Study on RHD and RHCE gene mRNA based on three generation sequencing technology / 中国输血杂志

【Objective】 To establish RH gene mRNA sequencing method based on nanopores sequencing and to explore the RHD and RHCE mRNA transcripts in D positive and Del individuals. 【Methods】 From March 2021 to May 2022, 5 RhD positive samples and 5 Del samples screened out by hospitals in Chengdu were sent to our laboratory for futher examination. The erythrocytes and buff coat were isolated, then DNA and RNA were extracted.All 10 samples were genotyped by PCR-SSP. After the mRNA was reversely transcribed into cDNA, the full-length mRNA of RHD and RHCE genes were simultaneously amplified by a pair of primers. Sanger sequencing and third-generation sequencing technology based on Nanopore were used to sequence the amplified products, and the types and expressions of different splices of RHD and RHCE gene mRNA transcripts were analyzed. 【Results】 The method established in this study can simultaneously amplify the full length transcripts of RHD and RHCE. Ten different RHD gene mRNA transcripts and nine RHCE gene mRNA transcripts were detected in 10 samples. RHD full-length transcript (RHD-201) can be detected in RhD Del type, but the expression amount was significantly lower than that in RhD positive samples. The expression amount of transcript RHD-207 (Del789) in Del samples was significantly higher than that in RhD positive samples. The transcript RHD-208 (Del8910+ 213) was only detected in RhD Del type individuals, and no significant difference was found between other RHD transcripts and all RHCE transcripts in the two phenotypes. 【Conclusion】 In this study, an analytical method for sequencing full-length transcript isomers of RHD and RHCE mRNA via the third generation was successfully established, and complex alternative splicing patterns were found in RHD and RHCE genes, providing a new method for the study of alternative splicing of blood group gene variants mRNA.

Effect of FTY720 on transfusion-related acute lung injury: a preliminary investigation / 中国输血杂志

【Objective】 To investigate the effects and mechanisms of different doses of fingolimod (FTY720) on non-antibody-mediated transfusion-related acute lung injury (TRALI). 【Methods】 A TRALI mouse model was constructed using lipopolysaccharide (LPS) pre-stimulation and platelets (Plt) of different storage days for second strike. The success of the modeling was determined by protein concentration in lung tissue homogenates, myeloperoxidase (MPo) activity, lung wet/dry weight ratio (W/D ratio), lung tissue damage score and pathological sections. Ceramide and sphingosine-1-phosphate (S1P) contents in platelets of different storage days were detected. FTY720 was administered 1 h after LPS injection to investigate the role of FTY720 in TRALI. The expression levels of vascular endothelial cadherin (VE-cadherin) and zonula occludens-1 (ZO-1) were analyzed by WB. 【Results】 Mice infused with stored 5-day Plt (d5Plt group) exhibited typical signs of TRALI, and the differences in lung tissue homogenate protein concentration (6 546.38±409.50) μg/mL, MPO activity (49.38±4.43) U/L, W/D ratio 4.79±0.21, and lung tissue damage score 7.24±0.38 from the rest of the groups were statistically significant (P<0.05). With the increase of platelet storage time, the ceramide content gradually increased and S1P content gradually decreased, and the ratio of the two was imbalanced. d5Plt showed statistically significant differences (P<0.01) in ceramide content (58.37±5.69) μmol/L and S1P content (149.81±4.86) nmol/L from the rest of the groups. After preventive administration of FTY720, 1 mg/kg FTY720 had no significant effect on TRALI mice, whose lung tissue homogenate protein concentration (6 170.26±545.50) μg/mL, MPO activity (45.97±4.79) U/L, W/D ratio 4.88±0.25, and lung tissue damage score 7.92±0.65 were significantly higher than those of the normal and LPS control groups (P<0.01). The low-dose (0.5, 0.2, and 0.1 mg/kg) FTY720 group alleviated lung injury, and its protein concentration, MPO activity, W/D ratio, and lung tissue injury score were significantly lower than those of the d5Plt group (P<0.05). Pathological sections also showed similar results. In terms of endothelial intercellular junction protein expression, the VE-cadherin expression levels in the 1 mg/kg FTY720 group were significantly lower than those in the normal and LPS control groups (P<0.05), and the VE-cadherin and ZO-1 expression levels in the low-dose (0.5, 0.2, and 0.1 mg/kg) FTY720 group were significantly higher than those in the d5Plt group (P<0.05), which tended to be normalized. 【Conclusion】 In this study, a TRALI mouse model was successfully established by one strike of LPS and two strikes of d5Plt. Low doses of FTY720 (0.5, 0.2, 0.1 mg/kg) were protective against TRALI, while high doses of FTY720 (1 mg/kg) may aggravate the symptoms of TRALI. This protective effect may be somewhat dependent on the expression of VE-cadherin and ZO-1.

