Current scenario and challenges of clinical pharmacists to implement pharmaceutical care in DRG/DIP payment hospitals in China: a qualitative interview study.

Purpose: The Diagnosis-Related Group (DRG) or Diagnosis-Intervention Packet (DIP) payment system, now introduced in China, intends to streamline healthcare billing practices. However, its implications for clinical pharmacists, pivotal stakeholders in the healthcare system, remain inadequately explored. This study sought to assess the perceptions, challenges, and roles of clinical pharmacists in China following the introduction of the DRG or DIP payment system. Methods: Qualitative interviews were conducted among a sample of clinical pharmacists. Ten semi-structured interviews were conducted, either online or face to face. Thematic analysis was employed to identify key insights and concerns related to their professional landscape under the DRG or DIP system. Results: Clinical pharmacists exhibited variable awareness levels about the DRG or DIP system. Their roles have undergone shifts, creating a balance between traditional responsibilities and new obligations dictated by the DRG or DIP system. Professional development, particularly concerning health economics and DRG-based or DIP-based patient care, was highlighted as a key need. There were calls for policy support at both healthcare and national levels and a revised, holistic performance assessment system. The demand for more resources, be it in training platforms or personnel, was a recurrent theme. Conclusion: The DRG or DIP system's introduction in China poses both opportunities and challenges for clinical pharmacists. Addressing awareness gaps, offering robust policy support, ensuring adequate resource allocation, and recognizing the evolving role of pharmacists are crucial for harmoniously integrating the DRG or DIP system into the Chinese healthcare paradigm.

A bibliometric and visualized in oral microbiota and cancer research from 2013 to 2022.

Numerous studies have highlighted the implication of oral microbiota in various cancers. However, no bibliometric analysis has been conducted on the relationship between oral microbiota and cancer. This bibliometric analysis aimed to identify the research hotspots in oral microbiota and cancer research, as well as predict future research trends. The literature published relating to oral microbiota and cancer was searched from the Web of Science Core Collection database (WoSCC) from 2013 to 2022. VOSviewer or Citespace software was used to perform the bibliometric analysis, focusing on countries, institutions, authors, journals, keywords and references. A total of 1516 publications were included in the analysis. The number of publications related oral microbiota and cancer increased annually, reaching its peak in 2022 with 287 papers. The United States (456) and China (370) were the countries with the most publications and made significant contributions to the field. Sears CL and Zhou XD were the most productive authors. The high frequency of keywords revealed key topics, including cancer (colorectal cancer, oral cancer), oral microbiota (Fusobacterium nucleatum, Porphyromonas gingivalis), and inflammation (periodontal disease). The latest trend keywords were F. nucleatum, dysbiosis, prognosis, tumor microenvironment, gastric microbiota, complications and survival, suggesting a new hotspot in the field of oral microbiota and cancer. Our study provides a comprehensive analysis of oral microbiota and cancer research, revealing an increase in publications in recent years. Future research directions will continue to focus on the diversity of oral microbiota impacted by cancers and the underlying mechanism connecting them, providing new ideas for targeted therapy of tumorigenesis.

Elucidation of unusual biosynthesis and DnaN-targeting mode of action of potent anti-tuberculosis antibiotics Mycoplanecins.

DNA polymerase III sliding clamp (DnaN) was recently validated as a new anti-tuberculosis target employing griselimycins. Three (2 S,4 R)-4-methylproline moieties of methylgriselimycin play significant roles in target binding and metabolic stability. Here, we identify the mycoplanecin biosynthetic gene cluster by genome mining using bait genes from the 4-methylproline pathway. We isolate and structurally elucidate four mycoplanecins comprising scarce homo-amino acids and 4-alkylprolines. Evaluating mycoplanecin E against Mycobacterium tuberculosis surprisingly reveals an excitingly low minimum inhibition concentration at 83 ng/mL, thus outcompeting griselimycin by approximately 24-fold. We show that mycoplanecins bind DnaN with nanomolar affinity and provide a co-crystal structure of mycoplanecin A-bound DnaN. Additionally, we reconstitute the biosyntheses of the unusual L-homoleucine, L-homonorleucine, and (2 S,4 R)-4-ethylproline building blocks by characterizing in vitro the full set of eight enzymes involved. The biosynthetic study, bioactivity evaluation, and drug target validation of mycoplanecins pave the way for their further development to tackle multidrug-resistant mycobacterial infections.

