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【Objective】 To investigate the preparation conditions of hemoglobin-based oxygen carrier (HBOC) at room temperature without oxygen isolation and the quality of poly-COHb. 【Methods】 Using human cord blood hemoglobin as raw material and glutaraldehyde as crosslinking agent, the experimental group selected hemoglobin concentration, glutaraldehyde to COHb molar ratio, reaction time and reaction temperature to do four factors and three levels of orthogonal test (n=3), to determine the best matching conditions and prepare three batches of poly-COHb, continuously.In the control, 3 batches of poly-Hb were prepared under low temperature and oxygen isolation.The contents of superlarge molecules, degree of polymerization, average molecular weight, methemoglobin and P50 of crosslinked products were compared between the two groups. 【Results】 The optimal matching conditions of poly-COHb were as follows: [Hb] 70g/L, the molar ratio of glutaraldehyde to COHb was 11∶1, the reaction time was 60 min, and the reaction temperature was 25℃.Comparison of poly-COHb and poly-Hb: The superlarge molecular content (%) was 0.67±0.51 vs 0.60±0.01 (P>0.05); degree of polymerization (%) and average molecular weight (kD) were 84.25±0.99 vs 54.16±5.12, and 235.27±13.50 vs 97.62±6.57, respectively.(P<0.05); methemohemoglobin ratio (%) was 2.40±0.66 vs 2.47±0.46 (P>0.05); P50 (mmHg) was 6.37±0.30 vs 14.20±1.09 (P<0.05). 【Conclusion】 Poly-COHb can be prepared at room temperature without oxygen isolation.Compared with poly-Hb, poly-COHb prepared under the experimental conditions can improve the polymerization degree and average molecular weight, and greatly reduce the difficulty of preparation.
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【Objective】 To study the red blood cell substitute preparation method of glutaraldehyde(GDA) and Bis(3, 5-dibromosalicyl) fumarate(DBBF) double polymerized, and also observe its curative effect. 【Methods】 The affecting factors of the crosslink of DBBF and human placental hemoglobin(Hb) were selected, including solution pH, inositol hexaphosphoric acid concentration, and molar ratios of DBBF/Hb. The changes of P50, Hill Co and methemoglobin(MetHb) content during the preparation process were studied and the molar ratio of GDA to DBBF-Hb were determined. The final product was obtained through the steps of ultrafiltration, filtration, and deoxygenation, etc., and the final product was subjected to a study on the curative effect of anti-hemorrhagic shock in rats. 【Results】 Under the condition of pH 8.0, the concentration of inositol hexaphosphoric acid was 10mM/L, the molar ratio of DBBF/hemoglobin was 6∶1, the content of dimer was(22.67±3.28)%, the content of tetramer was (74.64±7.05)%, and its P50 was (25.25±2.31) mmHg. As the molar ratio of DBBF/Hb increased, P50 and MetHb content would gradually increase, while the Hill Co will gradually decrease. The molar ratio of GDA and DBBF-Hb was 7∶1, the content of dimer was (8.23±0.79)%, the content of tetramer was (27.87±3.63)%, the content of super-macromolecule was (1.05±0.31)%. As the molar ratio of GDA and DBBF/Hb increased, MetHb content would increase, while the P50 and Hill Co would gradually decrease. The 24h survival rate of the product in rats with hemorrhagic shock was 83.33%(10/12), which was significantly higher than that in the negative control group[41.67%(5/12)], while similar with the positive control group [91.67%(11/12)]. 【Conclusion】 The red blood cell substitutes with GDA and DBBF double polymerized can effectively reduce the dimer content, which is more conducive to industrial production, and has a positive effect on the treatment of hemorrhagic shock.
