Objective:To examine the expression of transmembrane emp24 domain-containing protein 4(TMED4) in liver tissue of patients with hepatocellular carcinoma, and to investigate the effects of TMED4 gene on the proliferation and migration of hepatocellular carcinoma cells and related molecular mechanisms. Methods:The expression of TMED4 at protein level in liver cancer tissue and paracancerous tissue of patients with hepatocellular carcinoma were detected by Western blotting and immunohistochemical stainning, and the correlation between its expression and clinicopathological features was analyzed. The effects of TMED4 overexpression or knockdown on proliferation, migration and healing ability of hepatocellular carcinoma cells in vitro and in vivo were determined by cell proliferation test, Transwell test, wound healing test and subcutaneous tumor formation in nude mice. The molecular mechanism of TMED4 in regulating the biological behavior of hepatocellular carcinoma cells was preliminarily explored by pathway analysis. Independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:The results of Western blotting showed that the expression of TMED4 at protein level in hepatocellular carcinoma tissue was lower than that in paracancerous tissue(0.52±0.29 vs. 0.83±0.22), and the difference was statistically significant( t=2.54, P=0.022). The results of immunohistochemical examination indicated that the expression of TMED4 at protein level in liver cancer tissue was lower than that in paracancerous tissue(5.46±3.37 vs. 7.58±3.08), and the difference was statistically significant( t=3.49, P<0.001). The expression of TMED4 at protein level was significantly correlated with vascular invasion and Barcelona clinic liver cancer stage( χ2=6.83 and 4.20, P=0.009 and 0.040). The results of cell proliferation assay showed that the absorbance value of SMMC-7721 cells in TMED4 overexpression group was lower than that in control group(1.38±0.05 vs. 2.37±0.08), while the optical density value of HepG2 in TMED4 knockdown group was higher than that in control group(0.76±0.04 vs. 0.54±0.01), and the differences were statistically significant( t=18.23 and 8.85, both P<0.001). The results of Transwell test showed that the number of migrated SMMC-7721 cells in TMED4 overexpression group was less than that in control group(286.30±13.01 vs. 439.70±12.34), while the number of migrated HepG2 cells in TMED4 knockdown group was higher than that in control group(249.00±6.00 vs. 160.00±6.56), and the differences were statistically significant( t=14.81 and 17.34, both P<0.001). The wound healing test showed that the healing rate of SMMC-7721 cells in TMED4 overexpression group was lower than that in control group((0.21±0.01)% vs.(0.45±0.01)%), the healing rate of HepG2 cells in TMED4 knockdown group was higher than that in control group((0.46±0.01)% vs.(0.20±0.01)%), and the differences were statistically significant( t=200.10 and 30.46, both P<0.001). The results of subcutaneous tumor formation assay in nude mice showed that the growth rate of cells in TMED4 overexpression group was slower than that in control group. After cell inoculation for 6 weeks, the subcutaneous tumor volume of mice in TMED4 overexpression group was smaller than that in control group(27.36 mm 3(138.70 mm 3) vs. 1 741.62 mm 3(1 783.39 mm 3)), the tumor weight was lower than that in control group(0.06 g(0.14 g) vs. 1.46 g(1.09 g)), and the differences were statistically significant(both Z=-2.31, both P<0.001). The results of Western blotting showed that the expression of Snail at protein level in SMMC-7721 cells of the TMED4 overexpression group was lower than that of the control group(0.32±0.01 vs. 0.90±0.03), the protein level of Snail in HepG2 cells of TMED4 knockdown group was higher than that of control group(1.03±0.01 vs. 0.97±0.01), and the differences were statistically significant( t=28.49 and 12.31, both P<0.001). The results of real time fluorescent quantitative polymerase chain reaction showed that the expression of Snail at mRNA level in SMMC-7721 cells of TMED4 overexpression group was lower than that of control group(0.13±0.05 vs. 1.00±0.15), the expression of Snail at mRNA level in HepG2 cells of TMED4 knockdown group was higher than that of control group(1.25±0.32 vs. 0.21±0.14), and the differences were statistically significant( t=9.62 and 5.10, P<0.001 and P=0.007). Conclusion:TMED4 may affect the proliferation and migration of hepatocarcinoma cells by regulating the expression of Snail, and which is expected to become a potentially therapeutic target for hepatocellular carcinoma.

