Drug Sensitivity Testing Research Articles

Primary Pre-Extensively Drug-Resistant (Pre-XDR) Tuberculosis with Meningoencephalitis in Nineteen Years Old Young Woman : A Case Report

Introduction. Mycobacterium tuberculosis (MTB) is continually evolving in order to resist treatment. This has resulted in drug-resistant tuberculosis, which has a greater rate of therapy failure and mortality, as well as the necessity for other ways of disease transmission prevention. Pre-extensively drug-resistant tuberculosis (Pre-XDR TB) is caused by MTB strains that are resistant to isoniazid, rifampicin, and any fluoroquinolone medicines. Case Report. A 19-year-old HIV-negative immunocompetent female presented to our emergency department with dyspnea and severe headache. She also complained of a cough, a low-grade temperature, and weight loss. She has no relevant medical history and has never been exposed to tuberculosis. A chest X-ray revealed infiltrates with cavities and fibrosis in both lungs, while a brain CT suggested meningoencephalitis. We performed a sputum drug sensitivity test and discovered that this patient was resistant to rifampin, isoniazid, levofloxacin, and moxifloxacin. She was later diagnosed with Pre-XDR TB and was given a tailored lengthier regimen that included Bedaquiline, Cycloserin, Linezolide, Clofazimine, and Etambutol. Her symptoms improved significantly as a result of this program. Discussion. Primary pre-XDR TB in an immunocompetent patient is uncommon, and our case is even more unusual due to meningoencephalitis involvement. The treatment of pre-XDR TB requires a personalized regimen that is tailored to the patient's clinical status and comorbidities. The possibility of transmission via a positive contact was raised in this patient, prompting us to conduct a more thorough contact inquiry to prevent future spread. Conclusion. In our region, there is a risk of'silent' drug resistant tuberculosis. Early detection and treatment of such patients are critical to preventing uncontrollable pan-resistant TB. Successful management with cautious contact investigation will minimize the mortality rate and spread rate of drug-resistant TB. Keywords. Drug-Resistant Lung Tuberculosis, Drug-Resistant Meningoencephalitis Tuberculosis

Improving treatment plan and mental health in children with abdominal infection for broad-spectrum bacterial infections

BACKGROUND Pediatric abdominal infection is a common but serious disease that requires timely and effective treatment. In surgical treatment, accurate diagnosis and rational application of antibiotics are the keys to improving treatment effects. AIM To investigate the effect of broad-spectrum bacterial detection on postoperative antibiotic therapy. METHODS A total of 100 children with abdominal infection who received surgical treatment in our hospital from September 2020 to July 2021 were grouped. The observation group collected blood samples upon admission and sent them for broad-spectrum bacterial infection nucleic acid testing, and collected pus or exudate during the operation for bacterial culture and drug sensitivity testing; the control group only sent bacterial culture and drug sensitivity testing during the operation. RESULTS White blood cell count, C-reactive protein, procalcitonin, 3 days after surgery, showed better postoperative index than the control group (P < 0.05). The hospital stay in the observation group was significantly shorter than that in the control group. The hospitalization cost in the observation group was significantly lower than that in the control group, and the difference between the two groups was statistically significant (P < 0.05). CONCLUSION Early detection of broad-spectrum bacterial infection nucleic acids in pediatric abdominal infections can help identify pathogens sooner and guide the appropriate use of antibiotics, improving treatment outcomes and reducing medical costs to some extent.

Late Subcutaneous Infection Caused by Serratia marcescens Following Hyaluronic Acid Injection: A Case Report and Systemic Review.

The field of cosmetic filler injection has experienced rapid development over the past two decades, especially in facial augmentation utilizing hyaluronic acid (HA) fillers. Gram-negative bacteria are found to be the main pathogens of infective nodules after HA injection. The occurrence of cutaneous infections attributed to Serratia marcescens is exceedingly rare and predominantly noted in patients with compromised immune systems. To summarize the clinical features, diagnosis, and treatment of subcutaneous infection caused by Serratia marcescens following hyaluronic acid injection. A rare case of cutaneous Serratia marcescens infection following hyaluronic acid injection was presented. A comprehensive review of the published literature describing the management of skin infection caused by S. marcescens in immunocompetent patients was then conducted, which encompassed three case series and eight case reports published between 1999 and 2017. Data extraction included information on authors, gender, age, signs and symptoms, previous treatment, corresponding management strategies, and follow-up duration. Serratia marcescens were isolated in abscesses (n = 6, 35.29%), painful nodules (n = 2, 11.76%), ulcers (n = 6, 35.29%), and others (n = 3, 17.65%). In cases providing salvage plans (n = 11), quinolones were shown to be the most effective antibiotics for salvage, with eight full recoveries (72.73%), and trimethoprim-sulfamethoxazole was the second most useful antibiotic (18.18%). With the help of pathogen examination and drug-sensitive tests, sensitive aminoglycosides, quinolone (especially moxifloxacin), or TMP-SMX for at least 2 weeks can be considered as the first-line treatment of late subcutaneous infection caused by Serratia marcescens following hyaluronic acid injection.

