Volume 46, Issue 1

Urinary tract infections affect 150 million people worldwide, yet the diagnosis of this common infection is not straightforward. Misdiagnoses and incorrect prescriptions are frequent in the treatment of urinary tract infections; this also contributes to the increase in antibiotic resistance among pathogens. Present diagnostic practices take 2–3 days for pathogen identification and antibiotic susceptibility testing. New technologies are urgently needed for improved patient care as well as to promote antibiotic stewardship. An ideal new diagnostic technology will test clinical urine samples directly and identify the pathogen and determine its antibiotic susceptibilities within a few hours such that the patient can be prescribed the appropriate antibiotic treatment the same day. Screening tools, such as flow cytometers and new dipstick assays, can help with rapidly identifying negative samples and improving workflow and reducing costs. Several groups have made progress in optimizing mass spectrometry methods for direct urine processing, and there are also new multiplex PCR panels that are specific for UTI pathogens and antibiotic resistance. We also discuss several emerging technologies – microfluidics, biosensors, real-time microscopy systems, and sequence-based diagnostics – that show huge potential in delivering rapid results.

Objectives Liver transplantation (LT) can benefit the long-term survival of hepatocellular carcinoma (HCC) patients. We hypothesized that circulating tumor cell (CTC) levels and subtypes are intimately associated with metastasis status of HCC patients. This study was designed to test that compositive hematological indices including CTC can provide a prediction of post-LT metastasis. Methods Between 2017 and 2018, 37 HCC patients within Hangzhou criteria receiving LT were included for analysis. The 24-month follow-up was mainly conducted by outpatient and telephone. Blood samples were collected, and hematological indices were examined. The outcomes such as PFS, recurrence, metastasis, location of recurrence/metastasis, and number of metastases were recorded. Results The follow-up analysis showed that microvascular invasion (MVI) classification at the baseline is associated with metastasis. Next, α-fetoprotein (AFP) level was another useful indicator of postoperative metastasis, especially at the third or fourth month; the protein induced by vitamin K absence or antagonist-II (PIVKA-II) level three months after LT was significantly higher for those who had later metastasis. The mesenchymal CTC level at the 45th day was increased for in the metastasis group. Using two-ends Logistic regression, the calculated value MP (metastasis predictor, by above factors). Had an AUC of 0.858 in the ROC curve, with a cutoff value of 0.328. Conclusions In conclusion, microvascular invasion, AFP level at the third or fourth month, PIVKA-II level at the third month, and mesenchymal CTC level at day 45 were associated with post-LT metastasis. Using Logistic regression based on above variables, the two-year metastasis can be predicted with satisfactory sensitivity and accuracy.

Objectives This study aimed to evaluate the relationship between C-reactive protein/albumin (CRP/Alb), neutrophil/lymphocyte (NLR), platelet/lymphocyte (PLR) ratios and the Acute Physiology And Chronic Health Evaluation II (APACHE II) score and 28-day mortality among 400 patients admitted to internal medicine and anesthesia reanimation intensive care unit (ICU). Methods This prospective study included a total of 400 patients who were admitted to hospital internal medicine and anesthesia reanimation ICUs. Results The most common reasons for ICU admission were pneumonia (29.3%), gastrointestinal bleeding (10.3%), acute exacerbation of chronic kidney disease (CKD) (10.3%), and acute kidney injury (7.5%). The comparison of the laboratory findings with survival outcomes revealed that among the patients with acute exacerbation of CKD, the median NLR (p=0.043) and median CRP/Alb (p=0.021) were significantly higher in patients who died. For all of the patients, the APACHE II score was positively correlated with CRP (p<0.001) and CRP/Alb (p<0.001), negatively correlated with Alb (p<0.001), positively correlated with the NLR (p<0.001), and positively correlated with the PLR. Conclusions The APACHE II score was significantly correlated with the CRP/Alb ratio, NLR, and PLR. The NLR and CRP/Alb ratio were statistically associated with mortality in patients hospitalized for acute exacerbation of CKD.

