: Bacteroides, as one of the most abundant and diverse genera in the human gut, is regarded as a window into the study of gut microbiota-host interactions. Currently, CRISPR/Cas-based gene editing systems targeting Bacteroides have been widely applied, while the large size of Cas nucleases limits their potential application scenarios (such as in situ gut Bacteroides editing based on phage delivery). Therefore, this study aims to develop a compact and highly efficient genetic editing tool in Bacteroides., We developed a miniaturized CRISPR/Cas gene editing system for human gut Bacteroides. First, the editing capabilities of different miniaturized CRISPR/Cas systems, including AsCas12f, CasΦ2, and ISDge10, were evaluated in Bacteroides fragilis. Subsequently, the editing capability of AsCas12f was assessed across various Bacteroides species, and the size of this system was further optimized. The results demonstrated that the CRISPR/AsCas12f genome editing system exhibited the highest editing efficiency in B. fragilis. The CRISPR/AsCas12f system achieved efficient genome editing in B. fragilis, Bacteroides thetaiotaomicron, and Phocaeicola vulgatus. Furthermore, with a repair template of 500 bp homologous arms, the editing efficiency remained as high as 94.7%. In conclusion, CRISPR/AsCas12f can serve as a chassis tool enzyme for the development of Bacteroides-based miniature gene editors and derivative technologies, laying a foundation for the further development of gene editing technology for Bacteroides.
CRISPR-Cas Systems/genetics* ; Gene Editing/methods* ; Bacteroides/genetics* ; Humans ; Gastrointestinal Microbiome/genetics* ; Bacteroides fragilis/genetics*

Objective:To explore the active ingredients of shengxian and jinshuiliujun decoction with the method of network pharmacology, and to verify the experimental mechanism of its treatment of silicosis.Methods:In May 2023, the active ingredients and targets of drugs in shengxian and jinshuiliujun decoction were obtained through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. The target of silicosis disease was screened by databases such as Genecards, Disease Gene Network (DisGeNET), Comparative Toxicogenomics Database (CTD), etc. The screened drug targets and disease targets were intersected to obtain the target set of shengxian and jinshuiliujun decoction for the treatment of silicosis. Protein-protein interaction (PPI) network analysis was performed on the target set through STRING database, and core target genes were screened. GO enrichment analysis and KEGG pathway analysis of intersection genes were performed based on Metascape database, and molecular docking verification of key components and targets of shengxian and jinshuiliujun decoction was carried out. Twenty-four adult male SD rats with SPF grade were randomly divided into control group, model group and TCM intervention group, with 8 rats in each group. The dust-stained rat model was prepared by non-tracheal exposure of 1 ml silica suspension (50 mg/ml) in one go, and TCM intervention group was given shengxian and jinshuiliujun decoction[6 g/ (kg·d) ] on the second day. The CT of the lungs of each group was observed 28 days after the dust-stained rat model. Paraffin sections of rat lung tissues were prepared and stained with Hematoxylin-Eosin (HE) and Masson. Western blot was used to verify the expression of core target-related proteins in rat lung tissues after the intervention of shengxian and jinshuiliujun decoction for 28 days, and the differences in protein expression between groups were compared by one-way analysis of variance.Results:A total of 205 active ingredients and 3345 active compounds were selected from shengxian and jinshuiliujun decoction, corresponding to 281 targets, among which 240 targets were related to silicosis. Serine/threonine kinase 1 (AKT1), tumor protein p53 (TP53), tumor necrosis factor (TNF) and interleukin (IL) 6 may be the key targets of shengxian and jinshuiliujun decoction in the treatment of silicosis. Through enrichment analysis, 30 GO entries and 20 potential signaling pathways were screened according to P-value, including nuclear factor κB (NF-κB), mitogen-activated protein kinase (MAPK) and cancer signaling pathways. Molecular docking showed that the active compounds of shengxian and jinshuiliujun decoction had good binding with the core target proteins, and the strongest binding properties were beta-sitosterol and TNF-α (-10.45 kcal/mol). In animal experiments, the inflammatory infiltration and fibrosis of lung tissue of rats in TCM intervention group were significantly improved. Compared with control group, the levels of TNF-α, IL-1β, IL-6 and NF-κB in lung tissue of model group were significantly increased ( P<0.05). Compared with model group, the lung injury of rats in TCM intervention group was significantly improved, and the expressions of TNF-α, IL-1β, IL-6 and NF-κB were significantly decreased ( P<0.05) . Conclusion:Shengxian and jinshuiliujun decoction in the treatment of silicosis may play an anti-fibrosis role by inhibiting the NF-κB signal transduction pathway mediated by inflammatory factors such as TNF-α and IL-1β, which provides a reference for further exploring the material basis and mechanism of its action.

