Yinhuapinggan granule ameliorates lung injury caused by multidrug-resistant Acinetobacter baumannii via inhibiting NF-κB/NLRP3 pathway.

Yinhuapinggan granule (YHPG) is a traditional Chinese medicine prescription with rich clinical experience for the treatment of colds and coughs. The aim of this study is to investigate the protective effect of YHPG on multidrug-resistant (MDR) Acinetobacter baumannii (A. baumannii) infection in vivo and its potential anti-inflammatory mechanism. BALB/c mice were intranasally inoculated with MDR A. baumannii strain to establish the pneumonia infection model, and received intraperitoneally cyclophosphamide to form immunosuppression before attack. YHPG (6, 12 and 18 g/kg) was administered by gavage once a day for 3 consecutive days after infection. The protective effect of YHPG was evaluated by lung index, spleen index, thymus index, pathological changes of lung tissue and inflammatory factors (IL-1ß, IL-6 and TNF-α) in serum. The expression of key targets of NF-κB/NLRP3 signaling pathway in vivo was analyzed by immunohistochemistry, immunofluorescence, reverse transcription quantitative PCR (RT-qPCR) and Western blot. The results showed that YHPG improved the lung index and its inhibition rate, immune organ indexes and lung pathological changes in infected mice, and significantly reduced IL-1ß, IL-6 and TNF-α levels in serum. In addition, YHPG significantly down-regulated the mRNA and protein expression of NF-κB p65, NLRP3, ASC, Caspase-1, TNF-α, IL-6 and IL-1ß in mice lung tissue. The results of the current study demonstrated that YHPG has significant protective effects on mice infected with MDR A.baumannii, which may be related to the regulation of inflammatory factors and NF-κB/NLRP3 signaling pathway, indicating that YHPG has a wide range of clinical application value and provides a theoretical basis for its treatment of MDR A.baumannii infection.

Clinical and Molecular Characteristics of Carbapenemase-Producing E. coli Strains from Patients with Biliary System Diseases and Hematological Malignancies.

Purpose: This study aims to investigate the clinical and molecular characteristics of carbapenemase-producing E. coli strains (CPECO). Patients and Methods: We collected 38 non-repetitive CPECO strains, identified them using MALDI-TOF, and assessed their antimicrobial susceptibility via the VITEK-Compact II system. We gathered demographic and clinical patient data. Phenotypic assays were employed to detect carbapenemase types. Polymerase chain reaction (PCR) was utilized to identify the carbapenemase genes. Seven housekeeping genes were amplified and sequenced to determine the multilocus sequence typings (MLSTs). Results: These CPECO strains, primarily isolated from aseptic site and stool screening specimens, exhibited significant resistance to most clinical antibiotics, except for tigecycline and amikacin. Most patients had underlying medical conditions and underwent invasive procedures. There were significant differences among patients concerning the presence of malignancies, digestive system disorders, endoscopic retrograde cholangiopancreatography (ERCP) surgeries and abdominal drainage tubes. However, no significant differences were observed among patients regarding conditions, including hypertension, diabetes, respiratory diseases, urinary diseases and cardiovascular diseases, as well as invasive procedures such as deep venous catheterization, endotracheal intubation and gastrointestinal catheterization. Metallo-ß-lactamase was primarily responsible for carbapenem resistance, including blaNDM-5(24/38), blaNDM-1(5/38), blaNDM-9(1/38) and blaIMP-4(1/38). Additionally, 7 CPECO strains carried blaKPC-2. The distribution of CPECO sequence types (STs) was diverse, with seven strains being ST131, six strains being ST410, three strains each of ST1196 and ST10, although most STs were represented by only one strain. Conclusion: CPECO infections in patients with biliary system diseases may result from intestinal CPECO translocation, with ERCP surgery potentially facilitating this. Meanwhile, malignant tumor was found to be a significant factor affecting CPECO infections in patients with hematological diseases. blaNDM-5, blaNDM-1 and blaNDM-9 were primarily responsible for carbapenem resistance in CPECO strains. The emergence of carbapenem-resistant ST131 and ST410 strains should be alert to prevent the spread of carbapenem-resistant genes within high-risk epidemic clones.