Objective:To investigate the effect and possible molecular mechanism of mannan-binding lectin-associated serine protease (MASP) 1 on the biological behavior of hepatocellular carcinoma(HCC) cells.Methods:From January 10 to October 25, 2022, 20 pairs of fresh liver cancer tissue and adjacent (3 cm from cancer tissue) normal tissue samples of patients who underwent hepatic cancer resection at the Affiliated Hospital of Nantong University were collected, and the expression of MASP1 was analyzed. From March 1, 2012 to May 20, 2017, the cancer tissue and adjacent (3 cm from cancer tissue) normal tissue samples of 67 patients with HCC were collected from the tissue sample bank of the Department of Pathology, the Affiliated Hospital of Nantong University and the correlation between the expression level of MASP1 and clinical data of patients with HCC were analyzed. Human hepatoma cells lines SK-Hep1 and Hep3B were cultured, and MASP1 overexpression and MASP1 knockdown cell lines were constructed. SK-Hep1 negative control group, SK-Hep1 MASP1 overexpression group, Hep3B negative control group and Hep3B MASP1 knockdown group were set up. The effect of MASP1 on the proliferation of HCC cells was detected by subcutaneous tumor transplantation experiment in nude mice. The effect of MASP1 on the expression of epithelial-mesenchymal transition related proteins (N-cadherin, matrix metalloproteinase 9(MMP9), and E-cadherin), and the effects of MASP1 on the expression of phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) pathway related proteins and their phosphorylation levels were detected by Western blotting. Independent sample- t test, paired- t test and chi-square test were used for statistical analysis. Results:The results of immunohistochemical staining of 20 pairs of fresh tissue samples showed that the expression level of MASP1 in liver cancer tissue was lower than that in adjacent normal tissue (3.73±1.03 vs. 6.76±1.46), and the difference was statistically significant ( t=16.97, P<0.001). The correlation analysis of MASP1 expression level and clinical data of 67 patients with HCC revealed that the expression level of MASP1 was related to vascular invasion of the tumor, and the difference was statistically significant( χ2=5.20, P=0.023). The subcutaneous tumor transplantation experiment in nude mice showed that the volume and weight of subcutaneous tumor of mice in SK-Hep1 MASP1 overexpression group were lower than those of the SK-Hep1 negative control group((165.42±149.08) mm 3vs. (260.42±179.78) mm 3, (0.13±0.12) g vs. (0.18±0.12) g), and the differences were statistically significant ( t=5.15 and 3.89, both P<0.05). The results of Western blotting showed that the expression levels of N-cadherin and MMP9 in SK-Hep1 MASP1 overexpression group were lower than those of SK-Hep1 negative control group, while the expression level of E-cadherin was higher than that of SK-Hep1 negative control group (0.73±0.01 vs. 1.02±0.02, 0.40±0.01 vs. 0.69±0.01, 0.91±0.02 vs. 0.78±0.02), and the differences were statistically significant ( t=24.23, 36.87 and 9.27, all P<0.001). The expression levels of N-cadherin and MMP9 in Hep3B MASP1 knockdown group were higher than those in Hep3B negative control group, and the expression levels of E-cadherin was lower than that in Hep3B negative control group (1.04±0.01 vs. 0.31±0.01, 0.54±0.02 vs. 0.04±0.01, 0.56±0.01 vs. 0.93±0.01), and the differences were statistically significant ( t=163.20, 53.16, 60.74, all P<0.001). The expression levels of phosphoinositide PI3K, phosphoinositide Akt, and phosphoinositide mTOR of SK-Hep1 MASP1 overexpression group were lower than those of SK-Hep1 negative control group (0.59±0.01 vs.1.02±0.01, 0.64±0.01 vs. 1.12±0.02, 0.10±0.01 vs. 1.05±0.02); the expression levels of phosphoinositide PI3K, phosphoinositide Akt, and phosphoinositide mTOR of Hep3B MASP1 knockdown group were higher than those of Hep3B negative control group (0.96±0.01 vs. 0.55±0.01, 0.99±0.01 vs. 0.38±0.01, 0.93±0.02 vs. 0.06±0.01), and the differences were statistically significant ( t=40.87, 40.91, 87.30, 44.17, 107.50, 67.28, all P<0.001). Conclusions:The expression level of MASP1 is low in HCC tissue, and is significantly correlated with the poor prognosis of HCC patients and the occurrence of tumor vascular invasion. MASP1 may affect the proliferation and migration of liver cancer cells by regulating the PI3K/Akt/mTOR signaling pathway, suggesting that MASP1 may become a key gene in the treatment of HCC.