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High-quality donor organs is of significance for the success of organ transplantation. However, standard donors fail to meet the requirements of kidney transplantation due to the increasing quantity of patients with kidney failure. Marginal donor kidneys have been widely applied in clinical practice, which also poses challenges to the existing preservation methods of donor kidneys. Ischemia-reperfusion injury (IRI) is one of the critical factors affecting the early graft function after kidney transplantation. In addition, it exerts harmful effect upon the long-term survival of the graft. Current studies have demonstrated that hemoglobin-based oxygen carrier (HBOC) may reduce the IRI during kidney transplantation, effectively improve the preservation quality and prolong the preservation time of donor kidney. In this article, the research progress on HBOC in kidney transplantation was reviewed, aiming to provide reference for modifying the preservation method of donor kidney, improve the quality of donor kidney and enhance clinical prognosis of the recipients.
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【Objective】 To explore the protective effects of hemoglobin base on oxygen carries (HBOCs) with different oxygen affinity on isolated rat hearts. 【Methods】 Using Langendorff isolated heart perfusion model, 45 adult male SD rats (SPF grade), perfused with 30 min KH solution baseline, were randomly divided into sham operation group and control group: St. Thomas (STS) solution perfusion volume was 3mL/100g body weight; high P50 HBOCs group: [STS + high P50 HBOCs (P50=35.0 mmHg, 2.5 mg/100 g) product] perfusion volume was 3mL/100g body weight; medium P50 HBOCs group: [STS + medium P50HBOCs (P50=26.5.0 mmHg, 2.5 mg/100 g) product] perfusion volume was 3 mL/100 g body weight; low P50 HBOCs group: [STS + low P50 HBOCs (P50=11.0 mmHg, 2.5 mg/100 g) product] perfusion volume was 3mL/100g body weight, and the heart was arrested and placed in a 37℃ water bath to make the heart ischemic for 35 minutes, and then reperfused for 2 hours. The left ventricular development pressure (LVDevP), left ventricular end diastolic pressure (LVEDP), the rate of change of left ventricular pressure (LVPCR) and heart rate (HR) in the rat heart during reperfusion were observed and recorded. 1 min perfusion fluid from each rat in the basic and reperfusion phase was taken, and blood gas analyzer was used to measure the blood gas indexes of rats, and the myocardial injury marker enzymes [cardiac enzyme creatine kinase (CK-MB), lactate dehydrogenase (LDH) and the release of α-hydroxybutyrate dehydrogenase (α-HBDH)] were measured by ELISA kit. 【Results】 The cardiac function and the release of myocardial enzymes in the 5 groups of rats in the basal cardiac perfusion stage were similar (P>0.05). However, in the reperfusion stage, except for the insignificant changes in HR (P>0.05), the heart LVDevP (mmHg) of the three P50 HBOCs groups and the control group were 10.69±3.65 vs 8.50±2.88, 23.26 ±5.62 vs 8.50±2.88, 35.60±3.82 vs 8.50±2.88, LVEDP (mmHg) were 43.34±8.08 vs 54.64±7.42, 39.43±8.30 vs 54.64±7.42, 31.46±4.11 vs 54.64±7.42, dp/dt were 12.09±9.96 vs 6.09±0.98, 25.65±8.87 vs 6.08±0.98, 35.32±9.33 vs 6.09±0.98, -dp/dt were 17.53±11.28 vs 11.39±2.16, 28.80±13.70 vs 11.39±2.16, 43.36±3.83 vs 11.39±2.16, respectively (all P<0.05); the rebound situation and the release of CK-MB, LDH, and α-HBDH in the three P50 HBOCs groups were better than those in the control group (P<0.05). Among the three P50HBOCs products, the low P50HBOCs group had the best cardiac function indexes. The myocardial enzyme indexes of the high, medium and low HBOCs groups were CK-MB (ng/mL): 110.47±4.04, 90.2±2.46, 77.1±3.51; LDH (U/L): 162.23±7.71, 135.13±23.69, 92.20±4.21; a-HBDH (U/L): 228.00±8.03, 172.30±8.99, 131.00±2.02. 【Conclusion】 STS solution containing HBOCs products can improve the function of the reperfused heart at normal temperature ischemia for 35 min and 2 h reperfusion, and reduce heart damage. The STS solution containing low P50 HBOCs has the most obvious protective effect in rat isolated heart perfusion.