Polydatin prevent lung epithelial cell from Carbapenem-resistant Klebsiella pneumoniae injury by inhibiting biofilm formation and oxidative stress.

Carbapenem-resistant Klebsiella pneumoniae (CRKP) causes severe inflammation in various infectious diseases, such as bloodstream infections, respiratory and urinary tract infections, which leads to high mortality. Polydatin (PD), an active ingredient of Yinhuapinggan granule, has attracted worldwide attention for its powerful antioxidant, anti-inflammatory, antitumor, and antibacterial capacity. However, very little is known about the effect of PD on CRKP. In this research, we evaluated the inhibitory effects of PD on both the bacterial level and the bacterial-cell co-culture level on anti-biofilm and efflux pumps and the other was the inhibitory effect on apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP) after CRKP induction. Additionally, we validated the mechanism of action by qRT-PCR and western blot in human lung epithelial cells. Firstly, PD was observed to have an inhibitory effect on the biofilm of CRKP and the efflux pump AcrAB-TolC. Mechanically, CRKP not only inhibited the activation of Nuclear Factor erythroid 2-Related Factor 2 (Nrf-2) but also increased the level of ROS in cells. These results showed that PD could inhibit ROS and activate Nrf-2 production. Together, our research demonstrated that PD inhibited bacterial biofilm formation and efflux pump AcrAB-TolC expression and inhibited CRKP-induced cell damage by regulating ROS and Nrf-2-regulated antioxidant pathways.

Lung cancer patient with Tropheryma whipplei and Nocardia co-infection.

A 34-year-old male presented with lung shadow and was asymptomatic during medical examination. The patient had a prior history of thyroid tumors. Imaging manifestation showed a nodule in the medial segment of the right middle lobe, with partial obstruction of the distal bronchus within the lesion. Ground-glass and inflammatory nodules were observed in the anterior segment of the right upper lobe, as well as chronic inflammatory changes in the lower lobe of the right lung. Lung histopathological examination suggested invasive adenocarcinoma. A morphological examination of the bronchoalveolar lavage fluid revealed the presence of Tropheryma whipplei (TW) and Nocardia. Although TW infection has been reported in cancer patients, co-infection with Nocardia is a unique occurrence in this case. Opportunistic pathogens are common in immunocompromised patients but in this case, the patient was a young adult with normal immunity and an early-stage tumor with TW and Nocardia co-infection. We demonstrated the presence of rare microorganisms through imaging findings, combined with different staining methods of bronchoalveolar lavage fluid and lung tissue sections and evaluation of morphological characteristics. The aim of the present study was to provide early diagnosis and treatment of patients by improving microbial morphological detection.

UTILITY OF STREM-1 BIOMARKER AND HCP GENE FOR IDENTIFICATION OF ACINETOBACTER BAUMANNII COLONIZATION AND INFECTION IN LUNG.

ABSTRACT: Objective: Respiratory infections or colonization of Acinetobacter baumannii (Ab) are common in clinical practice but are treated differently. Early identification of Ab infection and colonization reduces the risk of antibiotic mismatch but objective laboratory indicators to distinguish between bacterial infections and colonization are lacking. To distinguish infection and colonization of Ab, we tested the role of two biomarkers, triggering receptor expressed on myeloid cells-1 (TREM-1) and hemolysin coregulated protein. Methods: A total of 96 inpatients with Ab were divided into infection and colonization groups. Blood samples were collected on days 1, 2, 3, 5, 8, and 10 and daily maximum body temperature was recorded. Polymerase Chain Reaction and Reverse Transcription Polymerase Chain Reaction were used to detect the presence and expression levels of the hcp gene in Ab clinical isolates. Results : sTREM-1 and procalcitonin (PCT) levels on days 1 to 10 and neutrophil classification (N%) on days 1 to 3 were different ( P < 0.05) in the infection group and colonization group. Receiver operating characteristic (ROC) curves showed significant differences in N% and sTREM-1 on days 2 and 3 ( P < 0.01). sTREM-1 had the highest AUC ROC on days 1, 2, and 3 of all the markers. On day 1, the ROC curve of "WBC&N%&PCT&sTREM-1" was statistically different from individual indices (white blood cell count, N%, and PCT; P < 0.05) and was equal to the ROC curve of sTREM-1 ( P > 0.05). Thirty five of 96 patients were classified as infection group and 61 as colonization group with hcp gene detection rates of 71.43% (25/35) and 31.15% (19/61), respectively. No differences in hcp gene presence and transcript levels were found between two groups ( P > 0.05). Conclusions: Dynamic monitoring of sTREM-1 and PCT is valuable in identifying Ab infection and colonization. sTREM-1 can be improved by combination with multiple biomarkers in the early stage for identification of infection and colonization. The hcp gene was more likely to be present in the infection cohort.

