Constructing an efficient and easy-to-operate multigene expression system is currently a crucial part of plant genetic engineering. In this study, a fragment carrying three independent gene expression cassettes and the expression unit of the gene-silencing suppressor protein(RNA silencing suppressor 19 kDa protein, P19) simultaneously was designed and constructed. This fragment was cloned into the commonly used plant expression vector pCAMBIA300, and the plasmid pC1300-TP2-P19 was obtained. Each gene expression cassette consists of different promoters, fusion tags, and terminators. The target gene can be flexibly inserted into the corresponding site through enzymatic digestion and ligation or recombination and fused with different protein tags, which provides great convenience for subsequent detection. The enhanced green fluorescent protein(eGFP) reporter gene was individually constructed into each expression cassette to verify the feasibility of this vector system. The results of tobacco transient expression and laser-confocal microscopy showed that each expression cassette presented independent and normal expression. Meanwhile, the three key enzyme genes in the betanin synthesis pathway, BvCYP76AD, BvDODA1, and DbDOPA5GT, were constructed into the three expression cassettes. The results of tobacco transient expression phenotype, protein immunoblotting(Western blot), and chemical detection of product demonstrated that the three exogenous genes were highly expressed, and the target compound betanin was successfully produced. The above results indicated that the constructed multigene expression system for plants in this study was efficient and reliable and can achieve the co-transformation of multiple plant genes. It can provide a reliable vector platform for the analysis of plant natural product synthesis pathways, functional verification, and plant metabolic engineering.
Nicotiana/metabolism* ; Genetic Vectors/metabolism* ; Gene Expression Regulation, Plant ; Plant Proteins/metabolism* ; Plants, Genetically Modified/metabolism* ; Genetic Engineering/methods* ; Green Fluorescent Proteins/metabolism* ; Gene Expression

Colorectal cancer(CRC) is one of the most common malignant tumors worldwide, primarily originating from recurrent inflammatory bowel disease(IBD). Therefore, blocking the inflammation-cancer transformation in the colon has become a focus in the early prevention and treatment of CRC. The inflammation-cancer transformation in the colon involves multiple types of cells and complex pathological processes, including inflammatory responses and tumorigenesis. In this complex pathological process, immune cells(including non-specific and specific immune cells) and non-immune cells(such as tumor cells and fibroblasts) interact with each other, collectively promoting the progression of the disease. In traditional Chinese medicine(TCM), inflammation-cancer transformation in the colon belongs to the categories of dysentery and diarrhea, with the main pathogenesis being cold and heat in complexity. This paper first elaborates on the complex molecular mechanisms involved in the inflammation-cancer transformation process in the colon from the perspectives of inflammation, cancer, and their mutual influences. Subsequently, by comparing the pathogenic characteristics and clinical manifestations between inflammation-cancer transformation and the TCM pathogenesis of cold and heat in complexity, this paper explores the intrinsic connections between the two. Furthermore, based on the correlation between inflammation-cancer transformation in the colon and the TCM pathogenesis, this paper delves into the importance of the interaction between inflammation and cancer. Finally, it summarizes and discusses the clinical and basic research progress in the TCM intervention in the inflammation-cancer transformation process, providing a theoretical basis and treatment strategy for the treatment of CRC with integrated traditional Chinese and Western medicine.
Humans ; Colon/pathology* ; Integrative Medicine ; Animals ; Cold Temperature ; Cell Transformation, Neoplastic/drug effects* ; Medicine, Chinese Traditional ; Hot Temperature ; Inflammation ; Drugs, Chinese Herbal/therapeutic use* ; Colonic Neoplasms/drug therapy*

OBJECTIVE: To observe the clinical effect of acupuncture at yinsanzhen combined with auricular point sticking on primary dysmenorrhea (PDM). METHODS: Sixty patients with PDM were randomly divided into an observation group and a control group, with 30 cases in each group. Patients in the observation group were treated with acupuncture at yinsanzhen combined with auricular point sticking. The acupuncture was given at yinsanzhen (Guanyuan [CV4] and bilateral Guilai [ST29], Sanyinjiao [SP6]) once daily for 5 consecutive days. Auricular point sticking was applied to gan (CO₁₂), shen (CO₁₀), neifenmi (CO₁₈), etc. every other day, alternated between ears, totaling 3 sessions. All treatments were started 5 days before menstruation. Patients in the control group were treated with ibuprofen sustained-release capsules on the first day of menstruation for 3 consecutive days. Both groups were treated for 3 menstrual cycles. The scores of Cox menstrual symptom scale (CMSS) and visual analogue scale (VAS) were compared between the two groups before and after treatment and at the second menstrual cycle after treatment completion (follow-up). The serum contents of prostaglandin (PG) F2α and PGE2 were detected before and after treatment, and the clinical effect and safety of the two groups were evaluated. RESULTS: After treatment and during follow-up, the CMSS severity and duration scores and VAS scores of the two groups were lower than those before treatment (P<0.05 ), and the scores in the observation group were lower than those in the control group (P<0.05). After treatment, the serum contents of PGF2α were decreased, and the contents of PGE2 were increased (P<0.05) in the two groups. The total effective rate of the observation group was 93.3% (28/30), which was higher than 80.0% (24/30) of the control group (P<0.05). There were no adverse reactions in both groups. CONCLUSION Yinsanzhen combined with auricular point sticking can effectively improve the pain symptoms, relieve the degree of pain and shorten the duration of pain in patients with PDM, which may play a therapeutic role by reducing the content of serum PGF2α and increasing the content of serum PGE2.
Humans ; Female ; Dysmenorrhea/therapy* ; Acupuncture Points ; Adult ; Young Adult ; Acupuncture, Ear ; Treatment Outcome ; Adolescent ; Acupuncture Therapy ; Combined Modality Therapy

