Diagnostic stewardship programme for urine culture: impact on antimicrobial prescription in a multi-centre cohort.

BACKGROUND: Restricting urine culture to patients with genuine urinary tract infection (UTI) reduces excessive antimicrobial prescription for asymptomatic bacteriuria. AIM: To evaluate the impact of urine culture diagnostic stewardship on antimicrobial consumption. METHODS: This quasi-study involved two general hospitals and 10 community clinics. In the pre-intervention (control) phase (25th November 2018-2nd February 2019), microscopy and culture results of all urine specimens were reported. In the post-intervention (study) phase (25th November 2019-2nd February 2020), urine cultures were processed and reported only if at least one of the following criteria were met: presence of white blood cells or bacteria on microscopy; patient from obstetrics, urology, paediatrics, oncology or renal transplant ward; specimen labelled as 'pregnancy', 'urological procedure', 'renal transplant' or 'neutropenic'; and ureteric, nephrostomy or suprapubic urine. For urine samples that did not fulfil these criteria, the microscopy results and a rejection comment were reported. FINDINGS: In total, 12,282 urine specimens were included in the intervention phase. Of these, 4757 (38.7%) specimens did not fulfil the screening criteria, and the microscopy result and a rejection comment were reported. One hundred and sixty-three (3.4%) of these non-reported urine cultures yielded significant bacterial growth, and the majority were Escherichia coli (N=58, 35.6%). Diagnostic stewardship was independently associated with lower antimicrobial consumption [adjusted odds ratio 0.76, 95% confidence interval (CI) 0.70-0.83, P<0.001] on multi-variable logistic regression across all healthcare settings. Diagnostic stewardship had no effect on patient mortality (adjusted hazard ratio=0.95, 95% CI 0.89-1.01, P=0.08). No patients with unreported urine culture developed bacteraemia from untreated UTI. CONCLUSION: Diagnostic stewardship of urine culture safely reduced excessive antimicrobial prescription for asymptomatic bacteriuria.

[The expression of P19ink4d in the pathogenesis and development of hearing loss].

Objective: This study aimed to investigate the P19ink4d expression in cochlea of mice model with noise induced hearing loss and the role of P19ink4d in the degeneration of inner ear cells. It also searched for P19ink4d gene alterations in patients with profound sensorineural deafness.Method: CBA/J mice were exposed to broad band noise at 101 dB SPL for 2 hours, auditory brainstem response (ABR) were examined to confirm noise lead to the permanent threshold shift. Immunohistochemical staining, Western blotting, and real-time polymerase chain reaction (PCR) were performed on cochlear tissues, to elucidate changes in P19ink4d expression in mice after noise exposure. For clinical evaluation, 400 children from unrelated families with severe or profound sensorineural hearing loss (SNHL) were recruited, genomic DNA was obtained from the patients and was subjected to DNA microarray to screen mutations in 4 most common genes. The sample that carried none of the common mutant alleles were subjected to PCR and sequenced to detect mutations in P19ink4d gene.Result: The ABR threshold shift of mice in the experimental group significantly increased after noise exposure and was higher than that in the null-noise group. The ABR of 1 day post noise was least among experimental groups and there is no statistical different between ABR of 7 days and 14 days post noise. The missing of outer hair cells occurred after noise exposure, while the inner hair cells hardly miss. It was found that the P19ink4d expression increased significantly in the inner ear cells 3 hours after noise exposure, then recovered in 24 hours. Western blot indicated that the amount of P19ink4d increased transitorily 3-6 h after the noise. However, no mutation existed within the coding exons of P19ink4d in the patients with profound sensorineural deafness.Conclusion: The results support the concept that P19ink4d may play an important role in the pathogenesis and development of noise induced hearing loss.

[Primary Ewing's sarcoma of the temporal bone: a rare case].

