Objective: To establish a "regular blood donor red blood cell gene database"(hereafter referred to as the "database") by applying molecular biology techniques for red blood cell antigens genotyping and utilizing information technology software, and to determine the significance and application value of this "database" in precise red blood cell transfusion. Methods: Fifteen antigens [C, c, E, e, M, N, S, s, Fy (a), Fy (b), Jk (a), Jk (b), Le (a), Le (b), P1] across six blood group systems (RHCE, MNS, FY, JK, Lewis and P1PK) were detected among 9 426 regular blood donors using the TaqMan-MGB method combined with an improved U-shaped microplate approach. With the assistance of information technology software, the "database" was integrated into the overall inventory management system of the blood supply chain. This enabled comprehensive management of regular blood donor and patient information, test results, specific antigen screening for regular blood donors, graded antigen matching between donors and patients, and rare blood type donor records. Results: The TaqMan-MGB method successfully detected paired antigens (C/c, E/e, M/N, S/s, Fy /Fy , Jk /Jk ) within a single reaction well using a standardized PCR amplification protocol. This method provided a reliable testing solution for clinical institutions and empowered blood collection and supply organizations with high-throughput screening capabilities. In the blood supply chain, genotyped red blood cells accounted for 13.2% (721/5 462 U) of the total inventory, with 95.34% (348/365) originating from donors who donated two units of blood. Moreover, the “database” fulfilled 94.06% (443/471 U) of compatible transfusion requirements from medical institutions and effectively managed rare blood type donors. Conclusion: The establishment of the "database" facilitated the transition of blood compatibility testing from traditional serological methods to molecular biology-based gold standard techniques, significantly advancing the implementation of precise transfusion strategies based on multi-antigen matching between donors and patients.

Objective:To study the relationship between the type of hypertriglyceridemia(HTG) and the severity and prognosis of acute pancreatitis (AP).Methods:A total of 112 AP patients with HTG diagnosed and treated in Bozhou People's Hospital from June 2022 to June 2024 were retrospectively selected as the study objects, the AP severity and complication rate of patients with different types of HTG were compared. According to the discharge status of AP patients, they were divided into poor prognosis group (26 cases) and good prognosis group (86 cases). Univariate and multivariate Logistic regression were used to analyze the risk factors of poor prognosis in HTG patients with AP.Results:The main types of HTG in AP patients were type Ⅰ, type Ⅳ and type Ⅴ. The proportion of severe disease in AP patients with type Ⅰ HTG was higher than that of other HTG types, there were statistical differences ( P<0.05). The incidence of pancreatic pseudocyst and infectious pancreatic necrosis in AP patients with different HTG types had no statistical differences ( P>0.05). The incidence of systemic inflammatory response syndrome and organ failure in AP patients with HTG type Ⅰ was higher than that with other HTG types, there were statistical differences ( P<0.05). The proportion of type I HTG and severe AP in the poor prognosis group were higher than those in the good prognosis group : 88.46%(23/26) vs. 68.60%(59/86), 76.92%(20/26) vs. 13.95%(12/86); the triglycerides (TG), very low-density lipoprotein (VLDL), total cholesterol (TC) and fasting plasma glucose (FPG) in the poor prognosis group were higher than those in the good prognosis group: (17.29 ± 3.48) mmol/L vs. (12.29 ± 2.45) mmol/L, (3.04 ± 0.46) mmol/L vs.(2.46 ± 0.68) mmol/L,(6.03 ± 1.13) mmol/L vs.(5.25 ± 1.23) mmol/L, (12.01 ± 1.67) mmol/L vs. (11.14 ± 1.82)mmol/L; while albumin and blood calcium were lower than those in the good prognosis group: (36.28 ± 1.53) g/L vs. (37.33 ± 1.65) g/L, (1.65 ± 0.57) mmol/L vs. (2.04 ± 0.72) mmol/L; there were statistical differences ( P<0.05). The results of single factor analysis showed that combined HTG type, AP severity, triglyceride (TG), very low density lipoprotein (VLDL), total cholesterol (TC), fasting blood glucose (FPG), albumin and blood calcium levels were risk factors affecting the prognosis of AP patients ( P<0.05). Multivariate Logistic regression analysis showed that combined HTG type, AP severity, TG, VLDL, TC, FPG, blood calcium and albumin levels were independent risk factors affecting the prognosis of AP patients ( P<0.05). Conclusions:The type of HTG is closely related to the severity and prognosis of AP patients. Among them, AP patients with type Ⅰ HTG have a higher rate of severe disease. The combination of type Ⅰ HTG, the severity of AP, and abnormal lipid metabolism indexes are all important independent risk factors affecting the prognosis of AP patients.

