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Aims Low caspofungin exposure is frequently encountered in patients with invasive candidiasis caused by Candida albicans. This study aimed to investigate the effects of caspofungin on C. albicans at sub-inhibitory concentrations. Methods and Results First, a comparative transcriptomics analysis was performed on C. albicans receiving caspofungin at sub-minimum inhibitory concentrations (sub-MICs). The results showed that caspofungin significantly changed the mRNA expression profile in DAY185, with DE-mRNAs enriched in the functions of cell wall biosynthesis, metabolism, etc. Subsequently, cellular fitness, cell aggregation, energy metabolism activity and the proportion of persister cells of C. albicans were quantitatively and/or qualitatively assessed after sub-MIC caspofungin exposure. No significant changes in cell fitness and aggregation formation were observed during treatment of C. albicans with sub-MIC caspofungin. In C. albicans aggregation treated with sub-MIC caspofungin, we observed a decrease in respiratory metabolism and an increase in persister cells; this effect was more pronounced in als1ΔΔ than in DAY185. Conclusions Pre-exposure to sub-MIC caspofungin suppresses C. albicans respiratory metabolism and promotes persister cell development. Significance and Impact of the Study Caspofungin should be used with caution in patients with C. albicans infections, as anti-infection therapy may fail due to persister cells.

Objective To study the effect of borneol combined with astragaloside IV and Panax notoginseng saponins (BAP) on promoting neurogenesis by regulating microglia polarization after cerebral ischaemia–reperfusion(CI/R) in rats. Methods A focal CI/R injury model was established. Evaluated the effects of BAP on ischaemic brain injury, on promoting neurogenesis, on inhibiting Inflammatory microenvironment and TLR4/MyD88/NFκB signalling pathway. A microglia oxygen-glucose deprivation reoxygenation (OGD/R) model was established that evaluated the effects of BAP on regulating the polarization of microglia and inflammatory microenvironment. Results BAP can inhibit the expression of TLR4, MyD88 and NFκB proteins, reduce IL-1β and increase IL-10, reduce M1 type microglia and increase M2 microglia. The proliferation of neural stem cells increased, synaptic gap decreased, synaptic interface curvature increased, expression of SYN and PSD95 proteins increased, which improved the neurological dysfunction and reduced the volume of cerebellar infarction and nerve cell injury. Conclusion BAP can reduce CI/R injury and promote neurogenesis, the effect is related to inhibition of the activation of TLR4/MyD88/NFκB, regulating the polarization of microglia from M1 type to M2 type and inhibition of inflammatory response.

Aimed at eddy current detection of defects in thin-walled stainless steel seamless pipes, an effective detection method for identifying defect types is proposed. First, the empirical mode decomposition (EMD) method is used to process the collected eddy current signals and obtain the principal intrinsic mode function (IMF) components of different defects. The Hilbert-Huang transform (HHT) is used to extract the frequency-domain features of the principal IMF components, which are combined with the time-domain features to form an effective defect feature vector. Then, principal component analysis (PCA) is used to reduce the dimensions of the defect feature vector and the redundant information is removed to obtain the principal component vector of the defect. Finally, two radial basis function (RBF) neural networks are used to identify and classify the defect types and three error evaluation indicators are selected to evaluate the performance of the classification network models.

Background Alterations in genes encoding chromatin regulatory proteins are prevalent in cancers and may confer oncogenic properties and molecular changes linked to therapy resistance. However, the impact of copy number alterations (CNAs) of the SWItch/Sucrose NonFermentable (SWI/SNF) complex on the oncogenic and immunologic properties has not been systematically explored across human cancer types. Methods We comprehensively analyzed the genomic, transcriptomic and clinical data of The Cancer Genome Atlas (TCGA) dataset across 33 solid cancers. Results CNAs of the SWI/SNF components were identified in more than 25% of all queried cancers, and tumors harboring SWI/SNF CNAs demonstrated a worse overall survival (OS) than others in several cancer types. Mechanistically, the SCNA events in the SWI/SNF complex are correlated with dysregulated genomic features and oncogenic pathways, including the cell cycle, DNA damage and repair. Notably, the SWI/SNF CNAs were associated with homologous recombination deficiency (HRD) and improved clinical outcomes of platinum-treated ovarian cancer. Furthermore, we observed distinct immune infiltrating patterns and immunophenotypes associated with SWI/SNF CNAs in different cancer types. Conclusion The CNA events of the SWI/SNF components are a key process linked to oncogenesis, immune infiltration and therapeutic responsiveness across human cancers.