SummaryEwing's sarcoma is a malignant, round cell tumor arising from the bones and primarily affecting children and adolescent. Involvement of the skull bones is rarely reported, constituting 1%-6% of the total Ewing's sarcoma cases. We describe a 33 years old male patient having Ewing sarcoma of the mastoid and petrous parts of temporal bone, whose clinical presentation mimicked mastoiditis with facial nerve palsy. We discuss the clinical and therapeutic course of an extensive primary Ewing sarcoma of the temporal bone and review this entity's literature in detail. The etiopathology of an acute peripheral facial palsy is often hard to identify. If the facial weakness starts together with symptoms suggesting an inflammatory process, the differential diagnosis may be focused first on diseases like herpes zoster oticus and a severe course of acute purulent otitis media. As an uncommon tumor of the temporal bone, physicians should consider Ewing's sarcoma in the differential diagnosis of children and adolescents who present with facial nerve paralysis. And in the case of ambiguous clinical findings, a surgical exposure of the middle ear is recommended.

[Analysis of nystagmus and medical history of 121 patients positive with positional test].

Objective: To observe the characteristics of positional nystagmus and clinical profile of patients with positive positional test, and to explore its possible pathogenesis.Method: One hundred and twenty-one patients with positive positional test in the vestibular function examination were enrolled in the Peking University International Hospital from January to June in 2017. According to the 2017 BPPV guidelines, patients with test positive positional nystagmus were divided into two groups: definite BPPV and the controversial syndrome. Analyses of gender, age and characteristics of nystagmus, with or without recurrent dizziness, headache, and motion sickness were undertaken between the two groups, as well as response to the repositioning maneuver. Result: Of the total 121 cases, 49 cases were diagnosed as definite BPPV, accounting for 40.5%, 72 cases as controversial syndrome, accounting for 59.5%. The proportion of women in the two group was 76.2% and 78.9%, respectively. The average age of definite BPPV and the controversial syndrome was 51.2±16.8 and 51.3±15.7, respectively.There were significant differences in nystagmus duration, spontaneous nystagmus and nystagmus after headshaking between the two groups by chi square test(P<0.01). The mean intensity of horizontal and vertical nystagmus in posterior semicircular canal BPPV was(10.2±7.4) °/s and(36.6±17.5) °/respectively. And the mean intensity of nystagmus in the strong and weak side in horizontal semicircular canal BPPV was(40.8±25.1) °/s and(20.7±11.1) °/respectively. The intensity of horizontal and vertical nystagmus of the controversial syndrome group was(7.2±7.7) °/s and(7.2±4.3) °/s respectively. The incidence of headache in the controversial syndrome group was significantly higher than that in the BPPV group, P=0.013. According to the guidelines, patients were evaluated one day after the initial treatment. The cure rate and effective rate of the definite BPPV group was 75%(36/48) and 87.5%(42/48),and was 0 and 30.4% in the controversial syndrome group respectively. Conclusion: The patients in controversial syndrome group have a preponderance of exhibiting positive nystagmus during positional test. Nystagmus were usually of low velocity and sustained. Most of these cases presented spontaneous nystagmus and headshaking induced nystagmus, as well as headache and lacunar infarctions in history, and the response to the repositioning maneuver were often poor. It may be related to vestibular migraine and central nervous system. The diagnosis of BPPV must be prudent.Both characteristics of nystagmus and medical history should be carefully analyzed to avoid overdiagnosis.

[The investigation rate and influence factors of tinnitus with chronic suppurative otitis media].

Objective:To investigate the incidence of tinnitus and its influencing factors in patients with chronic suppurative otitis media, and to provide clinical data for the study of the pathogenesis of tinnitus.Method:The clinical data of 77 patients with chronic suppurative otitis media who underwent modified radical mastoidectomy and tympanoplasty were investigated. When tinnitus and otitis media happened in the same side,then the tinnitus is judged to be otitis media related. Patients were further divided into otitis media related tinnitus and the no tinnitus groups. The differences of tinnitus severity, sleep disturbance, migraine (migraine features), snoring and gastroesophageal reflux were compared between the two groups in tinnitus occurrence and classification.Result:The incidence of otitis media related tinnitus was 55.8%(43/77). Most of the tinnitus happened(33/43) later than the occurrence of otitis media for several years or even decades. There were 43 cases of tinnitus associated with otitis media, and 31 cases without tinnitus. Between the groups, significant differences were observed in migraine features, and the P value is 0.011, while no significant differences were noticed in the severity of hearing loss and sleep disorders, snoring, gastroesophageal reflux. Ranking of otitis media related tinnitus was positively related to the degree of hearing loss, especially the bone conduction threshold. With Spearman rank correlation test,P values of mean value(250Hz,500Hz,1kHz,2kHz,4kHz), middle frequency (1kHz, 2kHz)and high frequency (4kHz) of bone conduction threshold were 0.010,0.019 and 0.003, and the correlation coefficients were 0.391,0.356 and 0.443, respectively.Conclusion:The occurrence of tinnitus in patients with otitis media may not be consistent with the time of otitis media, and theoretically later than the occurrence of otitis media is more reasonable. Whether tinnitus occurs in patients with otitis media is not related to sleep disorders, the degree of air conduction and bone conduction hearing loss, but is related to migraine features. The severity of tinnitus associated with otitis media is associated with bone conduction hearing loss.