Abstract A023: Patient-Derived Organoid Cultures for Personalized Therapies and Targeted Drug Screening Applications

Abstract Patient-derived organoids (PDOs) have emerged as a powerful tool for modeling human cancers due to their ability to closely mirror in vivo architecture and preserve the genetic landscape of tumors. They offer a crucial platform for studying disease progression and treatment responses. In this study, we present a comprehensive characterization of a unique collection of PDO cultures derived from pancreatic ductal adenocarcinoma (PDAC), a particularly aggressive malignancy often diagnosed in advanced stages with limited treatment options and poor prognosis. Our investigation explores morphological and genetic profiling of 10 PDAC PDOs, specifically identifying distinct drug-response patterns. The morphological characteristics of these PDO cultures were investigated by microscopy, while the mutational status was acquired by panel sequencing. Subsequently, PDOs were treated with a panel of 10 inhibitors, either individually or in combination. The response to these treatments was assessed through live-cell imaging (Incucyte®) to capture and quantify changes in organoid morphology and endpoint measurements of cellular viability using a CellTiter-Glo® assay. Our detailed in vitro drug sensitivity testing revealed a remarkable diversity in the responses of the PDAC PDOs to different chemotherapeutic agents, including those currently under clinical trials. Of notable significance is the depth of understanding we have achieved, allowing us to establish potential correlations between PDO morphology, genetic mutations, and drug reactions. This finding provides a key insight into the complex interplay of these factors and their impact on treatment outcomes, offering valuable insights into understanding individual tumor subtypes' sensitivity or resistance to treatments. The PDOs exhibited a spectrum of cancer-associated mutations, including KRAS, TP53, SMAD4, CDKN2A, and ARID1A, reflecting the heterogeneity observed in PDAC patients. Morphologically, the organoids displayed various growth patterns, ranging from cystic and regular structures to filled or unfilled, granular, dark, and light membrane structures. Drug treatments underscored the diverse and unique drug-response profiles within the PDOs, with some demonstrating sensitivity to multiple treatments. In contrast, others exhibited resistance, highlighting the intricate nature of drug responses within this model. Overall, our findings emphasize the ability of PDAC organoids to faithfully capture the complexity and heterogeneity of the disease, making them promising platforms for guiding personalized oncology approaches. The diverse drug response profiles observed in PDOs advocate leveraging this model to develop precision medicine strategies tailored to patients' characteristics, ultimately paving the way for more effective and personalized treatment regimens in the fight against PDAC. Citation Format: Constanza Tapia Contreras, Günter Schneider, Denise Müller, Kimia Mirzakhani, Karly Conrads, Silke Kaulfuß, Tim Beißbarth, Gabriela Salinas, Nils Beyer, Sophia Reinländer, Ulrix Sax, Elisabeth Heßmann, Volker Ellenrieder, Philipp Ströbel, Michael Ghadimi. Patient-Derived Organoid Cultures for Personalized Therapies and Targeted Drug Screening Applications [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: Advances in Pancreatic Cancer Research; 2024 Sep 15-18; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2024;84(17 Suppl_2):Abstract nr A023.

Establishment and characterization of a novel human gallbladder cancer cell line, GBC-X1

In this study, we successfully established a novel gallbladder cancer cell line, designated as GBC-X1, derived from a primary tumor of a gallbladder cancer patient. By comprehensively analyzing the cell line’s phenotype, molecular characteristics, biomarkers, and histological characteristics, we confirmed that GBC-X1 serves as a valuable model for investigating the pathogenesis of gallbladder cancer and developing therapeutic agents. GBC-X1 has been continuously cultured for one year, with over 60 stable passages. Morphologically, GBC-X1 exhibits typical features of epithelial tumors. The population doubling time of GBC-X1 is 32 h. STR analysis validated a high consistency between GBC-X1 and the patient’s primary tumor. Karyotype analysis revealed an abnormal hypertetraploid karyotype for GBC-X1, characterized by representative karyotypes of 98, XXXX del (4) p (12) del (5) p (21) der (10). Under suspension culture conditions, GBC-X1 efficiently forms tumor balls, while subcutaneous inoculation of GBC-X1 cells into NXG mice leads to xenograft formation with a rate of 80%. Drug sensitivity testing demonstrated that GBC-X1 is resistant to oxaliplatin and sensitive to 5-FU, gemcitabine, and paclitaxel. Immunohistochemistry revealed positive expression of CK7, CK19, E-cadherin, MMP-2, CD44, SOX2, and TP53 in GBC-X1 cells, weak positive expression of Vimentin, and a Ki67 positive rate of 35%. Our research highlights GBC-X1 as a novel gallbladder cancer cell line and emphasizes its potential as an effective experimental model for investigating the pathogenesis of gallbladder cancer and drug development.

