the system to compete binding sites of cyclodextrins with drugs, drug permeability can be improved by increasing the concentration of free drugs in the inclusion complex system. In this paper, a rapid in vitro screening method for competing agents of cyclodextrin inclusion complex is proposed based on the principle that good drug permeability is in accord with good cell uptake. The equilibrium constants between drugs and hydroxypropyl-beta-cyclodextrin (HPCD) were determined by phase equilibrium solubility method. Cinnarizine (CN) with a high equilibrium constant was selected as a competing agent, coumarin 6 (C6) and 9-octadecyl berberine (BD) with smaller equilibrium constants were selected as model drugs. Both changes of solubility and uptake by Caco-2 and A549 cells of C6 and BD were investigated different concentrations of CN to the HPCD solution of C6 and BD. The results showed that the uptake of C6 and BD increased in a CN concentration-dependent manner, and the solubility of C6 and BD in HPCD solution decreased with the prolongation of equilibrium time. It might be due to increased free drug concentrations that resulted from the competition of CN for drug binding sites with HPCD. In our study, in vitro cell uptake method was firstly used to validate the ability of CN as a competing agent to increase drug permeability (cell uptake). This method can be used for preliminarily screening of competing agents for drug-cyclodextrin inclusion complexes.

Evaluation and Comparison of Thromboelastography and Conventional Coagulation Tests for Blood Coagulation Function in Children with Henoch-Schönlein Purpura / 中国实验血液学杂志

OBJECTIVE@#To evaluate the coagulation function of children with Henoch-Schönlein purpura (HSP) by thromboelastography (TEG) and conventional coagulation tests (CCTs), and to explore the correlation and consistency of the 2 test methods.@*METHODS@#A total of 468 children with HSP were selected from January 2017 to December 2017 in Beijing Children's Hospital, Capital Medical University. The TEG and CCTs data were analyzed to evaluate coagulation function of children with HSP, meanwhile, the coagulation results were analysed the superiority of the 2 test methods was compared by Pearson correlation and Kappa consistency analysis.@*RESULTS@#There were no clinically significant abnormalities practically in HSP children by TEG and CCTs analysis, except for D-dimer level was elevated (t=9.15, P＜0.001). There were no significant changes for coagulation data from, sex comparison of HSP children (P＞0.05 all), but the coagulation reaction time (R), blood clot formation time (K), α-Angle, CI value, fibrinogen, D-dimer and anti-thrombin III in HSP children with different age groups showed difference (P＜0.05 all), and the blood in children aged 0-2 years old tended to be hypercoagulable. The TEG indexes demonstrated no significant difference in coagulation function of children with HSP each other (P＞0.05). However, CCTs data showed that the blood in children with severe kidney involvement were hypercoagulable. Comparision results of the correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children showed that R was weakly correlated with prothrombin time (PT), International Normalized Ratio (INR) and activated partial thromboplastin time (APTT). There were weak correlation between K, α-Angle and Fib (0.1＜|r|＜0.4 all). There was no obvious consistency between them each other (kappa＜0.4 all).@*CONCLUSION@#The overall changes in coagulation function in children with HSP are not obvious, but the hyperfibrinolysis in hypercoagulable state may exists. Furthermore, younger age and severe kidney involvement may cause hypercoagulation in HSP children. The weakly correlation and consistency of TEG and CCTs in detecting coagulation function of HSP children are furtherly confirmation, and the 2 test methods may be irreplaceable.

