Objective: To investigate prevalence of hepatitis E virus (HEV) infection among voluntary blood donors in Dalian and provide evidence for enhancing blood screening strategies. Methods: A total of 3 277 blood donor samples collected between December 2023 and February 2024 at Dalian Blood Center underwent routine blood screening (ALT, HBsAg, anti-HCV, HIV Ag/Ab, anti-TP, and HBV/HCV/HIV NAT). Subsequently, HEV antigen (Ag) was detected using chemiluminescence immunoassay (CLIA). HEV-Ag reactive samples were further tested for HEV RNA, IgM and IgG antibodies. Blood donors with repeated reactive HEV Ag results were followed up to clarify the status of infection. Results: Among the 3 277 blood donor samples, 6 (0.18%) were repeatedly reactive for HEV Ag. However, supplemental testing for HEV RNA, anti-HEV IgM, and anti-HEV IgG on these samples yielded non-reactive results. One of these six blood donors was successfully followed up. On day 218 after the initial detection of HEV Ag reactivity, HEV Ag, HEV RNA, HEV IgM and IgG antibody were found to be non-reactive. Conclusion: The reaction rate for HEV antigen screening among voluntary blood donors in Dalian is low. CLIA method for detecting HEV antigen is easy to operate and cost-effective, but demonstrates some false reactivity. Improving the specificity of the assay and combining it with nucleic acid testing (NAT) would be valuable for implementing a selective HEV screening strategy for blood donors.

Objective To observe the clinical efficacy and safety of cattle encephalon glycoside and ignotin injection combined with reduced glutathione injection and levodopa and benserazide hydrochloride tablets in the treatment of elderly patients with Parkinson's disease complicated with mild cognitive impairment(PD-MCI).Methods Elderly patients with PD-MCI were randomly divided into control group and treatment group.Two groups were treated with levodopa and benserazide hydrochloride tablet for anti-Parkinson's disease therapy,and on this basis,the control group was given intravenous drip of reduced glutathione 1.2 g(qd),and the treatment group was given intravenous drip of cattle encephalon glycoside and ignotin injection 10 mL(qd)on the basis of the control group.Both groups of patients were treated continuously for 4 weeks.The clinical efficacy,serum regulated upon activation,normal T cell expressed and secreted(RANTES),α-synuclein(α-syn),brain-derived neurotrophic factor precursor protein(pro-BDNF),catalase(CAT),total superoxide dismutase(T-SOD),cognitive function and quality of life were compared between both groups,and the adverse drug reactions were observed.Results In treatment group,48 cases were enrolled but 3 cases were lost,thus 45 cases were finally included in the analysis.In control group,48 cases were enrolled but 6 cases were lost,thus 42 cases were included in the analysis.After treatment,the total effective rates in treatment and control groups were 93.33％(42 cases/45 cases)and 76.19％(32 cases/42 cases)respectively(P＜0.05).After treatment,RANTES levels in treatment group and control group were(25.57±4.62)and(30.29±4.92)pg·mL-1;α-syn levels were(3.08±0.76)and(3.74±1.09)μg·L-1;pro-BDNF levels were(144.65±17.54)and(182.26±15.75)ng·mL-1;CAT levels were(168.54±10.64)and(160.48±9.74)kU·L-1;T-SOD levels were(123.75±20.54)and(110.18±22.66)kU·L-1;Montreal Cognitive Assessment Scale scores were(25.08±2.75)and(23.14±2.64)points;and 39-item Parkinson's Disease Quality of Life Questionnaire scores were(45.84±8.42)and(50.34±7.62)points,respectively.The differences of above indexes were statistically significant between two groups(all P＜0.05).The adverse drug reactions in treatment group were arrhythmia,dizziness,chills and gastrointestinal discomfort.The adverse drug reactions in control group were transient thrombocytopenia and arrhythmia.The total incidences of adverse drug reactions in treatment group and control group were 8.89％and 4.76％wthout significant difference(P＞0.05).Conclusion Cattle encephalon glycoside and ignotin injection combined with reduced glutathione injection and levodopa and benserazide hydrochloride tablets has exact clinical efficacy in the treatment of elderly patients with PD-MCI,and it can significantly enhance the neuroprotective and antioxidant effects and improve the cognitive function and quality of life of patients,and it does not increase the incidence rate of adverse drug reactions.