Knockdown of PGC1α suppresses dysplastic oral keratinocytes proliferation through reprogramming energy metabolism.

Oral potentially malignant disorders (OPMDs) are precursors of oral squamous cell carcinoma (OSCC). Deregulated cellular energy metabolism is a critical hallmark of cancer cells. Peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC1α) plays vital role in mitochondrial energy metabolism. However, the molecular mechanism of PGC1α on OPMDs progression is less unclear. Therefore, we investigated the effects of knockdown PGC1α on human dysplastic oral keratinocytes (DOKs) comprehensively, including cell proliferation, cell cycle, apoptosis, xenograft tumor, mitochondrial DNA (mtDNA), mitochondrial electron transport chain complexes (ETC), reactive oxygen species (ROS), oxygen consumption rate (OCR), extracellular acidification rate (ECAR), and glucose uptake. We found that knockdown PGC1α significantly inhibited the proliferation of DOKs in vitro and tumor growth in vivo, induced S-phase arrest, and suppressed PI3K/Akt signaling pathway without affecting cell apoptosis. Mechanistically, downregulated of PGC1α decreased mtDNA, ETC, and OCR, while enhancing ROS, glucose uptake, ECAR, and glycolysis by regulating lactate dehydrogenase A (LDHA). Moreover, SR18292 (an inhibitor of PGC1α) induced oxidative phosphorylation dysfunction of DOKs and declined DOK xenograft tumor progression. Thus, our work suggests that PGC1α plays a crucial role in cell proliferation by reprograming energy metabolism and interfering with energy metabolism, acting as a potential therapeutic target for OPMDs.

Application of Induced Pluripotent Stem Cells in Malignant Solid Tumors.

In the past decade, induced pluripotent stem cells (iPSCs) technology has significantly progressed in studying malignant solid tumors. This technically feasible reprogramming techniques can reawaken sequestered dormant regions that regulate the fate of differentiated cells. Despite the evolving therapeutic modalities for malignant solid tumors, treatment outcomes have not been satisfactory. Recently, scientists attempted to apply induced pluripotent stem cell technology to cancer research, from modeling to treatment. Induced pluripotent stem cells derived from somatic cells, cancer cell lines, primary tumors, and individuals with an inherited propensity to develop cancer have shown great potential in cancer modeling, cell therapy, immunotherapy, and understanding tumor progression. This review summarizes the evolution of induced pluripotent stem cells technology and its applications in malignant solid tumor. Additionally, we discuss potential obstacles to induced pluripotent stem cell technology.

Establishment of an orthotopic model of lung cancer by transthoracic lung puncture using tumor fragments.

Background: Orthotopic models of lung cancer have been widely utilized, and the purpose of this study was to demonstrate the viability of our proposed modified modeling approach. Methods: A total of 50 female BALB/c mice were implanted with 1×1×1 mm fragments of a tumor sample into the left lung lobe. After 2 months of observation, the mice were humanely euthanized through CO2 inhalation. The macroscopic specimens were photographed, and the most representative neoplastic lesions were collected for histological analysis. Small-animal positron emission tomography/computed tomography (PET/CT) scans were conducted on 6 randomly selected mice. Results: Local tumor formation, ipsilateral thoracic tissue infiltration, the contralateral chest wall, right lung metastases, and distant kidney metastases were observed in these models. Overall, the tumor development and metastasis rates were 60.86% (28/46) and 57.14% (16/28), respectively. The 3 mice that had a small-animal PET/CT scan developed a local tumor, but no distant metastases were observed. Conclusions: This modified method was deemed reliable, reproducible, minimally invasive, straightforward, and comprehensible; it might serve as the foundation for developing patient-derived orthotopic xenografts of lung cancer.

Impact of endoscopic thoracic R4 sympathicotomy combined with R3 ramicotomy for primary palmar hyperhidrosis.

