OBJECTIVE:Researches show that a combination of platelet-rich plasma and adipose-derived stem cells can accelerate the healing of skin lesions.However,systematic evidence for the combination of the two is still lacking.The purpose of this study was to assess the efficacy of a combination of two interventions in a clinical rodent skin wound model.METHODS:We searched PubMed,Embase,Cochrane,and CNKI and selected the studies of platelet-rich plasma,adipose-derived stem cell transplantation,or their combination on skin wounds in experimental animals published until July 2023.Wound healing and wound transformation growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor were used as indicators.RevMan 5.3 and Stata 15.0 were used to analyze the data.RESULTS:A total of 12 studies were included,of which 8 studies used rats as experimental subjects and 4 studies used mice as experimental subjects.The experimental group was treated with platelet-rich plasma combined with adipose-stem cell transplantation,and the control group was treated with platelet-rich plasma alone.The results of meta-analysis showed that the wound healing rate of the experimental group at 3,7,and 10 days after treatment was greater than that of the control group[SMD=2.65,95％CI(1.29,4.01),Z=3.81,P=0.0001;SMD=3.38,95％CI(2.47,4.30),Z=7.24,P<0.00001;SMD=2.62,95％CI(1.50,3.73),Z=4.61,P<0.00001].The wound healing time of the experimental group was shorter than that of the control group[SMD=-2.12,95％CI(-3.5,-0.74),P=0.003].The expression of transforming growth factor β,positive rate of CD31,expression of type Ⅰ collagen,and vascular endothelial growth factor in wound of experimental group were higher than those of control group[SMD=5.65,95％CI(1.22,10.08),Z=2.50,P=0.01;SMD=2.49,95％CI(1.96,3.02),Z=9.28,P<0.00001;SMD=3.44,95％CI(0.72,6.17),Z=2.48,P=0.01;SMD=2.38,95％CI(0.97,3.79),Z=3.30,P=0.0010].CONCLUSION:Our results show that platelet-rich plasma+adipose-derived stem cells combined treatment can improve the wound healing rate,shorten the wound healing time,and at the same time increase the expression of transforming growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor to accelerate healing.Due to the limitations of the model,more animal testing and clinical trials are needed.

OBJECTIVE To explore the effect and mechanism of Huiyang Shengji Decoction on wound healing in mice with dia-betic yin syndrome.METHODS SPF C57BL/6 mice were selected,and 24 diabetic ulcer models were established by intraperitoneal injection of streptozotocin and skin defect method.The mice were randomly divided into model group,low-dose,medium-dose and high-dose Huiyang Shengji Decoction groups,with 6 mice in each group.Six mice were selected to establish a common wound model as the blank group.The blank group and model group were treated with distilled water by intragastric administration every day,and the Huiyang Shengji Decoction groups were treated with Huiyang Shengji Decoction by intragastric administration every day.The wounds were photographed every day,and the wound healing rate was recorded.HE staining was used to observe the growth of granulation tis-sue in the wounds.Western blot was used to detect the expression of Axl,Tyro3 and Mertk proteins related to wound efferocytosis.Im-munohistochemistry was used to detect the expression of CD11c in wound tissue.qPCR was used to detect the mRNA expression of TNF-α,IL-1β,IL-10 and TGF-β1 in the wounds.TUNEL staining was used to calculate the number of apoptotic cells(AC)in the wounds.Mouse primary bone marrow dendritic cells(BMDC)were resuspended in 1640 medium containing 10%blank serum or 5%,10%,and 20%Huiyang Shengji Decoction-containing serum.After culturing in a cell culture incubator at 37℃and 5%CO2 for 24 h,the wound endocytosis-related receptors were detected by Western blot.BMDC and AC were co-cultured in vitro,and the phag-ocytic rate of BMDC phagocytosis of AC was detected by flow cytometry.RESULTS The wound healing in each dose group of Huiyang Shengji Decoction was significantly faster than that in the model group(P<0.05).Compared with the model group,the protein expres-sion of Axl and Tyro3 in the wound tissue of mice in the high dose group of Huiyang Shengji Decoction was significantly increased(P<0.01,P<0.000 1),and there was no significant difference in the expression of Mertk protein in the wound tissue of mice in the low,medium and high dose groups(P>0.05).The Huiyang Shengji Decoction significantly inhibited the expression of IL-1β and TNF-α mRNA in the wound tissue(P<0.05),promoted the expression of TGF-β1 and IL-10 mRNA(P<0.05),and the number of AC in the wound after treatment with Huiyang Shengji Decoction was less than that in the model group(P<0.05).In the in vitro experiment,compared with the model group,the protein expression of Axl and Mertk in BMDC in the 20%Huiyang Shengji Decoction-containing serum group was significantly increased(P<0.01,P<0.001),and the protein expression of Tyro3 in the 5%,10%,and 20%Huiy-ang Shengji Decoction-containing serum groups was higher than that in the model group(P<0.01,P<0.001).The phagocytic rate of AC in the Huiyang Shengji Decoction group was higher than that in the model group(P<0.05).CONCLUSION Huiyang Shengji Decoction can enhance the efferocytosis function of dendritic cells(DC),reduce AC aggregation in the wound,promote the disappear-ance of wound inflammation and tissue repair,and accelerate the healing of skin wounds.