ObjectiveTo understand the use of antibiotics in inpatients and the incidence and trend of hospital infections， to explore the implementation effect of comprehensive management measures， and to provide reference for hospitals to use antibiotics reasonably. MethodsBased on the hospital infection monitoring and management system， a retrospective analysis and comparison were conducted on the use of antibiotics， submission of microbial test samples， and incidence of hospital infections among inpatients in a tertiary hospital from 2012 to 2021. ResultsFrom 2012 to 2021， the use of antibiotics showed a downward trend， from 50.82% in 2012 to 41.29% in 2021. At the same time， the use rate of restricted and special antibiotics had also decreased， and the use rate of restricted and special antibiotics in patients without hospital infection was significantly lower than that in patients with hospital infection， and the microbial testing rate was also on the rise. The annual incidence rate of hospital infection was 0.69%‒1.92%， and the annual case-time prevalence rate was 0.79%‒2.17%. The annual average rate of the above two in 10 years was 1.18% and 1.34%， respectively. The results of the exponential smoothing model also showed that the utilization rate of antibiotics was decreasing and the incidence of nosocomial infection was stable. ConclusionLarge general hospitals took comprehensive management measures to strengthen the management of rational use of antibiotics， which led to a decline in the use rate of antibacterial drugs for inpatients and an increase in the rate of microbial examination. At the same time， the overall incidence of hospital infection was relatively stable， suggesting that the comprehensive management measures of antibacterial drugs in hospitals had achieved certain results. The current measures need to be optimized in the future to continuously improve the management level of rational use of antibacterial drugs.

Objective:To explore the active ingredients of shengxian and jinshuiliujun decoction with the method of network pharmacology, and to verify the experimental mechanism of its treatment of silicosis.Methods:In May 2023, the active ingredients and targets of drugs in shengxian and jinshuiliujun decoction were obtained through the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) database. The target of silicosis disease was screened by databases such as Genecards, Disease Gene Network (DisGeNET), Comparative Toxicogenomics Database (CTD), etc. The screened drug targets and disease targets were intersected to obtain the target set of shengxian and jinshuiliujun decoction for the treatment of silicosis. Protein-protein interaction (PPI) network analysis was performed on the target set through STRING database, and core target genes were screened. GO enrichment analysis and KEGG pathway analysis of intersection genes were performed based on Metascape database, and molecular docking verification of key components and targets of shengxian and jinshuiliujun decoction was carried out. Twenty-four adult male SD rats with SPF grade were randomly divided into control group, model group and TCM intervention group, with 8 rats in each group. The dust-stained rat model was prepared by non-tracheal exposure of 1 ml silica suspension (50 mg/ml) in one go, and TCM intervention group was given shengxian and jinshuiliujun decoction[6 g/ (kg·d) ] on the second day. The CT of the lungs of each group was observed 28 days after the dust-stained rat model. Paraffin sections of rat lung tissues were prepared and stained with Hematoxylin-Eosin (HE) and Masson. Western blot was used to verify the expression of core target-related proteins in rat lung tissues after the intervention of shengxian and jinshuiliujun decoction for 28 days, and the differences in protein expression between groups were compared by one-way analysis of variance.Results:A total of 205 active ingredients and 3345 active compounds were selected from shengxian and jinshuiliujun decoction, corresponding to 281 targets, among which 240 targets were related to silicosis. Serine/threonine kinase 1 (AKT1), tumor protein p53 (TP53), tumor necrosis factor (TNF) and interleukin (IL) 6 may be the key targets of shengxian and jinshuiliujun decoction in the treatment of silicosis. Through enrichment analysis, 30 GO entries and 20 potential signaling pathways were screened according to P-value, including nuclear factor κB (NF-κB), mitogen-activated protein kinase (MAPK) and cancer signaling pathways. Molecular docking showed that the active compounds of shengxian and jinshuiliujun decoction had good binding with the core target proteins, and the strongest binding properties were beta-sitosterol and TNF-α (-10.45 kcal/mol). In animal experiments, the inflammatory infiltration and fibrosis of lung tissue of rats in TCM intervention group were significantly improved. Compared with control group, the levels of TNF-α, IL-1β, IL-6 and NF-κB in lung tissue of model group were significantly increased ( P<0.05). Compared with model group, the lung injury of rats in TCM intervention group was significantly improved, and the expressions of TNF-α, IL-1β, IL-6 and NF-κB were significantly decreased ( P<0.05) . Conclusion:Shengxian and jinshuiliujun decoction in the treatment of silicosis may play an anti-fibrosis role by inhibiting the NF-κB signal transduction pathway mediated by inflammatory factors such as TNF-α and IL-1β, which provides a reference for further exploring the material basis and mechanism of its action.