Purpose To analyze the expression of SNAP23 in esophageal squamous cell carcinoma(ESCC)and its influence on invasion of ESCC cells.Methods A total of 41 cases of ESCC and paired normal adjacent tissue(NAT)were collected.The expression and localization of SNAP23 were detected by immunohistochemical EnVision method.The differences of SNAP23 expression levels between ESCC tissues and NAT tissues were compared to analyze the relationship between SNAP23 expression and clinicopathological characteristics.The expression level of SNAP23 in human immortalized esophageal epithelial cells SHEE and ESCC cell lines TE-1,TE-13 and KYSE150 were detected by Western blot.Lentiviral vector transfection technique was used to construct stable transfection cell lines with knock-down and overexpression of SNAP23 gene.The effect of SNAP23 on invasion of ESCC cell line TE-13 was evaluated by cell invasion experiment.Results The expression of SNAP23 in ESCC was significantly higher(53.7％,22/41)than that in the NAT group(19.5％,8/41),the difference was statistically significant(x2=10.303,P＜0.01).The expression level of SNAP23 in ESCC tissues was significantly correlated with maximum tumor diameter(x2=4.193,P＜0.05)and invasion depth(x2=7.264,P＜0.05),but was not correlated with patient gender,age,tumor site,tumor type,pathologic grade,vascular embolus,nerve invasion and lymph node metastasis(P＞0.05).Compared with human immortalized esophageal epithelial cells SHEE,SNAP23 was highly expressed in ESCC cell lines(P＜0.000 1).Compared with the sh-NT group,the invasion of ESCC cell line TE-13 was inhibited when SNAP23 expression was knocked down(P＜0.000 1);compared with the Vector group,the invasion of ESCC cell line TE-13 was enhanced after overexpression of SNAP23(P＜0.000 1).Conclusion SNAP23 is highly expressed in ESCC tis-sue and cell lines,and its expression level in ESCC tissues is positively correlated with tumor maximum diameter and invasion depth;SNAP23 promotes the invasion of ESCC cells.

Purpose To analyze the expression of SNAP23 in esophageal squamous cell carcinoma(ESCC)and its influence on invasion of ESCC cells.Methods A total of 41 cases of ESCC and paired normal adjacent tissue(NAT)were collected.The expression and localization of SNAP23 were detected by immunohistochemical EnVision method.The differences of SNAP23 expression levels between ESCC tissues and NAT tissues were compared to analyze the relationship between SNAP23 expression and clinicopathological characteristics.The expression level of SNAP23 in human immortalized esophageal epithelial cells SHEE and ESCC cell lines TE-1,TE-13 and KYSE150 were detected by Western blot.Lentiviral vector transfection technique was used to construct stable transfection cell lines with knock-down and overexpression of SNAP23 gene.The effect of SNAP23 on invasion of ESCC cell line TE-13 was evaluated by cell invasion experiment.Results The expression of SNAP23 in ESCC was significantly higher(53.7％,22/41)than that in the NAT group(19.5％,8/41),the difference was statistically significant(x2=10.303,P＜0.01).The expression level of SNAP23 in ESCC tissues was significantly correlated with maximum tumor diameter(x2=4.193,P＜0.05)and invasion depth(x2=7.264,P＜0.05),but was not correlated with patient gender,age,tumor site,tumor type,pathologic grade,vascular embolus,nerve invasion and lymph node metastasis(P＞0.05).Compared with human immortalized esophageal epithelial cells SHEE,SNAP23 was highly expressed in ESCC cell lines(P＜0.000 1).Compared with the sh-NT group,the invasion of ESCC cell line TE-13 was inhibited when SNAP23 expression was knocked down(P＜0.000 1);compared with the Vector group,the invasion of ESCC cell line TE-13 was enhanced after overexpression of SNAP23(P＜0.000 1).Conclusion SNAP23 is highly expressed in ESCC tis-sue and cell lines,and its expression level in ESCC tissues is positively correlated with tumor maximum diameter and invasion depth;SNAP23 promotes the invasion of ESCC cells.