Effect and mechanism of extracorporeal photopheresis on preventing acute graft versus host disease in ice / 中国输血杂志

【Objective】 To investigate the preventive effects of early apoptotic splenic mononuclear cells induced by extracorporeal photopheresis (ECP) on acute graft versus host disease (aGVHD) in mice and explore the underlying mechanisms. 【Methods】 1) Splenic mononuclear cells were extracted from C57BL/6 mice and treated with different concentrations of 8-MOP (50 ng/mL, 100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL). After treatment, irradiate the cells with 2 J/cm2 of ultraviolet light. Then, use the Annexin V-FITC/PI apoptosis detection kit to assess the early apoptosis rate of the cells and determine the optimal concentration of 8-MOP for the experiment.2) There were 35 SPF-grade female BALB/C mice (H-2Kd) aged 6-8 weeks. After whole-body irradiation with 8Gy X-rays, the mice were divided into five groups: sham irradiation group received intravenous infusion of 0.2 mL of normal saline, the syngeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of BALB/C mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the allogeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of C57BL/6 mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the aGVHD group received intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107) in 0.2 mL, the ECP prevention group received pre-transplant intravenous infusion of 0.2 mL of ECP-treated splenic mononuclear cells of C57BL/6 mice (including a cell count of 1×107 ) 48 hours before transplantation, and on the day of transplantation, intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107 ) in 0.2 mL.The preventive effects of ECP on aGVHD were observed, and the concentrations of IFN-γ, IL-2, TNF, IL-4 and IL-6 in mouse serum were measured using CBA. Th1 cell counts were determined by flow cytometry. 【Results】 Different concentrations of 8-MOP (50 ng/mL,100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL) were used to treat mouse splenic mononuclear cells. The early apoptosis rates (%), observed after treatment were as follows: (14.18±0.865) vs (16.76±0.407) vs (18.83±0.404) vs (19.27±0.404) vs (14.5±0.529). The appropriate concentration of 8-MOP was determined to be 200 ng/mL.In vivo experiment, the results showed that the aGVHD group had decreased survival rate, reduced body weight, and increased clinical scores compared to the syngeneic and allogeneic bone marrow transplantation groups (P<0.01), and the chimerism of bone marrow cells in mice after transplantation was over 90%. ECP significantly improved the survival rate of mice after transplantation, reduced clinical scores (P<0.05), and decreased the concentrations of Th1 cell cytokines in serum (P<0.05) and the counts of Th1 cells in the spleen (P<0.05). 【Conclusion】 ECP-induced early apoptotic single nuclear cells from the spleen can prevent the occurrence of aGVHD by reducing the Th1 response in mouse.

Factors affecting the efficacy of platelet transfusion and specificity of HLA-Ⅰ antibodies in tumor patients / 中国输血杂志

【Objective】 To analyze the factors affecting the efficacy of platelet transfusion and the specificity of HLA-Ⅰ antibodies in tumor patients. 【Methods】 Tumer patients applied for platelet transfusion from November 2019 to July 2021 in Sichuan Cancer Hospital province were screened for platelet antibodies. The transfusion efficacy was evaluated and its influencing factors were analyzed, Antibody positive patients were detected for HLA-Ⅰ antibodies. 【Results】 The positive rates of platelet antibody and HLA-Ⅰ antibody, as well as the platelet transfusion refractoriness rate were 35.2%(50/142), 32.4%(46/142) and 16.9%(24/142), respectively. The platelet refractoriness rate of platelet antibody positive patients was higher than that of platelet antibody negative patients [30.0%(15/50) vs 9.8%(9/92)] (χ2=9.428, P<0.05). Multivariate logistic results showed that gender (χ2=12.608, P<0.001, OR=3.800, 95%CI: 1.819-7.940) was an independent risk factor for platelet antibody production, and platelet antibody(χ2=8.648, P<0.05, OR=3.952, 95%CI: 1.581~9.878) was the independent risk factor for transfusion efficacy. The detection rates of strong positive, positive and weak positive HLA-I antibodies were 69.6% (32/46), 80.4% (37/46) and 97.8% (45/46), respectively. The antibodies with high detection rate were anti-B*15∶12, anti-B*57∶03, anti-B*57∶01, anti-B*13∶02, anti-B*49∶01, anti-B*50∶01, anti-A*25∶01, anti-B*15∶01, anti-B*15∶02 and anti-B*44∶02. There were 19 kinds of HLA-Ⅰ antibodies with MFI≥10 000, including anti-A*02∶01, anti-A*02∶03 and anti-A*02∶06. The CCI values were lower in HLA-Ⅰ antibody positive patients(n=46) than in HLA-I antibody negative patients(n=96) by Wilcoxon test (P<0.05). 【Conclusion】 Among tumor patients who received platelet transfusion, gender(female) was the risk factor for platelet antibody production. Platelet antibody positive patients were prone to platelet transfusion refractoriness. HLA-Ⅰ antibodies were the main platelet antibodies, suggesting that HLA-Ⅰ gene matching platelets could improve the efficacy of platelet transfusion.

