Objective To analyze the clinical characteristics and risk factors of vancomycin-resistant En-terococcus faecium(VRE-fm)isolated from urine,and to provide a scientific basis for the prevention and treatment of VRE-fm in clinic.Methods The clinical data and drug sensitivity results of 60 patients with VRE-fm strains isolated from urine in this hospital from January 2017 to June 2023 were retrospectively ana-lyzed.The patients were divided into the infection group(n=46)and the non-infection group(n=16)accord-ing to the status of urinary tract infection.Univariate and multivariate logistic regression were used to analyze the risk factors of urinary tract infection caused by VRE-fm.Results Among the 60 patients with VVRE fm strain isolated from urine,43 cases(71.7%)were female,36 cases(60.0%)were≥65 years old,42 cases(70.0%)had a history of indignant catheter,and the median length of hospitalization was 26.5 days.The re-sults of in vitro VRE-fm drug sensitivity test showed that the resistance rate of Teicolanin was 95%,and the resistance rate of linezolid and tigacycline were 1.7%.Multivariate logistic regression analysis showed that u-rinary sludge white blood cell count＞5/HP was an independent risk factor for VRE-fm urinary tract infec-tion(P＜0.05).Conclusion Urine-isolated VRE-fm infections were more common in female,older patients and were significantly associated with a history of indwelling catheters.

Objective To investigate the risk factors for carbapenem-resistant Acinetobacter baumannii(CRAB)bloodstream infection in intensive care units(ICU)and construct a nomogram prediction model.Methods A retrospective analysis was conducted on the medical records of patients with CRAB bloodstream(BSI)infection in the ICU ward of the First Afliated Hospital,Sun Yat-sen University from January 1,2018 to December 31,2023.Cox regression analysis was usedo determine the risk factors for death in patients with CRAB bloodstream infection and to construct a nomogram.prediction model.The predictive ability of the no-mogramprediction model was evaluated by drawing calibration curves,receiver operating characteristic(ROC)curves,and consistency index(C-index).Results Septic shock(HR=7.770,95％CI:1.852-32.593)was an independent risk factor for mortality in patients with CRAB bloodstream infections,and hospitalisation day＞14 d(HR=0.331,95％CI:0.165-0.665)was an independent protective factor for mortality in patients with CRAB bloodstream infections.A nomogram was constructed based on the above factors,with a C-index of 0.725(95％CI:0.652-0.798).The predictive efficacy of patient survival rates at 20 d and 30 d was 0.831(95％CI:0.752-0.910)and 0.826(95％CI:0.715-0.937),respectively.The calibration curve was well fit-ted.Conclusion Septic shock was an independent risk factor for mortality in patients with CRAB bloodstream infections,and hospitalisation days＞14 d was an independent protective factor for mortality in patients CRAB BSI infections.The nomogram prediction model constructed accordingly has good predictive value for the sur-vival rate of patients in the intensive care unit who undergo CRAB bloodstream infections.

Objective To explore the bloodstream infection of refractory drug-resistant Pseudomonas aeruginosa(DTR-PA)patients with clinical characteristics and risk factors.Methods Clinical data of patients with Pseudomonas aeruginosa bloodstream infection in the First Affiliated Hospital of Sun Yat-sen University from January 2017 to December 2022 were retrospectively analyzed.Multivariate Logistic regression analysis was used to explore the independent risk factors for DTR-PA bloodstream infection.Cox proportional hazards regression was used to explore the risk factors related to the death of Pseudomonas aeruginosa bloodstream in-fection.Results A total of 114 patients with Pseudomonas aeruginosa bloodstream infection were enrolled,with an average age of(51.05±17.55)years.The 28 d mortality was 14.0％(16/114),and 24 patients(21.1％,24/114)had DTR-PA.Age,ICU admission,diabetes,hypoproteinemia and septic shock were associ-ated with DTR-PA bloodstream infection(P＜0.05).Multivariate Logistic regression analysis showed that septic shock was an independent risk factor for DTR-PA bloodstream infection(P＜0.05).ICU admission was an independent risk factor for the death of patients with DTR-PA and Pseudomonas aeruginosa blood-stream infection(P＜0.05).Conclusion There are many risk factors for DTR-PA bloodstream infection.Septic shock and ICU admission are independent risk factors for DTR-PA bloodstream infection and death,respectively.