Preparation of Rat Spinal Synaptosome and Qualitative and Quantitative Test of Neurotransmitters / 中国康复理论与实践

Objective:To prepare rat spinal synaptosomes, and to conduct qualitative and quantitative analysis of various neurotransmitters in rat spinal synaptosomes to explore the physiological activities. Methods:Rat spinal synaptosomes were prepared by discontinuous Percoll density gradient centrifugation and their morphology and structure were observed under transmission electron microscopy. The properties and content of neurotransmitters in rat spinal synaptosomes were determined by ultra performance liquid chromatography-tandem mass spectrometry. Results:Under the experimental conditions, rat spinal synaptosomes were found to contain dopamine, norepinephrine, serotonin, homovanillic acid, 3,4-dihydroxybenzene, 5-hydroxyindoleacetic acid, methoxytyramine, tryptophan, γ-aminobutyric acid, kynurenine, 3-hydroxy kynurenine and 3-hydroxy-2-aminobenzoic acid, and so on. The corresponding neurotransmitter contents were detected, and the corresponding linear relationship and correlation coefficient were obtained. Conclusion:Discontinuous Percoll density gradient centrifugation could prepare rat spinal synaptosomes and detect neurotransmitters and their contents in rat spinal synaptosomes.

DNA hypermethylation of Fgf16 and Tbx22 associated with cleft palate during palatal fusion

Abstract Objective: Cleft palate (CP) is a congenital birth defect caused by the failure of palatal fusion. Little is known about the potential role of DNA methylation in the pathogenesis of CP. This study aimed to explore the potential role of DNA methylation in the mechanism of CP. Methodology: We established an all-trans retinoic acid (ATRA)-induced CP model in C57BL/6J mice and used methylation-dependent restriction enzymes (MethylRAD, FspEI) combined with high-throughput sequencing (HiSeq X Ten) to compare genome-wide DNA methylation profiles of embryonic mouse palatal tissues, between embryos from ATRA-treated vs. untreated mice, at embryonic gestation day 14.5 (E14.5) (n=3 per group). To confirm differentially methylated levels of susceptible genes, real-time quantitative PCR (qPCR) was used to correlate expression of differentially methylated genes related to CP. Results: We identified 196 differentially methylated genes, including 17,298 differentially methylated CCGG sites between ATRA-treated vs. untreated embryonic mouse palatal tissues (P<0.05, log2FC>1). The CP-related genes Fgf16 (P=0.008, log2FC=1.13) and Tbx22 (P=0.011, log2FC=1.64,) were hypermethylated. Analysis of Fgf16 and Tbx22, using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG), identified 3 GO terms and 1 KEGG pathway functionally related to palatal fusion. The qPCR showed that changes in expression level negatively correlated with methylation levels. Conclusions: Taken together, these results suggest that hypermethylation of Fgf16 and Tbx22 is associated with decreased gene expression, which might be responsible for developmental failure of palatal fusion, eventually resulting in the formation of CP.

Factors affecting thrombolysis in myocardial infarction myocardial perfusion frame count: insights of myocardial tissue-level reperfusion from a novel index for assessing myocardial perfusion / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Myocardial tissue-level perfusion failure is associated with adverse outcomes following ST-elevation myocardial infarction (STEMI) despite successful epicardial recanalization. We have developed a new quantitative index-thrombolysis in myocardial infarction (TIMI) myocardial perfusion frame count (TMPFC)--for assessing myocardial tissue level perfusion. However, factors affecting this novel index of myocardial perfusion are currently unknown.</p><p><b>METHODS</b>A total of 255 consecutive STEMI patients undergoing primary angioplasty were enrolled. Myocardial tissue level perfusion was assessed by TMPFC, which measures the filling and clearance of contrast in the myocardium using cine-angiographic frame counting. We differentiate three groups with two cut off values for TMPFC: a TMPFC of 90 frames was the upper boundary of the 95% confidence interval (CI) for the TMPFC observed in normal arteries, and a TMPFC of 130 was the 75th percentile of TMPFC.</p><p><b>RESULTS</b>STEMI patients with TMPFC > 130 frames (68 patients, 26.7%) had higher clinical and angiographic risk factor profiles as well as a higher 30-day MACE rate compared with those with TMPFC ≤ 90 frames and those with TMPFC > 90 and ≤ 130 frames. Multivariable analysis identified that the independent predictors of TMPFC > 130 frames were age ≥ 75 years (OR 2.08, 95%CI 1.21 to 3.58, P = 0.007), diabetes (OR 1.37, 95%CI 1.01 to 1.86, P = 0.042), Killip class ≥ 2 (OR 1.52, 95%CI 1.05 to 2.21, P = 0.027), and prolonged pain-to-balloon time (OR 1.73, 95%CI 1.07 to 2.79, P = 0.013). TMPFC > 130 frames was identified as the strongest independent predictor of 30-day major adverse cardiac event (MACE) (OR 2.77, 95%CI 1.21 to 6.31, P = 0.008), along with age ≥ 75 years (OR 2.19, 95%CI 1.11 to 4.33, P = 0.016), female gender (OR 1.67, 95%CI 1.03 to 2.70, P = 0.038), and Killip class ≥ 2 (OR 1.83, 95%CI 1.07 to 3.14, P = 0.021).</p><p><b>CONCLUSIONS</b>STEMI patients with poor myocardial perfusion assessed by TMPFC had higher risk factor profiles. Advanced age, diabetes, higher Killip class, and longer ischemia time were independent predictors of impaired TMPFC after primary percutaneous coronary intervention. These results emphasize that particular attention should be paid on myocardial microvascular reperfusion in STEMI patients with these risk factors.</p>