Patient-derived acellular ascites fluid affects drug responses in ovarian cancer cell lines through the activation of key signalling pathways.

Malignant ascites is commonly produced in advanced epithelial ovarian cancer (EOC) and serves as unique microenvironment for tumour cells. Acellular ascites fluid (AAF) is rich in signalling molecules and has been proposed to play a role in the induction of chemoresistance. Through in vitro testing of drug sensitivity and by assessing intracellular phosphorylation status in response to mono- and combination treatment of five EOC cell lines after incubation with AAFs derived from 20 different patients, we investigated the chemoresistance-inducing potential of ascites. We show that the addition of AAFs to the culture media of EOC cell lines has the potential to induce resistance to standard-of-care drugs (SCDs). We also show that AAFs induce time- and concentration-dependent activation of downstream signalling to signal transducer and activator of transcription 3 (STAT3), and concomitantly altered phosphorylation of mitogen-activated protein kinase kinase (MEK), phosphoinositide 3-kinase (PI3K)-protein kinase B (AKT) and nuclear factor NF-kappa-B (NFκB). Antibodies targeting the interleukin-6 receptor (IL6R) effectively blocked phosphorylation of STAT3 and STAT1. Treatments with SCDs were effective in reducing cell viability in only a third of 30 clinically relevant conditions examined, defined as combinations of drugs, different cell lines and AAFs. Combinations of SCDs and novel therapeutics such as trametinib, fludarabine or rapamycin were superior in another third. Notably, we could nominate effective treatment combinations in almost all conditions except in 4 out of 30 conditions, in which trametinib or fludarabine showed higher efficacy alone. Taken together, our study underscores the importance of the molecular characterisation of individual patients' AAFs and the impact on treatment resistance as providing clinically meaningful information for future precision treatment approaches in EOC.

A case of necrotic pneumonia caused by Streptococcus pneumoniae was diagnosed using a pneumonia antigen test in BALF: A case report.

Streptococcus pneumoniae is a common cause of community-acquired pneumonia. Currently, it is believed that many cases of pulmonary infection with negative results on pathogenic testing are caused by S. pneumoniae. There have been no reports of the detection of S. pneumoniae antigen in lung lavage fluid. An elderly male patient with suboptimal fasting blood glucose control and a history of liver abscess. Chest computed tomography (CT) revealed inflammatory lesions in both lungs with consolidation in the middle lobe of the right lung. After admission, we collected alveolar lavage fluid in a timely manner and performed pneumococcal antigen detection and etiological testing. Prompt testing for pneumococcal antigen in bronchoalveolar lavage fluid yielded a positive clinical outcome. Subsequent analysis via bacterial culture of sputum and next-generation sequencing (mNGS) of BALF definitively identified S. pneumoniae as the etiological agent. Following the analysis of drug sensitivity test results from the identified pathogens, adjustments were made to the antibiotic regimen, and appropriate pus puncture drainage was performed. Subsequently, the patient's condition improved, leading to discharge. The identification of S. pneumoniae antigen in bronchoalveolar lavage fluid may facilitate earlier and more precise diagnosis of pneumonia attributed to S. pneumoniae.

Isolation and identification of Acinetobacter lwoffii from the Chinese Giant Salamander (Andrias davidianus)

This study aimed to identify the primary pathogenic bacteria responsible for the mortality of the Chinese giant salamander (Andriasd davidianus). A pathogenic bacterium was isolated from a diseased Chinese giant salamander exhibiting typical symptoms under aseptic conditions and then identified by morphological examinations, biochemical analysis, and the sequence analysis of bacterial 16S rRNA. Artificial infection testing was then conducted to determine the pathogenicity of the isolated bacteria strain. Drug susceptibility tests were conducted using the agar diffusion method. The isolated pathogenic bacteria, named DN-2, was successfully identified as Acinetobacter lwoffii. The artificial infection showed that the typical symptoms of the disease could be replicated as the ones originally occurred, and this bacterium exhibited high pathogenicity to the Chinese giant salamander. In the Chinese giant salamander, the median lethal dosage (LD50) of A. lwoffii DN-2 for A. davidianus was determined to be 4.63×104 CFU/g. Drug sensitivity testing showed that these bacteria were highly sensitive to erythromycin, gentamicin, neomycin, streptomycin, midecamycin, ciprofloxacin, piperacillin, florfenicol, doxycycline, carbenicillin, and sulfanilamide. In summary, A. lwoffii was identified as the primary pathogen responsible for the demise of A. davidianus. Our study first presents how these bacteria harm Chinese giant salamanders.

