OBJECTIVE: CRISPR-Cas protects bacteria from exogenous DNA invasion and is associated with bacterial biofilm formation and pathogenicity. METHODS: We analyzed the type I-F CRISPR-Cas system of Legionella pneumophila WX48, including Cas1, Cas2-Cas3, Csy1, Csy2, Csy3, and Cas6f, along with downstream CRISPR arrays. We explored the effects of the CRISPR-Cas system on the in vitro growth, biofilm-forming ability, and pathogenicity of L. pneumophila through constructing gene deletion mutants. RESULTS: The type I-F CRISPR-Cas system did not affect the in vitro growth of wild-type or mutant strains. The biofilm formation and intracellular proliferation of the mutant strains were weaker than those of the wild type owing to the regulation of type IV pili and Dot/Icm type IV secretion systems. In particular, Cas6f deletion strongly inhibited these processes. CONCLUSION The type I-F CRISPR-Cas system may reduce biofilm formation and intracellular proliferation in L. pneumophila.
Legionella pneumophila/pathogenicity* ; CRISPR-Cas Systems ; Biofilms/growth & development* ; Phenotype ; Bacterial Proteins/metabolism* ; Gene Deletion

Objective To observe the efficacy and safety of Morinda officinalis oligosaccharides in the continuation treatment of mild and moderate depression.Methods An open,single-arm,multi-center design was adopted in our study.Adult patients with mild and moderate depression who had received acute treatment of Morinda officinalis oligosaccharides were enrolled and continue to receive Morinda officinalis oligosaccharides capsules for 24 weeks,the dose remained unchanged during continuation treatment.The remission rate,recurrence rate,recurrence time,and the change from baseline to endpoint of Hamilton Depression Scale(HAMD),Hamilton Anxiety Scale(HAMA),Clinical Global Impression-Severity(CGI-S)and Arizona Sexual Experience Scale(ASEX)were evaluated.The incidence of treatment-related adverse events was reported.Results The scores of HAMD-17 at baseline and after treatment were 6.60±1.87 and 5.85±4.18,scores of HAMA were 6.36±3.02 and 4.93±3.09,scores of CGI-S were 1.49±0.56 and 1.29±0.81,scores of ASEX were 15.92±4.72 and 15.57±5.26,with significant difference(P＜0.05).After continuation treatment,the remission rate was 54.59％(202 cases/370 cases),and the recurrence rate was 6.49％(24 cases/370 cases),the recurrence time was(64.67±42.47)days.The incidence of treatment-related adverse events was 15.35％(64 cases/417 cases).Conclusion Morinda officinalis oligosaccharides capsules can be effectively used for the continuation treatment of mild and moderate depression,and are well tolerated and safe.

Objective To investigate the efficacy and safety of Cyberknife in the treatment of elderly patients(aged≥75 years)with early stage non-small cell lung cancer(NSCLC),and to compare the results with those of patients aged<75 years.Methods We retrospectively analyzed 75 patients with early(T1-2N0M0)NSCLC admitted to the 960th Hospital of Jinan People's Liberation Army from January 2013 to October 2019.There were 32(42.7%)patients aged<75 years,and 43(57.3%)patients aged≥75 years.All patients were treated with 45-66Gy/3-8F,60%-85%isodose line as the prescription dose to cover planning target volume(PTV),and irradiation once a day and five times a week.The clinical efficacy,survival status and radiotherapy toxicity of the two groups were compared,and the factors affecting the efficacy of elderly patients were analyzed.Results The disease control rates of patients aged<75 and≥75 years were 96.9%and 93.0%,respectively(P>0.05).The 5-year local control rate(LC),progression-free survival(PFS)and cancer-specific survival(CSS)were 70.9%and 85.4%,58.5%and 54.4%,and 70.4%and 64.5%,respectively(P>0.05).However,the overall survival(OS)of patients aged≥75 years was significantly lower than that of patients aged<75 years,and the 5-year OS was 49.2%and 68.2%,respectively(P<0.05).There was no significant difference in the treatment complications between the two groups(P>0.05).Multivariate analysis showed that biologic effective dose(BED)was an independent factor affecting OS in patients aged≥75 years.Conclusion Stereotactic body radiotherapy with cyberknife is a safe and effective treatment for elderly patients with early stage NSCLC who are not suitable for surgery.