Effect of levosimendan on B-type natriuretic peptide levels in patients with advanced heart failure: a meta-analysis / 南方医科大学学报

<p><b>OBJECTIVE</b>To conduct a meta-analysis of the effect of levosimendan on B-type natriuretic peptide (BNP) levels and evaluate the therapeutic effect of levosimendan on advanced heart failure.</p><p><b>METHODS</b>A meta-analysis was performed on the selected data to analyze the effect of levosimendan on BNP levels.</p><p><b>RESULTS</b>Levosimendan decreased BNP by a mean of 337.66 [95%CI (-296.30, -379.02)] pg/ml 24 h after the administration, and by 259.92 [95%CI (-195.76, -324.08)] pg/ml at 48 h, and by 123.09 [95%CI(-53.32,-195.86)] pg/ml at 1 week. Levosimendan resulted in improvements of the cardiac function by about 29%, 22%, and 10% at 24 h, 48 h and 1 week after the administration.</p><p><b>CONCLUSION</b>Levosimendan produces favorable effects on the cardiac functions and BNP levels.</p>

Remifentanil preconditioning lowers cardiac troponin I levels in patients undergoing off-pump coronary artery bypass graft surgery / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the value of cardiac troponin I (cTnI) levels in assessing myocardial protection by remifentanil precondition against myocardial injury induced by off-pump coronary artery bypass (OPCAB).</p><p><b>METHODS</b>Twenty-four patients undergoing OPCAB were randomized into control and remifentanil preconditioning group (n=12). All the patients received pretreatment with oral diazepam (10 mg), intramuscular morphine (10 mg) and hyosine (0.3 mg). General anesthesia was induced with midazolam (0.08 mg/kg), etomidate (0.1-0.3 mg/kg), fentanyl (5-10 microg/kg), and rocuronium (1 mg/kg), and maintained with isoflurane inhalation and propofol infusion. Intermittent fentanyl and pipecuronium were given intravenously. In remifentanil preconditioning group, remifentanil (5 microg/kg in 50 ml normal saline) was infused in 10 min after anesthesia induction, and only NS was administered in the control group. Blood samples were obtained before and at 0, 2, 6, 24, and 48 h after the operation to determine serum cTnI levels.</p><p><b>RESULTS</b>In both of the two groups, the cTnI levels increased significantly at the postoperative time points (0, 2, 6, 24, and 48 h) as compared with those before the operation (P<0.05). The cTnI levels of remifentanil preconditioning group were markedly decreased after the operation in comparison with those of the control group (P<0.05).</p><p><b>CONCLUSION</b>Remifentanil preconditioning decreases the cTnI levels and reduces myocardial injury induced by OPCAB.</p>

Establishment of method collecting peripheral blood hematopoietic stem/progenitor cells from infants / 中国实验血液学杂志