[Relevant factors of tinnitus in the elderly: an analysis of 150 volunteers].

Objective:To evaluate and characterize tinnitus in elderly volunteers，try to find out the relevant factors that can affect the incidence of tinnitus.Method:The study included 150 elderly volunteers. All volunteers had taken the otology examination and pure tone audiometry. They were interviewed by the investigators who were trained together, using the same questionnaire. The characteristics of tinnitus and the relationship between all relevant factors and tinnitus were analyzed.Result:Average age was 71.4 years. There are now 77 patients with tinnitus(51.3%), of which 31 cases have sustained tinnitus for more than 3 months, accounting for 40.3% of existing tinnitus volunteers. There was negative correlation between tinnitus and age. There was positive correlation between tinnitus and hearing loss. Tinnitus and headache had no correlation(P>0.05).Conclusion:After reach a certain age(70 years old), the incidence of tinnitus decreased. Hearing loss might be the most dangerous factor.If the hearing loss was more serious, the incidence of the tinnitus became higher. Tinnitus in the elderly may be the result of a combination of factors.

CDKN2B deletion is essential for pancreatic cancer development instead of unmeaningful co-deletion due to juxtaposition to CDKN2A.

Pancreatic cancer is among the deadliest malignancies; however, the genetic events that lead to pancreatic carcinogenesis in adults remain unclear. In vivo models in which these genetic alterations occur in adult animals may more accurately reflect the features of human cancer. In this study, we demonstrate that inactivation of Cdkn2b (p15ink4b) is necessary for induction of pancreatic cancer by oncogenic KRASG12D expression and inactivation of Tp53 and Cdkn2a in adult mouse pancreatic ductal cells (P60 or older). KRASG12D overexpression in these cells activated transforming growth factor-ß signaling and expression of CDKN2B, which, along with CDKN2A, led to cellular senescence and protected cells from KRAS-mediated transformation via inhibition of retinoblastoma phosphorylation. These results show a critical role of CDKN2B inactivation in pancreatic carcinogenesis, and provide a useful adult animal model by genetic engineering via lentiviral delivery.

Characterization and expression of Megalobrama amblycephala toll-like receptor 22 involved in the response to Aeromonas hydrophila.

The toll-like receptors (TLR) tlr22 was identified and characterized for the first time in one of the economically most important freshwater fish species in China, Megalobrama amblycephala. The full-length cDNA (4039 bp) of M. amblycephala tlr22 contains an open reading frame of 2706 bp, encoding a 901 amino-acid long polypeptide. The putative polypeptide contains 16 leucine-rich repeat (LRR) motifs, an LRR C-terminal, a transmembrane region and a cytoplasmic toll-interleukin-1 receptor (TIR) domain. Phylogenetic analyses revealed that M. amblycephala Tlr22 shared the closest relationship with a grass carp ortholog. tlr22 was constitutively expressed in nine tissues and during 10 developmental stages studied, albeit with varying expression levels. Along with many pathological changes observed after Aeromonas hydrophila bacterium infection, tlr22 and myd88 mRNA were significantly upregulated in blood, head kidney, spleen and intestine, indicating that tlr22 is involved in the immune response. These results provide an insight into tlr22 regulation mechanisms in the innate immune response to bacterial infection.

Exosomes derived from human embryonic mesenchymal stem cells promote osteochondral regeneration.