Background: Endoscopic thoracoscopic sympathectomy (ETS) is the preferred method for treating primary palmar hyperhidrosis (PPH) that bears the risk of compensatory hyperhidrosis (CH) following surgery. The current study aims to evaluate the effectiveness and safety of an innovative surgical procedure of ETS. Methods: A survey of the clinical data of 109 patients with PPH who underwent ETS in our department from May 2018 to August 2021 was retrospectively conducted. The patients were organized into two groups. Group A underwent R4 sympathicotomy combined with R3 ramicotomy. Group B underwent R3 sympathicotomy. Patients were followed up to evaluate the safety, effectiveness and the incidence of postoperative CH of the modified surgical approach. Results: A total of 102 patients completed follow-up, and seven of the total enrolled patients were lost to follow-up, with a loss rate of 6% (7/109). Among these, Group A constitutes 54 cases, group B constitutes 48 cases, and the mean follow-up was 14 months (interquartile range 12-23 months). There was no statistically difference in surgical safety, postoperative efficacy, and postoperative quality of life (QoL) score between group A and group B (p > 0.05). The score of the psychological assessment was higher (p = 0.004) in group A (14.15 ± 2.06) compared to group B (13.30 ± 1.86). The incidence of CH in group A was lower than in group B (p = 0.019). Conclusion: R4 sympathicotomy combined with R3 ramicotomy is safe and effective for PPH treatment, along with a reduced incidence of postoperative CH rate and improved postoperative psychological satisfaction.

Investigation of oral health knowledge and attitudes towards oral health education among elementary school teachers in Zunyi.

OBJECTIVES: This study aimed to investigate the oral health knowledge of elementary school teachers and assess their attitude towards oral health education in Zunyi. METHODS: A total of 636 teachers from 10 primary schools in Zunyi were selected by stratified sampling, and their general information, oral health care habits, results of oral health knowledge questionnaire, and attitude towards oral health and oral health education were investigated. Data were statistically analyzed using SPSS 21.0. RESULTS: A total of 614 teachers answered the questionnaires. Only 8.8% brush their teeth for more than three minutes, 23.8% brush their teeth horizontally, 64.7% do not performteeth cleaning, and 78.2% do not use floss. Teachers have a weak understanding that six-year teeth are permanent, that pit and fissure sealing could prevent dental caries, and that dental floss could remove dental plaque. However, their attitude towards oral health and oral health education was found to be good. CONCLUSIONS: Schools could improve teachers' oral health know-ledge by organizing training and other activities. Teachers could also play an active role in leading and cultivating school-age children to establish good oral habits.

Photonic ADC-based scheme for joint wireless communication and radar by adopting a broadband OFDM shared signal.

We propose and demonstrate a novel, to the best of our knowledge, joint wireless communication and radar system based on a photonic analog-to-digital converter (PADC), which can receive broadband radio-frequency (RF) signals. Owing to this property, a broadband orthogonal frequency division multiplexing (OFDM) shared signal, which owns obvious advantages in communication applications, can be adopted to realize efficient data communication and high-performance target detection simultaneously. In the experiment, a communication rate of 6 Gbit/s is achieved. Inverse synthetic aperture radar (ISAR) imaging is demonstrated with a two-dimensional (2D) resolution of 3.97 cm × 2.94 cm. Finally, it is verified that high-accuracy radial resolution and high-speed communication can be maintained while increasing the pulse repetition period to detect remote target at around 374.6 m.

Automatic target recognition based on receiver optimization of photonic time-stretched coherent radar.

We demonstrate an automatic target recognition (ATR) scheme based on an improved photonic time-stretched coherent radar (PTS-CR). The reception apertures of the PTS-CR can cover the entire detection range by receiving the echo signal with high repetition rate pulses and increasing the amount of dispersion of the first dispersive medium in the receiver. Two channels with different stretching factors are simultaneously used to restore the signal delay information. Simulated and experimental results verify the feasibility of the new scheme. Finally, based on the improved receiving scheme, PTS-CR successfully performed ATR on four different targets placed on a rotating stage. Combining this with the training of the convolutional neural network (CNN), the recognition accuracy rate is 94.375%.

Self-Assembled Nanoparticles for Tumor-Triggered Targeting Dual-Mode NIRF/MR Imaging and Photodynamic Therapy Applications.