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【Objective】 To compare the similarities and differences of polymerized human cord hemoglobin (PolyCHb) and free hemoglobin (FHb) on partial coagulation indexes in vitro, so as to analyze the effect of PolyCHb on coagulation dysfunction. 【Methods】 Using normal saline, two concentrations of FHb and PolyCHb and 36% methemoglobin-containing PolyCHb to mix with fresh whole blood or plasma-rich plasma (PRP) in equal proportions, and incubate at 37°C for 30 minutes to detect prothrombin time (PT), activated partial thromboplastin time (APTT), coagulation factor Ⅱactivity (FⅡ∶C), coagulation factor Ⅴactivity (FⅤ∶C), coagulation factor Ⅷactivity (FⅧ∶C), coagulation factor Ⅸactivity (FⅨ∶C), von Willebrand factor (vWF) and platelet P-selectin (CD62P). 【Results】 1) NaCl group: PT(22.68±1.76) s; APTT(59.58±7.52) s; FⅡ∶C(45.91±3.27) %; FⅤ∶C(30.86±4.43) %; FⅧ∶C(41.32±12.94) %; FⅨ∶C(23.96±5.10) %; vWF (2.14±0.54) mg/L; CD62P(7.44±4.47) %. This group kept as a diluted control. 2) 2% FHb group compared with 7% FHb group: FⅧ∶C (42.16±12.31) %vs (56.64±12.22 ) % (P0.05). 3) There is no significant difference between 2% PolyCHb group and 7% PolyCHb group (P>0.05). 4) There is no significant difference between 2% FHb group and 2% PolyCHb group (P>0.05). 5) 7% FHb group compared with 7% PolyCHb group: PT(23.31±1.34)s vs (21.97±1.56)s (P0.05). 6) 7% PolyCHb group Compared with Met-PolyCHb group: APTT(55.43±5.43) s vs (46.33±4.86)s (P0.05). 【Conclusion】 The effect of PolyCHb on coagulation markers is different from FHb. At the concentration of this study, PolyCHb will not cause coagulation disorders. However, if the methemoglobin (MetHb) content is too high, it will activate the intrinsic coagulation pathway.
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Objective:To analyze the influential factors for inpatient care among residents in Shanghai and to provide evidence for health policy.Methods:Data were obtained from the Fifth National Health Services Survey,the independent variables were selected based on the framework of Andersen Model,and the two models were used to analyze the determinants-relevant hospitalization probability and the cost among residents in Shanghai.Results:The hospitalization rate among permanent residents over 18 years old was 6.1％ in 2013 in Shanghai.For the hospitalized population,its average expenses (median) were 10 000.00 yuan.Gender,age,education,marriage,smoking,drinking,exercise,the travel time,family annual income,Engel coefficient and the number of chronic diseases were the determinants of hospitalization probability.Gender,age,smoking,drinking,exercise,Engel coefficient,medicaid and level of medical institution were the influential factors for the hospitalization costs.Conclusion:The utilization of inpatient care among residents in Shanghai is determined by many factors.Thus,the corresponding health policies should be formulated for different populations.
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Objective To investigate the risk factors of delayed gastric emptying after pancreaticoduodenectomy (PD) for periampullary carcinoma.Methods The clinical data of 96 periampullary carcinoma patients who received PD with or without pylorus preservation at Zhongshan Hospital Fudan University from October 2010 to September 2013 were retrospectively analyzed.The risk factors associated with delayed gastric emptying were analyzed.The univariate and multivariate analysis were done using the chi-square test and Logistic regression model respectively.Results A total of 17 patients had postoperative delayed gastric emptying, with the incidence of 18％.The results of univariate showed that female sex, pancreatogastrostomy, increased intraoperative blood loss and transfusions and postoperative abdominal infection were risk factors of delayed gastric emptying (x2 =4.512, 6.412, 6.245, 6.695, 6.816, all P ＜ 0.05).The results of multivariate analysis revealed that only pancreatogastrostomy and postoperative abdominal infection were independent risk factors of delayed gastric emptying (OR =4.031 and 5.281).Conclusions Pancreatogastrostomy and postoperative abdominal infection indicate a higher incidence of delayed gastric emptying after pancreaticodudenectomy for periampullary carcinoma.
