Objective To analyze medication patterns and the targets and pathways of core drug combinations in treatment of palpitation.Methods The prescriptions of Li Yaping for treatment of palpitation from March 2023 to March 2024 were collected,and frequency counts of drugs'nature and flavour,channel tropism,and efficacy were performed.Apriori algorithm,association rules,and clustering analysis were carried out using SPSS Modeler 18.0 and SPSS 26.0.The core drugs and disease targets were searched,and gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis were performed on the targets of their therapeutic action for palpitation.Results A total of 220 prescriptions were collected,involving 192 flavors of traditional Chinese medicines,with a cumulative medication frequency of 3978 times,and 18 flavors of high-frequency medicines.The medicines were mainly tonics,sedative,and promoting blood circulation for removing blood stasis.The distribution of medicinal properties were mainly warm,cold and flat.The medicinal flavors were mainly sweet,bitter and pungent,and channel tropism were mostly heart,liver and spleen channel.Association rule analysis showed that Radix Angelicae Sinensis,Radix et Rhizoma Salviae Miltiorrhizae,Radix et Rhizoma Glycyrrhizae,Radix Ophiopogonis,and Radix Astragali were the core drugs.Cluster analysis showed that there was 3 cluster combinations.In the network pharmacology part,there were 181 targets intersected by drug combinations and diseases.KEGG analysis showed that the core drugs for palpitation mainly involved signaling pathways such as phosphoinositide 3-kinase/protein kinase B,hypoxia-inducible factor-1,mitogen-activated protein kinase,interleukin-17,etc.GO analysis obtained 1000 GO pathways,of which 760 were biological processes,93 were cellular components,and 147 were molecular functions.Conclusion In the treatment of palpitation,Li Yaping advocates benefiting qi and promoting yang,removing blood stasis and eliminating turbidity,and tranquilizing the mind,emphasizing the"two hearts in the same adjustment",and treating the heart and liver at the same time,taking into account the spleen and stomach,and the combination of core medicines can intervene in the course of palpitation through multi-components,multi-targets,and multi-pathways,which is of great significance for the treatment of palpitation in the clinical setting.

ObjectiveTo evaluate the efficacy and safety of topical Bisaitong （鼻塞通） in treating moderate-to-severe allergic rhinitis （AR）. MethodsA randomized， positive-controlled， non-inferiority clinical trial design was adopted. Totally， 108 cases of moderate-to-severe AR were randomly divided into Bisaitong group and mometasone furoate group，with 54 cases in each group. The Bisaitong group was treated with Bisaitong smeared at the nasal cavity twice a day， and the mometasone furoate group received inhalation of mometasone furoate nasal spray 100 μg in each nostril， once a day. Both groups were treated for 4 weeks and followed up after additional 4 weeks. Both groups were compared on the rhinoconjunctivitis quality of life questionnaire （RQLQ）， rhinoconjunctivitis total symptom score （RTSS）， visual analogue score （VAS） of sneezing， runny nose， nasal itching， nasal congestion degree， days of AR episodes at enrollment， after 2- and 4-week， and at follow-up. The peripheral blood eosinophil （EOS） count and percentage （EOS%）， serum eosinophil cationic protein （ECP）， serum dust mite， dermatophagoides farinae， and cockroach allergen-specific IgE （sIgE） levels were compared between groups at enrollment and after 4-week treatment. Drug overuse rate was calculated， and the safety was evaluated. The analysis of all efficacy outcomes was based on both full analysis set （FAS） and per-protocol set （PPS）. ResultsThe lower limit of the 95% confidence interval for the differences in RQLQ scores were greater than -0.6 measured after 2- and 4-week treatment and at follow-up compared to that measured at the enrollment in both groups， indicating of the Bisaitong group being non-inferior to the mometasone furoate group. There was no statistically significant difference between groups on RTSS score， VAS scores of sneezing， runny nose， nasal itching， nasal congestion degree and days of episodes at all timepoints （P＞0.05）， but each outcome changed significantly over time in both groups （P＜0.01）. The differences between groups in EOS count， EOS%， ECP levels， serum dust mite， dermatophagoides farinae， cockroach sIgE levels， and drug overuse rate were not statistically significant at enrollment and after 4-week treatment （P＞0.05）. Adverse events occurred in eight cases （15.10%） in the Bisaitong group and five cases （9.30%） in the mometasone furoate group， showing no significant difference between groups （P＞0.05）. ConclusionTopical Bisaitong is non-inferior to mometasone furoate nasal spray in the treatment of moderate to severe AR in terms of clinical symptom relief，reduction in the episodes， improvement of quality of life， and sound safety.