The aim of this study was to explore a safe method collecting peripheral blood stem/progenitor cell (PBSPC) from the infants of body weight less than 20 kg by using the COBE Spectra Blood Cell Separator through Auto-PBSC procedure. After washing tube by normal saline, one unit of irradiated RBC was infused into the apheresis set. When the collection terminated, only the concentrated RBC in the apheresis set was returned to the infant. The peripheral mononuclear cells (PBMNCs) and CD34+ cells were counted, the cell viability was determined. The results showed that 13 PBSPC collections were carried out successfully from 7 infants of body weight<20 kg. The average count of MNCs was 4.44x10(8)/kg [(3.46-6.45)x10(8)/kg], the CD34+ count was 2.20x10(6)/kg [(1.34-3.79)x10(6)/kg] and the cell viability was 98.45% (97%-100%) respectively. The vital signs of all the infants went smoothly during collection of PBSPCs. In conclusion, with the aid of COBE Spectra blood cell separator and other measures, the collection of PBSPCs from infants of body weight<20 kg is safe and effective, the PBMNCs containing enough PBSPC can be harvested for transplantation.

Frequency of ABO type IgM antibody in 276 infants younger than six months / 中国实验血液学杂志

The aim of study was to explore the frequency of ABO type IgM antibody in infants younger than six months. 309 hospitalized infants younger than six months were selected at first and their EDTA K(3) anticoagulant blood samples were taken. All the infants were divided into five groups: neonates within 1 week as group I; neonates aged 8 to 14 days as group II; neonates aged 15 days to 1 month as group III; infants aged two to 3 months as group IV and infants aged 4 to 6 months as group V. The monocolonal anti-A, anti-B serums, A cells, B cells and O cells were utilized to carried out the blood typing with tube test. The results indicated that from 309 samples tested 33 AB type sample were excluded. Out of the remains of 276 samples, 29 of 46 samples in group I were positive and with the ABO type consistent rate 63% (29/46); 41 of 64 samples in group II were positive and with the ABO type consistent rate 64% (41/64); 47 of 74 samples in group III were positive and with the ABO type consistent rate 63% (47/74); 28 of 45 samples in group IV were positive and with the ABO type consistent rate 62% (28/45); 40 of 47 samples in group V were positive and with the ABO type consistent rate 85%. It is concluded that the ABO type IgM antibody appear in most infants younger than six months and these IgM antibodies may be regarded as the important evidence for ABO typing in infants.

Etiologic analysis of severe neonatal upper respiratory tract obstruction / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the causes of severe upper respiratory tract obstruction in neonates.</p><p><b>METHODS</b>Forty seven cases with severe dyspnea history were reviewed and retrospective analysis performed. Clinical manifestation, direct laryngoscope, CT, X-ray barium meal examination results and the treatment process were recorded.</p><p><b>RESULTS</b>Among 47 neonates, 41 cases were congenital diseases, accounting for 87.2% (41/47). In 41 cases, 15 cases were congenital laryngeal stridor, accounting for 31.9%, involving 6 cases accompanied by the gastroesophageal reflux. Fourteen cases were congenital upper respiratory tract cyst (10 cases of lingual root cyst, 3 cases of epiglottis cyst, 1 case of laryngeal cyst), accounting for 29.8%, in which 13 cases were misdiagnosis as congenital laryngeal stridor. Others were 6 cases of acute membrane laryngotracheobronchitis, 3 cases of congenital main respiratory tract stenosis, 2 cases of congenital laryngeal webs, 2 cases of vocal cord paralysis, 3 cases of Pierre Robin syndrome, 2 cases of Cri-du-chat syndrome. Except for 3 in 47 cases whose parents refused treatment, dyspnea in other 44 cases were relieved with inhaling oxygen and drug treatment, 37 case received tracheal intubation and sputum suction, 19 cases received direct laryngoscope or self-retaining laryngoscope operation.</p><p><b>CONCLUSIONS</b>Congenital disease is the main cause of severe upper respiratory tract obstruction. For the neonates suffering from severe upper respiratory obstruction, finding out the cause and prompt treatment are essential for prognosis and reducing mortality.</p>