Objective:To observe the neuroanatomical properties of the projection pathway from the nucleus reuniens(Re)to the dorsal(dHC)and ventral(vHC)hippocampus.Methods:Adeno-associated virus SV40 was injected into the Re of C57BL/6 mice and the profile of downstream projection in the whole brain was examined and ranked.Retro-grade tracer Fluoro-Gold(FG)was injected into the whole hippocampus(wHC),dorsal hippocampus(dHC)or ven-tral hippocampus(vHC),and the distribution of the retrogradely labeled neurons and its relationship with calretinin(CR)in the Re were analyzed.Results:After injection of SV40 virus into the Re,we observed projection fibers or ter-minals in 18 areas across the brain.Among the areas,those to the hippocampus(HC)were mainly distributed in the dorsal and ventral lacunosum molecular layer,which ranked No.2 with regard to the projection intensity.Following injection of FG into the hippocampus,we observed that the retrogradely labeled neurons projecting to the wHC were densely distributed in the rostrocaudal segments of Re,with a more concentrated aggregation in the ventromedial part.FG+CR+double labeled neurons accounted for(47.47±0.07)％of FG labeled neurons,and for(67.13±0.10)％of CR+neurons in this case.The retrograde labeled neurons projecting to the dHC were sparse and mainly distributed in the dorsolateral part of Re.FG+CR+double labeled neurons accounted for(24.11±0.06)％of FG labeled neurons,and for(32.99±0.19)％of CR+neurons.The retrogradely labeled neurons projecting to the vHC were mainly distrib-uted in the ventralmedial part of Re.FG+CR+double labeled neurons accounted for(49.55±0.03)％of FG labeled neurons,and for(69.14±0.12)％of CR+neurons.Conclusion:Hippocampus is an essential target of the Re.The projections to the dHC and vHP differ in the location of projection neurons and the positive ratio with CR,which may re-flect the differential pathophysiological functions of the two pathways in vivo.

Objective:To explore the value of coronary artery plaque progression parameters based on coronary CT angiography (CCTA) in predicting the occurrence of major adverse cardiovascular events (MACE) in patients with non-obstructive coronary artery disease.Methods:The study included clinical, imaging, and prognosis (MACE) parameters of non-obstructive coronary artery disease patients who underwent CCTA at the First Affiliated Hospital of Nanjing Medical University from September 2010 to December 2022. Patients were grouped based on the occurrence of MACE, and differences in clinical data, plaque baseline, and progression parameters between the two groups were compared. Univariate and multivariate Cox regression analyses were employed to identify factors that could effectively predict the occurrence of MACE in patients. Models were constructed using plaque baseline parameters, plaque progression parameters, and a combination of both. The concordance index-time curve, net reclassification improvement and integrated discrimination improvement were used to evaluate the risk stratification ability of the models.Results:A total of 258 patients were included, of whom 62 cases experienced MACE during the follow-up period. In comparison to the MACE(-) group, patients in the MACE(+) group exhibited longer lesion length, greater degree of stenosis, larger plaque total volume, calcified plaque volume, non-calcified plaque volume, fibrous plaque volume, total plaque burden, lipid-rich plaque burden, higher peri-coronary adipose tissue attenuation index (FAI), and annual change of diameter stenosis(ΔDS/y). There were also more cases of coronary artery disease reporting and data system upgrades and non-obstructive progression to obstructive status ( P<0.05). Multivariate Cox analysis revealed that FAI, ΔDS/y, and non-obstructive progression to obstructive status were independent predictors of MACE occurrence. Concordance index-time curve results indicated that the combined model had a better predictive efficacy for MACE in patients with non-obstructive coronary artery disease compared to models based on plaque baseline parameters and plaque progression parameters. Conclusion:The plaque progression parameters and FAI based on CCTA have the potential to predict the high-risk population for MACE in patients with non-obstructive coronary artery disease, demonstrating good risk stratification value.

Objective:To observe the neuroanatomical properties of the projection pathway from the nucleus reuniens(Re)to the dorsal(dHC)and ventral(vHC)hippocampus.Methods:Adeno-associated virus SV40 was injected into the Re of C57BL/6 mice and the profile of downstream projection in the whole brain was examined and ranked.Retro-grade tracer Fluoro-Gold(FG)was injected into the whole hippocampus(wHC),dorsal hippocampus(dHC)or ven-tral hippocampus(vHC),and the distribution of the retrogradely labeled neurons and its relationship with calretinin(CR)in the Re were analyzed.Results:After injection of SV40 virus into the Re,we observed projection fibers or ter-minals in 18 areas across the brain.Among the areas,those to the hippocampus(HC)were mainly distributed in the dorsal and ventral lacunosum molecular layer,which ranked No.2 with regard to the projection intensity.Following injection of FG into the hippocampus,we observed that the retrogradely labeled neurons projecting to the wHC were densely distributed in the rostrocaudal segments of Re,with a more concentrated aggregation in the ventromedial part.FG+CR+double labeled neurons accounted for(47.47±0.07)％of FG labeled neurons,and for(67.13±0.10)％of CR+neurons in this case.The retrograde labeled neurons projecting to the dHC were sparse and mainly distributed in the dorsolateral part of Re.FG+CR+double labeled neurons accounted for(24.11±0.06)％of FG labeled neurons,and for(32.99±0.19)％of CR+neurons.The retrogradely labeled neurons projecting to the vHC were mainly distrib-uted in the ventralmedial part of Re.FG+CR+double labeled neurons accounted for(49.55±0.03)％of FG labeled neurons,and for(69.14±0.12)％of CR+neurons.Conclusion:Hippocampus is an essential target of the Re.The projections to the dHC and vHP differ in the location of projection neurons and the positive ratio with CR,which may re-flect the differential pathophysiological functions of the two pathways in vivo.