Objective Currently, coronavirus disease 2019 (COVID-19) has spread worldwide and become a global health concern. Here, we report a familial cluster of six patients infected with severe acute respiratory coronavirus 2 (SARS-CoV-2) in a northern Chinese region and share our local experience with regard the control of COVID-19. Methods The demographic data, clinical features, laboratory examinations, and epidemiological characteristics of enrolled cases were collected and analyzed. Two family members (Cases 1 and 2) had Hubei exposure history and were admitted to the hospital with a confirmed diagnosis of COVID-19; eight familial members who had contact with them during the incubation period underwent quarantine in a hospital. We closely followed up all the family members and analyzed their clinical outcome. Results Case 3 had negative SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) results but was suspected to have COVID-19 because of radiographic abnormalities. Cases 4 and 5 developed symptomatic COVID-19. Case 6 was considered an asymptomatic carrier as his SARS-CoV-2 RT-PCR result was positive. The other four family members with close contacts to COVID-19 patients had no evidence of SARS-CoV-2 infection. Conclusions Our findings suggest that COVID-19 has infectivity during the incubation period and preventive quarantine is effective for controlling an outbreak of COVID-19 infection.

Airway epithelial cells, the first barrier of the respiratory tract, play an indispensable role in innate immunity. Integrin β4 (ITGB4) is a structural adhesion molecule that is involved in the pathological progression of acute inflammatory diseases and is downregulated in asthmatic patients. Research has shown that endothelial ITGB4 has proinflammatory properties in acute lung injury (ALI). However, the role of epithelial ITGB4 in a murine ALI model is still unknown. This study investigated the role of ITGB4 in lipopolysaccharide (LPS)-induced ALI. We found that ITGB4 in the airway epithelium had remarkably increased after the introduction of LPS in vivo and in vitro. Then, we constructed airway epithelial cell-specific ITGB4 knockout (ITGB4-/- ) mice to study its role in ALI. At a time point of 12 h after the tracheal injection of LPS, ITGB4-/- mice showed increased macrophages (mainly M1-type macrophages) and neutrophil infiltration into the lungs; inflammation-related proteins including interleukin (IL)-6, tumor necrosis factor, and IL-17A were significantly elevated compared to their levels in ITGB4+/+ mice. Furthermore, we investigated the role of ITGB4 in the anti-inflammatory response. Intriguingly, in the ITGB4-/- + LPS group, we found significantly reduced expression of anti-inflammatory factors, including IL-10 messenger RNA (mRNA) and ARG-1 mRNA. We also observed that monocyte chemotactic protein (MCP-1) increased significantly both in vivo and in vitro. Airway epithelium activates macrophages, most likely driven by MCP-1, which we confirmed in the coculture of epithelia and macrophages. These phenomena indicate that ITGB4 in airway epithelial cells plays an important role in the process of inflammation and activation of macrophages in ALI. Overall, these data demonstrated a novel link between airway epithelial ITGB4 and the inflammatory response in LPS-induced ALI.