Objective:To retrospectively analyze the clinical and pathologic characteristics, response to treatment, survival, and prognosis of patients with primary large B-cell lymphoma of the central nervous system (PCNSLBCL) .Methods:Clinical and pathologic data of 70 patients with PCNSLBCL admitted to Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine from December 2010 to November 2022 were collected for retrospective analysis. Survival analysis was performed using the Kaplan-Meier method and log-rank test, and prognosis analysis was conducted using the Cox proportional hazards model.Results:Among 70 patients with PCNSLBCL, complete remission (CRs) were achieved in 49 (70.0% ) and partial remission in 4 (5.7% ) after the first-line induction therapy; the overall remission rate was 75.7%. The 2-year progression-free survival (PFS) rate was 55.8% and the median progression-free survival (mPFS) time was 35.9 months, whereas the 2-year overall survival (OS) rate was 79.1% with a median OS time not reached. After CR induced by first-line therapy, cumulative incidence of relapse (CIR) was lower in patients who had received auto-HSCT than in those who had not received consolidation therapy ( P=0.032), whose 2-year PFS rate was 54.4% and mPFS time was 35.9 months; comparatively, the 2-year PFS rate in patients having received oral maintenance of small molecule drugs reached 84.4% with a mPFS time of 79.5 months ( P=0.038). Multivariant analysis demonstrated that Class 3 in the Memorial Sloan-Kettering Cancer Center (MSKCC) prognostic model is an independent adverse prognostic factor of OS in patients with PCNSLBCL ( HR=3.127, 95% CI 1.057-9.253, P=0.039) . Conclusions:In patients with PCNSLBCL achieving CR after the first-line induction therapy, auto-HSCT as consolidation therapy would lead to a decreased CIR, and PFS time could be prolonged by oral maintenance of small molecule drugs. Class 3 MSKCC prognostic model is independently associated with poorer OS.

Objective:To investigate the effect of teaching practice under the concept of narrative medicine on improving the empathy level of interns in oral mucosal diseases.Methods:The interns of stomatology in the class of 2018 in School of Stomatology, Shanghai Jiao Tong University, were divided into narrative medicine teaching group (27 interns receiving narrative medicine concepts and methods before and during internship) and traditional teaching group (21 interns received patients directly under the guidance of teachers without the addition of narrative medicine concepts and methods). A questionnaire was used to collect the general information of students, and the Chinese version of Jefferson Scale of Physician Empathy (JSPE) (the version for medical students) was used to measure the empathy level of students. After the end of internship, a statistical analysis was performed for the scores of both groups, and a questionnaire survey was conducted to investigate the acceptance of internship under the guidance of narrative medicine among the students in the narrative medicine teaching group. GraphPad Prism 9.3.0 was used to perform the t-test and the rank sum test. Results:There was a significant difference in JSPE score between the traditional teaching group and the narrative medicine teaching group (92.26±8.23 vs. 104.20±15.65, t=2.70, P=0.005), and in addition, 88.89% (24/27) of the students in the narrative medicine teaching group were interested in participating in internship under the guidance of narrative medicine. However, there was no significant difference in the score of internship between the narrative medicine teaching group and the traditional teaching group (87.28±2.77 vs. 85.47±4.31, t=1.68, P=0.100). Conclusions:Incorporating the concepts and methods of narrative medicine into clinical teaching of oral mucosal diseases can significantly improve the empathy ability of interns and raise the awareness that empathy is as important as scientific literacy among students.