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Objective To compare the short-term efficacy of pylorus-preserving pancreaticoduodenectomy (PPPD) and standard pancreaticoduodenectomy (SPD) for the treatment of periampullary carcinoma.Methods The clinical data of 85 patients with periampullary carcinoma who were admitted to the Zhongshan Hospital of Fudan University from October 2010 to October 2012 were retrospectively analyzed.Forty-four patients who underwent PPPD were divided into the PPPD group and 41 patients who underwent SPD were divided into the SPD group.The pancreatic fistula(Grade B and above), biliary fistula, blood loss, intra-abdominal infection, delayed gastric emptying (DGE) and prognosis were analyzed.Patients were followed up by telephone interview and outpatient examination once every 3 months within postoperative 1 year and once every 6 months within postoperative 2-3 years till October 2014.Count data were analyzed using the chi-square test, measurement data with normal distribution were presented as M(Qn) and comparison was analyzed using the Mann-Whitney U test.The survival curve was drawn by the Kaplan-Meier method, and survival rate was analyzed using the Log rank test.Results The operation time was 195 minutes (180 minutes, 240 minutes) in the PPPD group and 210 minutes (180 minutes,300 minutes) in the SPD group, with a significant difference (Z =-2.090, P ＜ 0.05).The volume of intraoperative blood loss, intraoperative blood transfusion and duration of postoperative hospital stay were 200 mL(113 mL,288 mL), 0 mL(0 mL, 0 mL) and 17 days(12 days, 24 days) in the PPPD group, and 200 mL(150 mL, 325 mL),0 mL(0 mL, 400 mL) and 16 days(12 days, 30 days) in the SPD group respectively, with no significant differences between the 2 groups (Z =-1.185,-1.780,-0.533, P ＞0.05).There was no perioperative death and incidence of overall complication was 42.4％ (36/85) with pancreatic fistula, intra-abdominal infection and DGE as the top 3 common postoperative complications.The incidence of DGE was 20.5％ (9/44) in the PPPD group,which was significantly different from 4.9％ (2/41) in the SPD group (x2=4.571, P ＜ 0.05).The incidence of pancreatic fistula, biliary fistula, intra-abdominal infection, postoperative bleeding and 2 or more complications were 20.5 ％ (9/44), 2.3 ％ (1/44), 15.9％ (7/44), 4.5 ％ (2/44), 25.0％ (11/44) in the PPPD group, and 14.6％ (6/41), 4.9％ (2/41), 19.5％ (8/41), 7.3％ (3/41), 14.6％ (6/41) in the SPD group, respectively, showing no significant difference between the 2 groups (x2=0.495, 0.423, 0.295, 0.190, 1.425, P ＞ 0.05).Eighty-five patients were followed up for 6-47 months with a median time of 31 months, and postoperative overall 1-and 3-year survival rates were 95.3％ and 75.5％, respectively.The 1-and 3-year survival rates were 97.7％ and 78.9％ in the PPPD group, and 92.7％ and 71.7％ in the SPD group, respectively, with no significant difference in 3-year survival rate (x2=0.690, P ＞0.05).The 3-year overall survival rate was 80.5％ in patients without lymph node involvement (LNI) compared with 54.9％ in patients with LNI, showing a significant difference (x2=4.290, P ＜ 0.05).Conclusions Both PPPD and SPD have good short-term efficacy for periampullary carcinoma.There is no significant difference between PPPD and SPD concerning short-term survival rate of periampullary carcinoma.PPPD has shorter operation time, but has a higher postoperative DGE incidence.LNI is a significant prognostic factor for short-term survival of periampullary carcinoma.PPPD is not recommended while the lymph nodes are involved.