Objective: To investigate the precise detection methods for weak partial D type 15 and evaluate their implications for blood transfusion safety, along with the development of corresponding strategies. Methods: A combination of serological methods, including the microplate method, indirect antiglobulin tube method, and microcolumn gel card method, was employed to identify RhD-negative and RhD variant samples. RhD-negative samples were screened for the presence of RHD genes using whole-blood direct PCR amplification. Subsequently, RhD variant samples and RhD-negative samples containing RHD genes underwent full-coding-region sequencing of the RHD gene to confirm their genotypes. The genotyping results were further correlated with the serological test findings for comprehensive analysis. Results: Among 615 549 first-time healthy blood donors, 3 401 samples with an RhD-negative phenotype and 156 samples with RhD variant were identified. Of the 3 401 RhD-negative samples, 1 054 were found to harbor RHD genes. Gene sequencing analysis of the 156 RhD variants and the 1 054 serological negative samples revealed that 89 samples contained the RHD 15 (c. 845G>A) allele. Conclusion: The integration of serological testing methods and genotyping technologies for the precise determination of RhD blood type plays a critical role in ensuring the safety and compatibility of blood transfusions.

OBJECTIVE:Researches show that a combination of platelet-rich plasma and adipose-derived stem cells can accelerate the healing of skin lesions.However,systematic evidence for the combination of the two is still lacking.The purpose of this study was to assess the efficacy of a combination of two interventions in a clinical rodent skin wound model.METHODS:We searched PubMed,Embase,Cochrane,and CNKI and selected the studies of platelet-rich plasma,adipose-derived stem cell transplantation,or their combination on skin wounds in experimental animals published until July 2023.Wound healing and wound transformation growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor were used as indicators.RevMan 5.3 and Stata 15.0 were used to analyze the data.RESULTS:A total of 12 studies were included,of which 8 studies used rats as experimental subjects and 4 studies used mice as experimental subjects.The experimental group was treated with platelet-rich plasma combined with adipose-stem cell transplantation,and the control group was treated with platelet-rich plasma alone.The results of meta-analysis showed that the wound healing rate of the experimental group at 3,7,and 10 days after treatment was greater than that of the control group[SMD=2.65,95％CI(1.29,4.01),Z=3.81,P=0.0001;SMD=3.38,95％CI(2.47,4.30),Z=7.24,P<0.00001;SMD=2.62,95％CI(1.50,3.73),Z=4.61,P<0.00001].The wound healing time of the experimental group was shorter than that of the control group[SMD=-2.12,95％CI(-3.5,-0.74),P=0.003].The expression of transforming growth factor β,positive rate of CD31,expression of type Ⅰ collagen,and vascular endothelial growth factor in wound of experimental group were higher than those of control group[SMD=5.65,95％CI(1.22,10.08),Z=2.50,P=0.01;SMD=2.49,95％CI(1.96,3.02),Z=9.28,P<0.00001;SMD=3.44,95％CI(0.72,6.17),Z=2.48,P=0.01;SMD=2.38,95％CI(0.97,3.79),Z=3.30,P=0.0010].CONCLUSION:Our results show that platelet-rich plasma+adipose-derived stem cells combined treatment can improve the wound healing rate,shorten the wound healing time,and at the same time increase the expression of transforming growth factor β,CD31,type Ⅰ collagen,and vascular endothelial growth factor to accelerate healing.Due to the limitations of the model,more animal testing and clinical trials are needed.