Prognostic value of plasma brain natriuretic peptide and C-reactive protein in patients with acute coronary syndromes underwent percutaneous coronary intervention / 中华心血管病杂志

<p><b>OBJECTIVE</b>To evaluate the prognostic value of plasma brain natriuretic peptide (BNP) and C-reactive protein (CRP) in patients with acute coronary syndromes (ACS) underwent percutaneous coronary intervention (PCI).</p><p><b>METHODS</b>Patients with ACS underwent PCI in our hospital from December 2004 to September 2005 were included in this study. Plasma BNP (n = 189) and CRP (n = 141) were measured at a median of (34.2 +/- 16.3) hours from symptom onset, total mortality and the risk for major adverse cardiac events (MACE, including death, recurrent MI, recurrent angina, heart failure, readmission for any reason) at 30 days and at 3 months was analyzed.</p><p><b>RESULTS</b>Patients were divided into 4 groups according to their BNP levels (BNP <or= 100 ng/L, BNP > 100 ng/L to <or= 300 ng/L, BNP > 300 ng/L to <or= 600 ng/L and BNP > 600 ng/L) and the 3-month mortality was 0%, 1.4%, 7.7%, 48.3% and 3-month incidence of MACE was 7.9%, 17.1%, 57.7%, 79.3% respectively. Multivariate logistic regression analyses showed that the plasma BNP level predicted 30-day (r = 0.8515, P < 0.01) and 3-month (r = 0.9201, P < 0.01) mortality and 30-day (r = 0.7066, P < 0.01) and 3-month (r = 0.7090, P < 0.01) incidence of MACE independent of other known prognostic factors such as age, gender, family heredity, hypercholesterolemia diabetes, hypertension, smoking and LVEF. Patients were divided into 3 groups according to their CRP levels (CRP <or= 8.0 mg/L, CRP > 8.0 mg/L to <or= 32.0 mg/L, CRP > 32.0 mg/L) and 3-month mortality was 2.7%, 7.7% and 28.6% and 3-month incidence of MACE was 28.4%, 41.0% and 60.7% respectively. CRP predicted 30-day (r = 0.5882, P = 0.0044) and 3-month (r = 0.5235, P = 0.0038) mortality independent of traditional risk factors, and predicted 30-day (r = 0.2705, P = 0.0380) and 3-month (r = 0.2290, P = 0.0429) incidence of MACE after adjustment for patient age. CRP lost its predictive value after BNP was introduced into the model, while BNP was still an independent predictor for mortality and incidence of MACE at 30 days and 3 months in ACS patients underwent PCI.</p><p><b>CONCLUSION</b>Both plasma BNP and CRP are good predictors for early mortality and MACE incidence in ACS patients underwent PCI.</p>

The clinical value of sixteen-detector row computed tomography angiography for the assessment of coronary artery bypass graft / 中华心血管病杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the value of sixteen-detector row computed tomography angiography (CTA) for the assessment of coronary artery bypass graft (CABG).</p><p><b>METHODS</b>Sixty-two consecutive patients undergoing coronary artery bypass grafting were recruited. Among them, 6 patients were excluded from the study due to unfavorable control of heart rate. A total of 56 patients with 152 coronary artery bypass grafts (internal mammary artery, n = 48; saphenous venous grafts, n = 104) were examined by computed tomography angiography (CTA) with sixteen-detector row CT and by conventional invasive coronary angiography (CAG). All CT procedures were performed with retrospective electrocardiogram gating method. The patients' mean heart rate was 58 +/- 6 beats/minute. 120 ml of Visipaque 320 were continuously injected with the rate of 4.0 ml/sec during the procedure. The patency and the stenosis of coronary artery bypass grafts were evaluated by two experienced readers.</p><p><b>RESULTS</b>All the coronary artery bypass grafts were visualized by CTA, and all the proximal bypass anastomoses and 71% of the distal bypass anastomoses were also visualized by CTA. Furthermore, 29 occlusions and 13 significant stenoses of coronary artery bypass grafts were detected by CTA. The comparison of the results between CTA and CAG showed that among all the 42 occluded and stenosed coronary artery bypass grafts detected by CTA, 34 were confirmed by CAG; among all the 110 normal coronary artery bypass grafts detected by CTA, 108 were confirmed by CAG. There were 8 false positive and 2 false negative findings, resulting in a sensitivity of 94%, a specificity of 95%, a positive predictive value of 86%, and a negative predictive value of 99%.</p><p><b>CONCLUSION</b>Sixteen-detector row CTA technology may provide a reliable visualization and higher diagnostic accuracy of coronary artery bypass grafts lesions. This technique can be used as a noninvasive procedure for the diagnosis of suspected coronary artery bypass grafts dysfunction.</p>