Genomic Determinants of Pathogenicity and Antimicrobial Resistance of Nosocomial Acinetobacter baumannii Clinical Isolates of Hospitalized Patients (2019-2021) from a Sentinel Hospital in Hangzhou, China.

Purpose: Acinetobacter baumannii (A. baumannii or AB) is one of the most opportunistic, nosocomial pathogens threatening public healthcare across countries. A. baumannii has become a primary growing concern due to its exceptional ability to acquire antimicrobial resistance (AMR) to multiple antimicrobial agents which is increasingly reported and more prevalent every year. Therefore, there is an urgent need to evaluate the AMR knowledge of A. baumannii for effective clinical treatment of nosocomial infections. This study aimed to investigate the clinical distribution AMR phenotypes and genotypes, and genomic characteristics of A. baumannii isolates recovered from hospitalized patients of different clinical departments of a sentinel hospital to improve clinical practices. Methods: A total of 123 clinical isolates were recovered from hospitalized patients of different clinical departments during 2019-2021 to analyze AMR patterns, and further subjected to whole-genome sequencing (WGS) investigations. Multi-locus sequence typing (MLST), as well as the presence of antimicrobial-resistant genes (ARGs), virulence factor genes (VFGs) and insertion sequences (ISs) were also investigated from WGS data. Results: The results highlighted that A. baumannii clinical isolates had shown a high AMR rate, particularly from the intensive care unit (ICU), towards routinely used antimicrobials, ie, ß-lactams and fluoroquinolones. ST2 was the most prevalent ST in the clinical isolates, it was strongly associated to the resistance of cephalosporins and carbapenems, with blaOXA-23 and blaOXA-66 being the most frequent determinants; moreover, high carrier rate of VFGs was also observed such as all strains containing the ompA, adeF, pgaC, lpsB, and bfmR genes. Conclusion: Acinetobacter baumannii clinical isolates are mostly ST2 with high rates of drug resistance and carrier of virulence factors. Therefore, it requires measurements to control its transmission and infection.

Glycyrrhetinic acid protects against Multidrug-resistant Acinetobacter baumannii-induced lung epithelial cells injury by regulating inflammation and oxidative stress.

Glycyrrhetinic acid (GA) is a bio-effective component of Licorice. The GA is a monomer and the ingredient is an Oleanane-type pentacyclic triterpenes that has been used as a remedy for years. Due to the abuse of antibiotics, people pay attention to the emergence of Multidrug-resistant Acinetobacter baumannii (MDR-AB). As a conditional pathogen, MDR-AB causes severe infection, endangering human lives. Our previous studies found GA played an important role in Yinhua Pinggan, a Chinese medicine. However, whether GA could protect lung epithelium from MDR-AB-induced cell injury was elusive. Herein, we investigated the effects of GA on MDR-AB-infected A549 cells. The results showed GA had slightly antibacterial activity to MDR-AB in the GA (high concentration) but no impact on drug resistance genes. Notwithstanding, GA could reverse MDR-AB-induced cell apoptosis, hampered adhesion and invasion of MDR-AB to cells, and inhibit pro-inflammatory cytokines expression of IL-1ß, IL-6, and TNF. Besides, MDR-AB-induced reactive oxygen species, pro-oxidative protein malonaldehyde, and myeloperoxidase of cells were decreased by GA, while antioxidative proteins were recovered, showing antioxidative capacity of GA might play a critical role. The expressions of toll-like receptor (TLRs) - 1, 2, 4, 5, 6, and 9 were increased by MDR-AB infection, while GA reversed the tendency. Interestingly, GA inhibited MDR-AB induced myeloiddifferentiationfactor88 expression (MYD88), one downstream con-factors of TLRs, but no affection on Interferon regulatory Factor 3 (IRF3), the other one, indicating GA inhibited MDR-AB induced cell injury by impact TLR/MYD88 pathway to attenuate inflammation. Altogether, our results demonstrated that GA protects against MDR-AB-induced cell injury through its antioxidative and anti-inflammatory properties, which deserve further study in the future.