In this study, the self-assembling strategy was used to synthesize a therapeutic and diagnostic nanosystem for tumor-triggered targeting dual-mode near-infrared fluorescence (NIRF)/magnetic resonance (MR) imaging and photodynamic therapy applications. This theranostic nanosystem was synthesized based on the self-assembling of the short peptide (PLGVRGRGDC) and the gadolinium chelator (diethylenetriamine pentaacetic acid) functionalized amphiphilic DSPE-PEG2000, followed by loading with the insoluble photosensitizer therapeutic agent chlorin e6 (Ce6). The formed theranostic nanosystem can accumulate in the matrix metalloproteinase 2 (MMP2) rich tumor sites guided by the enhanced permeability and retention effect and MMP2-substrate peptide (PLGVR) targeting. After PLGVR was hydrolyzed in the tumor microenvironment by MMP2, the nanosystem was actively taken up by tumor cells via Arg-Gly-Asp (RGD) peptide-mediated internalization. With the coexistence of gadolinium and Ce6, the formed nanosystem can be used for both NIRF/MR dual-mode imaging and photodynamic therapy. These tumor-triggered targeting self-assembled nanoparticles with low cytotoxicity and high endocytosis efficiency can efficiently induce A549 cancer cell apoptosis under laser irradiation. Meanwhile, they possessed enhanced tumor-targeted NIRF/MR imaging ability and efficiently inhibited tumor growth with minimal side effects in mice bearing A549 lung cancer. Therefore, these self-assembled theranostic nanoparticles may have great potential for cancer clinical diagnosis and therapy.

Efficacy of probiotics in the management of halitosis: a systematic review and meta-analysis.

BACKGROUND: Halitosis is defined as a foul odour emitted from the oral cavity. Many interventions have been used to control halitosis from mouthwashes to chewing gums. Probiotics have been reported as an alternative method to alleviate halitosis. OBJECTIVE: The present study aimed to investigate the effect of probiotics on halitosis from a time perspective. DESIGN AND METHODS: This is a meta-analysis study performed in indexed databases up to February 2021. Randomised controlled trials that compared the effects of probiotics and placebo on primary outcomes (organoleptic (OLP) scores and volatile sulfur compound (VSC) levels) and secondary outcomes (tongue coating scores (TCS) and plaque index (PI)) were included. Data extraction and quality assessment were conducted independently by two reviewers. Publication bias and leave-one-out analyses were performed. RESULTS: The standardised mean difference (SMD) and 95% CI were calculated to synthesise data. The data were subgrouped and analysed in the short term (≤4 weeks) and long term (>4 weeks) based on the follow-up time. Seven articles were included in this meta-analysis. The primary outcomes, OLP scores (SMD=-0.58; 95% CI -0.87 to -0.30, p<0.0001) and VSC levels (SMD=-0.26; 95% CI -0.51 to -0.01, p=0.04), both decreased significantly in the probiotics group compared with the placebo group in the short term. However, a significant reduction was observed only in OLP scores (SMD=-0.45; 95% CI -0.85 to -0.04, p=0.03) in the long term. No significant differences were observed in secondary outcomes. There was no evidence of publication bias. The leave-one-out analysis confirmed that the pooled estimate was stable. CONCLUSIONS: According to the results of this work, it seems that probiotics (eg, Lactobacillus salivarius, Lactobacillus reuteri, Streptococcus salivarius and Weissella cibaria) may relieve halitosis in the short term (≤4 weeks). The results of the biased assessment, limited data and heterogeneity of the clinical trials included might reduce the reliability of the conclusions.

Mucin1 relieves acute lung injury by inhibiting inflammation and oxidative stress.

Acute lung injury/acute respiratory distress syndrome (ALI/ARDS) is a kind of diffuse inflammatory injury caused by various factors, characterized by respiratory distress and progressive hypoxemia. It is a common clinical critical illness. The aim of this study was to investigate the effect and mechanism of the Mucin1 (MUC1) gene and its recombinant protein on lipopolysaccharide (LPS)-induced ALI/ARDS. We cultured human alveolar epithelial cell line (BEAS-2B) and used MUC1 overexpression lentivirus to detect the effect of MUC1 gene on BEAS-2B cells. In addition, we used LPS to induce ALI/ARDS in C57/BL6 mice and use hematoxylin and eosin (H&E) staining to verify the effect of their modeling. Recombinant MUC1 protein was injected subcutaneously into mice. We examined the effect of MUC1 on ALI/ARDS in mice by detecting the expression of inflammatory factors and oxidative stress molecules in mouse lung tissue, bronchoalveolar lavage fluid (BALF) and serum. Overexpression of MUC1 effectively ameliorated LPS-induced damage to BEAS-2B cells. Results of H&E staining indicate that LPS successfully induced ALI/ARDS in mice and MUC1 attenuated lung injury. MUC1 also reduced the expression of inflammatory factors (IL-1ß, TNF-α, IL-6 and IL-8) and oxidative stress levels in mice. In addition, LPS results in an increase in the activity of the TLR4/NF-κB signaling pathway in mice, whereas MUC1 decreased the expression of the TLR4/NF-κB signaling pathway. MUC1 inhibited the activity of TLR4/NF-κB signaling pathway and reduced the level of inflammation and oxidative stress in lung tissue of ALI mice.