OBJECTIVE To explore the effect and mechanism of Huiyang Shengji Decoction on wound healing in mice with dia-betic yin syndrome.METHODS SPF C57BL/6 mice were selected,and 24 diabetic ulcer models were established by intraperitoneal injection of streptozotocin and skin defect method.The mice were randomly divided into model group,low-dose,medium-dose and high-dose Huiyang Shengji Decoction groups,with 6 mice in each group.Six mice were selected to establish a common wound model as the blank group.The blank group and model group were treated with distilled water by intragastric administration every day,and the Huiyang Shengji Decoction groups were treated with Huiyang Shengji Decoction by intragastric administration every day.The wounds were photographed every day,and the wound healing rate was recorded.HE staining was used to observe the growth of granulation tis-sue in the wounds.Western blot was used to detect the expression of Axl,Tyro3 and Mertk proteins related to wound efferocytosis.Im-munohistochemistry was used to detect the expression of CD11c in wound tissue.qPCR was used to detect the mRNA expression of TNF-α,IL-1β,IL-10 and TGF-β1 in the wounds.TUNEL staining was used to calculate the number of apoptotic cells(AC)in the wounds.Mouse primary bone marrow dendritic cells(BMDC)were resuspended in 1640 medium containing 10%blank serum or 5%,10%,and 20%Huiyang Shengji Decoction-containing serum.After culturing in a cell culture incubator at 37℃and 5%CO2 for 24 h,the wound endocytosis-related receptors were detected by Western blot.BMDC and AC were co-cultured in vitro,and the phag-ocytic rate of BMDC phagocytosis of AC was detected by flow cytometry.RESULTS The wound healing in each dose group of Huiyang Shengji Decoction was significantly faster than that in the model group(P<0.05).Compared with the model group,the protein expres-sion of Axl and Tyro3 in the wound tissue of mice in the high dose group of Huiyang Shengji Decoction was significantly increased(P<0.01,P<0.000 1),and there was no significant difference in the expression of Mertk protein in the wound tissue of mice in the low,medium and high dose groups(P>0.05).The Huiyang Shengji Decoction significantly inhibited the expression of IL-1β and TNF-α mRNA in the wound tissue(P<0.05),promoted the expression of TGF-β1 and IL-10 mRNA(P<0.05),and the number of AC in the wound after treatment with Huiyang Shengji Decoction was less than that in the model group(P<0.05).In the in vitro experiment,compared with the model group,the protein expression of Axl and Mertk in BMDC in the 20%Huiyang Shengji Decoction-containing serum group was significantly increased(P<0.01,P<0.001),and the protein expression of Tyro3 in the 5%,10%,and 20%Huiy-ang Shengji Decoction-containing serum groups was higher than that in the model group(P<0.01,P<0.001).The phagocytic rate of AC in the Huiyang Shengji Decoction group was higher than that in the model group(P<0.05).CONCLUSION Huiyang Shengji Decoction can enhance the efferocytosis function of dendritic cells(DC),reduce AC aggregation in the wound,promote the disappear-ance of wound inflammation and tissue repair,and accelerate the healing of skin wounds.

Objective:This study aimed to assess the infection status of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in healthy populations in China over the past decade and analyze the differences in CMV and EBV infection and related risk factors in healthy populations before and after the lifting of coronavirus disease 2019 (COVID-19) pandemic control measures.Methods:This study retrospectively analyzes the CMV and EBV infection status of 8 827 healthy donors who underwent prehematopoietic stem cell transplantation screening at Peking University People's Hospital from January 2014 to December 2023. Logistic regression analysis was conducted to determine the risk factors for CMV and EBV infection.Results:The CMV and EBV IgG positivity rates were 94.52% and 95.40% among the healthy donors, respectively, with no significant differences before and after the lifting of pandemic control measures (all P value>0.05). However, IgG antibody titers increased [CMV: (100.44±36.50) U/ml vs (109.98±36.31) U/ml, P<0.001; EBV: (281.57±226.79) U/ml vs (361.08±268.58) U/ml, P<0.001] after lifting the COVID-19 restrictions. However, the CMV IgM positivity rate remained unchanged. The EBV IgM positivity rate significantly increased after lifting measures (2.77% vs 6.29%, P<0.001), reaching 8.10% within 3 months. Further analysis of the factors affecting EBV IgM positivity revealed that gender ( OR=1.479, 95% CI 1.169-1.872, P=0.001), age[compared with the group younger than 18 years, the 18-50-year age group ( OR=0.584, 95% CI 0.421-0.820, P=0.002), the >50-year age group ( OR=0.389, 95% CI 0.248-0.610, P<0.001) ], and the lifting of COVID-19 restrictions ( OR=2.360, 95% CI 1.287-3.047, P<0.001) were independent factors influencing EBV IgM positivity in the general population. The EBV IgM positivity rate in individuals under 18 years old was not affected by gender or the lifting of COVID-19 restrictions when stratified by age group. Both genders ( OR=1.499, 95% CI 1.138 - 1.975, P=0.004) and the lifting of COVID-19 restrictions ( OR=2.608, 95% CI 1.940-3.507, P<0.001) were independent factors affecting EBV IgM positivity in the 18-50-year age group. The lifting of COVID-19 restrictions ( OR=2.222, 95% CI 1.101-4.484, P=0.026) was the sole independent factor affecting EBV IgM positivity in individuals over 50 years old. Conclusions:Previous infection rates of CMV and EBV are high in healthy populations in China, which increase with age. COVID-19 infection may increase EBV reactivation rates in healthy individuals, with a more pronounced effect on those aged >18 years.