Objectives The present study aimed to survey the prevalence of gestational diabetes mellitus (GDM) in Southern China and further to analyze the correlation between the prevalence of GDM and maternal age. Methods A retrospective cross-sectional study was carried out at the Southern Medical University Affiliated Maternal & Child Health Hospital of Foshan, Foshan, China between January and April 2020. Oral glucose tolerance tests (OGTT) was performed, using a 75 g glucose load and venous samples were drawn at 0 h, 1 h and 2 h at 24–28 weeks of gestation. GDM was diagnosed by International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria. Results The prevalence of GDM was 14.87% by IADPSG criteria. The incidence of GDM outcome increased and plasma glucose levels remained high among the age subgroups (<20, 20–24, 25–29, 30–34, 35–39, ≥40 years old) in pregnant women. Moreover, the levels of plasma glucose levels after OGTT kept rising among the pregnant women with non-gestational diabetes mellitus (non-GDM). Furthermore, pregnant women were inclined to have abnormal plasma glucose values at 1 h and 2 h than initial fasting plasma glucose (FPG) after OGTT as the age increased. Conclusions Our findings demonstrate that the incidence of GDM outcome and plasma glucose levels increase as the maternal age increase.

Objectives An anal fistula is an external abnormal anatomical connection between the rectum and the outer skin of the anus. Symptoms include anorectal pain, abscesses, perianal cellulitis, smelly or bloody drainage of pus, and, in some cases, difficulty controlling bowel movements. Diagnosis and evaluation of anal fistulas is crucial for prognosis and for the choice of the treatment method. In this study, we aimed to discover potential biomarkers from serum proteins for the prediction of anal fistulas. Methods Using antibody array technology, the expression of 40 proteins was simultaneously detected in serum samples from 13 patients with anal fistulas with chronic diarrhea, 14 patients with chronic diarrhea and six healthy volunteers. Differentially expressed proteins were subsequently validated by ELISA, with a sample population expanded to 30 patients with anal fistulas and chronic diarrhea, 30 patients with chronic diarrheas only and 20 healthy controls. Results Quantification analysis identified MIP-1α, MIP-1β and TNF-R1 with significant differential expression between the anal fistula with chronic diarrhea, chronic diarrhea only and healthy control groups. Bioinformatics analyses, including PCA and heat map creation, showed a clear separation between the three groups using the expression of MIP-1α, MIP-1β and TNF-R1. Validation by ELISA with the expanded sample population fistulas showed significant differential expression levels of MIP-1α, MIP-1β and TNF-R1, displaying accuracy rates of 0.898, 0.987 and 1.0 between the anal fistula with chronic diarrhea and healthy control groups. Accuracy rates between the anal fistula with chronic diarrhea and the chronic diarrhea only groups were 0.9768, 0.909 and 0.964, respectively. Conclusions These results suggest the feasibility of employing serum proteins MIP-1α, MIP-1β and TNF-R1 as potential biomarkers for rapid and convenient diagnosis of anal fistula in chronic diarrhea patients.

Objectives This study aimed to examine the antioxidant activity and antiinflammatory effects of ethanol extract of Polygonum senticosum (EPS) on lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods Antioxidant activity of EPS was assessed by radical-scavenging effects on ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals. Proinflammatory markers produced by LPS-induced RAW 264.7 macrophages were quantified to assess the antiinflammatory activity of EPS. Results Our results showed that EPS significantly increased FRAP and DPPH radical-scavenging activity. Additionally, EPS reduced LPS-induced proinflammatory mediators, such as nitric oxide (NO) and prostaglandin E 2 (PGE 2 ), along with proinflammatory cytokines, including tumor necrosis factor (TNF)-α and interleukin (IL)-1β, without significant cytotoxicity. EPS significantly downregulated the expression of inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α, and IL-1β in LPS-stimulated RAW 264.7 macrophages. Positive correlations were noted between FRAP and DPPH radical-scavenging activity and antiinflammatory capacity. Conclusions Our results indicate that EPS downregulates the expression of pro-inflammatory mediators such as NO, PGE 2 , and cytokines (IL-1β and TNF-α) in LPS-stimulated RAW 264.7 macrophage cells. Further research is needed for its use as a treatment for inflammation and related diseases.