Clinical Characteristics, Serotypes and Antimicrobial Resistance of Invasive Salmonella Infections in HIV-Infected Patients in Hangzhou, China, 2012-2023.

Developing countries, invasive Salmonella infections can cause considerable morbidity and mortality. There is a relative lack of data on coinfection with Salmonella in HIV-infected patients in Hangzhou, China. In this study, we manually collected case data of patients aged >18 years with HIV combined with invasive Salmonella infections admitted to Xixi Hospital in Hangzhou from January 2012 to August 2023 by logging into the Hospital Information System, and identified 26 strains of invasive Salmonella using a fully automated microbiological identification system and mass spectrometer. Serotypes were determined using Salmonella diagnostic sera based on the White-Kauffmann-Le Minor scheme. Drug sensitivity tests were performed using the automated instrumental method of the MIC method. A total of 26 HIV-infected patients with invasive Salmonella coinfections were identified over 11 years; Twenty-five of the 26 patients (96.2%) were males, with a mean age of 33.5 years (26.75, 46.75). The most common type of infection was bloodstream infection (92.3%). One patient also had concomitant meningitis and osteoarthritis, followed by pneumonia (7.7%). The presence of multiple bacterial infections or even multiple opportunistic pathogens was clearly established in 7 (26.9%) patients. Three (11.6%) patients were automatically discharged from the hospital with deterioration of their condition, and one (3.8%) patient died. Salmonella enteritidis was the most common serotype in 6 patients (23.2%), and Salmonella Dublin was the most common serotype in 6 patients (23.2%). Drug sensitivity results revealed multidrug resistance in a total of 8 (30.8%) patients. The clinical presentation of invasive Salmonella infection in HIV patients is nonspecific and easily masked by other mixed infections. A CD4+ count <100 cells/µL and comorbid intestinal lesions may be important susceptibility factors. Salmonella has a high rate of resistance to common antibiotics, and the risk of multidrug resistance should not be ignored.

Developing Patient-Derived 3D-Bioprinting models of pancreatic cancer

IntroductionPancreatic cancer (PC) remains a challenging malignancy, and adjuvant chemotherapy is critical in improving patient survival post-surgery. However, the intrinsic heterogeneity of PC necessitates personalized treatment strategies, highlighting the need for reliable preclinical models. ObjectivesThis study aimed to develop novel patient-derived preclinical PC models using three-dimensional bioprinting (3DP) technology. MethodsPatient-derived PC models were established using 3DP technology. Genomic and histological analyses were performed to characterize these models and compare them with corresponding patient tissues. Chemotherapeutic drug sensitivity tests were conducted on the PC 3DP models, and correlations with clinical outcomes were analyzed. ResultsThe study successfully established PC 3DP models with a modeling success rate of 86.96%. These models preserved genomic and histological features consistent with patient tissues. Drug sensitivity testing revealed significant heterogeneity among PC 3DP models, mirroring clinical variability, and potential correlations with clinical outcomes. ConclusionThe PC 3DP models demonstrated their utility as reliable preclinical tools, retaining key genomic and histological characteristics. Importantly, drug sensitivity profiles in these models showed potential correlations with clinical outcomes, indicating their promise in customizing treatment strategies and predicting patient prognoses. Further validation with larger patient cohorts is warranted to confirm their potential clinical utility.

Drug sensitivity tumor cell clusters in malignant peritoneal mesothelioma.

To explore the most effective adjuvant chemotherapy regimen for malignant peritoneal mesothelioma (MPM) through patient derived tumor-like cell clusters (PTC) drug sensitivity test. PTC were cultured in vitro with intraoperative specimens, and drug sensitivity test was performed to calculate the most effective chemotherapy regimen for MPM. The patients were divided into conventional and individualized chemotherapy group according to whether they received PTC drug testing. Univariate and multivariate analyses were conducted to identify independent prognostic factors. Among 186 MPM patients included, 63 underwent PTC culture and drug sensitivity test. The results showed that the most effective chemotherapy regimen was oxaliplatin + gemcitabine. After propensity score matching, a total of 64 patients were enrolled in the following study, including 32 patients receiving individualized chemotherapy guided by PTC drug results as group 1 and 32 patients receiving conventional chemotherapy as group 2. Survival analysis showed that the median OS of group 1 was not reached, significantly longer than that of group 2 (23.5 months) (p < 0.05). Compared with conventional chemotherapy, individualized chemotherapy guided by PTC drug sensitivity tests can prolong patient survival, and oxaliplatin + gemcitabine + apatinib could be the optimal adjuvant treatment regimen for MPM.