Objective To investigate the serological characteristics and molecular mechanism of an individual with Am phenotype.Methods The sample with ABO blood group discrepancy was confirmed by serological techniques.The full cod-ing and flanking regions of the ABO gene including intron 1 transcription factor binding site were identified through direct se-quencing of PCR-amplified products.PCR products of exon 6-7 were validated to isolate the ABO gene haplotypes by clo-ning and sequencing individual colonies.Bioinformatics software was used to analyze the structure of the mutant protein.Re-sults The serologic characteristics of ABO blood typing showed the rare Am phenotype.The c.467C/T and c.912C/A heter-ozygous sites in exon 7 were identified by direct sequencing analysis.Further TA cloning and sequencing revealed that the patient carried an ABO*O.01.01 allele and a novel ABO*A allele.The new allele sequence had one nucleotide alteration(C＞A)at position 912 on the background of the ABO*A1.02 allele.The new allele sequence has been included in the Gen-Bank database with the entry number JX489776.The c.912C＞A mutation was predicted to be"probably damaging"and"deleterious"by PolyPhen2 and PROVEAN algorithms,respectively.The free energy change(ΔΔG)value predicted it to have a destabilizing effect on the GTA protein.Meanwhile,modeling of the 3D structure predicted that the p.S304R amino acid substitution may alter the hydrogen bond of the GTA protein.Conclusion The p.S304R substitution of α-1,3-N-acetylgalactosaminyltransferase gene may reduce the antigen expression owing to a greatly destabilizing effect on the structure and function of the GTA protein.

OBJECTIVE To systematically evaluate the efficacy and safety of the four most common cell therapies， namely purified CD34+ （PCCs）， bone marrow mononuclear cells （BMMNCs）， bone marrow mesenchymal stem cells （BMMSCs） and peripheral blood mononuclear cells （PBMNCs） in the treatment of critical limb ischemia （CLI）. METHODS PubMed， Scopus， Embase， Cochrane Central Register of Controlled Trials （CENTRAL） and Web of Science databases were searched from the establishment of each database to June 2023 to collect randomized controlled trials （RCTs） comparing the efficacy and safety of four different cell therapies， namely PCCs， BMMNCs， BMMSCs and PBMNCs， with other cell therapies or standard therapy （ST） in the treatment of CLI. The outcomes indexes included amputation rate， ankle-brachial index （ABI）， transcutaneous oxygen partial pressure （TCPO2）， ulcer healing rate， pain-free walking distance （PFWD） and angiogenesis. After data extraction from clinical studies that met the inclusion criteria， the RoB 2.0 tool was used to assess the risk of bias， and Stata 15.0 software was used for statistical analysis. RESULTS Meta-analysis included 22 studies， involving 1 318 patients. The treatment groups involved 4 types of cell therapies， namely PCCs，BMMNCs， BMMSCs， and PBMNCs. Network meta-analysis showed that the amputation rates of the four cell therapies groups were lower than that of ST group， and only the difference in PBMNCs group was statistically significant（P＜0.05）. Four cell interventions were better than ST in improving ABI （P＜0.05）， and BMMNCs had the most significant effect on improving ABI. PBMNCs and BMMNCs groups had statistically significant differences in improving TCPO2， compared with ST group and BMMSCs group （P＜0.05）. Four cell interventions were better than ST in improving ulcer healing rate， among which BMMNCs group had no statistical difference with ST group （P＞0.05）； ulcer healing rates of the other three groups were higher than that of ST group （P＜0.05）， and those of PBMNCs and BMMSCs groups were significantly higher than that of BMMNCs group （P＜ 0.05）. BMMSCs group had a significantly better effect on improving the PFWD of patients than the ST group after transplantation， with statistical significance （P＜0.05）， but there was no significant difference in PBMNCs and BMMNCs groups compared with ST group （P＞0.05）. The three cell therapies of BMMSCs， BMMNCs and PBMNCs had a significantly better effect on angiogenesis than the ST group， and the BMMSCs group had a significantly better effect than the BMMNCs and PBMNCs groups， with statistical significance （P＜0.05）. CONCLUSIONS The four cell therapies can improve the prognosis of CLI patients to varying degrees. PBMNCs show the lowest amputation rate after transplantation and have the most significant effect on improving TCPO2 and improving the ulcer healing rate. BMMNCs possess the most significant effect on improving ABI. BMMSCs represent obvious advantages in PFWD and angiogenesis.