Objective:To study the relationship between the type of hypertriglyceridemia(HTG) and the severity and prognosis of acute pancreatitis (AP).Methods:A total of 112 AP patients with HTG diagnosed and treated in Bozhou People's Hospital from June 2022 to June 2024 were retrospectively selected as the study objects, the AP severity and complication rate of patients with different types of HTG were compared. According to the discharge status of AP patients, they were divided into poor prognosis group (26 cases) and good prognosis group (86 cases). Univariate and multivariate Logistic regression were used to analyze the risk factors of poor prognosis in HTG patients with AP.Results:The main types of HTG in AP patients were type Ⅰ, type Ⅳ and type Ⅴ. The proportion of severe disease in AP patients with type Ⅰ HTG was higher than that of other HTG types, there were statistical differences ( P<0.05). The incidence of pancreatic pseudocyst and infectious pancreatic necrosis in AP patients with different HTG types had no statistical differences ( P>0.05). The incidence of systemic inflammatory response syndrome and organ failure in AP patients with HTG type Ⅰ was higher than that with other HTG types, there were statistical differences ( P<0.05). The proportion of type I HTG and severe AP in the poor prognosis group were higher than those in the good prognosis group : 88.46%(23/26) vs. 68.60%(59/86), 76.92%(20/26) vs. 13.95%(12/86); the triglycerides (TG), very low-density lipoprotein (VLDL), total cholesterol (TC) and fasting plasma glucose (FPG) in the poor prognosis group were higher than those in the good prognosis group: (17.29 ± 3.48) mmol/L vs. (12.29 ± 2.45) mmol/L, (3.04 ± 0.46) mmol/L vs.(2.46 ± 0.68) mmol/L,(6.03 ± 1.13) mmol/L vs.(5.25 ± 1.23) mmol/L, (12.01 ± 1.67) mmol/L vs. (11.14 ± 1.82)mmol/L; while albumin and blood calcium were lower than those in the good prognosis group: (36.28 ± 1.53) g/L vs. (37.33 ± 1.65) g/L, (1.65 ± 0.57) mmol/L vs. (2.04 ± 0.72) mmol/L; there were statistical differences ( P<0.05). The results of single factor analysis showed that combined HTG type, AP severity, triglyceride (TG), very low density lipoprotein (VLDL), total cholesterol (TC), fasting blood glucose (FPG), albumin and blood calcium levels were risk factors affecting the prognosis of AP patients ( P<0.05). Multivariate Logistic regression analysis showed that combined HTG type, AP severity, TG, VLDL, TC, FPG, blood calcium and albumin levels were independent risk factors affecting the prognosis of AP patients ( P<0.05). Conclusions:The type of HTG is closely related to the severity and prognosis of AP patients. Among them, AP patients with type Ⅰ HTG have a higher rate of severe disease. The combination of type Ⅰ HTG, the severity of AP, and abnormal lipid metabolism indexes are all important independent risk factors affecting the prognosis of AP patients.