Evaluation of Clinical Distribution and Antimicrobial Resistance of Klebsiella Pneumoniae

Purpose: This paper aims to analyze the clinical distribution and drug resistance changes of Klebsiella Pneumoniae (KPN) from 2017 to 2021 in the Beijing Hospital of Integrated Traditional Chinese and Western Medicine to provide a reference for the clinical rational use of antibiotics. Methods: We collected Klebsiella Pneumoniae isolated from various clinical specimens in 2017-2021, analyzed the isolation rate, specimen distribution, and department distribution characteristics during the five years, and statistically analyzed their drug sensitivity tests and multiple drug resistance. Zhuhai Deere DL-96 full-automatic microbial analyzer was used for bacterial identification and drug sensitivity tests. The drug sensitivity test was interpreted according to the standards recommended by the American Clinical and Laboratory Standards Institute (CLSI). Results: A total of 1057 strains of Klebsiella Pneumoniae were identified between 2017 and 2021, with proportions of 18.6%, 15.7%, 15.4%, 15.1%, and 15.0% in each respective year. Specimen distribution included sputum (66.0%), urine (17.9%), throat swab (9.4%), secretion (2.4%), pus (0.7%), venous blood (0.6%), vaginal swab (0.4%), and other sources (2.6%). Distribution by the department revealed specimens originating from the respiratory department (21.2%), cardiology department (17.8%), neurology department (13.4%), oncology department (13.0%), nephrology department (12.2%), acupuncture department (10.1%), and other departments (12.3%). In terms of drug susceptibility testing, Klebsiella Pneumoniae exhibited high resistance rates to ceftriaxone, cefotaxime, ceftazidime, and ampicillin/sulbactam, with rates of 50.8%, 46.8%, 46.3%, and 43.6% respectively. Conversely, resistance rates to minocycline, amikacin, imipenem, and meropenem were relatively low, at 8.6%, 16.5%, 8.5%, and 9.4% respectively. Resistance rates to cefepime/- sulbactam and piperacillin/tazobactam were 29.9% and 28.9%, respectively, while cephalosporin resistance rates ranged from 36.1% to 50.8%. Regarding multidrug resistance, the detection rates of ESBL-producing Klebsiella Pneumoniae were 8.2%, 10.9%, 4.5%, 10.6%, and 6.4% from 2017 to 2021, with an average detection rate of 7.9%. The detection rates of CR-Kp were 12.1%, 11.7%, 5.8%, 9.9%, and 8.9% respectively, averaging 9.6% over the five-year period. Conclusion: The sputum specimen of Klebsiella Pneumoniae exhibits the highest detection rate among specimen distributions, signifying its significance as a pathogenic bacterium in respiratory tract infections. Notably, the respiratory department demonstrates the highest detection rate, underscoring the necessity to enhance the monitoring and management of Klebsiella Pneumoniae infections in respiratory patients. Over the past five years, our hospital has observed a decreasing trend in the overall drug resistance rate of Klebsiella Pneumoniae to 17 antibiotics. While imipenem and meropenem exhibit minimal resistance rates, these carbapenem antibiotics serve as crucial agents for treating gram-negative bacilli, particularly in critically ill patients, and are thus not recommended as first-line choices for routine clinical use. Conversely, minocycline, amikacin, ceftazidime/ sulbactam, and piperacillin/tazobactam showcase relatively low resistance rates, enabling their empirical use based on clinical experience. Combination therapy with other antibiotics is advised for amikacin. Conclusion: Nevertheless, cephalosporins display a relatively high resistance rate, necessitating a reduction in their clinical utilization. Regarding multidrug resistance, the detection of ESBLs-producing Klebsiella Pneumoniae (KP) and Carbapenem-Resistant KP (CR-Kp) has exhibited a declining trend over the past three years. Despite this positive trend, the issue of multidrug resistance in Klebsiella Pneumoniae remains severe, with instances of complete drug resistance reported. Clinicians are urged to judiciously administer antibiotics guided by drug sensitivity test results and resistance rate variations, restrict the use of broad-spectrum antibiotics, and manage the emergence and spread of ESBLs-producing and CR-Kp bacteria.

Analysis of prognosis and drug sensitivity in lung cancer patients based on the integration of 18 programmed cell death modes and single-cell sequencing data