Application of prone position in severe ARDS caused by pneumocystis jirovecii pneumonia after kidney transplantation / 器官移植

Objective To investigate the improvement of oxygenation after the treatment of prone position in patients with severe acute respiratory distress syndrome (ARDS) caused by pneumocystis jirovecii pneumonia (PJP) after kidney transplantation. Methods Clinical data of 5 cases of moderate and severe ARDS caused by PJP after kidney transplantation were analyzed retrospectively, and clinical characteristics, treatment regimen and prognosis were summarized. Results Clinical manifestations of 5 patients were fever, dry cough, chest tightness, shortness ofbreath,sweating and fatigue, and body temperature fluctuated between 38 ℃ and 39 ℃, percutaneous arterial oxygen saturation(SpO2) was gradually decreased, and respiratory distress symptoms were worsened. Pulmonary CT scan showed diffuse ground-glass shadow. After transfer to intensive care unit (ICU), immunosuppressive drugs were terminated, and all patients were given with compound sulfamethoxazole, caspofungin, low-dose glucocorticoids against pneumocystis jirovecii (PJ), oxygen therapy and other symptomatic supportive treatments. Four patients diagnosed with severe ARDS upon admission to ICU were treated in a prone position. One patient with moderate ARDS was not kept in a prone position. At 1 d after treatment in a prone position, partial pressure of arterial oxygen (PaO2) and oxygenation index were increased, whereas alveolar-arterial oxygen difference (A-aDO2) was decreased compared with before treatment (allP<0.05). Compared with 1 d after treatment, SpO2, PaO2 and oxygenation index were all increased, while A-aDO2 was decreased at 4 d after treatment (all P<0.05). Box diagram showed that oxygenation index showed an overall upward trend after prone-position treatment, whereas A-aDO2 showed an overall downward trend. The length of ICU stay of 5 patients was 14 (8, 29) d. All patients in a prone position did not develop complications, such as skin pressure sore, tube detachment and tube displacement, etc. Among 5 patients, 4 patients were mitigated, and 1 patient died of septic shock and multiple organ failure. Conclusions For both conscious and intubated patients, a prone position may significantly improve oxygenation and prognosis of patients with severe ARDS caused by PJP after kidney transplantation. Early diagnosis and accurate and standardized treatment play a pivotal role in enhancing cure rate.

Genotyping Characteristics of Human Fecal Escherichia coli and Their Association with Multidrug Resistance in Miyun District, Beijing / 生物医学与环境科学（英文）

OBJECTIVE@#To explore the genotyping characteristics of human fecal Escherichia coli( E. coli) and the relationships between antibiotic resistance genes (ARGs) and multidrug resistance (MDR) of E. coli in Miyun District, Beijing, an area with high incidence of infectious diarrheal cases but no related data.@*METHODS@#Over a period of 3 years, 94 E. coli strains were isolated from fecal samples collected from Miyun District Hospital, a surveillance hospital of the National Pathogen Identification Network. The antibiotic susceptibility of the isolates was determined by the broth microdilution method. ARGs, multilocus sequence typing (MLST), and polymorphism trees were analyzed using whole-genome sequencing data (WGS).@*RESULTS@#This study revealed that 68.09% of the isolates had MDR, prevalent and distributed in different clades, with a relatively high rate and low pathogenicity. There was no difference in MDR between the diarrheal (49/70) and healthy groups (15/24).@*CONCLUSION@#We developed a random forest (RF) prediction model of TEM.1 + baeR + mphA + mphB + QnrS1 + AAC.3-IId to identify MDR status, highlighting its potential for early resistance identification. The causes of MDR are likely mobile units transmitting the ARGs. In the future, we will continue to strengthen the monitoring of ARGs and MDR, and increase the number of strains to further verify the accuracy of the MDR markers.