Near-infrared light-responsive hybrid hydrogels for the synergistic chemo-photothermal therapy of oral cancer.

Light-stimulus-responsive therapies have been recognized as a promising strategy for the efficient and safe treatment of oral squamous cell carcinoma (OSCC). Hydrogels have emerged as a promising multifunctional platform combining localized drug delivery and sustained drug release with multimodal properties for combined OSCC therapy. However, inaccurate drug release and limited light-absorption efficiency have hindered their on-demand chemo-photothermal applications. To tackle these problems, an injectable and near-infrared (NIR) light-responsive hybrid system was developed by incorporating light-responsive mesoporous silica nanoparticles (MSNs) as doxorubicin (DOX) carriers into the IR820/methylcellulose hydrogel networks for chemophotothermal therapy. Under NIR radiation, the incorporated IR820, a new green cyanine dye, was excited to induce photothermal effects against tumor cells. Meanwhile, MSNs achieved self-degradation-controlled DOX release via the cleavage of diselenide bonds induced by reactive oxygen species. Through the combination of chemotherapy and phototherapy, a long-lasting synergistic anti-tumor effect was achieved in vitro and in vivo with less toxicity. These findings demonstrate the potential of light-responsive hydrogels as a multifunctional platform for accurate synergistic chemophotothermal treatment of OSCC.

A salivary microbiome-based auxiliary diagnostic model for type 2 diabetes mellitus.

OBJECTIVE: Studies have shown that oral microbiota composition is altered in type 2 diabetes mellitus, implying that it is a potential biomarker for diabetes. This study aimed at constructing a noninvasive auxiliary diagnostic model for diabetes based on differences in the salivary microbial community. DESIGN: Salivary microbiota from 24 treatment-naive type 2 diabetes mellitus patients and 21 healthy populations were detected through 16S rRNA gene sequencing, targeting the V3/V4 region using the MiSeq platform. Salivary microbiome diversity and composition were analyzed so as to establish a diagnostic model for type 2 diabetes. RESULTS: Salivary microbiome for treatment-naive type 2 diabetes mellitus patients was imbalanced with certain taxa, including Slackia, Mitsuokella, Abiotrophia, and Parascardovia that being significantly dominant, while the abundance of Moraxella was high in healthy controls. Diabetic patients exhibited varying levels of Prevotella nanceiensis and Prevotella melaninogenica which were negatively correlated with glycosylated hemoglobin and fasting blood glucose levels, as well as fasting blood glucose levels, respectively. Based on differences in salivary microbiome composition between diabetic and healthy groups, we developed a diagnostic model that can be used for the auxiliary diagnosis of type 2 diabetes mellitus with an accuracy of 80 %. CONCLUSIONS: These findings elucidate on the differences in salivary microbiome compositions between type 2 diabetic and non-diabetic populations, and the diagnostic model provides a promising approach for the noninvasive auxiliary diagnosis of diabetes mellitus.

[Potential application of human microbiomes in the diagnosis and treatment of type 2 diabetes mellitus].

Human microbiome refers to the total microorganism genetic information of human body surface and internal, which is closely related to human health and disease. Oral and gut microbiomes are the most diverse microbial communities, which can interact and play a role in the development of the disease, and can reflect the health and disease state in real time. Type 2 diabetes mellitus is a metabolic disorder caused by both genetic and environmental factors. Recent research has shown a link between microbes and diabetes. This article reviewed the latest research on the changes of oral and gut microbiomes in type 2 diabetes mellitus patients, which expects to provide a reference for exploring the development of the disease model for prediction, diagnosis and prognosis of type 2 diabetes mellitus based on human microbiome characteristics.