OBJECTIVE: Clinical and animal studies have demonstrated the efficacy of mesenchymal stem cell (MSC) therapies in cartilage repair. As the efficacy of many MSC-based therapies has been attributed to paracrine secretion, particularly extracellular vesicles/exosomes, we determine here if weekly intra-articular injections of human embryonic MSC-derived exosomes would repair and regenerate osteochondral defects in a rat model. METHODS: In this study, osteochondral defects were created on the trochlear grooves of both distal femurs in 12 adult rats. In each animal, one defect was treated with 100 µg exosomes and the contralateral defect treated with phosphate buffered saline (PBS). Intra-articular injections of exosomes or PBS were administered after surgery and thereafter weekly for a period of 12 weeks. Three unoperated age-matched animals served as native controls. Analyses were performed by histology, immunohistochemistry, and scoring at 6 and 12 weeks after surgery. RESULTS: Generally, exosome-treated defects showed enhanced gross appearance and improved histological scores than the contralateral PBS-treated defects. By 12 weeks, exosome-treated defects displayed complete restoration of cartilage and subchondral bone with characteristic features including a hyaline cartilage with good surface regularity, complete bonding to adjacent cartilage, and extracellular matrix deposition that closely resemble that of age-matched unoperated control. In contrast, there were only fibrous repair tissues found in the contralateral PBS-treated defects. CONCLUSION: This study demonstrates for the first time the efficacy of human embryonic MSC exosomes in cartilage repair, and the utility of MSC exosomes as a ready-to-use and 'cell-free' therapeutic alternative to cell-based MSC therapy.

[Analysis of tinnitus with sudden deafness].

Objective:To explore the pathogenesis of tinnitus.Method:Retrospective analysis of 192 patients with sudden deafness was done with the characteristics.Charactier of tinnitus and the factors that may affect tinnitus.Result:The intense of tinnitus is related with sex,χ²=14.704,P<0.01,and with age increased,the difference was more significant.The intense of tinnitus has not significant difference between age group with 45 years old(χ² =6.515,P>0.05) and the classification of sudden deafness and the degree of hearing loss(χ²=7.783,P>0.05,χ²=17.374,P>0.05).Conclusion:Tinnitus was the protection mechanism of the body,and the intensity was irrelevant to hearing loss.On one hand,there may be different modulate systems between different individual,on the other hand,women over 45 years old were more suffered from the severe tinnitus.

DDX17 (P72), a Sox2 binding partner, promotes stem-like features conferred by Sox2 in a small cell population in estrogen receptor-positive breast cancer.

We have previously demonstrated the existence of two phenotypically distinct cell subsets in estrogen receptor (ER)-positive breast cancer (BC) based on their differential response to a Sox2 reporter (SRR2), with reporter responsive (RR) cells being more tumorigenic and stem-like than reporter unresponsive (RU) cells. To delineate the molecular mechanisms underlying this phenotypic dichotomy, we tested our hypothesis that Sox2, which is a key regulator of the RR phenotype, is under the control of its binding partners. In this study, we focused on DDX17, known to be a transcription co-activator and found to be a Sox2 binding partner by liquid chromatography-mass spectrometry. Using immunoprecipitation, we confirmed the binding between DDX17 and Sox2, although this interaction was largely restricted to RR cells. While DDX17 was found in both the cytoplasm and nuclei in RU cells, it is confined to the nuclei in RR cells. siRNA knockdown of DDX17 in RR cells substantially decreased the Sox2-SRR2 binding and significantly decreased the SRR2 reporter activity without affecting the protein level of Sox2. Using ChIP-PCR, DDX17 knockdown also significantly decreased the binding of Sox2 to genomic SRR2, as well as 3 of its specific gene targets including MUC15, CCND1 and CD133. Correlating with these findings, siRNA knockdown of DDX17 significantly reduced soft agar colony formation and mammosphere formation in RR cells but not RU cells. To conclude, DDX17 is a Sox2-binding protein in ER-positive BC. In RR but not RU cells, DDX17 enhances the tumorigenic and stem-like features of Sox2 by promoting its binding to its target genes.

NPM-ALK mediates phosphorylation of MSH2 at tyrosine 238, creating a functional deficiency in MSH2 and the loss of mismatch repair.