Lung cancer is one of the most common malignant neoplasms worldwide. CD24 is a marker of tumor stem cells that plays an important role in tumorigenesis. Hsp70 is an important molecular chaperone. However, the co-expression and interaction of CD24 and Hsp70, as well as the significance for the prognosis of lung cancer are still unclear. The expression levels of CD24 and Hsp70 were detected by immunohistochemistry and their correlation was analyzed. The expression levels of CD24 mRNA and protein were examined using qRT-PCR and western blotting in SPCA1, A549, H1975, and H1650 cell lines. A CD24-overexpressing cell model was established. The interaction between CD24 and Hsp70 was verified by co-immunoprecipitation and western blotting. CD24 and Hsp70 expression were significantly higher in lung cancer tissues than in adjacent tissues (CD24: P = 0.008; Hsp70: P < 0.001). CD24 protein expression showed a positive correlation with lymph node metastasis, TNM stage, and vascular cancer thrombus. Hsp70 protein expression showed a positive correlation with differentiation, lymph node metastasis, and TNM stage. CD24 and Hsp70 high expression were also correlated with poor survival. The positive co-expression rate of CD24 and Hsp70 in lung cancer tissues was 52.7% (49/93). CD24 and Hsp70 expression in lung cancer were positively correlated (r = 0.368, P < 0.001), and co-immunoprecipitation was verified that both endogenous and exogenous CD24 co-precipitated with Hsp70 directly or indirectly. When Hsp70 inhibitor 17-AAG was added to A549 cells, Hsp70 and CD24 protein expression were significantly decreased. The present study demonstrated that CD24 and Hsp70 were highly expressed in lung cancer tissues, and associated with invasion, metastasis, and poor survival. Hsp70 may regulate CD24 expression. Co-expression of CD24 and Hsp70 may be a prognostic biomarker for lung cancer.

AIM: To investigate the relationship between neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and type 2 diabetic retinopathy (DR). METHODS: A total of 200 cases with type 2 diabetes mellitus were involved. All patients were divided into three groups according to the fundus examination: diabetes mellitus (DM, n=100), non-proliferative diabetic retinopathy (NPDR, n=62) and proliferative diabetic retinopathy (PDR, n=38). 100 healthy persons were selected as the normal control group (NCG, n=100). The related indicators, such as neutrophil count, lymphocyte count and platelet count, were measured. RESULTS: The value of NLR was significantly higher in PDR group patients than in NC group (1.81), DM group (1.76) and NPDR group (1.85) (P

Background Neu–Laxova syndrome (NLS) is an autosomal recessive genetic disease characterized by a variety of congenital malformations. It is a fatal developmental disorder caused by homozygous or compound heterozygous mutations in PHGDH, PSAT1, and PSPH. Method In this study, we aimed to characterize the clinical and molecular features of a fetus with NLS caused by novel heterozygous missense variants in PHGDH that were identified in his non-consanguineous parents. The fetus showed severe intrauterine growth restriction and abnormal ultrasound signs at 28 weeks of gestation. Fetal muscular tissue and peripheral blood samples from both parents were collected for whole-exome sequencing and Sanger sequencing. Results The clinical features were consistent with a diagnosis of NLS. Three missense mutations were detected in PHGDH in the fetus: c.488G > A(p.Arg163Gln) was carried by the father and c.607A > T(p.Ile203Phe) and c.743C > T (p.Ala248Val) were carried by the mother, resulting in a compound heterozygous mutation. Clinvar database suggests that c.743C > T may be benign variation, So we speculate that c.607A > T(p.Ile203Phe) is the pathogenic site carried by the mother. Conclusions This is the first report of PHGDH mutations in a patient with NLS in Northeast China. Thus, we detected a novel compound heterozygous pathogenic mutation in PHGDH, thereby expanding the molecular and phenotypic spectrum and providing strong support for the contribution of PHGDH to the pathogenesis of NLS.