Glycyrrhetinic acid prevents carbapenem-resistant Klebsiella pneumoniae-induced cell injury by inhibiting mitochondrial dysfunction via Nrf-2 pathway.

OBJECTIVES: Due to the abuse of antibiotics, the high reoccurrence of drug-resistance strains, such as carbapenem-resistant Klebsiella pneumoniae (CRKP), deteriorates CRKP-infected pneumonia in the clinic, suggesting it is necessary to find new alternatives. Glycyrrhetinic acid (GA), an active ingredient of Yinhuapinggan granule, has antioxidant & anti-inflammatory capacity. Little, however, is known about the effects of GA on CRKP-induced epithelial injury. METHODS: In this research, we examined the protective effects of GA against pulmonary epithelium damage caused by CRKP infection and potential molecular mechanisms. RESULTS: Our results noted GA significantly promoted cell survival and reduced pro-inflammatory cytokines production, during CRKP-induced human pulmonary epithelial cell. Mechanically, GA alleviated mitochondrial-damage-induced apoptosis amid CRKP infection by inhibiting mitochondrial damage. Additionally, we found GA inhibited the expression of pro-apoptotic proteins Cyto-c, the Bax, and Caspase-3 while increasing the expression of anti-apoptotic protein Bcl-2. Further exploration found GA could trigger Nrf-2 expression at both gene and protein levels, activating antioxidative proteins to diminish CRKP-induced oxidative stress. CONCLUSION: Together, our results demonstrated that GA was a promising candidate to alleviate CRKP-infected lung injury as well as a synergist to treat CRKP infection with antibiotics.

Correlations between maternal hepatitis B virus carrier status and Down's syndrome prenatal screening indicators and their effects on the screening results.

The aim of this study was to determine the correlation between maternal hepatitis B virus (HBV) carrier status and pregnancy-associated serum screening indicators, as well as their implications on the prenatal screening results of Down's syndrome (DS). This retrospective cohort study included two groups, namely the healthy gravidas group (n = 19804) and the maternal HBV carrier group (n = 792). Serum pregnancy-associated plasma protein A (PAPP-A), alpha-fetoprotein (AFP), and free beta subunit of human chorionic gonadotropin (free ß-hCG) levels, as well as the foetal nuchal translucency (NT) thickness, were measured. Multivariatebinary logistic regression analysis was used to evaluate the association between the HBV carrier status and prenatal screening biomarkers. The PAPP-A multiple of the medium (MoM) and free ß-hCG MoM in the first trimester were significantly higher in the HBV carrier group than in the control group (both P < .05). Multivariate binary logistic regression analysis showed that HBV carrier status was identified as a risk factor for PAPP-A and the intrahepatic cholestasis of pregnancy (ICP), with adjusted odds ratios (aOR) of 1.363 (1.216-1.527) and 3.255 (2.356-4.499), respectively. Pregnant women with HBV carrier status had higher influence on serum PAPP-A level and ICP, and the risk calculation algorithm for DS in HBV carriers should be corrected in the first trimester of pregnancy. IMPACT STATEMENTWhat is already known on this subject? The maternal serum levels of pregnancy-associated plasma protein A (PAPP-A), alpha-fetoprotein (AFP), and free beta subunit of human chorionic gonadotropin (free ß-hCG), as well as the foetal nuchal translucency (NT) thickness, have been collectively used in the prenatal screening of Down's syndrome (DS), Edward's syndrome (ES), and open neural tube defects (ONTD). However, many factors, including the maternal age; maternal weight; gestational age; race; history of smoking and so on can affect those serum biomarker levels. Our aim is to know whether there is a difference for HBV status to pregnancy-associated serum screening indicators hoes.What the results of this study add? The PAPP-A multiple of the medium (MoM) and free ß-hCG MoM in the first trimester were significantly higher in the HBV carrier group than in the control group (both p < .05). Multivariate binary logistic regression analysis showed that the PAPP-A and intrahepatic cholestasis of pregnancy (ICP) were risk factors for HBV carriers, with aORs of 1.363 (1.216-1.527) and 3.255 (2.356-4.499), respectively.What the implications are of these findings for clinical practice and/or further research? The PAPP-A MoM in maternal HBV carriers was significantly higher than that in healthy gravidas, and the risk calculation algorithm for DS in maternal HBV carriers should be corrected in the first trimester of pregnancy.