Notoginseng Radix et Rhizoma Powder Treats Dyslipidemia via PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo evaluate the clinical efficacy and safety of Notoginseng Radix et Rhizoma powder in treating dyslipidemia by regulating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodSixty patients with dyslipidemia （syndrome of combined phlegm and stasis） treated in the Third Affiliated Hospital of Henan University of Chinese Medicine from May 2021 to June 2022 were selected in this study and randomized into two groups according to the randomized， double-blind control principle. The control group was treated with Xuezhikang capsules + Notoginseng Radix et Rhizoma powder placebo and the observation group with Notoginseng Radix et Rhizoma powder + Xuezhikang capsules placebo for 6 weeks. The clinical efficacy， traditional Chinese Medicine （TCM） syndrome scores， and liver and kidney function indicators were evaluated at weeks 0， 3， and 6. Enzyme-linked immunosorbent assay （ELISA） was employed to determine the expression of vascular endothelial growth factor （VEGF）， kinase insert domain receptor （KDR）， epidermal growth factor （EGF）， and epidermal growth factor receptor （EGFR） in the peripheral serum. Quantitative Real-time PCR was employed to measure the mRNA levels of KDR， EGFR， PI3K， and Akt in the mononuclear cells of the peripheral blood. ResultThe observation group （83.33%） showed the total effective rate comparable to that of the control group （89.66%） and no adverse reactions. Compared with before treatment， the patients in the observation group showed decreased TCM syndrome score and serum levels total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C） and after being treated for 3 and 6 weeks （P<0.05）， the level of high-density lipoprotein cholesterol （HDL-C） showed an upward trend, but the difference was not statistically significant. After treatment， the two groups showed no significant differences. Compared with that before treatment, the mRNA expression of PI3K, Akt and EGFR in peripheral blood mononuclear cell and the expression of EGF, VEGF and KDR in serum of the observation group showed a downward trend with time, in which the mRNA expression of PI3K, Akt, VEGF and KDR decreased more significantly (P<0.05)，The expression levels of KDR mRNA and serum EGFR show a trend of first increasing and then decreasing.Compared with the control group after treatment, there was no statistically significant difference in mRNA expression of PI3K, Akt, EGFR, and KDR, as well as serum levels of EGF, EGFR, VEGF, and KDR between the two groups of patients at the same time point. ConclusionNotoginseng Radix et Rhizoma powder is safe and effective in correcting dyslipidemia. It may inhibit the PI3K/Akt signaling pathway by down-regulating the expression of VEGF/KDR to lower the blood lipid level.

Study on protective effect and mechanism of Modified Yupingfeng nasal spray on nasal mucosal injury in AR model rats / 中国药房

OBJECTIVE To study protective effect and potential mechanism of Modified yupingfeng nasal spray （YPF+） on nasal mucosal injury in allergic rhinitis （AR） model rats. METHODS AR model was induced by ovalbumin （OVA） and randomly divided into model group， YPF+group （50 µg/side，twice a day）， positive control group （Mometasone furoate aqueous nasal spray， 50 µg/side，once a day）； the blank group was set up， with 10 rats in each group. Administration groups were given relevant medicine， and blank group and model group were given equivalent normal saline for consecutive 4 weeks. Thirty minutes after last medication， the behavioral scores of rats were recorded， and the pathological changes of their nasal mucosa tissue were observed. The level of reactive oxygen species （ROS） in nasal mucosa tissue was detected. The protein and mRNA expressions of nucleotide- binding oligomerization domain-like receptor protein 3 （NLRP3），caspase-1，gasdermin D （GSDMD） were detected； the contents of interleukin-1β （IL-1β） and IL-18 in serum were also determined. RESULTS Compared with blank group， in model group， the nasal mucosa tissue structure was disordered， inflammatory cells infiltrated seriously， and lamina propria vascular dilation was visible； its behavioral score and pathological score， the level of ROS， protein and mRNA expressions of NLRP3， caspase-1 and GSDMD， serum contents of IL-1β and IL-18 in nasal mucosa tissue were increased significantly （P＜0.05）. Compared with model group， the symptoms of nasal mucosal injury in rats of each drug group were improved to varying degrees， and the above indicators were significantly reduced （P＜0.05）. CONCLUSIONS YPF+ may improve nasal mucosal injury of rats， relieve AR symptoms such as sneezing， itchy nose， runny nose， the mechanism of which may be associated with reducing the production of ROS in nasal mucosa and downregulating NLRP3/caspase-1/ GSDMD pathway.