Salvia miltiorrhiza is a perennial herb of the genus Salvia(Lamiaceae). As one of the earliest medicinal plants to undergo molecular biology research, it has gradually become a model plant for molecular biology of medicinal plants. With the gradual analysis of the genome of S. miltiorrhiza and the biosynthetic pathways of its main active components tanshinone and salvianolic acids, the transcriptional regulation mediated by transcription factors and related regulatory proteins has gradually become a new research focus. Due to the lack of scientific and unified naming of transcription factors and different research indexes in different literature, this paper systematically sorted out the transcription factors in different literature with the genomes of DSS3 from selfing for three generations and bh2-7 from selfing for six generations as reference. In total, 73 transcription factors and related regulatory proteins belonging to 13 gene families were identified. The effects of overexpression or gene silencing experiments on tanshinone and salvianolic acids were also analyzed. This study unified the identified transcription factors, which laid a foundation for further constructing the regulatory networks of secondary metabolites and insect or stress resistance and improving the quality of medicinal materials by using global transcriptional regulation engineering.
Salvia miltiorrhiza/chemistry* ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant ; Transcription Factors/metabolism* ; Abietanes/metabolism*

The prevalence of Class III malocclusion varies among different countries and regions. The populations from Southeast Asian countries (Chinese and Malaysian) showed the highest prevalence rate of 15.8%, which can seriously affect oral function, facial appearance, and mental health. As anterior crossbite tends to worsen with growth, early orthodontic treatment can harness growth potential to normalize maxillofacial development or reduce skeletal malformation severity, thereby reducing the difficulty and shortening the treatment cycle of later-stage treatment. This is beneficial for the physical and mental growth of children. Therefore, early orthodontic treatment for Class III malocclusion is particularly important. Determining the optimal timing for early orthodontic treatment requires a comprehensive assessment of clinical manifestations, dental age, and skeletal age, and can lead to better results with less effort. Currently, standardized treatment guidelines for early orthodontic treatment of Class III malocclusion are lacking. This review provides a comprehensive summary of the etiology, clinical manifestations, classification, and early orthodontic techniques for Class III malocclusion, along with systematic discussions on selecting early treatment plans. The purpose of this expert consensus is to standardize clinical practices and improve the treatment outcomes of Class III malocclusion through early orthodontic treatment.
Humans ; Malocclusion, Angle Class III/classification* ; Orthodontics, Corrective/methods* ; Consensus ; Child

Cemental tear is a rare and indetectable condition unless obvious clinical signs present with the involvement of surrounding periodontal and periapical tissues. Due to its clinical manifestations similar to common dental issues, such as vertical root fracture, primary endodontic diseases, and periodontal diseases, as well as the low awareness of cemental tear for clinicians, misdiagnosis often occurs. The critical principle for cemental tear treatment is to remove torn fragments, and overlooking fragments leads to futile therapy, which could deteriorate the conditions of the affected teeth. Therefore, accurate diagnosis and subsequent appropriate interventions are vital for managing cemental tear. Novel diagnostic tools, including cone-beam computed tomography (CBCT), microscopes, and enamel matrix derivatives, have improved early detection and management, enhancing tooth retention. The implementation of standardized diagnostic criteria and treatment protocols, combined with improved clinical awareness among dental professionals, serves to mitigate risks of diagnostic errors and suboptimal therapeutic interventions. This expert consensus reviewed the epidemiology, pathogenesis, potential predisposing factors, clinical manifestations, diagnosis, differential diagnosis, treatment, and prognosis of cemental tear, aiming to provide a clinical guideline and facilitate clinicians to have a better understanding of cemental tear.
Humans ; Dental Cementum/injuries* ; Consensus ; Diagnosis, Differential ; Cone-Beam Computed Tomography ; Tooth Fractures/therapy*

Although surgical resection remains to be the best treatment strategy for stageⅠnon-small cell lung cancer (NSCLC), percutaneous thermal ablation offers an important option for patients who are unable to undergo surgical resection. Currently, there are three main thermal ablation methods used in the treatment of lung cancer, including radiofrequency ablation (RFA), microwave ablation (MWA) and argon-helium cryoablation (AHC). With the improvement of technique and the accumulation of experience in the treatment of lung cancer, some limitations are disclosed in the initial application of RFA, such as heat sink effect, skin burns and rapid carbonization. These shortcomings have been overcome in the development of MWA and AHC. The feasibility and safety of thermal ablation for the treatment of lung cancer has been demonstrated and its efficacy has been significantly improved (especially for the tumour diameter≤3 cm). This article will focus on the application and recent research developments of these ablation techniques in the treatment of lung cancer.