Objective To determine the anti-cataract effects of coconut water (CW) in vivo and in vitro, and to explore the potential pathogenic mechanism. Methods In this study, 48 male Sprague-Dawley rats were randomly divided into 4 groups: control (CO), diabetic (DM), diabetic treated with CW (DM + CW), and diabetic treated with Glibenclamide (DM + Gli). Except for the CO group, in the other three groups, intraperitoneal injection of STZ (60 mg/kg) was conducted to establish diabetic models. The experiment was conducted for 20 weeks. The slit-lamp examination was undertaken during the period of experiment (20 weeks), and then, all rats were sacrificed. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) in the left lens were measured by using biochemical assays. The right lens was used for pathological analysis. The rat lens epithelial cells (LECs) were cultured in vitro and the subcultured cell were divided into four groups, namely the normal glucose group (5 mmol /L glucose, Group I), the high glucose group (40 mmol/L glucose, Group II), high glucose +5% CW group (Group III), and high glucose +10% CW group (Group IV). LECs were cultured under the conditions as described above for 48 h. Cell proliferation and the morphological changes were observed with interted phase contrast microscope.The level of cell apoptosis was determined by flow cytometry. the level of SOD, MDA and GSH-Px were also detected. Results The lens opacity index decreased in diabetic rats, and LECs apoptosis ratio also decreased in high glucose environments that received CW. Under treatment with CW, reduced MDA level and elevated activities of SOD and GSH-Px were detected, both in vivo and in vitro experiments. The increased severity of cataract and LECs apoptosis were noted in diabetic rats that received normal water, while CW markedly mitigated the enhanced cataract severity and the reduction of LECs induced by diabetes mellitus. Conclusion CW is a functional food that can protect the lens from diabetic cataract. The possible underlying mechanism may be partly explained via the decreased oxidative stress in lens. However, further research needs to be conducted to indicate the pathogenic mechanism of anti-diabetic effects of CW.

To compare the clinical manifestations of pertussis in children of different ages and different immunization statuses in Wenzhou, and to explore the limitations of diagnostic criteria for pertussis.The clinical data of 288 children diagnosed with pertussis at Yuying Children's Hospitalthe Second Affiliated Hospital of Wenzhou Medical University from October 2017 to December 2019 were retrospectively analyzed. The clinical characteristics of children of different ages and different immunization statuses were analyzed. Their clinical data were compared to relevant diagnostic criteria of pertussis in children of different ages according to theAmong the 288 children, 124 cases (43.06%) were 3 months old or younger, and 164 cases (288, 56.94%) were3 months old. Among patients≤3 months of age, cyanosis, three-depression sign, face redness, dyspnea and peripheral blood lymphocyte ratio were significantly higher than those of patients3 months of age. They also had higher incidence of pneumonia, higher proportion of developing severe pertussis, and longer stay at the hospital. All these findings showed statistically significant difference (Pertussis in children ≤3 months of age in Wenzhou City were more serious, showing higher rate of diagnosis conforming to the recommended clinical diagnostic criteria than that in children3 months old. The WBC threshold in routine blood test of ≤3 months old could be lowered appropriately and the current diagnostic criteria still needed improvement.

Summary Background Laparoscopic surgery in patients with liver cirrhosis (CL) is considered to be challenging. Recent studies have shown that laparoscopic liver resection (LLR) is more beneficial of reduced operative stress and postoperative complications in patients with CL. Aim A meta-analysis was done to review the currently available published data comparing LLR for patients with CL versus those non-cirrhosis of the liver (NCL). Methods The electronic databases of PubMed, Wiley, Web of Science, Embase, and the Cochrane Library were searched from date of inception to January 29, 2018. Studies reporting a comparison of outcomes and methods of LLR in CL and NCL groups were included. The studies were evaluated using the modified Newcastle-Ottawa Scale. Results A total of 1573 patients from six cohort studies were included in final analysis. The CL group had a slightly shorter operative time compared with the NCL group (weighted mean difference [WMD], 18.78 min shorter; 95% confidence interval [CI], −43.54–5.98; P = 0.14) and delayed hospital stay (WMD, 1.26 days longer; 95% CI, −0.05–2.56; P = 0.06). Blood loss, blood transfusion rate, mortality, and conversion rate did not differ significantly between the groups. Conclusions LLR is safe and feasible in the CL compared with the NCL groups. Our present review indicates that LLR should be considered when selecting surgery for patients with CL.