Abstract OBJECTIVE Oxcarbazepine(OXC) is an antiepileptic drug, which is metabolized to the active 10-monohydroxy derivative(MHD) after oral administration. The half-life period of MHD in children is significantly shorter than that in adults, and the clearance is increased by 30% to 160% compared with that in adults, which indicates that the pharmacokinetics(PK) of MHD in children is obviously different from that in adults, while adults and children exhibit different levels of expression of metabolism enzymes and transporter proteins with the same genotype. At present, there is no study describing the influence of genetic polymorphism of PK-related enzymes on MHD plasma concentrations in children with epilepsy. This study investigates whether the polymorphism of metabolic enzymes and transporter genes have significant effects on MHD plasma concentrations in children with epilepsy in China, so as to provide the reference for individualized application of OXC in pediatric patients. METHODS The plasma samples from pediatric patients with epilepsy aged 0-14 years old at the First Affiliated Hospital of Fujian Medical University who received OXC were prospective collected from June 2021 to June 2023. The MHD blood concentrations of the patients were measured using enzyme amplified immunoassay, and the metabolic enzyme genes UGT2B7 802T>C, UGT1A9 I399C>T, as well as the transporter genes ABCB1 3435C>T and ABCB2 1249G>A polymorphism were detected using dideoxy chain-termination method in epilepsy children. According to Hardy Weinberg's law of genetic balance, the theoretical values of genotype frequency of the patients were calculated, and a Chi-Square test method was used to compare whether there was a significant difference between the theoretical value and the measured value, to examine whether the genotype of the patients included in the study is accordance with the law of genetic balance. One-way ANOVA statistical method was used to analyze the correlation of the four single nucleotide polymorphisms, daily maintenance dosage of OXC, and MHD blood concentrations. Subsequently, Fisher's least significant difference(LSD) test was performed. LSD test is a pairwise comparison of the differences between the mean values of each group, calculated based on the standard error and degrees of freedom to obtain the minimum significant difference between each two groups, while P<0.05 indicated that the difference was significant. RESULTS In this study, 161 trough concentrations were collected from children with epilepsy. The genotype of the included population conformed to the genetic balance law, which indicated that the included patients were representative for the population. Unite analysis of variance showed a significant correlation between the transporter gene ABCB1 3435C>T and MHD blood drug concentration(P<0.05). Subsequently, Fisher's minimum significant difference test was conducted, and MHD plasma concentrations of patients carrying the ABCB1 3435C>T mutation allele were significantly higher than that of non-carriers. No significant association was found between the four single nucleotide polymorphisms and the daily maintenance dosage of OXC, and no significant impact of the other metabolic enzyme and transporter genetic polymorphisms on MHD plasma concentrations was found. CONCLUSION The results of research shows that the ABCB1 3435C>T polymorphism significantly affect the MHD blood concentration of pediatric patients with epilepsy, and the effects of UGT2B7 802T>C, UGT1A9 I399C>T and ABCB2 1249G>A genetic polymorphisms on MHD blood concentration and daily maintenance dosage of OXC are not found. The results suggest that MHD blood concentrations are significantly increased by affecting the expression of the encoded MDR1 transporter protein after ABCB1 3435C>T site mutation, which also may increase the risk of adverse reactions of OXC. The genetic polymorphisms of ABCB1 3435C>T can be detected in children with epilepsy when taking OXC, and the dosage can be adjusted appropriately for patients with genetic mutations. The results of this study can provide the reference for the individualized administration of OXC in clinic.