Slide platelet aggregation test used as a monitor for patient treated with anti-pletelet drugs / 中国实验血液学杂志

The aim was to verify the effectiveness of slide platelet aggregation test (SPAT) to monitor the inhibition effect of anti-platelet drugs. A group of eight healthy volunteers was examined for SPAT value and T(50) (time necessary for reaching 50% of total aggregation) induced by ADP, arachidonic acid (AA) and cationic propyl gallate (c-PG) respectively before and after administration of ASA in dose of 100 mg/day for 3 days. The group of 41 inpatients at the Department of Cardiovascular Disease treated with anti-platelet drugs and the group of 327 healthy blood donors were also examined for SPAT. The SPAT value of healthy volunteer samples stored at room temperature were measured hourly for four hours. The results showed that: (1) no significant difference was detected between the T(50) before and after ASA administration in health volunteer group when ADP was used as inducer, but a significant difference was detected in this group when AA or c-PG was used as inducer. There was significant linear correlation between SPAT value and T(50) induced by c-PG in health volunteer group before and after administration of ASA (r = 0.998, P = 0.000); (2) there was no significant difference between the SPAT value of health volunteer group before administration of ASA and the SPAT value of health blood donors group (P = 0.853), but there was a significant difference between the SPAT values before and after administration of ASA in health volunteer group (P = 0.000). There was significant difference when the SPAT value of the inpatients treated with anti-platelet drugs was compared with that of healthy blood donor group and with that of health volunteer group before and after administration of ASA (P = 0.000). The cut-off value of SPAT in health blood donor group was 44.6 +/- 11.7 seconds, reference value was from 21.1 seconds to 68.0 seconds; (3) there was no significant difference between SPAT values when platelets samples stored at room temperature for 1, 2, 3, 4 hours (P = 0.815). In conclusion, SPAT can rapidly monitor the inhibition effect of anti-platelet drugs and SPAT may have the similar clinic value with T(50) induced by c-PG.

Collection of peripheral blood stem cells from ABO incompatible allogeneic donors by using blood cell separator / 中国实验血液学杂志

To evaluate the yield of the blood cell separator for collection of peripheral blood stem cells (PBSC) from ABO major and (or) minor incompatible allogeneic donors and the feasibility of PBSC component infusion to the recipients without removal of erythrocytes or plasma, the Cobe Spectra (Version 6.1) blood cell separator was utilized to collect PBSC component from 9 allogeneic donors. Of all the donors, 4 were ABO major incompatible, 2 were minor incompatible and the other 3 were both major and minor incompatible to corresponding recipients. In each cycle, different amount of PBSC component was harvested, and the variable volume plasma chased the cells into the bag was adjusted according to the ABO incompatibility. The nucleated cell count, percentage of mononuclear cell, number of CD34(+) cell and percentage of viable cell (trypan blue excluding rate) in the component were detected. At the time of infusion, a series of protective measures to the renal function of recipients were taken. The results showed that apheresis was twice performed on these eight donors to collected enough PBSC for transplantation or cryopreservation, except one apheresis was enough for cell amount needed by transplantation, as the donor's body weight was much heavier than that of the recipient. Altogether 17 apheresises were performed, the mean yield of nucleated cells was 3.77 x 10(10), in which 97% to 99% were mononuclear cells (MNC). The harvested number of CD34(+) cell was 8.62 x 10(6)/kg. All the trypan blue exclusion rate was 100%. In ABO major incompatible or both major and minor incompatible component, there were 8 - 10 ml packed erythrocytes; in ABO minor incompatible component, there were 80 - 120 ml of plasma. These components were infused into the recipients without removal of erythrocytes or plasma and no haemolytic reaction was observed in any recipient, and their hematopoietic functions soon recovered. Results suggest that enough PBSC can be acquired by using blood cell separator Cobe Spectra (Version 6.1), with the modified separation factors, and the collected PBSC component can be safely infused into the ABO incompatible recipients without removal of erythrocytes or plasma.