Objective: Programmed cell death (PCD) has therapeutic potential for a variety of malignant tumors, including lung cancer. In this study, we used PCD and bioinformatics to construct a prognostic model for lung cancer and explore new therapeutic strategies. Methods: Multiple bioinformatics algorithms (co-expression analysis, univariate Cox’s analysis, multivariate Cox’s analysis, and cross-validation) were used to screen PCD-related genes and construct a risk model. Lung cancer patients were divided into training and testing groups in a ratio of 7:3. The prognostic model was validated by comparing the risk scores of the high-risk and low-risk groups using receiver operating characteristic (ROC) curves, nomograms, and independent prognostic analyses. In addition, PCD patterns were classified and compared in terms of survival time, immune microenvironment and pathway regulation using consensus clustering methods with the validation of principal component analysis (PCA). Single-cell RNA sequencing (scRNA-seq) analysis was applied to validate screened PCD-related genes in this risk model. Results: Twelve risk genes were identified, including BIK, CDCP1, CHEK2, FADD, GLS2, IL33, ITPRIP, KRT8, MELK, MMP9, PTGIS and TRIB3, to construct prognostic risk model. In the process of lung cancer, the most significantly up-regulated gene was TRIML2, and the most down-regulated gene was GLS2. ROC curves, nomograms, and independent prognostic analyses confirmed the accuracy of risk model to predict the prognosis of lung cancer, indicating that it can be regarded as an independent prognostic model. In the immune cell infiltration, we found that patients with an increased M0 macrophage had a poorer prognosis. Drug sensitivity testing after reliable risk modeling identified three molecularly targeted drugs for lung cancer patients in the high-risk group, namely, Staurosporine, Luminespib and Docetaxel. scRNA-seq results further analyzed the reliability of ITPRIP and KRT8 as prognostic targets. Conclusion: This study identified twelve PCD-related genes and constructed an accurate risk model based on bioinformatics analysis, which can be used for prognostic prediction and design of clinical treatment strategies.

Exploring the Antimicrobial Resistance Profile of Salmonella typhi and Its Clinical Burden.

Background: Typhoid fever caused by Salmonella enterica serovar Typhi (S. typhi) continues to pose a significant risk to public health in developing countries, including Pakistan. This study investigated the epidemiological factors linked to suspected and confirmed S. typhi infections in Peshawar's hospital population. Methodology: A total of 5735 blood samples of patients with suspected enteric fever were collected from September 2022 to November 2023. S. typhi infection was confirmed using microbiological culture of blood samples, biochemical-based tests, and DNA-sequencing methods. Drug sensitivity testing on cultures was conducted as per the CLSI guidelines. Chi-square tests were used to analyze the clinical and epidemiologic characteristics of 5735 samples stratified by S. typhi infection status, and risk factors were assessed by applying logistic regression models to estimate odds ratios (ORs). Results: The number of confirmed typhoid fever cases in this hospital-based study population was 691 (/5735, 12.0%), more prevalent in males (447/3235 13.8%) and children (0-11 years) (429/2747, 15.6%). Compared to children, the risk of S. typhi infection was lower in adolescence (adjusted OR = 0.52; 95% CI: 0.42-0.66), adulthood (19-59 years; aOR = 0.30; 95% CI: 0.25-0.38), and older adulthood (aOR = 0.08; 95% CI: 0.04-0.18) (p < 0.001). Compared to males, the risk of S. typhi infection was lower in females (aOR = 0.67; 95% CI = 0.56-0.80; p = 0.002). Living in a rural residence (compared to urban) was associated with a higher risk of infection (aOR = 1.38; 95% CI: 1.16-1.63; p = 0.001), while access to a groundwater source (compared to municipal water supply) led to a lower risk (aOR = 0.56; 95% CI: 0.43-0.73; p = 0.002). Vaccination demonstrated a robust protective effect (aOR = 0.069; 95% CI = 0.04-0.11, p = 0.002). For those with typhoid infections, clinical biomarker analysis revealed the presence of leucopenia (65/691, 9.4%), thrombocytopenia (130/691, 18.8%), and elevated alanine aminotransferase (ALT) (402/691, 58.2%) and C-reactive protein (CRP) (690/691, 99.9%) levels. Worryingly, among the positive S. typhi isolates, there was a high prevalence of drug resistance (653/691), including multidrug-resistant (MDR 82/691, 11.9%) and extensively drug-resistant types (XDR, 571/691, 82.6%). Conclusions: This study highlights the importance of age, sex, locality, water source, and vaccination status in shaping the epidemiological landscape of S. typhi in the Peshawar district. It implies that expanding vaccination coverage to the broader population of Khyber Pakhtunkhwa province, particularly in the district of Peshawar, would be beneficial.

A study on factors influencing delayed sputum conversion in newly diagnosed pulmonary tuberculosis based on bacteriology and genomics

Conversion of sputum from positive to negative is one of the indicators to evaluate the efficacy of anti-tuberculosis treatment (ATT). We investigate the factors associated with delayed sputum conversion after 2 or 5 months of ATT from the perspectives of bacteriology and genomics. A retrospective study of sputum conversion in sputum positive 1782 pulmonary tuberculosis (PTB) was conducted from 2021 to 2022 in Beijing, China. We also designed a case-matched study including 24 pairs of delayed-sputum-conversion patients (DSCPs) and timely-sputum-conversion patients (TSCPs), and collect clinical isolates from DSCPs before and after ATT and initial isolates of TSCPs who successfully achieved sputum conversion to negative after 2 months of ATT. A total of 75 strains were conducted drug sensitivity testing (DST) of 13 anti-TB drugs and whole-genome sequencing (WGS) to analyze the risk factors of delayed conversion and the dynamics changes of drug resistance and genomics of Mycobacterium tuberculosis (MTB) during ATT. We found TSCPs have better treatment outcomes and whose initial isolates show lower levels of drug resistance. Clinical isolates of DSCPs showed dynamically changing of resistance phenotypes and intra-host heterogeneity. Single nucleotide polymorphism (SNP) profiles showed large differences between groups. The study provided insight into the bacteriological and genomic variation of delayed sputum conversion. It would be helpful for early indication of sputum conversion and guidance on ATT.