Objectives Unidentified SARS-CoV-2 infections among hospital staff can become a major burden for healthcare systems worldwide. We hypothesized that the number of previous SARS-CoV-2 infections among hospital employees is substantially higher than known on the basis of direct testing strategies. A serological study was thus performed among staff of Marburg University Hospital, Germany, in May and June 2020. Methods Anti-SARS-CoV-2 antibody titers were measured by spike protein (S1)-specific IgG ELISA (Euroimmun) and by nucleoprotein-(NCP) specific total antibody CLIA (Roche). Selected sera were analyzed by SARS-CoV-2 neutralization test. Participants provided questionnaires regarding occupational, medical, and clinical items. Data for 3,623 individuals (74.7% of all employees) were collected. Results Individuals reactive to both S1 and NCP were defined as seropositive; all of those were confirmed by neutralization test (n=13). Eighty-nine samples were reactive in only one assay, and 3,521 were seronegative. The seroprevalence among hospital employees at Marburg University Hospital was 0.36% (13/3,623). Only five of the 13 seropositive employees had reported a positive SARS-CoV-2 RT-PCR test result. Conclusions Usage of a single S1-specific assay highly overestimated seroprevalence. The data provided no evidence for an increased risk for a SARS-CoV-2 infection for staff involved in patient care compared to staff not involved in patient care.

Objectives Random mutations and recombinations are the main sources for the genetic diversity in SARS-CoV-2, with mutations in the SARS-CoV-2 spike (S) receptor binding motif (RBM) representing a high potential for the emergence of new putative variants under investigation (VUI) or variants of concern (VOC). It is of importance, to measure the different circulating SARS-CoV-2 lineages in order to establish a regional SARS-CoV-2 surveillance program. We established whole genome sequencing (WGS) of circulating SARS-CoV-2 lineages in order to establish a regional SARS-CoV-2 surveillance program. Methods We established a surveillance program for circulating SARS-CoV-2 lineages by performing whole genome sequencing (WGS) in SARS-CoV-2 isolates. Specimens were collected over a period of 5 months from three different sites. Specimens were collected from both patients suffering from COVID-19 and from outpatients without any clinical signs or symptoms; both in a tertiary university hospital, and two private laboratories within an urban area of eastern part Germany. Results Viral WGS from the 364 respiratory specimens with positive SARS-CoV-2 RT-PCR comprised 16 different SARS-CoV-2 lineages. The majority of the obtained sequences (252/364=69%) was assigned to the variant of concern (VOC) Alpha (B.1.1.7). This variant first appeared in February in our samples and quickly became the dominant virus variant. All SNP PCR results could be verified using WGS. Other VOCs found in our cohort were Beta (B.1.351, n=2) and Delta (B.1.617.2, n=1). Conclusions Lineage analysis revealed 16 different virus variants among 364 respiratory samples analyzed by WGS. The number of distinct lineages dramatically decreased over time in leaving only few variants, in particular, the VOC Alpha or B.1.1.7. By closer inspection of point mutations, we found several distinct mutations of the viral spike protein that were reported to increase affinity or enable immune escape. Within a study period of only 5 months, SARS-CoV-2 lineage B.1.1.7 became the dominant lineage in our study population. This study emphasizes the benefit of SARS-CoV-2 testing by WGS. The increasing use of WGS to sequence the entire SARS-CoV-2 genome will reveal additional VUIs and VOCs with the potential to evade the immune system and, thus, will be a promising tool for data mining of relevant information for epidemiological studies. SARS-CoV-2 lineage monitoring using WGS is an important surveillance tool for early detection of upcoming new lineages of concern.