Objective To observe the promoting effect of pulsed shortwave therapy on wound healing in pediatric patients with upper limb burns and explore its underlying mechanisms.Methods A total of 40 children with deep Ⅱ ° upper limb burns treated in our hospital from June 2021 to June 2023 were enrolled,and randomly divided into a control group and an observation group,with 20 cases in each group.The control group received conventional dressings,and the observation group received conventional dressings combined with pulsed shortwave therapy.The wound healing time,first appearance of skin islands on the wound,and initial number of skin islands were observed and recorded using a camera.The proportion of granulation tissue area in the wound was observed using eKare inSight 3D.The positive rate of wound bacteria was detected by collecting wound secretions for bacterial culture examination.Blood flow perfusion on the wound was measured using a blood flow meter.Results The observation group showed significantly faster wound healing and initial appearance of epithelialization(P＜0.05),and obviously higher rate of granulation tissue growth than the control group(P＜0.05).Moreover,the observation group demonstrated better control of bacterial infection in the wounds than the control group(P＜0.05).Blood flow measurement results revealed no significant increase in blood flow perfusion after pulsed shortwave therapy in the observation group(P＞0.05).However,with the elapse of time,both groups exhibited an increasing trend in blood flow perfusion(P＜0.05).Conclusion Pulsed shortwave therapy effectively promotes wound healing in pediatric patients with burns,by facilitating necrotic tissue clearance and granulation tissue regeneration,which may be associated with its non-thermal effects.

Based on the UHPLC-Q-Exactive-MS metabonomics technology, the effect of Hippocampus kuda Bleeker on the life span of Drosophila melanogaster was studied, and the change rule of endogenous metabolites in the aging process of Drosophila melanogaster after the intervention of Hippocampus kuda Bleeker japonicus was explored to clarify the anti-aging mechanism of Hippocampus. The natural aging model of Drosophila melanogaster was used. Different doses of raw Hippocampus and fried Hippocampus were given to observe the effects on the life span, climbing ability, sexual activity, and antioxidant enzyme activity of Drosophila melanogaster. Based on UHPLC-Q-Exactive-MS metabolomics technology, the metabolic profile of the aging Drosophila melanogaster was analyzed using metabonomics technology to explore the mechanism of Hippocampus kuda Bleeker delaying the aging of Drosophila melanogaster. The results showed that raw Hippocampus and crispy Hippocampus (1, 4 mg·mL^-1) could significantly prolong the average life span, median life span and maximum life span of male fruit flies, and significantly improve the climbing ability and sexual vitality of fruit flies. Catalase (CAT) and aldehyde content were increased, while malonaldehyde (MDA) content was decreased. Through metabonomics technology, it was identified that the Hippocampus can significantly recall 16 metabolites and participate in the biosynthesis of phenylalanine, tyrosine and tryptophan, starch and sucrose metabolism, tyrosine metabolism, cysteine and methionine metabolism, and histidine metabolism. The anti-aging mechanism is related to amino acid metabolism and sugar metabolism, which provides a substantial scientific basis for the development and utilization of Hippocampus and clarifying its role in senile diseases. The animal experiment of this study was approved by the Ethics Committee of Shanxi University (approval number: SXULL2021028).

Stable isotope tracer metabolomics tracks and analyzes the whole metabolic process of the body through the tracer atoms, which belongs to the frontier technology in the field of biomedicine. This technology is of great significance and value for explaining the pathogenesis of diseases, finding biomarkers of diseases and drug action targets. Taking the mechanism of glucose catabolism disorder in depression as an example, this paper systematically expounds the stable isotope tracer metabolomics technology and its application. The research idea of stable isotope tracer metabolomics based on unmarked metabolomics was put forward, and the research strategy of biological significance interpretation from four dimensions of metabolite isotope abundance, key metabolic enzymes, metabolic flow direction and metabolite flow was given, which broke through the bottleneck of stable isotope tracer metabolomics research technology based on overall animal experiment, and provided scientific basis for the promotion and application of this technology.