Prevention and control of vessel catheter associated infections: a harsh and long way to go / 中国实用护理杂志

Vascular access devices are regarded as double-edged swords since being used as lifesaving accesses widely as well as causing vessel catheter associated infections and other complications.In severe cases, it may even endanger life. This paper expounded the importance and necessity of vessel catheter associated infections prevention and control from several aspects such as national policy promotion, medical insurance payment innovation and clinical management demand, analyzed the key links of vessel catheter associated infections prevention and control and the research status quo here and abroad based on the life cycle of the catheter, and pondered the future directions considering the current difficulties and blind spots.

A novel method for electroencephalography background analysis in neonates with hypoxic-ischemic encephalopathy / 中国当代儿科杂志

OBJECTIVES@#To explore a new method for electroencephalography (EEG) background analysis in neonates with hypoxic-ischemic encephalopathy (HIE) and its relationship with clinical grading and head magnetic resonance imaging (MRI) grading.@*METHODS@#A retrospective analysis was performed for the video electroencephalography (vEEG) and amplitude-integrated electroencephalography (aEEG) monitoring data within 24 hours after birth of neonates diagnosed with HIE from January 2016 to August 2022. All items of EEG background analysis were enrolled into an assessment system and were scored according to severity to obtain the total EEG score. The correlations of total EEG score with total MRI score and total Sarnat score (TSS, used to evaluate clinical gradings) were analyzed by Spearman correlation analysis. The total EEG score was compared among the neonates with different clinical gradings and among the neonates with different head MRI gradings. The receiver operating characteristic (ROC) curve and the area under thecurve (AUC) were used to evaluate the value of total EEG score in diagnosing moderate/severe head MRI abnormalities and clinical moderate/severe HIE, which was then compared with the aEEG grading method.@*RESULTS@#A total of 50 neonates with HIE were included. The total EEG score was positively correlated with the total head MRI score and TSS (rs=0.840 and 0.611 respectively, P<0.001). There were significant differences in the total EEG score between different clinical grading groups and different head MRI grading groups (P<0.05). The total EEG score and the aEEG grading method had an AUC of 0.936 and 0.617 respectively in judging moderate/severe head MRI abnormalities (P<0.01) and an AUC of 0.887 and 0.796 respectively in judging clinical moderate/severe HIE (P>0.05). The total EEG scores of ≤6 points, 7-13 points, and ≥14 points were defined as mild, moderate, and severe EEG abnormalities respectively, which had the best consistency with clinical grading and head MRI grading (P<0.05).@*CONCLUSIONS@#The new EEG background scoring method can quantitatively reflect the severity of brain injury and can be used for the judgment of brain function in neonates with HIE.

Compilation and evaluation of gastrointestinal endoscopy satisfaction scale / 中南大学学报(医学版)

OBJECTIVES@#Gastrointestinal endoscopy plays an important role in the diagnosis and treatment of gastrointestinal diseases. The satisfaction degree of gastrointestinal endoscopy can directly affect the patient's compliance and further impact the treating effect. At present, there is no scale to evaluate the satisfaction degree of gastrointestinal endoscopy in China. This study aims to develop a satisfaction scale of gastrointestinal endoscopy suitable for national conditions and to evaluate its reliability and validity, which provides a tool for clinic to evaluate patients' satisfaction with gastrointestinal endoscopy.@*METHODS@#The original gastrointestinal endoscopy satisfaction scale was compiled by literature review, consulting senior endoscopists and experts. Through the first round of survey about 120 patients, the original scale was analyzed and modified according to the results to get the gastrointestinal endoscopy satisfaction scale (formal scale). The formal scale was used to conduct the second round of survey about 200 patients. The reliability and validity of the scale were analyzed and evaluated according to the survey results.@*RESULTS@#The reliability of the original scale was good but the validity was poor. The formal scale had 2 dimensions and 10 items, the Cronbach's alpha and split-half reliability were 0.889 and 0.823. The structure validity index χ2/df was 2.513, root mean square error of approximation (RMSEA) was 0.094, goodness of fit index (GFI) was 0.914, adjusted goodness of fit index (AGFI) was 0.861, comparative fit index (CFI) was 0.946, normed fit index (NFI) was 0.915. The aggregate validity was general, the discriminative validity was good, and the direct score of patients was strongly correlated with the total score of the scale.@*CONCLUSIONS@#The gastrointestinal endoscopy satisfaction scale has good reliability and validity, which can be used as a tool to evaluate patients' satisfaction with gastrointestinal endoscopy in China.

