Review on platelet-rich plasma preparation and its quality evaluation / 中国输血杂志

Platelet-rich plasma (PRP) currently has been widely used in various medical fields, such as tissue regeneration, wound healing, scar repair, skin and hair regeneration etc..PRP is rich in platelets, growth factors and other blood components, which can effectively promote tissue repair and healing. However, there is no optimal preparation method and unified standard of composition ratio for PRP, so its clinical application value has not been satisfactorily interpreted yet. In this paper, the preparation and quality standard of PRP were reviewed to provide basis for standardization of RPP in clinical application.

Blood coagulation function before and after peripheral blood stem cell mobilization and collection / 中国输血杂志

【Objective】 To study the changes of blood coagulation function of donors before and after peripheral blood stem cell(PBSC)mobilization and collection, so as to evaluate the safety of the current scheme. 【Methods】 30 donors who received PBSC mobilization and collection in Zhujiang Hospital from October 2018 to October 2020 were enrolled. After mobilization by G-CSF, the correlation between coagulation function, blood routine indexes and TEG indexes of donors was analyzed, and the influence of PBSC mobilization and collection on coagulation function of donors was evaluated. 【Results】 The TEG indexes R(min), K(min), α(°), MA(mm) and CI before and after PBSC collection were 6.12±1.18 vs 7.25±2.16, 1.98±0.41 vs 2.45±0.64, 62.82±4.98 vs 57.3±6.67, 60.93±3.26 vs 55.37±4.41, and -0.31±1.40 vs -2.32±2.18, respectively(P<0.05), suggesting that there was no risk of hypercoagulability after PBSC mobilization and collection. The peak values of WBC (×109/L), Plt (×109/L) and Hb (g/L) were 62.02, 357 and 162, respectively, which indicated that the blood routine indexes after PBSC mobilization and collection were in the safe range. After PBSC collection, the CI value of 26.7% (8/30) donors was less than -3, showing hypocoagulability. 【Conclusion】 The current mobilization and collection scheme of PBSC has little effect on the coagulation function. Most of the donors had no risk of hypercoagulability, but a few showed a trend of hypocoagulability after PBSC collection.

Multivariate analysis of peripheral blood mononuclear cells from children donors / 中国输血杂志

【Objective】 To analyze the characteristics of peripheral blood mononuclear cells (PBMC) collection in healthy children, and explore the factors affecting collection efficiency (CE). 【Methods】 The PBMC data, involving 70 episodes of apheresis from 42 children during January 2017 and June 2020 were retrospectively analyzed All children were collected in Zhujiang Hospital of Southern Medical University. 【Results】 Multiple linear regression analysis showed that mononuclear cells (MNC) in donor collections from healthy children were positively correlated with anticoagulant dosage, lymphocyte count and monocyte count (P<0.05), meanwhile, negatively correlated with age and platelet count. The PBMC CE was negatively correlated with age, platelet count, and processed whole blood volume (P<0.05). CD34+ cells (×107 /kg)was negatively correlated with age, meanwhile, positively correlated with numbers of collection and processed whole blood volume(r=-0.79). No statistical differences in red blood cell count, platelet count, lymphocyte count, monocyte count of healthy child donors were notable before versus after apheresis. 【Conclusion】 MNC can be collected effectively in children of different ages. The PBMC collection efficiency was related to age. Meanwhile, the higher the lymphocytes and monocytes were before apheresis, the more MNC were collected. The efficiency of MNC collection would decrease when the apheresis volume of the children exceeded their total blood volume twice. However, the absolute value of CD34+ cells in the final yields would increase.

Influencing factors and timing prediction of hematopoietic stem cell collection in peripheral blood / 中国输血杂志

【Objective】 To explore the effects of blood routine parameters on the peripheral blood hematopoietic stem cell collection of healthy donors, and predict collection timing based on these parameters. 【Methods】 The blood routine parameters pre-donation and the total number of mononuclear cells post-donation of 249 donors who applied blood cell separator to collect peripheral blood hematopoietic stem cells in our hospital from January 2018 to August 2020 were collected. Taking total nucleated cells of circulating blood per litre as the main evaluation index, and its collection with blood routine parameters pre-collection was analyzed. The relevant influencing factors were analyzed using multiple linear regression analysis. The blood routine parameters of healthy donors who donated peripheral blood hematopoietic stem cells in our hospital from September 2020 to October 2020 were substituted into the equation to obtain the predicted values, which were then compared with the actual values obtained from actual product using t test for verification. 【Results】 The analysis showed that the parameters of Hb, RBC, Hct, leukocyte count, neutrophil, lymphocyte, monocyte and Plt were statistically correlated with the total number of mononuclear cells of circulating blood per liter volume (P<0.05). There was a linear relationship between lymphocyte, monocyte, Plt and leukocyte count and the total number of mononuclear cells of circulating blood per liter. The total number of mononuclear cells of circulating blood per liter was set to (Y), and the variables such as lymphocyte (X1), monocyte (X2), Plt (X3), leukocyte count (X), and neutrophil were used as dependent variables for multiple linear regression, and the equation was: Y=9.814+ 3.131X1+ 1.666X2+ 0.020X3+ 0.124X4. There was no statistical difference between the predicted value and the calculated value (P>0.05). 【Conclusion】 The blood routine parameters of lymphocyte, monocyte, platelet count and leukocyte count of donors before collection can effectively predict the collection efficiency, therefore help predict the collection time.