Objective To study the pharmacokinetics and bioequivalence of two formulations of rasagiline mesylate tablets in healthy subjects under fasting and fed conditions.Methods The two-period,two-sequence,crossover study design was adopted in the fasting study.Thirty-six subjects were enrolled and given either test preparation or reference preparation 1 mg respectively in two periods.After collecting plasma samples,the plasma concentration of rasagiline was determined by liquid chromatography-tandem mass spectrometry(LC-MS/MS)and the bioequivalence was evaluated using the average bioequivalence(ABE)method.The four-period,two-sequence,fully replicate crossover study design was adopted in the fed study.Forty-eight subjects were enrolled and given the test preparation or the reference preparation at a dose of 1 mg twice respectively in four periods.According to the degree of intra-individual variation of Cmax,AUC0-t and AUC0-∞,the equivalence was evaluated using the reference-scaled average bioequivalence and ABE method,respectively.Results In the fasting study,the pharmacokinetic parameters of rasagiline of the test and reference preparation were as follow:Cmax were(9.70±3.14)and(9.62±3.85)ng·mL-1,AUC0-t were(6.03±1.47)and(6.02±1.95)ng·h·mL-1,AUC0-∞ were(6.13±1.51)and(6.12±1.97)ng·h·mL-1.The 90％confidence interval(CI)of the geometric mean ratio(GMR)were 94.11％-118.06％,99.22％-107.74％and 99.16％-107.44％for Cmax,AUC0-t and AUC0-∞,respectively,which were within the acceptance criteria of 80.00％-125.00％.In the fed study,the pharmacokinetic parameters of rasagiline of the test and reference preparation were as follow:Cmax were(3.00±1.92)and(3.52±1.77)ng·mL-1,AUC0_t were(5.02±1.20)and(5.06±1.20)ng·h·mL-1,AUC0-∞ were(5.11±1.23)and(5.14±1.22)ng·h·mL-1.The 90％CI of GMR were 96.99％-101.19％and 97.17％-101.41％for AUC0-t and AUC0-∞,which were within the acceptance criteria of 80.00％-125.00％.The 95％upper confidence bound of Cmax for were less than"0",and the point estimate of GMR were within the acceptance criteria of 80.00％-125.00％.The incidence of adverse events in fasting and fed studies was 22.86％and 22.92％,respectively,and all adverse events were moderate to mild.Conclusion The two rasagiline mesylate tablets were bioequivalent,and both the formulations were well tolerated.

The clinical tumor therapy was greatly challenged due to the complex characteristics of tumor microenvironment, however, which also provide arena for novel therapeutic strategies. In this study, poly(2-ethyl-2-oxazoline)-poly(lactic acid)-SS-poly(β-amino ester (PEOz-PLA-SS-PBAE) triblock copolymers with pH and GSH double response were synthesized, polymer micelles were prepared by thin film hydration method for loading of silybin to improve its antitumor activity. The critical micelle concentration was determined by pyrene fluorescence method as 1.8 μg·mL^-1. The particle size was 155.30 ± 1.80 nm as determined by dynamic light scattering, with polydispersity index of 0.168 ± 0.004. The drug loading and entrapment efficiency of the micelles were determined by HPLC as (5.48 ± 0.04)% and (68.52 ± 0.48)%, respectively. The in vitro drug release profiles showed that the micelles have low pH sensitivity and high GSH responsiveness, and exhibited sustained release profiles. The good biocompatibility of the material was proved by measuring the hemolysis rate and cytotoxicity of the blank micelle. The cytotoxicity and apoptosis rate of tumor cells showed that the drug loaded PEOz-PLA-SS-PBAE micelles had significant inhibitory effect and apoptosis-inducing effect on MDA-MB-231 cells. The results of wounding healing assay and Transwell invasion test showed that the drug loaded PEOz-PLA-SS-PBAE micelles could significantly inhibit the metastasis of MDA-MB-231 cells. The PEOz-PLA-SS-PBAE drug-loaded micelles prepared in this study have good inhibitory effect on tumor growth and anti-tumor metastasis in vitro, which lays the foundation for the further application of silybin.