【Objective】 To investigate the clinical manifestations, diagnosis, differential diagnosis and management strategies of thrombocytopenia caused by tirofiban. 【Methods】 The basic clinical data, platelet count changes and treatment course of 7 patients with acute coronary syndrome who used tirofiban resulting in severe thrombocytopenia during their hospitalization in our hospital from December 2021 to March 2023 were collected, and their individual and common characteristics were analyzed. 【Results】 Platelet counts were in the normal range in all 7 patients on admission. Six of the patients had thrombocytopenia occurring from 3 to 16 hours after tirofiban use, and one patient was detected at 34 h of tirofiban use. Their minimum platelet count ranged from (1-11)×10⁹/L. All 7 cases discontinued tirofiban and other antithrombotic drugs, and the platelet count increased to 50×10⁹/L in 6 patients in 2 to 4 days after stopping the drug and gradually returned to the normal range. During this period, there were no bleeding or acute thrombotic events, and no platelet transfusion was conducted. Five patients resumed antithrombotic therapy when the platelet count returned to (20-50)×10⁹/L, 1 patient underwent elective coronary artery bypass grafting (CABG) surgery when the count rose above 50×10⁹/L. One patient had bleeding manifestations after thrombocytopenia and required limited-duration CABG surgery, so 3 U platelet transfusion and immunoglobulin treatment were performed consecutively. CABG surgery was performed when the platelet count increased to 76×10⁹/L. The differential diagnosis of the cause of thrombocytopenia was performed in all seven patients, and other causes of thrombocytopenia, such as heparin, were excluded. 【Conclusion】 Tirofiban can cause acute severe or extremely severe thrombocytopenia. Routine platelet count testing at 6 hours after medication can prevent serious adverse events by discontinuing tirofiban promptly after thrombocytopenia occurs. At the same time, it is determined whether to perform platelet transfusion based on whether the patient has bleeding and the risk of bleeding, and the timing of resuming antithrombotic treatment is determined based on the recovery of platelet count and the risk of thrombosis.

Objective To establish reference intervals for serum soluble transferrin receptor(sTfR)and sTfR/log ser-um ferritin index(sTfR/lgSF)in apparently healthy adults in the Wuhan area,so as to provide reference for clinical diagno-sis and treatment of iron deficiency and iron-deficiency anemia.Methods A total of 273 individuals from the Wuhan Aisa General Hospital,including health examination participants and blood donors,were selected to measure sTfR,other iron metabolism indicators and high sensitivity C-reactive protein(hsCRP).The sTfR/lgSF values were calculated and reference intervals for sTfR and sTfR/lgSF were established using the percentile method(P2.5 to P97.5).Spearman correlation anal-ysis was used to evaluate the relationships between sTfR,sTfR/lgSF,and other iron metabolism indicators,as well as hsCRP.Results The sTfR levels(M,mg/L)between males and females(1.01 vs 1.07)were not statistically significant(P＞0.05),but the sTfR/lgSF levels if males were significantly lower than those in females(0.45 vs 0.62)(P＜0.05).There was no significant difference in sTfR(M,mg/L)and sTfR/lgSF(M)among different age groups,with values of 1.07 vs 1.02 vs 1.00 and 0.52 vs 0.53 vs 0.51,respectively(P＞0.05).The reference interval for STfR was(0.72-1.68)mg/L,the sTfR/lgSF reference interval was(0.31-0.88)for males,and(0.37-1.19)for females.Correlation analysis showed no correlation between sTfR,sTfR/lgSF and hsCRP(r=0.043,P＞0.05;r=-0.064,P＞0.05),while serum ferritin(SF),serum iron(SI),transferrin saturation(TSAT)were correlated with hsCRP(r=0.128,P＜0.05;r=-0.195,P＜0.01;r=-0.173,P＜0.01).There was no correlation between sTfR and SF(r=-0.115,P＞0.05),while sTfR/lgSF was significantly correlated with and SF(r=-0.685,P＜0.01).Conclusion Preliminary reference intervals for serum sTfR and sTfR/lgSF in apparently healthy adults in the Wuhan has been established.sTfR and sTfR/lgSF are not affected by inflammatory factors and are significant for identifying iron deficiency in anemia patients with elevated serum ferritin caused by inflammation.

To investigate the hypoglycemic effects of ethyl acetate extract of Alismatis Rhizoma (AREEA) on type 2 diabetes mellitus,the model of type 2 diabetic rats was induced by high-fat diet feeding for 4 weeks and then intraperitoneal injection of a low dose of streptozotoin (STZ).Rats without the above-mentioned treatment were selected as the normal control group.The diabetic rats were randomly divided into 5 groups:the model control group,low doses (20 mg/kg),medium doses (50 mg/kg),high doses (100 mg/kg) of AREEA groups and positive control-metformin group (100 mg/kg).After four weeks,oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were performed,respectively.12 hours after the last administration,the levels of serum glucose,glycated hemoglobin(HbA1 c),insulin (Ins),total cholesterol (TC),triglyceride (TG),high density lipoprotein (HDL-C),low density lipoprotein (LDL-C),superoxide dismutase (SOD),malondialdehyde (MDA),glutathione (GSH-Px),tumor necrosis factor-or (TNF-α),interleukin-6 (IL-6),insulin-mediated tyrosine of IRS-1 and Akt phosphorylation in adipose tissue were determined.In addition,the pathological changes of pancreas were examined.After administration for 4 weeks,all doses of AREEA significantly reduced the fasting blood glucose of type 2 diabetic rats (P ＜0.05).In the high doses group of AREEA,the levels of GLU,HbA1c,TC,TG,MDA,TNF-α and IL-6 in serum were decreased significantly,and the levels of SOD and GSH-Px were increased (P ＜ 0.05).These results suggest that AREEA has the therapeutic effect on type 2 diabetes-related symptoms by metabolic regulation of glucose and lipids.

