Objective: To evaluate the safety and efficacy of therapeutic whole blood exchange combined with lymphoplasmapheresis in the treatment of refractory autoimmune hemolytic anemia (AIHA). Methods: A retrospective analysis was performed on the clinical data of AIHA patients who underwent therapeutic whole blood exchange combined with lymphoplasmapheresis at our hospital from March 2022 to May 2025. Efficacy was assessed by comparing changes in hemoglobin, platelet count, and bilirubin levels before and after treatment. Safety was evaluated by analyzing vital signs before and after the procedure, parameters during the exchange, and adverse reactions. Results: A total of 12 AIHA patients were enrolled, completing 19 exchange procedures. The number of procedures per patient ranged from 1 to 3. The median treatment duration was 67 (65-73) minutes, with a median exchange volume of 2 025 (1 851-2 121) mL, comprising 4.5 (4-6) units of red blood cells and 1 350 (1 200-1 400) mL of plasma. Ten patients achieved partial remission, one achieved complete remission, and one showed no response, yielding an response rate of 91% (11/12). After a single session, hemoglobin increased significantly by 17.58±9.85 g/L (P<0.01), while platelets counts decreased by 45 (17.5, 79)×10 /L (P<0.05), and both systolic and diastolic blood pressure showed a significant elevation (P<0.05). However, no statistically significant differences were observed in total bilirubin, indirect bilirubin, white blood cell count, or heart rate. During the procedures, 4 adverse reactions occurred in 3 patients: one child experienced severe heart rate fluctuation twice consecutively, and two adults developed plasma allergies. All reactions resolved spontaneously without pharmacological intervention. Conclusion: The combination of therapeutic whole blood exchange and lymphoplasmapheresis appears to be a safe and effective treatment for refractory AIHA patients.

【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.

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.

Gastroenteropancreatic neuroendocrine tumor G3(GEP-NET G3) is a novel subtype of neuroendocrine neoplasms proposed in 2019,which has unique biological behavior characteristics. However,there are still many challenges and controversies in its diagnosis and treatment. There are obvious differences between GEP-NET G3 and neuroendocrine carcinoma (NEC) in genetic alterations and molecular profiles. The most frequently mutated genes in NET G3 are MEN1,DAXX/ATRX,while in NEC,TP53 and Rb are the most frequently mutated genes. Currently,the mainstream view is that NET G3 and NEC are two distinct diseases with different genetic backgrounds,and NET G3 will not develop into NEC. Several clinical and pathological factors should be considered to distinguish GEP-NET G3 and NEC,which including patients′ medical history,histopathological morphology of neoplasms,Ki-67 index,immunohistochemical results of TP53,Rb,DAXX/ATRX and other markers. Multidisciplinary treatment,including radical resection,chemotherapy,targeted therapy,peptide receptor radionuclide therapy,immunotherapy should be applied in patients with GEP-NET G3. Overall,given its relatively indolent biological behavior,the therapeutic strategy should be more actively. Although the cure strategy of NET G3 has many similarities with NET G1/2,it is completely different from NEC.

Gastroenteropancreatic neuroendocrine tumor G3(GEP-NET G3) is a novel subtype of neuroendocrine neoplasms proposed in 2019,which has unique biological behavior characteristics. However,there are still many challenges and controversies in its diagnosis and treatment. There are obvious differences between GEP-NET G3 and neuroendocrine carcinoma (NEC) in genetic alterations and molecular profiles. The most frequently mutated genes in NET G3 are MEN1,DAXX/ATRX,while in NEC,TP53 and Rb are the most frequently mutated genes. Currently,the mainstream view is that NET G3 and NEC are two distinct diseases with different genetic backgrounds,and NET G3 will not develop into NEC. Several clinical and pathological factors should be considered to distinguish GEP-NET G3 and NEC,which including patients′ medical history,histopathological morphology of neoplasms,Ki-67 index,immunohistochemical results of TP53,Rb,DAXX/ATRX and other markers. Multidisciplinary treatment,including radical resection,chemotherapy,targeted therapy,peptide receptor radionuclide therapy,immunotherapy should be applied in patients with GEP-NET G3. Overall,given its relatively indolent biological behavior,the therapeutic strategy should be more actively. Although the cure strategy of NET G3 has many similarities with NET G1/2,it is completely different from NEC.

【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.

【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.

Case-oriented Workshop training mode is a professional training method which has been highly praised by anesthesiologists in recent years. It is of great significance to stimulate the participation of anesthesiologists and strengthen the application of the extended focused assessment with sonography for trauma (EFAST) in perioperative anesthesia management. The training of EFAST is divided into two stages: the first stage is about the relevant knowledge of EFAST and watching standardized operations on site, and the second stage is for each trainee to practice the operations under the guidance of the teacher until they can complete them independently. The EFAST adopts Workshop training mode, and the trainees can complete the target area scanning in about 90 seconds on average. It can maximize the training effectiveness and comprehensively improve the trainees' mastery of knowledge and skills in an all-round way. It is a training mode worthy of promotion in anesthesiology department.

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.

OBJECTIVETo construct a universal, highly attenuated orf virus expression vector for exogenous genes using green fluorescent protein (GFP) as the reporter gene.

METHODSThe 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.

RESULTSThe 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.

CONCLUSIONGreen 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.

Cells, Cultured ; Endothelial Cells ; metabolism ; Gene Deletion ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Orf virus ; Plasmids ; Transfection