Challenges of SARS-CoV-2 Omicron Variant and appropriate countermeasures.

The Omicron (B.1.1.529) variant was first reported in South Africa and rapidly spread worldwide in early November 2021. This caused panic in various countries, so it is necessary to understand Omicron Variant. This paper summarizes omicron variant-related research achievements. Studies have shown that Omicron Variant contains many mutations that make it more infectious and transmissible. At the same time, immune escape is also caused, resulting in reduced efficacy of existing vaccines, increased risk of reinfection, treatment failure or reduction of monoclonal antibody therapies, and detection failure. However, current data indicate that Omicron Variant causes mild clinical symptoms and few severe cases and deaths. Omicron Variant is valid for a range of nonpharmaceutical interventions against SARS-CoV-2. Improving diagnostic accuracy and enabling timely isolation and treatment of diagnosed cases is also critical to interrupting the spread of omicron variants. COVID-19 vaccine boosters could undoubtedly help control Omicron spread and infection. However, developing a vaccine specific to Omicron Variant is also imminent.

Effect of Hcp Iron Ion Regulation on the Interaction Between Acinetobacter baumannii With Human Pulmonary Alveolar Epithelial Cells and Biofilm Formation.

Acinetobacter baumannii is a type of bacterial nosocomial infection with severe drug resistance. Hemolysin co-regulated protein (Hcp) is a marker of activated type VI secretion system (T6SS), a key secretory system that promotes Gram-negative bacteria colonization, adhesion, and invasion of host cells. Hcp is also regulated by iron ions (Fe). In this study, an ATCC17978 hcp deletion strain (ATCC17978Δhcp), an hcp complement strain (ATCC17978Δhcp+ ), and an A. baumannii-green fluorescent protein (GFP) strain were constructed and used to investigate the role of hcp in bacterial adhesion to cells (human pulmonary alveolar epithelial cells (HPAEpiC)) and biofilm formation. Our results indicate that the inhibitory concentrations of the three A. baumannii strains (ATCC17978 wild type, ATCC17978Δhcp, and ATCC17978Δhcp+) were drug-sensitive strains. A. baumannii hcp gene and iron ions might be involved in promoting the formation of a biofilm and host-bacteria interaction. Iron ions affected the ability of A. baumannii to adhere to cells, as there was no significant difference in the bacterial numbers when assessing the adhesion of the three strains to HPAEpiC in the presence of iron ion concentrations of 0 µM (F = 3.1800, p = 0.1144), 25 µM (F = 2.067, p = 0.2075), 100 µM (F = 30.52, p = 0.0007), and 400 µM (F = 17.57, p = 0.0031). The three strains showed significant differences in their ability to adhere to HPAEpiC. The numbers of bacteria adhesion to HPAEpiC were ATCC17978Δhcp>ATCC17978Δhcp+>ATCC17978 in descending order. Hcp gene was positively regulated by iron ions in the bacteria-cells' co-culture. It is speculated that the effect of iron ions on the interaction between A. baumannii and HPAEpiC might be related to the transport function of hcp and bacterial immune escape mechanisms.

Serum Small Proline-Rich Protein 2A (SPRR2A) Is a Noninvasive Biomarker in Gastric Cancer.