【Objective】 To understand the current situation of blood components distribution in domestic prefecture-level blood stations through analyzing the components distribution data of 24 prefecture-level blood stations in China. 【Methods】 The data of components distribution of 24 blood stations from 2017 to 2020 as well as the data of blood deployment of 24 blood stations from 2019 to 2020 were collected and analyzed. 【Results】 From 2017 to 2020, positive annual growth in red blood cells, plasma and cryoprecipitate was observed in 22, 19 and 15 out of the 24 blood stations, and the annual growth median rate of above three components was 5.24%, 3.80% and 3.25%, respectively. Among the 24 prefecture-level blood stations, 23 carried out the preparation of cryoprecipitate. 【Conclusion】 The distribution of red blood cells, cryoprecipitate and plasma in prefecture-level blood stations is increasing year by year. However, there is a overstock of plasma, and most blood stations need blood employment.

Objective:To investigate the clinical effects of glycated hemoglobin, fasting blood glucose and serum C-peptide combined screening for gestational diabetes screening and its clinical significance.Methods:A total of 40 pregnant women with gestational diabetes mellitus (observation group) and 40 pregnant women (control group) in Peking University Medical Lu Zhong Hospital from January 2016 to September 2019 were enrolled. The changes of glycated hemoglobin, fasting blood glucose, and serum C-peptide were compared between the two groups, and the efficacy of combined screening for gestational diabetes was evaluated.Results:The glycated hemoglobin of the pregnant women in the observation group was significantly higher than that in the control group [(8.12 ± 1.14)% vs. (5.23 ± 0.15)%], the serum C-peptide of the pregnant women in the observation group was lower than that in the control group [(1.19 ± 0.28) nmol/L vs. (1.61 ± 0.22) nmol/L], and there were significant differences ( P<0.05). There was no significant difference in fasting blood glucose between the two groups ( P>0.05). When glycosylated hemoglobin, fasting blood glucose and serum C-peptide index was screened combinedly, the sensitivity, specificity, positive predictive value, negative predictive value and the areas under the receiver operation characteristic (ROC) curve were 98.65%, 86.50%, 88.42%, 95.45%, and 0.943, respectively. Compared with all other individual indicators, the difference was statistically significant ( P<0.05). Conclusions:The combined detection of glycosylated hemoglobin, fasting blood glucose and serum C-peptide can more accurately diagnose gestational diabetes mellitus at an early stage.

Two proteins of similar molecular weight (named as ASPR-C-1 and ASPR-C-2) from the crude drug of Angelica sinensis were purified and characterized by 80% ammonium sulfate precipitation, Sephadex G-50 gel filtration chromatography, and DEAE-Sepharose anion exchange chromatography. The molecular weight of ASPR-C-1 and ASPR-C-2 on SDS-PAGE was 17.33 kDa and 17.18 kDa, respectively. They were mainly monomeric in solution, but partially formed dimers and they were glycoproteins with glycosyl content of 2.6% and 8.2%, respectively. Both ASPR-C-1 and ASPR-C-2 were identified to be members of pathogenesis-related 10 family of proteins by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and have ribonuclease activities with the specific activity of 73.60 U/mg and 146.76 U/mg, respectively. The optimum pH of the two isoforms was similar, at about 5.6, while their optimum temperatures were different. The optimum temperature of ASPR-C-1 was 50 ℃, and that of ASPR-C-2 was 60 ℃. Both isoforms presented highest thermal stability at 60 ℃. However, ASPR-C-2 was more thermotolerant than ASPR-C-1. The latter was rapidly inactivated and retained only about 20% residual activity while the former still maintained about 80% of its original activity at a higher treatment temperature (80 to 100 ℃). In addition, Fe²⁺ had an activating effect on the ribonuclease activities of two isoforms while Ca²⁺, Mg²⁺, Zn²⁺, Mn²⁺, Ag⁺, Cu²⁺, EDTA (Elhylene diamine tetraacetic acid), dithiothreitol and sodium dodecylsulphate showed different degrees of inhibition of the enzyme activities. Our findings provide a foundation for further research on the biological function of PR-10 protein from Angelica sinensis.
Angelica sinensis ; Chromatography, Gel ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Hydrogen-Ion Concentration ; Kinetics ; Molecular Weight ; Protein Isoforms ; Temperature