Objective:To investigate the rationality and surgical errors in countermeasures against difficult removal of screws so as to provide reference for standardization of technical procedures.Methods:A retrospective study was conducted to analyze the data of 99 patients who had encountered difficult removal of screws in operative removal of internal fixation at Department of Orthopaedics, Wuxi NO.9 People's Hospital Affiliated to Suzhou University from January 2018 to May 2022. There were 62 males and 37 females with an average age of 38.8±14.7 years. Their internal fixation time ranged from 7 months to 11 years. The irrationality was defined as insufficient preoperative preparation or a countermeasure that failed to follow the surgical indications or scientific principles of minimal injury or priority of simplicity. A surgical error was defined as unnecessary injury, failed removal or complications related to operation. Cases of irrationality and surgical errors were analyzed to find associations between them.Results:The operative removal was successful in 92 cases and failed in 7 cases. Of the patients who experienced difficult removal of screws, irrationality was found in 26.3% (26/99) and a surgical error or errors occurred in 28.3% (28/99). In the patients with countermeasure irrationality, the incidence of surgical errors was 53.9% (14/26) while in those without countermeasure irrationality, the incidence of surgical errors was 19.2% (14/73), showing a statistically significant difference ( χ2=11.360, P<0.001). In the patients with surgical errors, the incidence of countermeasure irrationality was 64.3% (18/28) while in the patients without surgical errors, the incidence of countermeasure irrationality was and 11.3% (8/71), showing a statistically significant difference ( χ2=29.148, P<0.001). In the patients with failed removal, the rate of countermeasure irrationality was 85.71% (6/7) while in those with successful removal, the rate of countermeasure irrationality was 21.7% (20/92), showing a statistically significant difference ( χ2=13.748, P<0.001). Conclusions:Close relationships exist between countermeasure irrationality, surgical errors and failed removal. The higher proportion of countermeasure irrationality, the higher possibility of surgical errors. Therefore, following the rationality principle may avoid or reduce surgical errors in difficult removal of screws.

Objective:To investigate the individual and combined effects of calf circumference and vitamin D levels on all-cause mortality risk in community-dwelling older adults based on the Chinese Longitudinal Healthy Longevity Survey.Methods:The calf circumference was measured in the baseline survey in 2012 and 2014.Low calf circumference was defined as <34 cm for men or 33 cm for women at screening.Vitamin D deficiency was defined as plasma 25-hydroxyvitamin D[25(OH)D]levels <50 nmol/L.All participants were followed up until 2018, when death outcomes and survival time were collected.Cox proportional hazard regression models were used to analyze the effects of calf circumference and 25(OH)D levels on the risk of all-cause mortality.Results:A total of 3 052 older adults were included in the analysis, of which 1 960(64.22%)had low calf circumference and 2 245(73.56%)had vitamin D deficiency.After 10 559.9 person-years of follow-up period, 1 312 death events were recorded.After adjusting for sociodemographic characteristics, physical activities, cognitive function, and multiple chronic diseases, calf circumference and 25(OH)D levels were negatively associated with the risk of all-cause mortality(both P<0.05). In the combined analysis, compared with the normal group, the risk of death was highest in the participants with both low calf circumference and vitamin D deficiency, which was higher than those with low calf circumference or vitamin D deficiency alone, with a hazard ratios( HR)(95% CI)of 2.51(1.81-3.45), 1.71(1.22-2.42)and 1.53(1.09-2.15), respectively.There was a significant additive interaction between low calf circumference and vitamin D deficiency on mortality(RERI>0). Conclusions:Low calf circumference and vitamin D deficiency are associated with higher mortality.Older adults with combined conditions had a even higher risk of death.Attention should be paid to joint screening and comprehensive intervention for older adults with both low calf circumference and vitamin D deficiency.