Research progress in strigolactones and application prospect in medicinal plants / 中国中药杂志

Strigolactones(SLs) are a class of sesquiterpenoids derived from the carotenoid biosynthesis pathway with the core carbon skeleton consisting of tricyclic lactone(ABC tricyclic ring) and α,β-unsaturated furan ring(D ring). SLs are widely distributed in higher plants and are symbiotic signals between plants and Arbuscular mycorrhiza(AM), which play key roles in the evolution of plant colonizing terrestrial habitats. As a new type of plant hormone, SLs possess such important biological functions as inhibiting shoot branching(tillers), regulating root architecture, promoting secondary growth, and improving plant stress resistance. Therefore, SLs have attracted wide attention. The biological functions of SLs are not only closely related to the formation of "excellent shape and quality" of Chinese medicinal materials but also have important practical significance for the production of high-quality medicinal materials. However, SLs have been currently widely studied in model plants and crops such as Oryza sativa and Arabidopsis thaliana, and few related studies have been reported on SLs in medicinal plants, which need to be strengthened. This review focused on the latest research progress in the isolation and identification, biological and artificial synthesis pathways, biosynthesis sites and transport modes, signal transduction pathways and mechanisms, and biological functions of SLs, and prospected the research on the regulation mechanism of SLs in the growth and development of medicinal plants and their related application on targeted regulation of Chinese herbal medicine production, which is expected to provide some references for the in-depth research on SLs in the field of Chinese medicinal resources.

Mechanisms of Coreopsis tinctoria Nutt in the treatment of diabetic nephropathy based on network pharmacyology analysis of its active ingredients / Mecanismos de Coreopsis tinctoria Nutt. En el tratamiento de la nefropatía diabética basado en el análisis farmacológico de red de sus principios activos

SUMMARY: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) can protect diabetic kidneys, but the mechanisms are unclear. This work is to investigate the potential mechanisms of C. tinctoria Nutt. in the treatment of diabetic nephropathy based on network pharmacology analysis of its active ingredients. Twelve small molecular compounds of C. tinctoria Nutt. and targets related to diabetic nephropathy were docked by Discovery Studio 3.0. DAVID database was used for GO enrichment and KEGG pathway analysis. Cytoscape 3.6.1 was used to construct active ingredient-target network. Cell viability was detected with MTT. Glucose consumption was analyzed with glucose oxidase method. Protein expression was measured with Western blot and immunofluorescence. Electron microscopy observed autophagosomes. The core active ingredients of C. tinctoria Nutt. included heriguard, flavanomarein, maritimein, and marein. Twenty-one core targets of the 43 potential targets were PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK, and ABL2. These 21 core targets were significantly enriched in 50 signaling pathways. Thirty- four signaling pathways were closely related to diabetic nephropathy, of which the top pathways were PI3K/AKT, insulin, and mTOR, and insulin resistance. The enriched GO terms included biological processes of protein phosphorylation, and the positive regulation of PI3K signaling and cytokine secretion; cellular components of cytosol, extracellular region, and extracellular space; and molecular function of protein kinase activity, ATP binding, and non-membrane spanning protein tyrosine kinase activity. In vitro experiments found that marein increased the expression of phosphorylated AKT/AKT in human renal glomerular endothelial cells of an insulin resistance model induced by high glucose, as well as increased and decreased, respectively, the levels of the microtubule-associated proteins, LC3 and P62. C. tinctoria Nutt. has many active ingredients, with main ingredients of heriguard, flavanomarein, maritimein, and marein, and may exert anti-diabetic nephropathy effect through various signaling pathways and targets.