The vast majority of anaplastic lymphoma kinase-positive anaplastic large cell lymphoma (ALK+ALCL) tumors express the characteristic oncogenic fusion protein NPM-ALK, which mediates tumorigenesis by exerting its constitutive tyrosine kinase activity on various substrates. We recently identified MSH2, a protein central to DNA mismatch repair (MMR), as a novel binding partner and phosphorylation substrate of NPM-ALK. Here, using liquid chromatography-mass spectrometry, we report for the first time that MSH2 is phosphorylated by NPM-ALK at a specific residue, tyrosine 238. Using GP293 cells transfected with NPM-ALK, we confirmed that the MSH2(Y238F) mutant is not tyrosine phosphorylated. Furthermore, transfection of MSH2(Y238F) into these cells substantially decreased the tyrosine phosphorylation of endogenous MSH2. Importantly, gene transfection of MSH2(Y238F) abrogated the binding of NPM-ALK with endogenous MSH2, re-established the dimerization of MSH2:MSH6 and restored the sensitivity to DNA mismatch-inducing drugs, indicative of MMR return. Parallel findings were observed in two ALK+ALCL cell lines, Karpas 299 and SUP-M2. In addition, we found that enforced expression of MSH2(Y238F) into ALK+ALCL cells alone was sufficient to induce spontaneous apoptosis. In conclusion, our findings have identified NPM-ALK-induced phosphorylation of MSH2 at Y238 as a crucial event in suppressing MMR. Our studies have provided novel insights into the mechanism by which oncogenic tyrosine kinases disrupt MMR.

Novel chimpanzee adenovirus-vectored respiratory mucosal tuberculosis vaccine: overcoming local anti-human adenovirus immunity for potent TB protection.

Pulmonary tuberculosis (TB) remains to be a major global health problem despite many decades of parenteral use of Bacillus Calmette-Guérin (BCG) vaccine. Developing safe and effective respiratory mucosal TB vaccines represents a unique challenge. Over the past decade or so, the human serotype 5 adenovirus (AdHu5)-based TB vaccine has emerged as one of the most promising candidates based on a plethora of preclinical and early clinical studies. However, anti-AdHu5 immunity widely present in the lung of humans poses a serious gap and limitation to its real-world applications. In this study we have developed a novel chimpanzee adenovirus 68 (AdCh68)-vectored TB vaccine amenable to the respiratory route of vaccination. We have evaluated AdCh68-based TB vaccine for its safety, T-cell immunogenicity, and protective efficacy in relevant animal models of human pulmonary TB with or without parenteral BCG priming. We have also compared AdCh68-based TB vaccine with its AdHu5 counterpart in both naive animals and those with preexisting anti-AdHu5 immunity in the lung. We provide compelling evidence that AdCh68-based TB vaccine is not only safe when delivered to the respiratory tract but, importantly, is also superior to its AdHu5 counterpart in induction of T-cell responses and immune protection, and limiting lung immunopathology in the presence of preexisting anti-AdHu5 immunity in the lung. Our findings thus suggest AdCh68-based TB vaccine to be an ideal candidate for respiratory mucosal immunization, endorsing its further clinical development in humans.

Gene expression analysis of pretreatment biopsies predicts the pathological response of esophageal squamous cell carcinomas to neo-chemoradiotherapy.

BACKGROUND: Neoadjuvant chemoradiotherapy (neo-CRT) followed by surgery has been shown to improve esophageal squamous cell carcinoma (ESCC) patients' survival compared with surgery alone. However, the outcomes of CRT are heterogeneous, and no clinical or pathological method can currently predict CRT response. In this study, we aim to identify mRNA markers useful for ESCC CRT-response prediction. PATIENTS AND METHODS: Gene expression analyses were carried out on pretreated cancer biopsies from 28 ESCCs who received neo-CRT and surgery. Surgical specimens were assessed for pathological response to CRT. The differentially expressed genes identified by expression profiling were validated by real-time quantitative polymerase chain reaction (qPCR), and a classifying model was built from qPCR data using Fisher's linear discriminant analysis. The predictive power of this model was further assessed in a second set of 32 ESCCs. RESULTS: The profiling of the 28 ESCCs identified 10 differentially expressed genes with more than a twofold change between patients with pathological complete response (pCR) and less than pCR (

Suppressor of cytokine signaling 6 (SOCS6) promotes mitochondrial fission via regulating DRP1 translocation.