Background Single intrauterine death in twin pregnancy has become a relatively frequent complication of twin pregnancy. Preterm delivery is one of the complications with high incidence, and is closely related to the prognosis of the surviving fetus. However, studies about preterm delivery related to single intrauterine death are limited. The purpose of this paper is to analyze the factors associated with preterm birth in twins after single intrauterine death.Methods This was a case-control study of single intrauterine death in twins delivered at Shengjing Hospital over a 12-year period, from January 1, 2008, to November 30, 2020. We classified these cases into the preterm birth group and full-term birth group according to gestational age at birth. We selected age, gestational age at Single intrauterine death, and parity, etc, as related factors of the preterm birth group. Logistic regression analyses were performed to determine the relevance and importance of related factors and their association with preterm birth. We divided the preterm birth group into four smaller groups according to gestational age at single intrauterine death (14-19 weeks, 20-24 weeks, 25-30 weeks, and 31-36 weeks) and compared them. To exclude human intervention, we analyzed the spontaneous delivery cases as well.Results The analysis included 175 twins (mean [SD] gestational age at single intrauterine death, 27.0 [5.8] weeks; mean [SD] gestational age at birth, 33.8 [3.2] weeks ) with single intrauterine death. A total of 139 (79.4%) were cases of preterm birth. A significant risk factor associated with preterm birth was gestational age at single intrauterine death (odds ratio [OR], 2.89; 95% CI, 1.90-4.39). We found that the 14- to 19-week age group had the oldest gestational age at birth (37.25 [1.29]weeks ). The length of gestation in 14-19 weeks group, 20-24 weeks group, 25-30 weeks group and 31-36 weeks group were 20.5 weeks, 11.0 weeks, 4.77 weeks and 1.12 weeks respectivelyConclusion The fetus’ gestational age at single intrauterine death was a risk factor for preterm birth in their twin. The length of gestation was inversely proportional to increasing gestational age at single intrauterine death.

To explore the main active components and effects of cell cycle regulation mechanism of Astragali radix (AR) and Angelicae sinensis radix (ASR) on oxidative damage in vascular endothelial cells, a model of oxidative damage in human umbilical vein endothelial cells (HUVECs) induced by oxidized low-density lipoprotein (ox-LDL) treatment was developed. Based on the “knock-out/knock-in” model of the target component, cell viability, intracellular reactive oxygen species (ROS), and lactate dehydrogenase (LDH) leakage were assessed by Cell Counting Kit-8 assay, fluorescent probe 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), and colorimetric assay, respectively, to evaluate the protective effect of the active components of AR and ASR (astragaloside IV (AS IV), astragaloside I (AS I), formononetin (FRM), calycosin (CAL), calycosin-7-O-β-D glucoside (CLG), and ferulic acid (FRA)) against oxidative damage. The cell cycle and expression of genes encoding cyclins and cyclin-dependent kinases (CDKs) were observed using flow cytometry and quantitative real-time polymerase chain reaction. The results showed that the combination of active components (ACC) significantly inhibited the decrease in cell viability as well as the increase in ROS and LDH release in HUVECs induced by ox-LDL treatment. AS IV and FRM promoted the proliferation of HUVECs but the proliferation index was decreased in the AS I and FRA groups; this inhibitory effect was counteracted by the ACC. The ACC reduced and increased the proportion of positive cells in G1 and S phases, respectively, followed by the upregulation of cyclin A (CCNA), cyclin E (CCNE), and CDK2 mRNA expression and downregulation of cyclin B (CCNB), cyclin D1 (CCND1), CDK1, CDK4, and CDK6 mRNA expression, which significantly mitigated inhibition of HUVECs proliferation induced by ox-LDL treatment. Taken together, AS IV, AS I, FRM, CAL, CLG, and FRA were the primary pharmacodynamic substances of AR and ASR that alleviated oxidative injury in HUVECs. ACC mitigated the upregulation of intracellular ROS levels and LDH release induced by ox-LDL treatment, which promoted the cell cycle procession of HUVECs by regulating the expression of genes encoding cyclins and CDKs and thus preventing oxidative damage in HUVECs.