Dayuanyin (DYY) has been shown to reduce lung inflammation in both coronavirus disease 2019 (COVID-19) and lung injury. This experiment was designed to investigate the efficacy and mechanism of action of DYY against hypoxic pulmonary hypertension (HPH) and to evaluate the effect of DYY on the protection of lung function. Animal welfare and experimental procedures are approved and in accordance with the provision of the Animal Ethics Committee of the Institute of Materia Medica, Chinese Academy of Medical Science. Male C57/BL6J mice were randomly divided into 4 groups: control group, model group, DYY group (800 mg·kg^-1), and positive control sildenafil group (100 mg·kg^-1). The animals were given control solvents or drugs by gavage three days in advance. On day 4, the animals in the model group, DYY group and sildenafil group were kept in a hypoxic chamber containing 10% ± 0.5% oxygen, and the animals in the control group were kept in a normal environment, and the control solvent or drugs continued to be given continuously for 14 days. The right ventricular systolic pressure, right ventricular hypertrophy index, organ indices and other metrics were measured in the experimental endpoints. Meantime, the expression levels of the inflammatory factors in mice lung tissues were measured. The potential therapeutic targets of DYY on pulmonary hypertension were predicted using network pharmacology, the expression of nuclear factor kappa B (NF-κB) signaling pathway-related proteins were measured by Western blot assay. It was found that DYY significantly reduced the right ventricular systolic pressure, attenuated lung injury and decreased the expression of inflammatory factors in mice. It can also inhibit hypoxia-induced activation of NF-κB signaling pathway. DYY has a protective effect on lung function, as demonstrated by DYY has good efficacy in HPH, and preventive administration can slow down the disease progression, and its mechanism may be related to inhibit the activation of NF-κB and signal transducer and activator of transcription 3 (STAT3) by DYY.

To investigate the effect of long-term indwelling gastric tube on the prevention and treatment of esophageal stenosis after endoscopic submucosal dissection (ESD) for esophageal circumferential superficial cancer, data of patients with esophageal circumferential superficial cancer who underwent ESD in the First Affiliated Hospital of Nanjing Medical University from January 2018 to December 2021 were retrospectively analyzed. There were 15 patients with gastric tube placement (GTP) after ESD (the GTP group ), and 23 patients without GTP (the non-GTP group). The general information, lesion location, pathological stage, postoperative complications, degree of esophageal stenosis (water intake), pain conditions, number of hospitalizations and medical expenses were compared between the two groups. The results showed that there was no significant difference in age, gender, lesion location or postoperative pathological stage between the two groups ( P>0.05). Compared with the non-GTP group, the rate of water intake in the GTP group was significantly higher (11/15 VS 6/23, P<0.05), the frequency of pain was less in the GTP group (7.3±3.1 times VS 10.7±3.6 times, t=3.00, P<0.05), and the number of hospitalizations and the medical expenses after ESD to before and after stent placement were significantly lower in the GTP group than those in the non-GTP group ( P<0.05). There were no significant differences in the incidence of delayed bleeding and perforation, or time of the first stenosis after ESD between the two groups ( P>0.05). The results of the study initially showed that long-term indwelling gastric tube after ESD can reduce the degree of esophageal stenosis with good safety for esophageal circumferential superficial lesions.

BACKGROUND: Clinical outcomes are poor if patients with acute heart failure (AHF) are discharged with residual congestion in the presence of renal dysfunction. However, there is no single indication to reflect the combined effects of the two related pathophysiological processes. We, therefore, proposed an indicator, congestion and renal index (CRI), and examined the associations between the CRI and one-year outcomes and the incremental prognostic value of CRI compared with the established scoring systems in a multicenter prospective cohort of AHF. METHODS: We enrolled AHF patients and calculated the ratio of thoracic fluid content index divided by estimated glomerular filtration rate before discharge, as CRI. Then we examined the associations between CRI and one-year outcomes. RESULTS: A total of 944 patients were included in the analysis (mean age 63.3 ± 13.8 years, 39.3% women). Compared with patients with CRI ≤ 0.59 mL/min per kΩ, those with CRI > 0.59 mL/min per kΩ had higher risks of cardiovascular death or HF hospitalization (HR = 1.56 [1.13-2.15]) and all-cause death or all-cause hospitalization (HR = 1.33 [1.01-1.74]). CRI had an incremental prognostic value compared with the established scoring system. CONCLUSIONS In patients with AHF, CRI is independently associated with the risk of death or hospitalization within one year, and improves the risk stratification of the established risk models.