Mitochondria are highly motile organelles that constantly undergo fission and fusion. Impairment of mitochondrial dynamics is associated with mitochondrial dysfunction and is frequently linked to the pathogenesis of neurodegenerative diseases and cancer. We have previously shown that biallelic inactivation of the suppressor of cytokine signaling 6 (SOCS6) gene is a frequent event in human gastric cancer. In this study, we recapitulated the event of SOCS6 loss using a Lentivirus-based knockdown approach, and demonstrated the linkage between SOCS6 depletion and the suppression of programmed cell death. SOCS6 promotes intrinsic apoptosis, with increased Bax conformational change, mitochondrial targeting, and oligomerization. Most importantly, SOCS6 is targeted to mitochondria and induces mitochondrial fragmentation mediated through an increase in DRP1 fission activity. Here, we show that SOCS6 forms complex with DRP1 and the mitochondrial phosphatase PGAM5, attenuates DRP1 phosphorylation, and promotes DRP1 mitochondrial translocation. Based on mutation analyses, SOCS6-mediated apoptosis is tightly coupled to its ability to induce mitochondrial fission. This study demonstrates an important role for SOCS6 in modulating mitochondrial dynamics and apoptosis.

Differentially imprinted innate immunity by mucosal boost vaccination determines antituberculosis immune protective outcomes, independent of T-cell immunity.

Homologous and heterologous parenteral prime-mucosal boost immunizations have shown great promise in combating mucosal infections such as tuberculosis and AIDS. However, their immune mechanisms remain poorly defined. In particular, it is still unclear whether T-cell and innate immunity may be independently affected by these immunization modalities and how it impacts immune protective outcome. Using two virus-based tuberculosis vaccines (adenovirus (Ad) and vesicular stomatitis virus (VSV) vectors), we found that while both homologous (Ad/Ad) and heterologous (Ad/VSV) respiratory mucosal boost immunizations elicited similar T-cell responses in the lung, they led to drastically different immune protective outcomes. Compared with Ad-based boosting, VSV-based boosting resulted in poorly enhanced protection against tuberculosis. Such inferior protection was associated with differentially imprinted innate phagocytes, particularly the CD11c(+)CD11b(+/-) cells, in the lung. We identified heightened type 1 interferon (IFN) responses to be the triggering mechanism. Thus, increased IFN-ß severely blunted interleukin-12 responses in infected phagocytes, which in turn impaired their nitric oxide production and antimycobacterial activities. Our study reveals that vaccine vectors may differentially imprint innate cells at the mucosal site of immunization, which can impact immune-protective outcome, independent of T-cell immunity, and it is of importance to determine both T-cell and innate cell immunity in vaccine studies.

Aberrant expression and biological significance of Sox2, an embryonic stem cell transcriptional factor, in ALK-positive anaplastic large cell lymphoma.

Sox2 (sex-determining region Y-Box) is one of the master transcriptional factors that are important in maintaining the pluripotency of embryonic stem cells (ESCs). In line with this function, Sox2 expression is largely restricted to ESCs and somatic stem cells. We report that Sox2 is expressed in cell lines and tumor samples derived from ALK-positive anaplastic large cell lymphoma (ALK(+)ALCL), for which the normal cellular counterpart is believed to be mature T-cells. The expression of Sox2 in ALK(+)ALCL can be attributed to nucleophosmin-anaplastic lymphoma kinase (NPM-ALK), the oncogenic fusion protein carrying a central pathogenetic role in these tumors. By confocal microscopy, Sox2 protein was detectable in virtually all cells in ALK(+)ALCL cell lines. However, the transcriptional activity of Sox2, as assessed using a Sox2-responsive reporter construct, was detectable only in a small proportion of cells. Importantly, downregulation of Sox2 using short interfering RNA in isolated Sox2(active) cells, but not Sox2(inactive) cells, resulted in a significant decrease in cell growth, invasiveness and tumorigenicity. To conclude, ALK(+)ALCL represents the first example of a hematologic malignancy that aberrantly expresses Sox2, which represents a novel mechanism by which NPM-ALK mediates tumorigenesis. We also found that the transcriptional activity and oncogenic effects of Sox2 can be heterogeneous in cancer cells.