Background::Non-smokers account for a large proportion of lung cancer patients, especially in Asia, but the attention paid to them is limited compared with smokers. In non-smokers, males display a risk for lung cancer incidence distinct from the females—even after excluding the influence of smoking; but the knowledge regarding the factors causing the difference is sparse. Based on a large multicenter prospective cancer screening cohort in China, we aimed to elucidate the interpretable sex differences caused by known factors and provide clues for primary and secondary prevention.Methods::Risk factors including demographic characteristics, lifestyle factors, family history of cancer, and baseline comorbidity were obtained from 796,283 Chinese non-smoking participants by the baseline risk assessment completed in 2013 to 2018. Cox regression analysis was performed to assess the sex difference in the risk of lung cancer, and the hazard ratios (HRs) that were adjusted for different known factors were calculated and compared to determine the proportion of excess risk and to explain the existing risk factors.Results::With a median follow-up of 4.80 years, 3351 subjects who were diagnosed with lung cancer were selected in the analysis. The lung cancer risk of males was significantly higher than that of females; the HRs in all male non-smokers were 1.29 (95% confidence interval [CI]: 1.20-1.38) after adjusting for the age and 1.38 (95% CI: 1.28-1.50) after adjusting for all factors, which suggested that known factors could not explain the sex difference in the risk of lung cancer in non-smokers. Known factors were 7% (|1.29-1.38|/1.29) more harmful in women than in men. For adenocarcinoma, women showed excess risk higher than men, contrary to squamous cell carcinoma; after adjusting for all factors, 47% ([1.30-1.16]/[1.30-1]) and 4% ([7.02-6.75]/[7.02-1])) of the excess risk was explainable in adenocarcinoma and squamous cell carcinoma. The main causes of gender differences in lung cancer risk were lifestyle factors, baseline comorbidity, and family history.Conclusions::Significant gender differences in the risk of lung cancer were discovered in China non-smokers. Existing risk factors did not explain the excess lung cancer risk of all non-smoking men, and the internal causes for the excess risk still need to be explored; most known risk factors were more harmful to non-smoking women; further exploring the causes of the sex difference would help to improve the prevention and screening programs and protect the non-smoking males from lung cancers.

Objective To study the composition and concentration of atmospheric particulate pollutants in four seasons in the industrial and clean living areas， and to provide a scientific basis for the strategy of controlling industrial pollution and atmospheric environment. Methods An industrial area dominated by the automobile industry in Shanghai and a relatively clean living area were selected. Samples were collected simultaneously in both areas and continuously for 7 days in the middle of each season. The composition and concentration of PM_2.5 were determined， and the ecological risk of heavy metals in PM_2.5 was evaluated by the potential ecological risk index method. Results We found PM_2.5 concentration was associated with seasonal changes. The PM_2.5 concentration in living areas was the highest in winter， followed by spring， and the lowest in summer. The PM_2.5 concentration in industrial areas was the highest in spring， followed by winter， and the lowest in summer. The heavy metals in PM_2.5 were the same， including Al， Cr， Mn， Ni， As， Cd， Hg and Pb. The content of Cr， Cd and Pb in PM_2.5 in the industrial area is significantly higher than that in the living area. The potential ecological hazard coefficient of PM_2.5 heavy metal Cd in the industrial zone was the highest， up to 189.47， and it was the main component of the total potential ecological hazard index of heavy metals. According to the total potential risk grade of heavy metals， the heavy metal Cd in the industrial area had different degrees of ecological harm with seasonal changes. The ecological harm degree of heavy metal Cd was the highest in winter， high in spring and autumn， and low in summer. Conclusion Although the concentration of PM_2.5 in the industrial area is not higher than that in the living area， the content of Cr， Cd and Pb in the PM_2.5 in the industrial area is significantly higher than that in the living area. The concentration of PM_2.5 in the industrial area is mainly related to seasons， industrial production and human factors. The potential ecological harm coefficient of heavy metal Cd in PM_2.5 is the highest in comparison with other heavy metals such as Cr， Hg and Pb， and it is the main component of the total potential ecological harm index ofheavy metals.