OBJECTIVE: Since early diagnosis is very important for treating gastric cancer (GC), we aimed to detect serum small proline-rich protein2A (SPRR2A) to verify its diagnostic value for GC patients. METHODS: Serum samples were collected from 200 patients with GC, 100 patients with gastritis, 40 patients with rectal cancer (RC), 50 patients with colon cancer (CC), and 100 healthy controls. An enzyme-linked immunosorbent assay (ELISA) detection kit was applied to measure serum SPRR2A concentration. The correlations between serum SPRR2A and carcinoembryonic antigen (CEA), clinical pathological parameters of GC, and receiver operating characteristic (ROC) curve were also analyzed. RESULTS: The median serum SPRR2A concentration in GC patients was significantly higher than those in healthy controls and gastritis or colorectal cancer patients (P < 0.001). Serum SPRR2A concentration at a cut-off value of 80.7 pg/ml yielded an AUC of 0.851, with 75.7% sensitivity and 74.5% specificity for discriminating GC patients from healthy people. The AUC for the serum SPRR2A concentration combined with the CEA concentration was 0.876, with 79.7% sensitivity and 78.7% specificity. Similarly, serum SPRR2A discriminated GC patients from gastritis patients with an AUC of 0.820, with 90.5% sensitivity and 61.7% specificity. The AUC for the serum SPRR2A concentration combined with the CEA concentration was 0.848, with 87.8% sensitivity and 68.1% specificity. The serum SPRR2A levels in GC patients were associated with lymph node metastasis and the tumor-node-metastasis (TNM) stage (P < 0.05). There was an obvious difference in serum SPRR2A expression between GC patients before and after surgery (P < 0.0001). CONCLUSION: These results suggest that serum SPRR2A can be used as an effective marker for GC.

Characterization of the IncX3 Plasmid Producing bla NDM-7 From Klebsiella pneumoniae ST34.

Carbapenemase-producing Klebsiella pneumoniae has been a major clinical threat worldwide because therapeutic options are limited. Although New Delhi metallo-ß-lactamase (NDM) is an important carbapenemase responsible for carbapenem resistance, it is uncommon in carbapenemase-producing K. pneumoniae in China. In this study, we described strain HZW25, an NDM-7-producing K. pneumoniae strain belonging to sequence type 34 (ST34). HZW25 exhibited resistance to all ß-lactams tested but was susceptible to aminoglycosides and fluoroquinolones. The whole genome of HZW25 was sequenced with Pacific Biosciences RSII SMRT technology. HZW25 was composed of one chromosomal DNA and four plasmids, and the resistance genes of HZW25 were all located on the chromosome, except bla NDM-7 was located on a conjugative plasmid belonging to type IncX3 designated P4. The results of conjugation and transformation experiments showed that bla NDM-7 could be horizontally transferred successfully from the donor strain, HZW25, to the recipient strains, E. coli J53 and E. coli DH5α. The NDM variant transposable elements of the bla NDM-7-harboring plasmid P4 were the ISL3 and IS3000 families. The upstream region of bla NDM-7 contained ΔISAba125, which was inserted near the IS5 or ΔIS5 sequence. Our study is the first report of metallo-ß-lactamase NDM-7 in a carbapenemase-producing K. pneumoniae strain with ST34 in China. The emergence of NDM-producing K. pneumoniae would be troublesome during treatment using ceftazidime-avibactam. Therefore, the rapid and accurate identification of carbapenemase-producing K. pneumoniae is necessary.

Effect of calcium on the interaction of Acinetobacter baumannii with human respiratory epithelial cells.

BACKGROUND: Investigating the factors that influence Acinetobacter baumannii(Ab) adhesion/invasion of host cells is important to understand its pathogenicity. Metal cations have been shown to play an important role in regulating the biofilm formation and increasing the virulence of Ab; however, the effect of calcium on host-bacterial interaction has yet to be clarified. Here, the dynamic process of the interaction between Ab and human respiratory epithelial cells and the effect of calcium on host-bacterial interaction were explored using microscopic imaging, quantitative PCR and real time cellular analysis (RTCA). RESULTS: The concentration of calcium, multiplicity of infection and co-culture time were all demonstrated to have effects on host-bacterial interaction. A unique "double peak" phenomenon changed to a sharp "single peak" phenomenon during the process of Ab infection under the effect of calcium was observed in the time-dependent cell response profiles. Moreover, calcium can increase Ab adhesion/invasion of epithelial cells by regulating the expression of Ab-related genes (ompA, bfmRS, abaI). CONCLUSIONS: Effective control of calcium concentrations can provide new approaches for the prevention and treatment of multi-drug resistant Ab.