Objective:To investigate the efficacy and safety of plasma exchange(PE) in the treatment of autoimmune hemolytic anemia in children.Methods:The data from 8 hospitals in China during November 2014 to April 2017 were collected, and the clinical characteristics of PE in children with AHA were analyzed retrospectively.Results:A total of 21 children with AHA were included in the study, including 17 cases from PICU and 4 cases from pediatric kidney ward, with 11 boys and 10 girls, and the median age was 3.64(0.25, 11.10)years old, and median hospital stay was 12(4, 45)days.There were 15 cases(71.4%) with infection, 2 cases(9.5%)with autoimmune diseases, 4 cases(19.0%) with unknown.Consciousness disturbance occurred in 4 patients before replacement and recovered to normal after PE.The volume of blood decreased in two cases(9.5%) and completely relieved.There were 20 cases of anemia (95.2%), 15 cases were normal after PE, and 5 cases were improved.Jaundice occurred in 18 cases (85.7%), 12 cases were normal after PE, 6 cases were improved.Hepatosplenomegaly was found in 11 cases, 10 cases were normal after PE, 1 case was improved.After PE, the hemoglobin and red blood cell count increased, while the total bilirubin, indirect bilirubin, urea nitrogen and lactate dehydrogenase decreased, there were significant differences between pre-and post-replacement ( P<0.05). Only 1 case had allergic reaction, which was improved after symptomatic treatment, and PE was continued.After PE, 2 cases (9.5%) had complete remission, 16 cases (76.2%) had partial remission and 3 cases (14.3%) had been discharged. Conclusion:PE therapy can obviously improve the clinical symptoms and laboratory indexes of children with AHA who have failed to respond to conservative treatment.It can be used as a treatment measure for children with severe AHA and has a good safety.

【Objective】 To investigate the effect of optimized preoperative hemoglobin (Hb) level on clinical outcome in patients undergoing coronary artery bypass grafting (CABG). 【Methods】 Retrospective analysis was performed on patients who were selected to receive CABG from April 2020 to August 2021 in our hospital. Preoperative basic data, perioperative blood transfusion volume, blood transfusion rate, acute liver function impairment, renal function impairment (AKI), ICU stay, length of hospital stay, and in-hospital mortality of patients, meeting the inclusion criteria, were collected. According to the perioperative red blood cell transfusion, the optimal preoperative Hb threshold was calculated by receiver operating characteristic curve (ROC). According to the threshold, all patients were divided into two groups, and the blood transfusion volume and clinical outcomes of the two groups were compared to evaluate the predictive value of the optimal threshold of Hb. 【Results】 A total of 915 patients who met the inclusion criteria were enrolled in the study. The optimal threshold for predicting red blood cell transfusion rate by calculating preoperative Hb value by ROC curve was 118 g/L for males and 116g/L for females. Group A: Hb≤ threshold (n=293) was divided into the red blood cell transfusion group A1 and the red blood cell non-transfusion group A2. Group B: Hb>threshold (n=622) was divided into the red blood cell non-transfusion group B1 and no red blood cell non-transfusion group B2. The risk factors for perioperative red blood cell transfusion were age (OR=1.033 874, 95%CI 1.000 4-1.068 3, P<0.01), gender (female) (OR=3.268 5, 95%CI 2.353 1-4.540 0, P<0.01), BMI (OR=0.927 8, 95%CI 0.883 3-0.974 4, P<0.01), chronic renal insufficiency (CKD) (OR=2.041 1, 95%CI 1.347 8-3.091 0, P<0.01). Preoperative Hb≤ threshold (OR=3.517 4, 95%CI 2.502 1-4.944 7, P<0.01) was an independent risk factor for perioperative red blood cell transfusion. Perioperative red blood cell transfusion in patients with preoperative anemia further increases the incidence of postoperative complications (acute liver injury, AKI) and length of ICU stay. 【Conclusion】 Preoperative Hb≤ threshold can effectively predict perioperative red blood cell transfusion in patients with CABG, and increase the risk of postoperative acute liver injury, AKI, prolonged ICU stay and hospital stay. Optimizing the preoperative Hb level in CABG patients, increasing the Hb level to 118 g/L in males and 116 g/L in females can reduce the incidence of perioperative red blood cell transfusion and postoperative complications.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.