Objective:To investigate the reference ranges for thyroid function and its influencing factors in preterm infants at 14 d after birth.Methods:This retrospective study involved 514 preterm infants who met the inclusion criteria in Affiliated Hospital of Inner Mongolia Medical University from January 1, 2019 to December 31, 2021. They were divided into three group according to their gestational age [early premature group (26-31 +6 weeks, n=153), middle premature group (32-33 +6 weeks, n=129) and late premature group (34-36 +6 weeks, n=232)] or birth weight (BW) [<1 500 g group ( n=129), 1 500-2 000 g group ( n=120) and ≥2 000 g group ( n=265)]. Venous blood samples were collected from the infants at 14 d after birth and their thyroid function was determined by chemiluminescence immunoassay. The reference values of free triiodothyronine (FT 3), free thyroxine (FT 4) and thyroid stimulating hormone (TSH) were calculated based on the values of 95% confidence intervals ( CI) and expressed as percentiles in the range from P2.5 to P97.5. Mann-Whitney U test or Kruskal-Wallis H test was used to compare those thyroid hormone levels between groups. Spearman correlation analysis was used to study the correlation of gestational age or birth weight with FT 3, FT 4 and TSH levels. The factors influencing the levels of thyroid hormones were analyzed by multiple linear regression. Results:The reference ranges for FT 3, FT 4 and TSH were 1.53-3.72 pg/ml, 0.81-1.91 ng/dl and 1.32-7.80 μIU/ml in the early premature infants, 1.74-4.16 pg/ml, 0.90-2.82 ng/dl and 0.63-7.64 μIU/ml in middle prematures and 2.07-4.88 pg/ml, 1.09-2.27 ng/dl and 1.14-7.06 μIU/ml in late prematures. The reference ranges for the above three indexes were 1.53-4.06 pg/ml, 0.81-1.83 ng/dl and 1.14-7.84 μIU/ml in premature infants with BW<1 500 g, 1.67-3.98 pg/ml, 0.88-2.97 ng/dl and 0.94-7.64 μIU/ml in those whose BW between 1 500 g and 2 000 g and 1.91-4.75 pg/ml, 1.09-2.31 ng/dl and 1.14-6.32 μIU/ml in those whose BW≥2 000 g. Multiple linear regression showed that the level of FT 3 was positively correlated with gestational age ( β=0.119, P<0.05) and birth weight ( β=1.950×10 －4, P<0.05); that of FT 4 was positively correlated with gestational age only ( β=0.031, P<0.05); and TSH level was negatively correlated with birth weight ( β=－4.250×10 －4, P<0.05). Conclusions:Gestational age and birth weight are the factors influencing thyroid function in preterm infants at 14 d after birth. Evaluation of thyroid function with FT 4 and TSH should based on the references ranges of different gestational age and birth weight .