A Proteomic Network Approach across the Kidney Stone Disease Reveals Endoplasmic Reticulum Stress and Crystal-Cell Interaction in the Kidney.

Crystal-cell interactions are a vital step toward kidney stone formation. However, its mechanisms remained unclear. Here, a protein-protein interaction (PPI) network analysis of a kidney stone revealed that the proteins were enriched in a posttranslational protein modification process in the endoplasmic reticulum (ER). The in vitro study showed that the markers of ER stress, including Bip and CHOP, were upregulated, PERK and ATF6 were activated, and XBP-1 mRNA was spliced. An ER stress-specific protein, caspase-12, was activated in the apoptotic cells induced by calcium oxalate monohydrate (COM) crystals. The treatment with tunicamycin, an ER stress inducer, promoted the crystal-cell adhesion assayed by atomic absorption, reduced cell viability assayed by MTT, and downregulated the expression of proteins involved in the crystal formations. The treatment with salubrinal, an ER stress inhibitor, reversed the above effects for both tunicamycin and COM crystals. The aforementioned main observations were supported by in vivo study. These data demonstrated that ER stress was an essentially biological process of crystal-cell interactions. Our findings suggest that blocking ER stress may become a potential approach to preventing a kidney stone.

Vitamin K1 Inhibition of Renal Crystal Formation through Matrix Gla Protein in the Kidney.

BACKGROUND AND OBJECTIVES: Vitamin K (VK) plays a major role in modifying the binding of calcium in bones and blood vessels. Understanding the effect of VK on crystal formation in the kidney would contribute to advancing the treatment and prevention of kidney stones. METHODS: Rats were treated with vitamin K1 (VK1) for 8 weeks. VK1 levels were detected and crystal formation were observed. HK2 cells were exposed to calcium oxalate monohydrate crystals. Apoptosis and cell viability were detected. Crystal deposition was analyzed using atomic absorption assay. The adenovirus vectors expressing matrix Gla protein (MGP) and siMGP were constructed to elucidate the effect and mechanism of VK1 on crystal formation. MGP expression in vivo and in vitro was analyzed by Western blot. The mRNA levels of monocyte chemoattractant protein-1 (MCP-1) and collagen I was measured by semiquantitative RT-PCR. RESULTS: The concentrations of VK1 in whole blood and kidney tissues rose under treatment with VK1. Crystal formation was inhibited from the second to the 6th week, the frequency and quality of crystal formation decreased significantly, and the location of crystal formation was limited to a greater extent in the rats treated by VK1 compared to the control group. Warfarin treatment in the crystals-exposed HK2 cells significantly increased the number of crystals adhering to cells and the number of apoptotic cells and reduced cell viability. VK1 treatment reversed warfarin's above influence. VK1 inhibited the upregulations of MCP-1 and collagen I in kidney tissues under crystal load. VK1 treatment increased MGP expression in vivo and in vitro, and MGP is necessary for VK1 to play a role in crystal deposition in cells. CONCLUSIONS: VK1 treatment can inhibit the formation of renal crystals in vivo. VK1 increases MGP expression and functions through MGP to reduce crystal deposition in cells and provide cell protection. Our findings suggest that VK1 treatment could be a potential strategy for the treatment and prevention of nephrolithiasis.

Clinical Significance of Interleukin-6 in the Diagnosis of Sepsis and Discriminating Sepsis Induced by Gram-negative Bacteria.