Short rib‑polydactyly syndrome type III (SRPS3) is a lethal perinatal skeletal disorder consisting of polydactyly and multi‑system organ abnormalities. To further assess the pathogenicity of two pairs of compound heterozygotes and to search for novel molecular etiology, X‑rays and hematoxylin and eosin staining were conducted in three cases: Two retrospective samples and a newly identified patient with SRPS3. In addition, next‑generation sequencing was used to evaluate a fetus with SRPS3. Typical radiological features of the three cases included a long, narrow thorax with short ribs, shortened long bones, spurs at the metaphysis of the long bones and congenital bowing of the femurs. The present study also observed atypical histopathological changes, together with the absence of proliferation and abundance of retaining cartilage in the primary spongiosum. In addition, two novel compound heterozygous variants were identified in the dynein cytoplasmic 2 heavy chain 1 (

Purpose. The objective of the current study was to investigate whether hcp plays a role in the process of Acinetobacter baumannii infection and to examine clinically relevant factors that may affect hcp expression. Methodology. Seventy-seven A. baumannii isolates from patients with a respiratory infection at the Second Affiliated Hospital and Yuying Childrens Hospital of Wenzhou Medical University (Wenzhou, China) were included in this study. PCR was performed to screen for the presence of hcp. Quantitative real time polymerase chain reaction (qRT-PCR) was carried out to examine the expression of hcp. Results. A total of 77.9 % (60 of 77) of the A. baumannii clinical isolates possessed the hcp gene. Expression of hcp was found to be strain-specific and associated with the infection status. Higher gene expression of hcp was found for invasive A. baumannii isolates causing an infection relative to the colonization group, and for the same strain at a post-infection status compared with that prior to infection. Acid environment was also found to be a trigger of hcp gene expression. Conclusion. The type VI secretion system and hcp predominate in A. baumannii causing respiratory infections. Expression of hcp is regulated by the infection status and acid environment, and might play a role in the process of triggering infection by the colonizer.

Agriculture is a primary contributor of greenhouse gas emissions and thus plays an important role in global climate change. Assessments of the carbon footprints of agricultural products are therefore helpful in climate change mitigation. The objective of this study was to provide a quantitative estimate of the carbon footprint of spring maize (Zea mays L.) production under different cultivation patterns. Four cultivation patterns were assessed in this study: traditional pattern, optimal pattern, super-high-yield pattern, and high-yield and high-efficiency pattern. The results showed that the greenhouse gas emissions from the agricultural inputs were 3225.2, 3152.9, 4557.2, and 4259.7 kg CO2-eq ha−1 for the traditional, optimal, super-high-yield, and high-yield and high-efficiency patterns, respectively, in the spring maize production process. Fertilizers, including chemical fertilizers and organic fertilizer, predominated among the contributors of total greenhouse gas emissions from agricultural inputs, accounting for 66.8, 65.9, 76.1, and 74.4% among the traditional, optimal, super-high-yield, and high-yield and high-efficiency patterns, respectively. The carbon footprint per yield among different cultivation patterns ranged from 0.48 kg CO2-eq kg−1 in the traditional pattern to 0.64 kg CO2-eq kg−1 in the high-yield and high-efficiency pattern, with intermediate values for the super-high-yield and optimal patterns. The N2O from soil and fertilizer application was the greatest contributor to the carbon footprint in spring maize production. Overall, higher yield and lower carbon footprint were concurrently observed under the super-high-yield pattern in the current study. Moreover, a reduction in the rate of fertilizers may provide a potential solution for reducing the carbon footprint of spring maize.