Vitamin K(VK)occurs naturally in two forms, including vitamin K 1(VK 1)and vitamin K 2(VK 2). The function of VK is related to the number of its side chains.In addition to the function of coagulation, VK is also gradually found to be involved in bone metabolism, which is closely related to the occurrence of adult osteoporosis.The rapid growth and development in childhood is a period of extremely vigorous bone metabolism.The role of VK in children′s bone metabolic diseases has been gradually recognized in recent years.However, there are still few clinical studies on the level of VK in children of different ages and how VK participates in children′s bone metabolic diseases.This article reviews the types and metabolism of VK, the mechanism of VK regulating bone metabolism, the level of VK and its role in bone metabolic diseases, so as to provide new understanding and ideas for the prevention and treatment of bone health in children.

Objective:To analyze the clinical characteristics of lung cancer associated acute ischemic stroke (LCA-AIS) and atrial fibrillation associated acute ischemic stroke (AFA-AIS).Methods:From January 1, 2014 to December 31, 2018, 46 patients diagnosed with LCA-AIS (LCA-AIS group)in Hainan Hospital of Chinese PLA General Hospital were selected, and 46 patients diagnosed with AFA-AIS (AFA-AIS group) were matched according to age and sex.The general situation, laboratory test results and imaging results of the two groups were analyzed.Results:(1) The neurological deficit symptoms in AFA-AIS group were more serious than those in LCA-AIS group; there was significant difference in National Institutes of Health Stroke Scale (NIHSS) score and the Modified Rankin Scale (mRs) score between the two groups ( P=0.001, P=0.003). (2)The D-D polymer concentration in LCA-AIS group was significantly higher than that in AFA-AIS group ( P<0.001), but the hemoglobin, erythrocyte count and hematocrit were significantly lower than those in AFA-AIS group (all P<0.001). (3)There was no significant difference in imaging classification and the number of infarct basins between LCA-AIS group and AFA-AIS group ( P>0.05). LCA-AIS patients was more likely to have poly-period acute ischemic lesions ( P=0.015), while AFA-AIS had significantly larger infarct diameter and more likely to be complicated with acute hemorrhagic stroke or bleeding ( P<0.001). Conclusions:The clinical characteristics of LCA-AIS and AFA-AIS are similar, so it is necessary to distinguish LCA-AIS from AFA-AIS in combination with neurological impairment, laboratory tests and imaging findings to avoid misdiagnosis.

OBJECTIVETo discover the techniques for ex vivo generation and cryopreservation of erythroid progenitor cells (EPCs)derived from umbilical cord blood (UCB)mononuclear cells (MNCs).

METHODSUCB was chosen as the source of EPCs. Erythrocytes were precipitated by hydroxyethyl starch (HES). MNCs were separated by Ficoll density gradient centrifugation. Erythroid progenitor cell were generated from MNC ex vivo in suspension culture supplemented with stem cell growth factor, insulin growth factor, erythropoietin, Fms- liketyrosinekinase ligand, transferrin and dexamethasone. Cell maturation was evaluated by morphologic analysis and CD71/CD235a expression profiling. In vitro induced cells were cryopreserved using different cryopreservation media. The cell survival rate, phenotype and proliferation curves were detected after cell thawing.

RESULTSWith the extension of culture time, the total number of cells increased significantly accompanied with the elevation of CD71 and CD235 positive populations. After 14- day inducing, the cells reached to approximately 110 times of the starting number with the cell viability as (88.92±0.95)%. The percentages of cell surface markers were (86.77±9.11)% for CD71 and (64.47±16.67)% for CD71/CD235, respectively. With the extension of inducing time, wright- Giemsa staining showed that the middle erythroblasts appeared mostly at day 10, and the late erythroblasts were seen at day 14. The red pellets were present at day 14, which indicated the more production of hemoglobin. Colony forming assay showed that erythroid colonies at induction day 7 were higher than that for non-induced cells (326.00±97.96vs 61.60±20.03 per 2 000 cells). With the extension of culture time, the number of erythroid colonies decreased. Induced EPCs were preserved with different cryopreservation solutions, in which 10% DMSO were better than 5% DMSO. Additionally, 10% DMSO + 2% HSA showed no different with 10% DMSO + 5% HSA. Combined 50% plasma with 2% HSA was more effective.

CONCLUSIONSThis non- serum culture media could effectively induced and expanded EPCs, and 10% DMSO + 2% HSA + 50% plasma appeared to be a desirable cryopreservation solution for EPCs from UCB.

Cell Culture Techniques ; Cell Differentiation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Erythroblasts ; cytology ; Erythroid Precursor Cells ; cytology ; Fetal Blood ; cytology ; Humans ; Leukocytes, Mononuclear ; cytology ; Umbilical Cord