Determination and comparison of anti-SARS antibody in children and adults / 中国实验血液学杂志

To investigate the positive rate of anti-SARS antibody in children and adults without SARS, 197 paediatric patients under 14 years old from inpatient and outpatient department of our hospital, 156 healthy children pupils from primary school, 453 adult patients over 18 years old from inpatient and outpatient department of our hospital and other 502 healthy adult blood donors were selected. Anti-SARS antibodies were determined by anti-SARS specific antibody detection kit and ELISA method. The results showed that both the positive rates of IgG antibody in paediatric patients and healthy children were about 2% (4/197 and 3/156), while the positive rates in adult patients and healthy adults were about 0.2% (1/453 and 1/502). The difference between the positive rates of children and adults was significant (chi(2) = 11.61, P < 0.001). IgM antibody was negative in all the samples. It is concluded that the anti-SARS IgG antibody positive rate in children was obvious higher than that in adults. This may be the cause why the cases with SARS in children is much less than in adults.

Qualitative analysis of batch preparing cryopreserved fresh platelet rich plasma / 中国实验血液学杂志

To evaluate the efficiency and effectiveness of batch preparing cryopreserved fresh platelet-rich plasma (cryo-FPRP) derived from the volunteer donors, platelet count (Plt), mean platelet volume (MPV), platelet distribution width (PDW), plasma pH, plasma lactic acid concentration, and lactic dehydrogenase (LDH) concentration, germiculture, CD62p positive rate, PAC-1 positive rate, and the fluorescence intensity of platelet GPIb-IX-V were detected in ACD whole blood, fresh platelet-rich plasma (FPRP), FPRP with 5% dimethyl sulphoxide DMSO (DMSO-FPRP), and thawed cryopreserved FPRP (cryo-FPRP); the procoagulant activity of FPRP and cryo-FPR was determinated. The results showed that (1) 70 percentage of platelet were separated from the whole blood into FPRP, and the counts of residual erythrocyte and leucocyte were below 1 x 10(9), and below 1 x 10(7) per unit respectively. (2) The plasma pH, lactic acid concentration and PAC-1 positive rate retained a stable level during the preparing, storing and thawing process. (3) Plasma LDH concentration, platelet CD62p positive rate and GPIb-IX-V concentration in platelet surface were enhanced significantly after being frozen and thawing. (4) The plasma clotting time induced by cryo-FPRP were significantly shorter than that induced by FPRP. It is concluded that: (1) The batch platelet preparing process can efficiently obtain platelet from whole blood donated by volunteer, and the process didn't activate the platelet. (2) Cryopreservation can prevent lactic acid accumulation, pH reduce and activation of GPIIb/IIIa. (3) The membrane of partial platelets are affected by freezing and thawing. (4) The density of GPIb-IX-V complexes in platelet surface and its procoagulant activity are enhanced significantly after the FPRP freezing and thawing process.