A flexible dicarboxylic acid, 4,4’-{[1,3-phenylenebis-(methylene)]bis(oxy) dibenzoic acid (H2L), in combination with 1,10-phenanthroline (phen) was used to reacted with Zn(II) ions under hydrothermal conditions, successfully affording a new metal-organic framework of [Zn(L)(phen)(H2O)]n (1). Luminescence sensing property investigations indicate that, compound 1 is sensitive detector of Fe3+ ions based on quenching of luminescence. Its treatment activity on the child allergic purpura was assessed and the specific mechanism of the compound was explored at the same time. First of all, the content of the inflammatory cytokine released into the skin tissue was measured with ELISA detection kit after the compound treatment. Apart from this, the activation ability of the AMPK inflammatory signaling pathway was measured with real time RT-PCR.

Rotavirus (RV) is the predominant cause of infantile gastroenteritis with multiple pathogenic factors, among which enterotoxin NSP4 is the most significant factor. NSP4 has been shown to induce elevation of the intracellular calcium concentration, alteration of the cytoskeleton organization, and cytopathic effect among other processes. However, increasing evidence suggests that RVs can escape from the gastrointestinal tract and invade other organs and tissues to cause extra-intestinal diseases. In this study, we investigated whether NSP4 has a pathogenic effect on extra-intestinal cells and examined possible molecular mechanisms in vitro. Our results showed that NSP486-175 has important functions in increasing intracellular Ca2+ concentration, altering actin cytoskeleton organization and inducing cellular damage in H9c2(2-1) cells. Blockade of the integrin α2 receptor using a specific antibody attenuated the increase of intracellular Ca2+ concentration and alleviated the observed cytopathic effects, suggesting that integrin α2 may be a receptor for NSP486-175. Collectively, these results indicate that extracellular NSP486-175 can induce elevation of the intracellular Ca2+ concentration, cause cytotoxic changes, and disrupt the actin cytoskeleton in H9c2(2-1) cells, which may constitute a possible mechanism for RV extra-intestinal pathogenesis.

Objectives Older antimicrobials such as fosfomycin are being considered as alternative agents in the treatment of drug-resistant organisms. However, there are limited data on the usefulness of fosfomycin against carbapenem-resistant Klebsiella pneumoniae (CRKP). This study aimed to investigate the prevalence of fosfomycin resistance and associated mechanisms in CRKP. Methods A total of 99 clinical CRKP isolates were collected in the Second Affiliated Hospital of Wenzhou Medical University (Wenzhou, China) between January 2017 and June 2018. Fosfomycin susceptibility testing was performed by the agar dilution method. Carbapenemase and fosfomycin resistance genes were detected by PCR. Analysis of the murA, glpT, uhpT, uhpA, ptsI and cyaA genes was performed by PCR and sequencing of four fosfomycin-resistant fosA3-negative CRKP strains. Conjugation experiments were employed to determine the mobility of the fosA3 gene. Results Of the 99 CRKP isolates, fosfomycin-non-susceptibility was detected in 48 (48.5%) isolates, among which the fosA3 gene was detected in 44. Among the four fosfomycin-resistant fosA3-negative CRKP isolates, one isolate possessed a single nucleotide insertion and deletion mutations as well as 219 nucleotide substitution mutations in murA, two isolates possessed deletion or mutation of large DNA fragments in glpT, and one isolate possessed a fragment insertion sequence in glpT. Transfection into Escherichia coli J53 via plasmid conjugation was successful for 19 (43.2%) of the 44 fosA3-positive CRKP isolates. Conclusion The fosA3 gene is the primary mechanism of fosfomycin resistance in CRKP and can be transmitted widely by plasmid in hospitals. Mutations in murA and glpT were found in fosfomycin-resistant fosA3-negative CRKP.

A parental diagnosis was performed for an unborn foetus of a healthy couple, who was due for ultrasound detection of multiple malformations and abnormal amniotic fluid karyotypes. For an accurate diagnosis, routine G-banding analysis and next generation sequencing (NGS) were carried out. Finally, conventional cytogenetic analysis suggested that the foetus had a karyotype of47,XX,+mar[52]/46,XN, meanwhile NGS also revealed a partial tetrasomy of 27.84Mb from 4q26-q31.21 (117,385,735-145,225,759), and G-banding analysis excluded the couple to have carried the 4q26-q31.21 duplication. We have identified a