Deep vein thrombosis(DVT)is a common peripheral vascular disease in clinical practice.The lack of precise and efficient early diagnostic techniques renders it susceptible to being overlooked or misdiagnosed,and therefore,identifying trustworthy biomarkers is a major issue that has to be resolved.In this study,the endogenous metabolites in the urine of DVT rats were screened by metabolomics technology based on gas chromatograph-mass spectrometry(GC-MS)and the characteristic metabolites were identified by multiple feature selection algorithms and multivariate statistical analysis,for the development of a machine learning-based diagnostic model for DVT.The urine samples in metabolic cage in the thrombus development phase(between 48 and 72 h)of rats were collected,which was used as the models for inferior vena cava ligation.The metabolic profiles of the control group and DVT were obtained using the GC-MS method.A total of 176 kinds of endogenous metabolites were identified in rat urine through comparison with the FiehnLib database,26 kinds of differential metabolites associated with DVT were screened through a combination of the Mann-Whitney U test and orthogonal partial least squares discriminant analysis(OPLS-DA),and 13 kinds of significant metabolites strongly correlated with DVT were further evaluated in conjunction with various machine learning feature selection techniques.For DVT diagnosis,machine learning models such as Gaussian Naive Bayes(GNB),support vector machine(SVM),logistic regression(LR),and linear discriminant analysis(LDA)were developed.The diagnostic model constructed using 13 kinds of key metabolites demonstrated excellent accuracy and stability,and surpassed the predictive performance of the models utilizing 176 kinds of metabolites and 26 kinds of differential metabolites,as evidenced by examination and comparison of each model's efficacy.The study showed that the integration of multiple feature selection algorithms for analyzing metabolite information in DVT rat urine was capable of effectively identifying reliable potential markers of DVT.Furthermore,the developed machine learning model offered a novel technical approach for the automated diagnosis of DVT.

Objective To investigate the distribution and antimicrobial susceptibility of clinically isolated bacteria from intensive care units(ICUs)in hospitals of Hunan Province Antimicrobial Resistance Surveillance System from 2012 to 2021.Methods According to China Antimicrobial Resistance Surveillance System,data of clinically isolated bacterial strains and antimicrobial susceptibility testing results of bacteria from ICUs reported by all member units of Hunan Province Antimicrobial Resistance Surveillance System were analyzed with WHONET 2022 software.Results From 2012 to 2021,the total number of bacteria isolated from ICUs of member units of the Hunan Province Antimi-crobial Resistance Surveillance System was 5 777-22 369,with Gram-negative bacteria accounting for 76.1％-78.0％annually.Staphylococcus aureus ranked first among isolated Gram-positive bacteria each year.The top 5 bacteria among Gram-negative bacteria were Acinetobacter baumannii,Klebsiella pneumoniae,Escherichia coli,Pseudo-monas aeruginosa,and Stenotrophomonas maltophilia.Detection rate of methicillin-resistant Staphylococcus aureus showed a downward trend year by year.No Staphylococcus spp.were found to be resistant to vancomycin,teico-planin and linezolid.Detection rates of vancomycin-resistant Enterococcus faecalis and vancomycin-resistant Entero-coccus faecium were 0.6-1.1％and 0.6％-2.2％,respectively.Resistance rates of Escherichia coli and Kleb-siella pneumoniae to imipenem were 3.1％-5.7％and 7.7％-20.9％,respectively.Resistance rates of Pseudo-monasaeruginosa and Acinetobacter baumannii to imipenem were 24.6％-40.1％and 76.1％-80.9％,respective-ly.Detection rates of carbapenem-resistant Pseudomonas aeruginosa declined year by year.Acinetobacter baumannii maintained high susceptibility to polymyxin B,with resistance rate＜10％.Conclusion Antimicrobial resistance of bacteria from ICUs is serious.Carbapenem-resistant Enterobacteriales has an upward trend after 2019.It is nece-ssary to strengthen the surveillance of bacterial resistance and carry out multidisciplinary collaboration.