Objective To investigate the diagnostic and prognostic value of serum exosomal exocirc_0023461 in coronary artery disease(CAD)related acute myocardial infarction(AMI).Methods From December 2019 to January 2022,383 patients with CAD related AMI(AMI group),200 pa-tients with chronic stable CAD but no AMI(control group),and 200 healthy individuals identified with physical examination(healthy group)were recruited in our hospital.The serum exocirc_0023461 level was determined by real-time quantitative fluorescence polymerase chain reaction.The correlation of serum exosomal circ_0023461 level with clinicopathological features and oxida-tive stress indicators was analyzed.Results The serum level of exocirc_0023461 was significantly higher in the AMI group than the control group and healthy group[3.54(1.39,9.82)vs 0.86(0.62,1.23)and 0.65(0.41,0.79),P＜0.01].ROC curve analysis showed that when the serum level of exocirc_0023461 ≥1.31,its AUC value for the diagnosis of AMI was 0.857(95％CI:0.827-0.887),with a sensitivity and specificity of 78.90％and 83.50％,respectively.Kaplan-Meier survival curve displayed that the survival time was significantly shorter in the high level AMI patients without MACE than those with low level(X2=19.390,Plog rank＜0.01).Multivariate Cox regression analysis revealed that age,peripheral artery disease and serum exocirc_0023461 were independent predictors of MACE occurrence in AMI patients during follow-up(P＜0.05,P＜0.01).Pearson correlation analysis indicated that serum exocirc_0023461 level was negatively correlated with serum GPX and SOD levels(r=-0.395,r=-0.193,P＜0.01),and positively correlated with serum MDA level(r=0.194,P＜0.01).Conclusion Serum exocirc_0023461 may be a potential biomarker for the diagnosis and prognosis of CAD-related AMI,and its mechanism seems to be associated with its regulating oxidative stress and thus affecting myocardial injury.

ObjectiveTo compare and evaluate the improvement degree of spermatogenic dysfunction mice at different recovery periods after cyclophosphamide modeling. MethodsForty-eight male ICR mice aged 4-5 weeks with the body weight of approximately 18-20 g were randomly divided into three control groups and three model groups, with 8 mice in each group. Each mouse of three model groups was intraperitoneally injected with 60 mg/kg cyclophosphamide continuously from the 1^st to 7^th day of the experiment, while each mouse of three control groups was intraperitoneally injected with the corresponding volume of normal saline. Then these mice were continued to be fed for another 7, 14 and 21 days after cyclophosphamide injection, respectively. A corresponding control group was set for each model group. The mice in each group were sacrificed after blood collection through orbital veins at corresponding time points. Testis, epididymis and seminal vesicle were taken and weighed, and their reproductive organ indexes were calculated. Histopathological changes of testis and epididymis were compared after HE staining.Sperm quality analysis was used to determine sperm-related indexes. Serum reproductive hormone content, testicular oxidative stress level and testicular signature enzyme activity were detected by ELISA and related kits.Results Compared with the control group, on the 7^th, 14^th and 21^st day after cyclophosphamide treatment, the testicular index of mice in the model group decreased significantly (P<0.01). The epididymis index decreased significantly on the 7^th and 14^th day, and the seminal vesicle index decreased obviously on the 7^th and 21^st day (P<0.05). And the histopathological damage of testis and epididymis of the model group gradually alleviated over time. On the 7^th and 14^th day after cyclophosphamide treatment, the sperm count of the model group declined remarkably (P<0.01), the serum testosterone (T) level reduced (P<0.05), the malonaldehyde (MDA) content of testis increased significantly (P<0.01), the content of reduced glutathione (GSH) and superoxide dismutase (SOD) decreased obviously (P<0.05),the lactic dehydrogenase (LDH) activity of testis reduced obviously (P<0.05), the gamma-glutamyl transpeptidase (γ-GT) activity increased significantly (P<0.05), the latter two of which are important testicular signature enzymes. Therein on the 7^th day after cyclophosphamide treatment, the sperm motility decreased significantly (P<0.001), the rate of sperm malformation increased obviously (P<0.05), the serum levels of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) increased notably (P<0.01). Nevertheless on the 21^st day after cyclophosphamide treatment, the sperm-related indexes, the content of serum reproductive hormone, the level of testicular oxidative stress and the activity of testicular signature enzyme did not change significantly (P＞0.05). ConclusionThe reproductive toxicity in mice was more apparent on the 7^th day after intraperitoneal injection with 60 mg/kg cyclophosphamide for seven days, at which time the more desirable spermatogenic dysfunction model of mice could be established. However, with the prolongation of the recovery period, the indexes of spermatogenic dysfunction in mice gradually recovered and approached the normal level on the 21^st day after cyclophosphamide treatment.