Gut microbiota at different stages of HBV infection / 实用医学杂志

Objective To investigate the correlation between gut microbiota and hepatitis B virus(HBV) infection at different stage. Methods 84 patients aged 25 ~ 40 were enrolled in the study,the ALT,AST and HBV antigens and antibodies qualitatively detected. Then they were divided into four groups based on HBV related antigens and antibodies qualitative detection according to guideline for chronic HBV infection:48 in the protective antibody positive group where vaccine injection was performed for producing protective antibodies ,14 in the HBV recovery group 9 in chronic HBV infection group And 13 in antibody negative group where vaccine injection was done. Their fecal samples were collected and total DNA were extracted and subjected to enterobacterial repetitive intergenic consensus sequence polymerase chain reaction(ERIC-PCR). After ERIC-PCR,gel electrophoresis and genetool software were used to analyse the differences in the amount of bands and the position of main bands between groups. Results The amounts of bands in the antibody negative and chronic HBV infected groups were significantly smaller than the protective antibody group and HBV recovery group. The position of main band in the protective antibody group and HBV recovery group were wildly spread within groups ,and most main bands in the HBV recovery group were above 282 bp. The main bands in the antibody negative and chronic HBV infected groups were centralized in 60 ~ 70 bp. Conclusions Compared with the protective antibody group and HBV recovery group ,the richness of gut microbiota in the antibody negative and chronic HBV infected groups are significantly decreased and lack of microbiota above 200 bp in ERIC-PCR.

Rapid selection of recombinant orf virus expression vectors using green fluorescent protein / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.</p><p><b>METHODS</b>The flanking regions of the ORFV132 of orf virus DNA were amplified by PCR to construct the shuttle plasmid pSPV-132LF-EGFP-132RF. The shuttle plasmid was transfected into OFTu cells and GFP was incorporated into orf virus IA82Delta 121 by homologous recombination. The recombinant IA82Delta121-V was selected by green fluorescent signal. The deletion gene was identified by PCR and sequencing. The effects of ORFV132 knockout were evaluated by virus titration and by observing the proliferation of the infected vascular endothelial cells in vitro.</p><p><b>RESULTS</b>The recombinant orf virus IA82Delta121-V was obtained successfully and quickly, and the deletion of ORFV132 did not affect the replication of the virus in vitro but reduced its virulence.</p><p><b>CONCLUSION</b>Green fluorescent protein is a selectable marker for rapid, convenient and stable selection of the recombinant viruses. Highly attenuated recombinant orf virus IA82Delta121-V can serve as a new expression vector for exogenous genes.</p>

Prevalence of chromosome 9 abnormalities among pediatric specimens / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To perform cytogenetic analysis for children, especially newborns suspected for chromosome abnormalities.</p><p><b>METHODS</b>Peripheral blood or born marrow specimens were respectively cultured in proper media. Karyograms were analyzed following G-banding.</p><p><b>RESULTS</b>Of 154 blood specimens, numerical chromosomal abnormalities were identified in 20 patients, which included 19 with trisomy 21. Structural aberrations were identified in 13 patients, among which chromosome 9 aberrations were seen in 6 cases. These included 3 inversions, 1 deletion, 1 insertion and 1 duplication. All aberrations were located in pericentromere region of chromosome 9 with clinical manifestations including congenital heart disease, peculiar facial appearance, paralysis, dysplasia and/or movement disorder. Chromosome polymorphisms were found in 20 patients, most of which had absence of satellites or variation of heterochromatin on chromosome 9. Of 10 bone marrow specimens from children suspected for acute leukemia, chromosome abnormalities were identified in 5 patients.</p><p><b>CONCLUSION</b>Cytogenetic analysis is useful for children featuring multiple congenital abnormalities. Chromosome 9 abnormalities and their clinical relevance should attract more attention.</p>

The change of HBeAg expression in patients infected hepatitis B virus with pre-C signal enzyme cleavage site mutation / 中华检验医学杂志

Objective To study HBeAg change in patients infected hepatitis B virus(HBV) with pre-C signal enzyme cleavage site mutation. Methods Mutation in pre-C signal enzyme cleavage site was detected by PCR-RFLP. The PreC/C gene with mutation was amplified by PCR and was cloned to EB viral eukarotic expression vector. Then transfect the vector with wild type or mutant PreC/C gene to HepG2 cell. SEAP reporter system was used to monitor the efficiency of transfection. HBeAg and its precursor in the supernatant and HepG2 cell were detected by ELISA and Western blot. Results HBeAg was positive in the supernatant of wild type and negative control in T1862 vaniant by ELISA. In HepG2 cell transfected with wild type, three proteins were detected by Western blot, they were HBeAg(17 000) and two HBeAg precursor(22 000 and 25 000). And in HepG2 cell transfected T1862 vaniant, only two HBeAg precursor was detected. The precursor in cells transfected withT1862 vaniant were significantly stronger than cells transfected with wild type. Conclusion Mutation in pre-C signal enzyme cleavage site may affect the decoration of HBeAg, which may cause great of HBeAg precursor locating in cells and lead to HBeAg negative in serum of patients infected with HBV.