Incorporating Tumor Genomic Profiling and Circulating Tumor Cell-derived Organoids into Clinical Practice for Gastrointestinal Cancers.

Precision medicine aims to revolutionize healthcare by tailoring treatment regimens. This study aimed to integrate comprehensive tumor genomic profiling (CTGP) by targeted-gene panel sequencing and drug screening by circulating tumor cell-derived organoids (CTOs) into clinical practice for the treatment of gastrointestinal (GI) cancers. Nine patients with various GI cancers underwent CTGP and CTO drug sensitivity testing. CTGP results guided targeted therapy and immunotherapy, while CTO drug sensitivity predicted response to chemotherapy and targeted agents. The drug recommendations from two platforms were correlated with the treatment response to the suggested medications retrospectively. Five patients received therapies aligned with CTGP, including HER2-targeted treatment, immunotherapy, and BRAF/MEK dual inhibition, showing positive responses. CTO drug sensitivity predicted progression under regorafenib (low potential benefit) and good response to chemotherapy with high potential benefit. The combination of CTGP and CTO drug sensitivity may exhibit significant correlation with clinical outcomes during treatment with candidate drugs, demonstrating a sensitivity of 79% and an accuracy of 75%. This encompasses various treatment modalities, such as chemotherapy, targeted therapy, and immunotherapy. The present investigation elucidated the integration of CTGP and CTO drug sensitivity screening into clinical practice in a complementary manner, showcasing a significant correlation between treatment response and testing outcomes. Additional prospective evaluation of these two testing modalities in a large cohort is warranted to confirm whether the inclusion of CTO drug sensitivity screening confers enhanced survival benefits compared to utilizing CTGP alone.

Profile of drug-resistant tuberculosis post universal drug susceptibility testing in Jabalpur division, Madhya Pradesh: study

Abstract Background: Drug-resistant tuberculosis (DR-TB) can be a menace for TB elimination, making it crucial to identify the drivers of this multifaceted challenge. Now, as drug sensitivity results are available at the commencement of the treatment, DRTB patients can be delineated earlier rather than once landing into treatment failure. Objectives: The study was done to assess the sociodemographic and behavioral profile of DR-TB patients and its association with the drug-resistant pattern following the introduction of universal drug sensitivity testing in the program. Materials and Methods: A cross-sectional study was conducted at the DRTB center, Jabalpur. Record review was done for registered patients through the NIKSHAY portal, notification, laboratory registers, and PMDT treatment cards, for the period January 2018 to December 2019 excluding the transferred-out cases. Patients were contacted for any missing information. Data were analyzed using IBM SPSS version 20. The Chi-square test was applied for statistical significance considering P ≤ 0.05 to be significant at 95% confidence level. Results: Among 201 patients, the majority were males (62.2%) of the younger (18–30 years) age group (42.3%), residing below the poverty line (62.2%) in urban slums (40.8%). There was a preponderance of tobacco users (88.5%), whereas 22.9% were alcoholics. Nutritional status revealed 59.7% to be underweight, whereas 9% were overweight. Review of drug-resistant profile had shown multidrug resistance (MDR) to be most common (70.6%), followed by H resistance (13.4%) and others (MDR + fluoroquinolone [FQ] [10.4%], extensively drug-resistant [3.5%], H + FQ/second-line injectable [SLI]/LZ [1.5%], MDR + SLI [0.5%]). Belonging to lower socioeconomic strata, residing in urban slums, and being engaged in a labor class occupational profile carries a statistically significant association with MDR TB. Conclusion: MDR is the most common pattern of DR TB in the study population, proportionally higher in urban slum dwellers.