GJ-4 is crocin enrichments extracted from Gardenia jasminoides J. Ellis, and our previous studies have shown that GJ-4 significantly improved learning and memory impairment induced by Aβ in mice. Herein, a memory deficit model was developed by injecting okadaic acid (OA) into the lateral ventricle of mice, and the neuroprotection and underlying mechanism of GJ-4 on neuronal injury caused by Tau hyperphosphorylation were investigated. The Animal Care & Welfare Committee, Institute of Materia Medica, CAMS & PUMC has approved all procedures (No.00000318). GJ-4 at different doses was intragastric administration to mice for 16 days. Step-down test and Morris water maze test showed that GJ-4 could significantly improve OA-induced memory impairment in mice, and reduced the loss of Nissl bodies in the hippocampus of mice. GJ-4 could also decrease the phosphorylation level of Tau protein at Ser396, Thr231 and Ser404 via increasing protein phosphatase 2A (PP2A) activity and inhibiting glycogen synthase kinase-3β (GSK-3β) activity. Besides, further researches indicated that GJ-4 could inhibit the level of oxidative stress in the brain of OA mice, reduce neuronal apoptosis and inhibit the neuroinflammation mediated by activation of astrocytes in the hippocampus of mice, and eventually achieve its effects in improving learning and memory impairment in mice. According to these findings, we anticipated that GJ-4 might be a potential therapeutic drug for Alzheimer's disease.

【Objective】 To validate the performance of a nucleic acid testing(NAT) system for blood screening in the high-altitude Nagqu region of Tibet, in order to assess the capability of NAT in high-altitude areas and further enhance blood safety. 【Methods】 Various methods were employed to evaluate the analytical sensitivity, reproducibility, ability to prevent cross-contamination, and comparison between different NAT systems. 【Results】 The NAT system in the Nagqu region of Tibet achieved a 100% detection rate for high-concentration HBV DNA and HIV-1 RNA samples, and over 90% for medium-concentration samples. PROBIT analysis revealed the lower limits of detection (LOD) for HBV DNA and HIV-1 RNA to be 8.29 IU/mL (95% CI, 5.88~20.55 IU/mL) and 40.52 IU/mL (95% CI, 30.26~85.92 IU/mL), respectively. For HCV RNA genotype 2a, the LOD was 97.14 IU/mL (95% CI, 71.00~182.67 IU/mL), all of which were lower than the declared minimum detectable concentrations in the instructions. Reproducibility analysis demonstrated a 100% level of consistency within the system. Cross-contamination performance verification showed a strong ability to resist cross-contamination. Comparative analysis of repeated testing of low-concentration HBV DNA samples and multi-system testing in plain areas revealed consistency rates of 77.78%(14/18) and 77.27%(17/22), respectively, indicating certain differences between the NAT system in Nagqu region and other systems. 【Conclusion】 The NAT system exhibited excellent performance in blood screening at high altitudes. The results of performance validation in high-altitude blood screening NAT systems were largely consistent with those in plain areas, providing a reliable basis for enhancing blood safety in high-altitude regions.

【Objective】 To investigate the prevalence of hepatitis D virus in Dalian blood donors. 【Methods】 The samples reactive to HBV in blood screening were selected with the following confirmed results: 1)HBsAg+ &HBV DNA+ ; 2)HBsAg+ & HBV DNA-; 3)HBsAg-&HBV DNA+ ; 4)NAT-yield uncertain. Qualified samples in routine blood screening were additionally tested with anti-HBc+ and anti-HBs+. All samples selected were tested HDV IgG further. Initial reactive samples would be tested by another HDV IgG assay and HDV IgM assay. HDV IgG positive was confirmed when samples were reactive to two HDV IgG assays. 【Results】 None HDV antibodies were detected among 1 344 unqualified samples (507 HBsAg+ &HBV DNA+, 33 HBsAg+ &HBV DNA-, 477 HBsAg-&HBV DNA+ and 327 NAT-yield uncertain samples) or 766 qualified samples (397 anti-HBc+ and 369 anti-HBs samples) in blood screening. 【Conclusion】 The prevalence of HDV infections among Dalian blood donors eligible in pre-donation screening seemed extremely low. However, for areas with high HBV prevalence, the risk of blood safety caused by OBI co-infection with HDV should not be ignored.