RESUMEN: Coreopsis tinctoria Nutt. (C. tinctoria Nutt.) puede proteger riñones diabéticos, sin embargo los mecanismos son desconocidos. Este trabajo se realizó para investigar los potenciales mecanismos de C. tinctoria Nutt. en el tratamiento de la nefropatía diabética basado en el análisis de farmacología en red de sus principios activos. Doce compuestos moleculares pequeños de C. tinctoria Nutt. y los objetivos relacionados con la nefropatía diabética fueron acoplados por Discovery Studio 3.0. La base de datos DAVID se utilizó para el enriquecimiento GO y el análisis de la vía KEGG. Se usó Cytoscape 3.6.1 para construir una red de ingrediente-objetivo activa. La viabili- dad celular se detectó mediante MTT. El consumo de glucosa se analizó con el método de glucosa oxidasa. La expresión proteica fue determinada mediante Western blot e inmunofluorescencia. En la microscopía electrónica se observó autofagosomas. Los principales ingredientes activos de C. tinctoria Nutt. incluyeron heriguard, flavanomarein, maritimin y marein. Veintiún de los 43 objetivos potenciales fueron PYGM, TLR2, RAF1, PRKAA2, GPR119, INS, CSF2, TNF, IAPP, AKR1B1, GSK3B, SYK, NFKB2, ESR2, CDK2, FGFR1, HTRA1, AMY2A, CAMK4, GCK y ABL2. Estos 21 objetivos principales se enriquecieron significativamente en 50 vías de señalización. Treinta y cuatro vías de señalización estuvieron estrechamente relacionadas con la nefropatía diabética, de las cuales las principales vías fueron PI3K/ AKT, insulina y mTOR, y resistencia a la insulina. Los términos GO enriquecidos incluyeron procesos biológicos de fosforilación proteica, la regulación positiva de la señalización de PI3K y la secreción de citoquinas; componentes celulares del citosol, región extracelular y espacio extracelular; y la función molecular de la actividad de la proteína quinasa, la unión de ATP y la actividad de la proteína tirosina quinasa que no se extiende por la membrana. Los experimentos in vitro encontraron que la mareína aumentaba la expresión de AKT/AKT fosforilada en células endoteliales glomerulares renales humanas en un modelo de resistencia a la insulina inducida por niveles elevados de glucosa, así como aumentaron y disminuyeron respectivamente, los niveles de las proteínas asociadas a los microtúbulos, LC3 y P62. C. tinctoria Nutt. tiene muchos principios activos, con ingredientes principales de heriguard, flavanomarein, maritimain y marein, y puede ejercer un efecto de nefropatía antidiabética a través de distintass vías de señalización y objetivos.

Effect of trehalose on platelets stored at 4℃ in vitro / 中国输血杂志

【Objective】 To investigate the effect of trehalose added to M-Sol on platelet preservation at 4℃ in vitro. 【Methods】 Seven bags of ABO homotype concentrated platelets were randomly selected and divided into 7 groups according to storage temperature, preservation medium and trehalose concentration: 22℃+ PRP group, PRP group, M-Sol+ PRP group, M-Sol+ PRP+ 1 g/L trehalose group, M-Sol+ PRP+ 5 g/L trehalose group, M-Sol+ PRP+ 15 g/L trehalose group and M-Sol+ PRP+ 25 g/L trehalose group. PRP group and M-Sol preservation groups were stored at 4℃. Plt, PDW, MPV, maximum aggregation, CD62p positive rate, GLU and LAC concentration were detected on the 1st, 3rd, 5th and 7th day after preservation, and the changes of GLU and LAC concentration within 7 days were calculated. 【Results】 With the extension of preservation time, Plt decreased in all groups, and there was no significant difference among groups at the same time (P>0.05). PDW and MPV increased in all groups. When preserved to the 7th day, the PDW 10.22±0.43(fL) and MPV 8.74±0.40(fL) of M-Sol+ PRP+ 5 g/L trehalose group were the lowest, which was significantly different from that of 22℃+ PRP group (P0.05). During the preservation period, the maximum aggregation degree of each group decreased gradually. Except for the PRP group, the maximum aggregation degree of the M-Sol+ PRP+ 5g/L trehalose group was the highest(27.29±6.62), which was significantly higher than that of the 22℃+ PRP group on the 7th day after preservation (P<0.05). On the 7th day after preservation, the positive rate of CD62p in M-Sol+ PRP+ 5g/L trehalose group was the lowest(15.46±2.46), and there was significantly different compared with other groups (P<0.05). 【Conclusion】 Adding appropriate concentration of trehalose to M-Sol can inhibit platelet activation at 4℃ and reduce platelet storage lesion.

Current research of RBC alloantibodies associated with blood transfusion and its susceptibility factors / 中国输血杂志

Blood transfusion, an important auxiliary means of clinical treatment, however, is not absolutely safe and risk-free as the transfusion-associated RBC alloantibodies being a potential risk. The yielding rate of RBC alloantibodies in Chinese Han population is about 0.2%, and the researches concerning its production mechanism is particularly critical due to its important clinical significance in patients′ future blood transfusion or pregnancy. This paper reviews the current research status of red blood cell alloantibodies associated with blood transfusion and its susceptibility factors.

