Study on the indeterminate results of characterization and verification of HIV antibody from Western blot test / 中华流行病学杂志

Objective To study the serological characterization of indeterminate Western blot(WB)results of HIV antibody and to find a new way to verify the HIV antibody indeterminate results and provide references for editing"National Guideline for Detection of HIV/AIDS".Methods All of the 42 subjects who were confirmed as indeterminate HIV antibody in People'Libaretion Amry HIV Confirmation Laboratory from 2005 to 2006,were collected.Line immunoassay.HIV viral load test and HIV-1 p24 were tested and followed up for 3-6 months'to compare the changes of WB bands patterns.Results (1)For the 42 individuals with indeterminate HIV antibody.a total of 8 different patterns of bands were found in WB test including 45.2% of them were p24 monoband,30.9% were gp160 monoband,11.9% were gp160 with p24,2.4%(only one case)were gp160gp120 ±,gp41p24,p24p17,gp41 or gp120respectively.It was noticed that the most patterns of common bands with indeterminate results were p24 monoband.gpl60 monoband and gpl60 with p24.which composed 88.0% of the whole indeterminate WB band patterns.(2)Twenty three cases had been followed up for more than 3 months with 22 giving no WB band image change and were confirmed as HIV sero-negative.The other one with case gp160 and p24 had developed to more bands in the period of 77 days follow-up with more bands,including gpl60,gp120,p66,p31,p24 and p17,showed up and was confirmed as HIV primary infection.(3)Line immunoassay was applied to all of those 23 cases who had been followed up and the results showed that only one serological change was found and the case was confirmed to be HIV-positive.Among the other 22 cases without serological changes.16 cases were proved to be HIV-negative,6 cases were still indeterminate.The specificitv was 72.7%.P24 antigen test showed negative in all the 23 cases,including the case which later was confirmed as HIV-positive.Of all the 23 originally indeterminate cases,viral loads were tested in 7 eases.Positive result was found in the case which was proved later to be HIV-positive.No viral loads were detected in the other 6 cases(＜LDL).Conclusion The most common band patterns of indeterminate HIV antibody were mainly p24 monoband,gp160 monoband or with p24.Most of them (95.6%)were not infected by HIV,the bands showed up in WB test and demonstrated as non-specific reactions.Line immunoassay could determine about 70% of the indeterminate reactions.Results from viral load test also suggested that it was an efficient method to discriminate indeterminate results.With these two techniques,HIV serology could be diagnosed without 3 months'follow-up in primary infection which gave indeterminate WB results.

Inhibition of L-arginine and cilostazol on activation of platelets in vitro / 中国实验血液学杂志

The purpose of study was to investigate the effects of L-arginine and cilostazol on platelet-activation and aggregation reserve in vitro, so as to provide proof for selecting reversible activation-inhibitors for platelets lyophilization. Activation and function of platelets were investigated by using flow cytometry with the CD62p and PAC-1 expression and re-expression after being activated by thrombin, and by means of platelet aggregation reaction to thrombin, ADP and propyl gallate, as well as coagulation activity of platelets. The results showed that expression of CD62p and PAC-1 increased after being pretreated. Both L-arginie and cilostazol could inhibit CD62p and PAC-1 expression and related with their concentrations. Cilostazol had an intensive inhibition effect on expressions of CD62p and PAC-1 induced by thrombin, and the inhibition increased when concentration augmented. L-arginine had the same effects on PAC-1, but had no effects on CD62p. L-arginine and cilostazol inhibited aggregation induced by thrombin, ADP and propyl gallate, and the inhibitions were related directly with dosage. When L-arginine concentration was higher or equal to 15 mmol/L, or cilostazol concentration was in range of 1 - 4 mmol/L, the aggregation time were prolonged so much or even no aggregation. It is concluded that when L-Arginine concentration is 5 mmol/L and 10 mmol/L, platelet activation can be inhibited, but aggregation ability and characters keep intact. Concentration at 5 mmol/L may be the best. 1 mmol/L of cilostazol can inhibit activation in vitro and retain part of platelet ability of aggregation and reexpression.

Experimental study on lyophilization of platelets / 中国实验血液学杂志

The aim of this study was to search a procedure of platelet lyophilization and find a way of long-term storage of human platelets at normal temperature with smaller size and lighter weight, to be convenient to transport at long distance thus to meet the demands in accidents and war time. Human platelets were pretreated by freezing, the first and the second desiccation, and were added with reversible activation-inhibitors of platelets, DMSO and trehalose, then were rehydrated. At the same time, the recovery rate of platelets, platelet maximal aggregation induced by thrombin, coagulation of platelets, CD62p expression and PAC-1 expression were assayed. The results indicated that the recovery rate of the platelets was 56.29%. The platelet maximal aggregation induced by thrombin had no significant difference between lyophilized platelets and the fresh platelet-rich plasma (FPRP), but the aggregation of platelets induced by ADP or propyl gallate was decreased by 49.34% and 26.25%. Coagulation of the lyophilized platelets was not significantly different from FPRP. CD62p expression of the lyophilized platelets (42.36%) was higher than that in FPRP while PAC-1 expression was 2.12%. CD62p re-expression rate induced by thrombin was 50.88% and PAC-1 re-expression was 54.55%. It is concluded that the ability of recovered lyophilized platelets added with reversible activation-inhibitors, DMSO and trehalose to aggregate and coagulate has showed no significant difference as compared with FPRP. The reversible activation-inhibitors can decrease CD62p expression of lyophilized platelets, and may enhance their survival ability and prolongate survival time. Therefore the efficiency of lyophilizing platelets can be improved based on this freeze-drying procedure.

Inhibiting effect of adenosine on platelet activation in vitro / 中国实验血液学杂志

This study was aimed to investigate the inhibiting effects of adenosine on platelet in vitro in order to select functional protectants for platelets before lyophilization. Platelet membrane surface CD62P expression was assayed by flow cytometer (FCM). Platelet aggregations induced by restocetin, thrombin, ADP and propyl gallate were detected by APACT-2. The results showed that platelets membrane surface CD62P expression increased significantly after pre-treating of freeze-drying. 0.75 mmol/L adenosine could inhibit CD62P expression in a dose-dependent manner. Adenosine could inhibit platelet aggregation induced by propyl gallate, but no action on restocetin. When adenosine concentration was 1.0 mmol/L or higher, the aggregation induced by thrombin was significantly restrained. When concentration of adenosine was 0.75 mmol/L, platelet activation resulted from retreating could be inhibited and platelet aggregation induced by restocetin and thrombin were not affected markedly. It is concluded that adenosine can be one of the functional protectants and activation inhibitors in vitro for platelet cryo-preservation.

A method for Rhesus box test / 中国实验血液学杂志

To study the method for Rhesus box test and its significance, the sequence specific primers of upstream, downstream and hybrid Rhesus boxes were designed according to RhD gene sequence; the upstream, downstream and hybrid Rhesus boxes were determined by PCP-SSP and mismatched PCR. The results showed that this method was confirmed by DNA Standard test. It was shown that in unrelative RhD positive individuals RHD(+)/RHD(-), RHD(+)/RHD(+) genotype accounted for 9.00%, 91.00% respectively, and in RhD negative individuals RHD(+)/RHD(-), RHD(+)/RHD(+), RHD(-)/RHD(-) genotype were 26.14%, 3.92%, 69.94% respectively. It is concluded that the method of Rhesus box test was confirmed to be reliable and can be used for the identification of RhD haplotype gene structure, as well as for study on inheritance, clinical transfusion and neonatal hemolytic diseases.