OBJECTIVE: To observe the impact of energy saving light,incandescent light and circadian light on the ethology of depressive rats and explore its possible mechanism on affecting the secretion of melatonin. METHODS: Thirty rats aged 6weeks were randomly selected from 40 specific pathogen free health female SD rats after they adapted to the living environment,depressive rat models were established in the rats by bilateral ovariectomy combined with isolated living and chronic unpredictable mild stress stimulation at the age of 11-14 weeks. Then these 30 ovariectomized rats were randomly divided into 3 intervention groups,including an energy saving light group,an incandescent light group and a circadian light group,with 10 rats in each group. The rats in these 3 groups were given specific experimental light intervention for 3 weeks respectively at the age of 17 weeks. The other 10 rats were raised in conventional environment as the control group. Their body weights were measured at the age of 17,19,20 and 21 weeks. The ethology tests were carried out by sucrose preference test and the open-field test at the age of 7,14 and 20 weeks respectively. The melatonin levels in peripheral blood of 7 time points from 19: 30 to 8: 30 were measured in the rats at age of 21 weeks. One rat in each group at every time point was randomly selected for examination. RESULTS: At the age of 17 weeks before light-intervention,the body weights of rats in 4 groups showed no significant difference( P > 0. 05). After light-intervention,at the age of 17-20 weeks,the body weights of rats in 3 intervention groups were gradually increased with the increase of age( P < 0. 05).There was no significant difference between body weights of rats at the age of 21 weeks and those at the age of 20 weeks in each group( P > 0. 05). At age of 7 weeks,no significant differences were found in sucrose consumption and standing scores among these 4 groups( P > 0. 05). After the depressive models were established,at the age of 14 weeks before light-intervention,in rats of these 3 intervention groups,the sucrose consumption and standing scores were lower than those of the control group( P < 0. 05),and there was no significant difference found in the above 2 indexes among these 3intervention groups( P > 0. 05). At the age of 20 weeks after light-intervention,the sucrose consumption and standing scores were not significantly different from each other among the 4 groups( P > 0. 05). The peak levels of melatonin in the peripheral blood of rats in these 3 intervention groups were higher than that in the control group. The peak levels onsets of melatonin in peripheral blood of rats in the circadian light group and the energy saving light group were earlier or 2 hours delayed compared to that of control group,while it was similar between the incandescent light group and control group.CONCLUSION: The circadian light,the energy saving light and the incandescent light are similarly effective in improving the behaviors of depressive rats. The circadian light can delay the onset of peak level of melatonin in peripheral blood.

Objective To analyze the characteristics of clinical manifestations, risk factors, therapies and acute outcomes in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage. Methods Seventy-five patients with cerebral venous sinus thrombosis were included in the study. According to the radiological findings on the brain image, patients were divided into two subgroups:cerebral hemorrhage group and non-hemorrhage group. The demo?graphic data, potential risk factors, clinical manifestations, radiological features, therapeutic strategies and acute out?comes were compared between two subgroups, and high risk factors were also analyzed. Results There were seventy-five patients with cerebral venous sinus thrombosis in the present study. Twenty-eight patients of them (37.2%) had cerebral hemorrhage whereas the remaining forty-seven patients (62.7%) did not have cerebral hemorrhage. Pregnancy/puerperi?um were significantly higher in patients with cerebral hemorrhage (with vs without;28.6%vs. 6.4%, P=0.015), while in?fection was markedly higher in patients without cerebral hemorrhage (with vs without;7.1% vs. 29.8%, P=0.021). Head?ache (92.9% vs. 70.2%, P=0.021), unconsciousness (25.0% vs. 6.4%,P=0.034), seizures (53.6% vs. 19.1%, P=0.002) and motor deficits (35.7% vs. 12.8%, P=0.019) were more common in patients with cerebral hemorrhage. Moreover, mul?tiple sinus involvement (1.4% vs. 44.7%, P=0.024) was significantly higher and the acute outcomes(mRS≥3: 46.4%vs.17.0%, P=0.006)were poorer in patients with cerebral hemorrhage. Binary Logistic analysis showed that pregnancy/pu?erperium (P=0.004) and multiple sinus involvement were positively, whereas infection was negatively correlated with cere?bral venous sinus thrombosis and hemorrhage ( P=0.007;P=0.03). Conclusions Pregnancy/puerperium, headache, uncon?sciousness, seizures, motor deficits and multiple sinus involvement are more frequently in patients with cerebral venous sinus thrombosis and hemorrhage, and the acute outcomes are poorer in patients with cerebral venous sinus thrombosis complicated by cerebral hemorrhage.

OBJECTIVETo gain insights into the role of forkhead box protein 3 (Foxp3) in the pathogenesis of hepatocellular carcinoma (HCC) by performing a comparative analysis of Foxp3 mRNA expression and promoter methylation status in HCC and normal liver tissues.

METHODSThirty-nine HCC and 13 normal liver tissue specimens were evaluated by real-time quantitative PCR and pyrosequencing to measure the expression of Foxp3 mRNA and determine the methylation status of its promoter, respectively. Statistical analyses of the data were conducted by rank-sum test and Spearman's rank correlation coefficient test.