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Objective To study the optimization process of matching conditions using glutaraldehyde (GDA) as crosslinking agent of hemoglobin based oxygen carriers (HBOCs),to further reduce the average molecular weight and the content of super-weight molecular,and improve the conversion ratio of polymerization.Methods The orthogonal designs were done on the basis of the previous single influencing factor research of human placenta hemoglobin crosslinking GDA.Three factors were selected including molar ratio of GDA to hemoglobin,mass concentration of hemoglobin and the rate of the feeding GDA.Results The molar ratio of GDA to hemoglobin is the most important influencing factor on the molecular weight distribution of polymerized hemoglobin,followed by the mass concentration of hemoglobin and the rate of feeding GDA.When analyzing the impact on the mean molecular weight,there were significant differences between mean molecular weight corresponding to different molar ratios of GDA to hemoglobin (P＜0.05),while there was no statistical significance between mean molecular weight corresponding to different mass concentrations of hemoglobin and the rates of feeding GDA (P＞0.05).When analyzing the impact on the effective conversion ratio,there were significant differences between effective conversion ratios corresponding to different molar ratios of GDA to hemoglobin and different mass concentrations of hemoglobin (P＜0.05),while there were no statistical significances between effective conversion ratios corresponding to different rates of feeding GDA (P＞0.05).When analyzing the impact on the content of super-weight molecular,there were significant differences between content of super-weight molecular corresponding to different molar ratios of GDA to hemoglobin,while there were no statistical significances between content of super-weight molecular corresponding to different mass concentrations of hemoglobin and different rates of feeding GDA.Conclusions The optimal matching conditions of hemoglobin polymerization process were determined by orthogonal designs.
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Objective To promote the diagnostic accuracy of intrapancreatic accessory spleen (IPAS).Methods The clinical data of 10 cases of IPAS admitted in Fudan University Zhongshan Hospital from Apr 2005 to Dec 2013 were retrospectively analyzed.Results There were ten cases of IPAS confirmed pathologically.Only 1 of the ten cases was diagnosed correctly and definitely with IPAS preoperatively.The other 9 cases were misdiagnosed with benign or malignant pancreatic tumors,including nonfunctional neuroendocrine neoplasms in 5 cases,pancreatic neuroendocrine cancers in 3 and pancreatic intraductal adenocarcinoma in one.All the nine misdiagnosed patients has no specific symptoms or laboratory indexes.All the IPASs located in the tail of the pancreas with the mean diameter (1.3 ±0.2) cm(0.8-2.5 cm).7 cases of IPAS show strikingly similar dynamic enhancement to the spleen on the CT scans and/or MRI.Accessory spleen around the splenic hilum was found in five cases.Conclusions Morphological study plays an important role in the diagnosis and IPAS carries parallel dynamic enhancement to the spleen on CT scans and/or MRI.IPAS should be considered as a differential diagnosis while the lesion is no more than 2.5 cm in diameter and when other accessory spleens were shown around the splenic hilum.
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Five influencing factors of the polymerization process of glutaraldehyde with hemoglobin were studied in cluding the approach of feeding glutaraldehyde, hemoglobin concentration, the molar ratio of glutaraldehyde and hemoglobin, reaction time and temperature, in order to reduce the average molecular weight and to improve the effective polymerization ratio. The results showed that the optimal process reduced the average molecular weight of hemoglobin based oxygen carriers (HBOCs) from (350.20 +/- 35.45)kD to (158.60 +/- 8.70)kD and improved the effective polymerization ratio from 53.27% +/- 4.95% to 69.50% +/- 3.70%. When the experiments expanded 30 folds, the results of the effective polymerization ratio and the average molecular weight mentioned above could be achieved.