Prognostic significance of exportin-5 in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the third leading cause of cancer-related deaths worldwide. As liver cancer often presents no noticeable symptoms in its early stages, most patients are diagnosed at an advanced stage, complicating treatment. Therefore, the identification of new biomarkers is crucial for the early detection and treatment of HCC. Research on exportin-5 (XPO5) could offer new avenues for early diagnosis and improve treatment strategies. To explore the role of XPO5 in HCC progression and its potential as a prognostic biomarker. This study assessed XPO5 mRNA expression in HCC using The Cancer Genome Atlas, TIMER, and International Cancer Genome Consortium databases, correlating it with clinical profiles and disease progression. We performed in vitro experiments to examine the effect of XPO5 on liver cell growth. Gene Set Enrichment Analysis, Kyoto Encyclopedia of Genes and Genomes, and Gene Ontology were used to elucidate the biological roles and signaling pathways. We also evaluated XPO5's impact on immune cell infiltration and validated its prognostic potential using machine learning. XPO5 was significantly upregulated in HCC tissues, correlating with tumor grade, T-stage, and overall survival, indicating poor prognosis. Enrichment analyses linked high XPO5 expression with tumor immunity, particularly CD4 T cell memory activation and macrophage M0 infiltration. Drug sensitivity tests identified potential therapeutic agents such as MG-132, paclitaxel, and WH-4-023. Overexpression of XPO5 in HCC cells, compared to normal liver cells, was confirmed by western blotting and quantitative real-time polymerase chain reaction. The lentiviral transduction-mediated knockdown of XPO5 significantly reduced cell proliferation and metastasis. Among the various machine learning algorithms, the C5.0 decision tree algorithm achieved accuracy rates of 95.5% in the training set and 92.0% in the validation set. Our analysis shows that XPO5 expression is a reliable prognostic indicator for patients with HCC and is significantly associated with immune cell infiltration.

A depression-related lncRNA signature predicts the clinical outcome and immune characteristics of gastric cancer

BackgroundDepression and long non-coding RNA (lncRNA) have been reported to be associated with tumor progression and prognosis in gastric cancer (GC). This study aims to build a GC risk classification and prognosis model based on depression-related lncRNAs (DRLs). MethodsTo develop a risk model, we performed univariate Cox regression and least absolute shrinkage and selection operator (LASSO) regression analyses using RNA sequencing data of GC from The Cancer Genome Atlas (TCGA) and depression-related genes (DRGs) from previous studies. Kaplan-Meier analysis, receiver operating characteristic (ROC) curve analysis, nomogram construction, pathway enrichment analysis, assessment of immunological features, and drug sensitivity testing were conducted using a series of bioinformatics methods. ResultsSeven DRLs were identified to build a prognostic model, whose robustness was verified in an internal cohort. Subsequent prognostic analyses found that high risk scores were associated with worse overall survival (OS). Univariate and multivariate analyses revealed that the risk score could be used as an independent prognostic factor. Furthermore, the ROC curve indicated that the risk score had higher diagnostic efficiency than traditional clinicopathological features. The calibration curve confirmed the accuracy and reliability of the nomogram. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses showed that there were differences in digestive system and nervous system-related pathways between the high- and low-risk groups. Results of tumor mutational burden (TMB) and tumor immune dysfunction and exclusion (TIDE) analyses indicated that patients in the low-risk group had a better response rate to immunotherapy. Finally, the results of drug sensitivity analysis showed that risk score could influence sensitivity to EHT 1864 in GC. ConclusionWe have successfully developed and verified a 7-DRL risk model which can assess the prognosis and immunological features and guide individualized therapy of GC patients.

Hainanenin-1, an oncolytic peptide, triggers immunogenic cell death via STING activation in triple-negative breast cancer

BackgroundIn triple-negative breast cancer (TNBC) therapy, insufficient tumor infiltration by lymphocytes significantly hinders the efficacy of immune checkpoint inhibitors. We have previously demonstrated that Hainanenin-1 (HN-1), a host defense peptide (HDP) identified from Hainan frog skin, induces breast cancer apoptosis and boots anti-tumor immunity via unknown mechanism.MethodsWe used in vitro experiments to observe immunogenic cell death (ICD) indicators in HN-1-treated TNBC cell lines, a mouse tumor model to verify HN-1 promotion of mice anti-tumor immune response, and an in vitro drug sensitivity test of patient-derived breast cancer cells to verify the inhibitory effect of HN-1.ResultsHN-1 induced ICD in TNBC in a process during which damage-associated molecular patterns (DAMPs) were released that could further increase the anti-tumor immune response. The secretion level of interleukin 2 (IL-2), IL-12, and interferon γ in the co-culture supernatant was increased, and dendritic cells (DCs) were activated via a co-culture with HN-1-pretreated TNBC cells. As a result, HN-1 increased the infiltration of anti-tumor immune cells (DCs and T lymphocytes) in the mouse model bearing both 4T1 and EMT6 tumors. Meanwhile, regulatory T cells and myeloid-derived suppressor cells were suppressed. In addition, HN-1 induced DNA damage, and double-strand DNA release in the cytosol was significantly enhanced, indicating that HN-1 might stimulate ICD via activation of STING pathway. The knockdown of STING inhibited HN-1-induced ICD. Of note, HN-1 exhibited inhibitory effects on patient-derived breast cancer cells under three-dimensional culture conditions.ConclusionsCollectively, our study demonstrated that HN-1 could be utilized as a potential compound that might augment immunotherapy effects in patients with TNBC.