RESULTSThe HCC specimens showed significantly higher mRNA expression of Foxp3 (vs. normal liver tissues, Z =-2.770, P =0.0056). Moreover, the HCC specimens showed significant hypomethylation of the Foxp3 promoter site A (vs. normal liver tissues, Z =2.118, P =0.0339), and the Foxp3 mRNA level was negatively correlated with the methylation of site A (rs =-0.344, P =0.046). None of the other four sites in the Foxp3 promoter showed a significant difference in methylation, and the overall methylation was not significantly different between the HCC and normal liver tissues.

CONCLUSIONOverexpression and low methylation of Foxp3 may be involved in the oncogenic and progression processes of HCC.

Carcinoma, Hepatocellular ; metabolism ; pathology ; CpG Islands ; DNA Methylation ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Promoter Regions, Genetic ; RNA, Messenger ; genetics

BACKGROUNDBacteremia remains a significant cause of morbidity and mortality after kidney transplantation. This study was conducted to investigate whether the polymorphisms of tumor necrosis factor (TNF)-β, interleukin (IL)-1β, and IL-1 receptor antagonist (IL-1ra) gene predicted the susceptibility to bacteremia within the first 6 months after kidney transplantation.

METHODSSubjects comprised 82 infected kidney transplant recipients and 60 non-infected kidney transplant recipients. Bacteremia was diagnosed in 16 of the 82 infected recipients. Genomic DNA from these 142 kidney transplant recipients was extracted from peripheral blood leukocytes. Regions containing the NcoI polymorphic site at position +252 of TNF-β gene and the AvaI polymorphic site at position -511 of IL-1β gene were amplified by polymerase chain reaction (PCR) and subsequently digested with NcoI and AvaI restriction enzymes, respectively. The polymorphic regions within intron 2 of IL-1ra gene containing variable numbers of a tandem repeat (VNTR) of 86 base pairs were amplified by PCR.

RESULTSGenotypic and allelic frequencies were similar between infected recipients and non-infected ones. Individual locus analysis showed that recipient TNF-β and IL-1ra gene polymorphisms were not associated with the presence of bacteremia (P = 0.684 and P = 0.567, respectively). However, genotype analysis revealed that recipient IL-1β-511CC genotype was strongly associated with susceptibility to develop bacteremia (P = 0.003). Recipient IL-1β-511CC genotype (odds ratio 5.242, 95% confidence intervals 1.645-16.706, P = 0.005) independently predicted the risk for bacteremia within the first 6 months after kidney transplantation.

CONCLUSIONSThese findings indicate a critical role of IL-1β gene polymorphisms in susceptibility to bacteremia after kidney transplantation, which may be useful to screen for patients at higher risk for post-transplant bacteremias. Thus, the identified individuals can benefit from preventive treatment and a less potent immunosuppressive regimen.

Adolescent ; Adult ; Bacteremia ; genetics ; Female ; Genotype ; Humans ; Interleukin 1 Receptor Antagonist Protein ; genetics ; Interleukin-1 ; genetics ; Kidney Transplantation ; Lymphotoxin-alpha ; genetics ; Male ; Middle Aged ; Multigene Family ; genetics ; Polymorphism, Genetic ; genetics ; Young Adult

OBJECTIVESTo investigate the clinical features of femoral neck fractures and analyze related causes.

METHODSThe clinical data of patients with femoral neck fractures from June 2002 to August 2009 were retrospectively analyzed. The gender, age, fracture side, fracture type, basic social data, activities before injury, injury causes and treatment were analyzed.

RESULTSA total of 219 patients (106 male and 113 female) was analyzed. All patients were divided into children group (age < 16 years), adult group (age ranged from 16 to 60 years) and older group (> 60 years). There were 5 patients (2.3%) in the children group, 81 patients (37.0%) in the adult group and 133 patients (60.7%) in the older group. There were 11 patients (5.0%) with Garden I fractures, 32 patients (14.6%) with Garden II fractures, 90 patients (41.1%) with Garden III fractures and 86 patients (39.3%) with Garden IV fractures. Fall damage and traffic injury were the main injury types. Home and public place were the main injury sites.

CONCLUSIONSThe incidence of femoral neck fracture shows the highest in the old persons. The male patients with femoral neck fractures are more than female patients in children and adult group, while the male patients with femoral neck fractures are less than female patients in older group. The dominant fractures type according to Garden classification is Garden III fractures in children and adult groups, but Garden IV fractures in older group. Fall damage and traffic injury are the main injury types. Home and public place are the main injury sites.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; Female ; Femoral Neck Fractures ; diagnosis ; diagnostic imaging ; epidemiology ; Humans ; Male ; Middle Aged ; Radiography ; Retrospective Studies ; Young Adult

OBJECTIVETo analyze the characteristic of bacterial infections, and the relationship between antibiotics treatment and bacterial infections after liver transplantation, and to prevent antibiotic-resistant bacterial infections.