BACKGROUND: To discuss the clinical significance of interleukin (IL)-6 in the differential diagnosis of sepsis and its capability of differentiating the sepsis induced by Gram-negative bacteria from that induced by Gram-positive bacteria. METHODS: A total of 379 children with sepsis were involved in this study to form the case group, and their C-reactive protein (CRP), procalcitonin (PCT) and IL-6 levels before antibiotics and after recovery were checked. Receiver operating characteristic curve was applied to evaluate the significance of CRP, PCT and IL-6 in the differential diagnosis of sepsis and their capability of differentiating the sepsis induced by Gram-negative bacteria from that induced by Gram-positive bacteria. RESULTS: When these 3 indicators were applied to the differential diagnosis of sepsis, the area under the curve (AUC) of IL-6, PCT and CRP was 0.881, 0.877 and 0.754, respectively. The combination of IL-6 and PCT presented highest diagnostic efficiency. CRP, PCT and IL-6 levels in children with sepsis induced by Gram-negative bacteria were significantly higher than those in children with sepsis induced by Gram-positive bacteria. CONCLUSIONS: CRP, IL-6 and PCT are applicable to the differential diagnosis of sepsis and differentiating the sepsis induced by Gram-negative bacteria from Gram-positive bacteria. Appropriate combinations of these indicators are capable of increasing differential diagnosis efficiency. These indicators can be used as markers of antibiotics usage, but whether they can be used as markers to withdraw antibiotics is still needed to be observed.

Optimization of multiplex quantitative polymerase chain reaction based on response surface methodology and an artificial neural network-genetic algorithm approach.

Multiplex quantitative polymerase chain reaction (qPCR) has found an increasing range of applications. The construction of a reliable and dynamic mathematical model for multiplex qPCR that analyzes the effects of interactions between variables is therefore especially important. This work aimed to analyze the effects of interactions between variables through response surface method (RSM) for uni- and multiplex qPCR, and further optimize the parameters by constructing two mathematical models via RSM and back-propagation neural network-genetic algorithm (BPNN-GA) respectively. The statistical analysis showed that Mg2+ was the most important factor for both uni- and multiplex qPCR. Dynamic models of uni- and multiplex qPCR could be constructed using both RSM and BPNN-GA methods. But RSM was better than BPNN-GA on prediction performance in terms of the mean absolute error (MAE), the mean square error (MSE) and the Coefficient of Determination (R2). Ultimately, optimal parameters of uni- and multiplex qPCR were determined by RSM.

Draft Genome Sequence of Carbapenem-Resistant Klebsiella pneumoniae XPY20 Collected from a Bloodstream Infection Patient.

Bloodstream infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) strains have been a severe problem with high clinical costs and high mortality rates. The blaKPC-2-producing CRKP strain XPY20 was collected from the blood of a patient. The genome characteristics and antimicrobial resistance mechanisms were determined using next-generation sequencing.

Simultaneous emergence and rapid spread of three OXA-23 producing Acinetobacter baumannii ST208 strains in intensive care units confirmed by whole genome sequencing.

Carbapenem-resistant Acinetobacter baumannii (CRAB) is a common nosocomial bacterial pathogen with limited treatment options. CRAB outbreaks are disastrous for critically ill patients. This study investigated carbapenemase-produced A. baumannii outbreaks in a tertiary hospital. Although multiple outbreaks were suggested by pulse-field gel electrophoresis, the genetic lineages and evolution between these isolates were not clear. To investigate the genomic epidemiology of these outbreaks and to reveal possible transmission routes, whole genome sequences (WGS) were compared and analyzed. From the WGS data, thirty isolates had the same sequence type (ST208); acquired resistance genes and chromosome resistant genes were detected and were responsible for multidrug resistance. A phylogenetic tree of single-nucleotide polymorphisms (SNPs) compared to the earliest index isolate found that three outbreaks had emerged and disseminated simultaneously. Of these, <10 SNPs were detected within the cluster, whereas at least 600 SNPs were found between the clusters. The probable transmission routes of outbreaks were generated combined with the genetic distance of isolates and patient epidemiological data. In conclusion, WGS was a convenient and accurate monitoring method for genomic epidemiologic investigation of outbreaks, and the genomic surveillance of multidrug-resistant bacterial pathogens would be a powerful warning system for the surveillance and prevention of outbreaks.