Effect of audiovisual distraction on the management of dental anxiety in children: A systematic review.

BACKGROUND: Audiovisual distraction, a non-pharmacological intervention, has been used to manage dental anxiety in prior clinical trials. AIM: Synthesize the available evidences to evaluate the efficacy of audiovisual distraction techniques on the management of dental anxiety in children. DESIGN: Electronic databases (PubMed, Cochrane Central Register of Controlled Trials, and Embase) were searched. We included randomized controlled trials (RCTs), and methodological quality of included trials was assessed using the Cochrane Collaboration's criteria. Information on reported anxiety, pain, behaviors, vital signs (including blood pressure, oxygen saturation, and pulse rate), and children satisfaction was analyzed. RESULTS: Nine studies were included for a systematic review, and none of them had low risk of bias. Significant differences in anxiety were found. According to the study, a majority of results indicated a significant difference in pain and behavior between the audiovisual and control group. Three studies reported children in the audiovisual group preferred usage of an audiovisual device for future dental visits. No significant differences could be found regarding blood pressure. CONCLUSIONS: There is some low-quality evidence suggesting that the usage of audiovisual distraction during dental treatment may relieve children's dental anxiety.

In Vitro CRISPR/Cas9 System for Efficient Targeted DNA Editing.

UNLABELLED: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, an RNA-guided nuclease for specific genome editing in vivo, has been adopted in a wide variety of organisms. In contrast, the in vitro application of the CRISPR/Cas9 system has rarely been reported. We present here a highly efficient in vitro CRISPR/Cas9-mediated editing (ICE) system that allows specific refactoring of biosynthetic gene clusters in Streptomyces bacteria and other large DNA fragments. Cleavage by Cas9 of circular pUC18 DNA was investigated here as a simple model, revealing that the 3'→5' exonuclease activity of Cas9 generates errors with 5 to 14 nucleotides (nt) randomly missing at the editing joint. T4 DNA polymerase was then used to repair the Cas9-generated sticky ends, giving substantial improvement in editing accuracy. Plasmid pYH285 and cosmid 10A3, harboring a complete biosynthetic gene cluster for the antibiotics RK-682 and holomycin, respectively, were subjected to the ICE system to delete the rkD and homE genes in frame. Specific insertion of the ampicillin resistance gene (bla) into pYH285 was also successfully performed. These results reveal the ICE system to be a rapid, seamless, and highly efficient way to edit DNA fragments, and a powerful new tool for investigating and engineering biosynthetic gene clusters. IMPORTANCE: Recent improvements in cloning strategies for biosynthetic gene clusters promise rapid advances in understanding and exploiting natural products in the environment. For manipulation of such biosynthetic gene clusters to generate valuable bioactive compounds, efficient and specific gene editing of these large DNA fragments is required. In this study, a highly efficient in vitro DNA editing system has been established. When combined with end repair using T4 DNA polymerase, Cas9 precisely and seamlessly catalyzes targeted editing, including in-frame deletion or insertion of the gene(s) of interest. This in vitro CRISPR editing (ICE) system promises a step forward in our ability to engineer biosynthetic pathways.

Highly efficient editing of the actinorhodin polyketide chain length factor gene in Streptomyces coelicolor M145 using CRISPR/Cas9-CodA(sm) combined system.

The current diminishing returns in finding useful antibiotics and the occurrence of drug-resistant bacteria call for the need to find new antibiotics. Moreover, the whole genome sequencing revealed that the biosynthetic potential of Streptomyces, which has produced the highest numbers of approved and clinical-trial drugs, has been greatly underestimated. Considering the known gene editing toolkits were arduous and inefficient, novel and efficient gene editing system are desirable. Here, we developed an engineered CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein) combined with the counterselection system CodA(sm), the D314A mutant of cytosine deaminase, to rapidly and effectively edit Streptomyces genomes. In-frame deletion of the actinorhodin polyketide chain length factor gene actI-ORF2 was created in Streptomyces coelicolor M145 as an illustration. This CRISPR/Cas9-CodA(sm) combined system strikingly increased the frequency of unmarked mutants and shortened the time required to generate them. We foresee the system becoming a routine laboratory technique for genome editing to exploit the great biosynthetic potential of Streptomyces and perhaps for other medically and economically important actinomycetes.