METHODS86 liver transplant recipients were retrospected. Different indexes including limited daily dose, the frequency of medication, drug use index were used to evaluate the rationality of the use of antibiotics, three-dimensional test was used to explore extended-spectrum beta-lactamase and AmpC enzyme of Gram-negative bacteria.

RESULTSThe major pathogens of infection after liver transplantation were Enterococcus faecalis, Enterobacter cloacae, fungi and E. coli. Pre-operative antibiotic utilization rate was 83.7%, it was mainly a single use of antibiotics; After- operative antibiotic usage was 100.0%, it was mainly joint use of two or three antibiotics; The top 3 antibiotics used were cephalosporins, the combined enzyme inhibitors and penicillin. Antibiotics with drug utilization index (DUI) more than 1.1 included ampicillin and Lalin proxy. 43.3% and 31.8% of Gram -Negative bacteria produced ESBLs and AmpC, respectively, while 21.3% Gram -Negative bacteria produced two enzymes.

CONCLUSIONThere is high incidence of bacterial infections after liver transplantation. The use of antibiotics is high dose, high-frequency and reasonable; High resistance of bacterial infections was prone to develop and the prevention of the high resistance of bacterial infections is very important.

Adolescent ; Adult ; Aged ; Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Bacterial Infections ; drug therapy ; etiology ; microbiology ; Child ; Child, Preschool ; Drug Resistance, Bacterial ; Female ; Gram-Negative Bacteria ; drug effects ; enzymology ; isolation & purification ; Gram-Positive Bacteria ; drug effects ; enzymology ; isolation & purification ; Humans ; Infant ; Liver Transplantation ; adverse effects ; methods ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Postoperative Complications ; drug therapy ; epidemiology ; microbiology ; Retrospective Studies ; Young Adult ; beta-Lactamases ; biosynthesis

Recombineering, a new genetic engineering technology based on high efficiency in vivo homologous recombination, can be used in target DNA knock-in, knock-out and gene cloning. In the process of gene subcloning mediated by Recombineering technique, high-quality target DNA fragments were difficult to obtain using in vitro overlapping PCR,therefore the efficiency of in vivo homologous recombination was severely interrupted. To solve this problem, some technology improvements have been established based on the principle of Red recombinases. The PCR DNA fragments of egfp and kan genes with complementary sequences on the end of each fragment were co-introduced into a pcDNA3.1 vector and Red recombinases containing E. coli DY331 host cells by electroporation. A recombinant plasmid pcDNA3.1-egfp-kan was screened directly by antibiotic marker. The positive rates can reach to 45%. The EGFP gene expression of pcDNA3.1-egfp-kan can be observed by transient transfection of 293 eukaryotic cells.
Bacteriophage lambda ; enzymology ; genetics ; DNA, Recombinant ; genetics ; Electroporation ; Escherichia coli ; genetics ; Gene Fusion ; genetics ; Genetic Engineering ; methods ; Green Fluorescent Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Recombinases ; genetics ; metabolism ; Recombination, Genetic

OBJECTIVE: To examine the infection and bacteria resistance of Helicobacter pylori (H.pylori) to clarithromycin and furazolidone,to determine whether the antibiotic resistance is primary or secondary, and to decide if a new H.pylori infection plays a role in eradication failures. METHODS: Twenty one H.pylori had been isolated from human biopsy specimens, and antimicrobial susceptibility testing was performed. DNA fingerprints were generated using random amplification polymorphic DNA (RAPD) to determine the identity of H.pylori before and after the eradication therapy. RESULTS: Eight bacteria resisted against clarithromycin, and one against furazolidone, with the resistant rates 38.1% and 4.8% respectively. The number of primary antibiotic resistance, secondary resistance and new infection was 1 for each. CONCLUSION Resistance to clarithromycin is more common compared with that to furazolidone. Development of primary and secondary resistance to clarithromycin occurs as a rule in eradication failures. New H.pylori infection plays a role in eradication failures.
Anti-Bacterial Agents ; pharmacology ; therapeutic use ; Clarithromycin ; pharmacology ; therapeutic use ; DNA Fingerprinting ; DNA, Bacterial ; analysis ; genetics ; isolation & purification ; Drug Resistance, Bacterial ; Furazolidone ; pharmacology ; therapeutic use ; Helicobacter Infections ; drug therapy ; microbiology ; Helicobacter pylori ; drug effects ; genetics ; Humans ; Microbial Sensitivity Tests ; Random Amplified Polymorphic DNA Technique