Effects of preparation and quality control of cryopreserved platelets / 中国实验血液学杂志

The purpose of this study was to establish a set of techniques for cryopreservation of platelets with dimethylsulphoxide (DMSO) to insure high quality of cryopreserved platelet. The methods were as following: (1) DMSO was filtered in stead of being sterilized before infusion into the bag with platelets. (2) The whole blood was centrifuged immediately after blood collection and the attached tube was tied on the top of the bucket. (3) The related centrifugal force was 480 x g, the accelerating and braking grades of the centrifuge for acceleration and deceleration were 9 and 4 respectively. (4) The flow rate of platelet rich plasma (PRP) could not be too high, 80 - 100 ml PRP should be harvested at 1 minute or so. The infusion rate of DMSO into the PRP was 1 ml/min. After the infusion of DMSO, the PRP bag must be put into the -80 degrees C ultra low freezer at once to make the product to be freezed as soon as possible. The cryopreserved platelet should be thawed in the cycling warm water at the temperature of 38 - 40 degrees C. (5) After thawing of platelet, the platelet, red blood cell and white blood cell were counted, and the bacteria culturing, tests for HBsAg, anti-HCV, anti-HIV, TP and ALT were carried out. The results showed that altogether 14 800 units of cryopreserved platelets were prepared including 80 units collected with blood cell separator, of which quality control was accomplished in 300 units. The manually collected platelet mean count >/= 2.4 x 10(10)/unit, while the apheresis platelet count >/= 2.5 x 10(11)/unit. The yield was over 70%. The contaminated red and white blood cells were </= 1 x 10(9) and </= 1 x 10(7)/unit respectively. All the bacteria cultures were negative, while tests for HBsAg, anti-HCV, anti-HIV and TP were negative too. The ALT values were all in normal range. The transfusion of cryopreserved platelets showed obvious effect of haemostasis. In conclusion, the cryopreserved platelets prepared with this method were of high quality and efficaciousness in haemostasis clinically.

Micro-column gel indirect anti-globulin technique for screening and identification of irregular antibody / 中国实验血液学杂志

The purpose of this study was to design an antibody screening method based on the micro-column gel indirect anti-globulin technique (MGIAT), using pooled cells and plasma, by comparison with the conventional indirect anti-globulin technique (CIAT) combined with a two-stage papain technique, and to explore the feasibility of the use of plasma instead of serum as test material. The samples of blood recipients in our hospital were screened for irregular antibody using pooled test cells. Screening of the antibodies was identified both by MGIAT and CIAT combined papain technique respectively. The results showed that the irregular erythrocyte antibodies were detected in 20 cases from 5,000 recipients screened by MGIAT, using pooled cells, the positive rate was 0.4%. The specificity of 20 cases of irregular antibodies was as follows: 2 cases of anti-D, 8 cases of anti-E, 1 cases of anti-C, 2 cases of anti-c, 2 cases of anti-Mi(a), 2 cases of anti-Jk(a), 1 case of anti-Le(a) and 2 cases of anti-Fy(a). Antibody was detected from 19 cases using CIAT. Anti-Le(a) was detected with adding complement from Le(a-b-) person. Only 13 cases antibody were found by papain technique. It was concluded that irregular antibody screening by MGIAT using pooled cells can take place of the CIAT combining with papain technique in clinical application. Plasma is superior to serum in antibody screening test.

Detection of the anti-SARS-coronavirus specific antibody levels in 156 SARS patients / 中国实验血液学杂志

The objective of this study was to explore the development of IgG and IgM against SARS CoV and characteristics of changes of antibody titers in patients with severe acute respiratory syndrome (SARS) and to search the opportunity for collecting specific anti-serum from convalescent patients with SARS. The anti-SARS-coronavirus specific antibody levels in 156 SARS patients were measured with ELISA. The results showed that the total positive rates of IgG and IgM were 75.6% and 41.7% respectively, and the negative rate of both IgG and IgM was 23.7%. The average titers of IgG and IgM antibody in positive samples were 18.23 +/- 24.72 and 2.18 +/- 1.13, respectively. There was no significant correlation between the titers of IgG/IgM and sex, age, course of diseases and duration of body temperature recovery. It was concluded that not all SARS patients could produce the anti-SARS-coronavirus specific antibody. The titers of the anti-body are diversified even if the antibodies have been emerged in them. In order to obtain effective anti-serum, the titers of antibody must be tested just before collection of convalescent serum, and it ensures the therapeutic effect and provides a measurable index for clinical transfusion.