Mechanism and transfusion strategy of immunogenic platelet transfusion refractoriness / 中国输血杂志

Platelet transfusion is an irreplaceable method to treat patients with platelet reduction in Hematology Department and Oncology Department. However, immunogenic platelet transfusion refractoriness is a huge challenge to patients who require repeatedly platelet transfusion. Therefore, mechanisms and reasonable transfusion strategies should be formulated to improve immunogenic platelet transfusion outcomes. This article aims to review the research progress and prospects concerning mechanisms and solutions of immunogenic platelet transfusion refractoriness, and provide rational transfusion strategies for platelet transfusion refractoriness patients, thus improving platelet transfusion outcomes.

Advances in extracorporeal photopheresis / 中国输血杂志

Extracorporeal photopheresis(ECP) is a bidirectional cellular immunomodulatory therapy based on leukapheresis, which can mediate not only immunopotentiation but also immunosuppression. Clinically, ECP has been observed to have good efficacy in the treatment of cutaneous T cell lymphoma(CTCL), graft versus-host disease(GVHD) and solid organ transplant rejection, also the US Food and Drug Administration(FDA) has approved ECP for the treatment of CTCL. The treatment guidelines for GVHD in the US and Europe also include ECP, however, there is a lack of relevant guidelines in China. Although the exact mechanism of ECP is not fully explained, recent studies provide a theoretical basis for a further understanding of the bidirectional regulation of ECP.The initial hypothesis was the combination of 8-methoxypsoralen(8-MOP) and ultraviolet A(UVA) to induce apoptosis of immune cells. As the research progressed, this idea was transformed into the differentiation of monocyte to dendritic cell(DC), cytokine alteration and the regulatory T cell(Treg) stimulation.In this article, the current exploration of the immunomodulatory mechanism in ECP and its clinical application were reviewed, also the latest molecular mechanism of ECP mediated immunopotentiation and immunosuppression was comprehensively analyzed, meanwhile the further promotion and clinical application of ECP in China had been prospected.

Extracellular vesicles in the blood components and their roles in transfusion-related immunomodulation / 中国输血杂志

Extracellular vesicles (EVs) with a variety of bioactive molecules can be continuously secreted during the storage of blood components, which can promote cell-cell interaction and mediate cell-to-cell signal transmission. EVs are widely involved in inflammation, metabolism and immunization. In recent years, studies have reported that EVs in the blood components can not only cause transfusion-related lung injury through activating neutrophils, but also act on monocytes to stimulate inflammatory response. EVs can play different regulatory effects on the immune system of patients. In this review, the role and mechanism in transfusion-related immunomodulation (TRIM) of EVs in different blood components have been summarized.

Genetic background of RhD negative blood donors / 中国输血杂志

【Objective】 To analyze the genetic background of RhD-negative blood donors by detecting RHD and RHCE genes of those donors. 【Methods】 From March 2021 to May 2022, the blood samples of RhD-negative blood donors, who had been screened out by RhD primary screening and confirmatory experiments in the Yaan Blood Center, were firstly identified whether the RHD allele was completely deleted, then whether there were deletions in 10 exons of non-RHD allele complete deletion samples, finally, the remaining samples without RHD alleles and exon deletions were further analyzed by DNA sequencing. RHCE gene was detected by SSP-PCR method. 【Results】 Among the RHD gene test results of 104 RhD-negative samples, 65 cases were completely deleted (d/d), 33 were RHD partially deleted (one allele deletion), and 6 were without RHD gene deletion. The RHD alleles of 33 samples with partial deletion were detected by 10 exons, 13 had partial exon deletion, with genotype as RHD*D-CE(3-9)-D/d and phenotype as RhD negativity, and the remaining 20 samples had no exon deletion. The exon sequencing results of the non-deletion samples showed RHD*1227A/RHD*1227A in 6 samples, RHD*1227A/d in 19, RHD*3A/d in 1; both of the last two were considered Del by ISBT. The RHCE gene test results showed that all cc genotype blood donors were RhD true negative, while Del blood donors had no cc genotype. 【Conclusion】 Through the genetic background study of RhD negative blood donors, it is found that there is a high proportion of Del with weak expression of RhD antigen, whether this blood type affects clinical blood safety needs further researches.

Research progress of evidence-based medicine in clinical practice of plasmatransfusion / 中国输血杂志

Plasma is widely used in clinical, but the reliable evidence-based medical evidences that can guide clinicians to properly use plasma are limited, and inappropriate use may even cause deterioration of the disease and serious adverse reactions. Based on the relevant international blood transfusion guidelines and published clinical trial studies, this paper aims to summarize the evidence-based basis of plasma in clinical applications, discuss the safety and efficacy of plasma applications under different conditions, and provide assistance to clinical practice and scientific research of plasma in the future.