OBJECTIVE: To assess the association of 5A/6A polymorphism in the promoter region of MMP3 gene with the stability of extracellular matrix of atherosclerotic plaque. METHODS: Clinical data of 776 consecutive patients undergoing percutaneous coronary intervention (PCI) was reviewed. MMP3 gene polymorphisms and serum level of MMP3 for the second admission were collected. The target gene fragment containing MMP3 promoter region was transfected into HepG2 vector cells. The influence of the polymorphism on the expression of the MMP3 gene was determined in vitro. RESULTS: Compared with the first admission data, the proportion of mutant MMP3 genotypes (5A/5A+5A/6A) was significantly higher in patients with acute myocardial infarction (AMI) compared with the control group (37.6% vs. 24.9%, P<0.01). 64.1% of the patients carrying the 5A allele had AMI, whereas only 50.11% of those carrying the 6A allele had AMI (P<0.01). The proportion of wild-type MMP3 genotype (6A/6A) was significantly higher in the stenotic group compared with the non-restenosis group (79.5% vs. 66.5%, P<0.01). Restenosis has occurred in 9.5% of patients harboring the 5A allele compared with 16.2% in those carrying the 6A allele (P<0.01). In addition, serum level of MMP3 in the restenosis group was significantly lower than that of the non-restenosis group (P<0.01). In vitro studies confirmed that the expression of pGL2-Basic/6A was significantly lower than that of pGL2-Basic/5A. CONCLUSION The 5A/6A polymorphism in the promoter region of the MMP3 gene may influence its transcriptional activity and impact on the degradation or push-up of extracellular matrix, resulting in a difference in the stability of atherosclerosis plaques, which in turn may induce different pathological processes in AMI or restenosis after stenting.
Case-Control Studies ; Extracellular Matrix ; Genetic Predisposition to Disease ; Genotype ; Humans ; Matrix Metalloproteinase 3 ; genetics ; Percutaneous Coronary Intervention ; Plaque, Atherosclerotic ; genetics ; Polymorphism, Genetic ; Promoter Regions, Genetic

The fusion of cell permeable peptide TAT and bifunctional antioxidant enzymes, GST (Glutathione sulfur transferase)-TAT-SOD1 (Cu, Zn superoxide dismutase), is an intracellular superoxide scavenger. Compared with SOD1-TAT, GST-TAT-SOD1 has better protective effect on oxidative damage but less transduction efficiency. A novel cell permeable bifunctional antioxidant enzymes with the fusion of GST, SOD1 and polyarginine R9 was constructed for higher transduction efficiency. The full nucleotide sequence of SOD1-R9 was synthesized and inserted into the prokaryotic expression vector pGEX-4T-1 with the GST tag. After the successful construction of the prokaryotic expression vectors of GST-SOD1-R9, the recombinant vector was then transformed into Escherichia coli BL21 (DE3) and the GST-SOD1-R9 fusion protein was produced with the induction of IPTG. The soluble expression of GST-SOD1-R9 fusion protein was combining with the induction temperature and time. The best soluble expression was obtained with the induction temperature of 25 ℃ and the induction time of 11 h. The fusion protein was purified through the combination of 80% ammonium sulfate precipitation and affinity chromatography using glutathione agarose, and verified by SDS-PAGE and special enzymatic activity. The thermal and pH stability of GST-SOD1-R9 fusion protein were analyzed and the SOD and GST activity of fusion protein were proved to be well maintained under physiological conditions. Finally, the transduction efficiency of GST-SOD1-R9 fusion protein was proved to be better than GST-TAT-SOD1 fusion protein (P<0.05). These works establish a foundation for further study of the protective effect of GST-SOD1-R9 fusion protein against oxidative damage.