Objective To evaluate the short-term outcomes and hospital costs of robot-assisted thoracic surgery (RATS) versus video-assisted thoracoscopic surgery (VATS) for non-small cell lung cancer. Methods The clinical data of patients who underwent lobectomy or sublobar resection for non-small cell lung cancer completed by the same operator in the Department of Thoracic Surgery, Gansu Provincial Hospital from June 2016 to June 2022 were retrospectively analyzed. According to the surgery approach, the patients were divided into a RATS group and a VATS group. The clinical data of the two groups were compared. Results A total of 516 patients were enrolled. There were 254 patients in the RATS group, including 103 males and 151 females, with a mean age of 60.0±4.2 years, and 262 patients in the VATS group, including 126 males and 136 females, with a mean age of 59.5±4.3 years. All patients in both groups successfully completed radical lung cancer surgery with no perioperative death. In terms of intraoperative bleeding (63.4±13.3 mL vs. 92.5±23.5 mL), postoperative drainage time (4.1±0.9 d vs. 4.7±1.2 d), postoperative hospital stay time (5.6±1.1 d vs. 6.7±1.4 d), number of lymph nodes dissected (17.9±2.1 vs. 13.9±1.4) and groups of lymph nodes dissected (5.4±0.8 groups vs. 4.4±1.0 groups), the RATS group had an advantage, and the difference was statistically significant (P<0.05). In terms of operative time and total postoperative chest drainage, the VATS group had an advantage, and the difference was statistically significant (P<0.05). There was no statistical difference between the two groups in terms of postoperative complications or intraoperative conversion to thoractomy (P>0.05). The total hospitalization, surgical and total consumables costs of the RATS group were higher than those in the VATS group (P<0.05). In terms of other costs and consumables costs (one-time costs of purely high-value consumables after deduction of robot-specific costs), the VATS group was higher than the RATS group (P<0.05). Conclusion RATS offers technical and short-term efficacy advantages, but comes with the disadvantage of high costs. Thoracic surgeons can make full use of the features of the robotic surgery system, exploiting its potential to continuously improve and optimize techniques and reduce the use of high-value consumables, thus achieving efficiency and cost reductions and allowing robotic surgery to reach more patients.

Objective To systematically review the efficacy and safety of cap-assisted endoscopy in the treatment of esophageal foreign bodies. Methods PubMed, Web of Science, The Cochrane Library, EMbase, CNKI and Wanfang databases were searched by computer for relevant literatures on cap-assisted endoscopy and traditional endoscopy for esophageal foreign bodies from inception to November 2022. The quality assessment of the literature was conducted using the Newcastle-Ottawa Scale (NOS). Meta-analysis was performed using RevMan 5.4.1. Results Finally, 27 studies were collected, including 17 randomized controlled trials, 2 cohort studies and 8 case-control studies, with a total of 3 619 patients. NOS scores of all studies were ≥7 points. Meta-analysis results showed that compared with traditional endoscopic treatment, the success rate of removing esophageal foreign bodies in the cap-assisted endoscopy group was higher (OR=14.43, 95%CI 10.64 to 19.55, P<0.000 1), postoperative complications were fewer (OR=0.30, 95%CI 0.23 to 0.38, P<0.000 1), patients' tolerance was better (OR=4.07, 95%CI 2.95 to 5.60, P<0.000 1), intraoperative visual field clarity was better (OR=12.00, 95%CI 7.29 to 19.76, P<0.000 1) and operative time was shorter (SMD=−1.83, 95%CI −2.31 to −1.34, P<0.000 1). Conclusion Cap-assisted endoscopy for esophageal foreign bodies is an effective and safe method, worthy of further promotion and application in clinical practice.

Objective Tissue uptake and distribution of nano-/microplastics was studied at a single high dose by gavage in vivo.Methods Fluorescent microspheres (100 nm, 3 μm, and 10 μm) were given once at a dose of 200 mg/(kg·body weight). The fluorescence intensity (FI) in observed organs was measured using the IVIS Spectrum at 0.5, 1, 2, and 4 h after administration. Histopathology was performed to corroborate these findings.Results In the 100 nm group, the FI of the stomach and small intestine were highest at 0.5 h, and the FI of the large intestine, excrement, lung, kidney, liver, and skeletal muscles were highest at 4 h compared with the control group (P < 0.05). In the 3 μm group, the FI only increased in the lung at 2 h (P < 0.05). In the 10 μm group, the FI increased in the large intestine and excrement at 2 h, and in the kidney at 4 h (P < 0.05). The presence of nano-/microplastics in tissues was further verified by histopathology. The peak time of nanoplastic absorption in blood was confirmed.Conclusion Nanoplastics translocated rapidly to observed organs/tissues through blood circulation;however, only small amounts of MPs could penetrate the organs.