Objective:To investigate the predictive value of inflammatory markers for the risk of stroke-associated infection (SAI) in patients with anterior circulation large vessel occlusive stroke who received endovascular therapy.Methods:Patients with anterior circulation large vessel occlusive stroke received endovascular treatment in Nanjing First Hospital, Nanjing Medical University from 2016 to 2020 were retrospectively enrolled. The clinical data of SAI group and non-SAI group were compared. Multivariate logistic regression analysis was used to screen the independent influencing factors of SAI, and then the predictive nomogram was established according to these influencing factors to verify its clinical application efficiency. Results:A total of 409 patients were enrolled during the study. Their age was 71.3±11.7 years, and 250 were male (61.1%). The median baseline Naitonal Institutes of Health Stroke Scale (NIHSS) score was 16. One hundred and nineteen patients (29.1%) received intravenous thrombolysis, 376 (91.9%) were successfully recanalized after endovascular therapy, and 293 (71.6%) developed SAI. Univariate analysis showed that age, atrial fibrillation ratio, NIHSS score at admission, fasting blood glucose, triglyceride, high sensitivity C reactive protein (hs-CRP), leukocyte count, neutrophil count, neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio (PLR) were significantly higher than those in the non-SAI group (all P<0.05). Multivariate logistic regression analysis showed that age, NIHSS score at admission, fasting blood glucose, hs-CRP, leukocyte count, neutrophil count and NLR were the independent influencing factors of SAI ( P<0.05). Receiver operating characteristics curve analysis showed that the predictive value of multiple inflammatory markers (hs-CRP, leukocyte count, neutrophil count and NLR) for SAI was significantly better than that of the single inflammatory marker ( P<0.01). The area under the curve was 0.782 (95% confidence interval 0.719-0.846), and the predictive sensitivity and specificity were 80.6% and 64.5% respectively. Decision curve analysis showed that compared with the traditional indicators, the predictive nomogram based on inflammation related indicators (hs-CRP, leukocyte count and NLR) had a higher net profitability for predicting SAI. Conclusion:The hs-CRP, leukocyte count and NLR can be used to predict the risks of SAI in patients with acute ischemic stroke receiving endovascular therapy.

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.
Gene Editing ; Aspergillus niger/genetics* ; CRISPR-Cas Systems/genetics* ; Plasmids/genetics*

Objective: To understand lunch satisfaction and leftovers of Changsha compulsory education schools under different supply modes served by school canteens and specialized enterprises,and to provide scientific basis for the government to formulate school lunch improvement policies. Methods: Multistage stratified cluster sampling method was used to random select 2 203 students from 8 primary and secondary schools in Changsha who were administered with questionnair survey in April to May of 2018. Results: For school lunch, the overall rate of satisfaction was 78.8%.The overall lunch satisfaction of female and primany school students were higher than that of male and junior high school students respectively,and the difference was statistically significant(P<0.05). Totally 76.0% of students had the highest satisfaction in food hygiene and the lowest satisfaction rate was 60.0% in food taste. By comparing different supply modes, the overall satisfaction rate of meal quantity was higher (47.8%) under school canteens, and that of dining environment was relatively high (43.7%) under specialized enterprises. "Don-t like some food" (63.8%) was the main reason for students leftovers under the two supply modes. The lunch leftover rate of specialized enterprises (37.8%) was significantly higher than that of school cafeteria (30.6%)(χ 2=12.81，P<0.05). Conclusion The satisfaction of food taste and the rate of lunch leftovers need to be further improved. School canteens environment and management should be strengthened, as well as the flexibility and communication under specialized enterprises and family-school communication. The joint efforts by school and family are needed for healthy eating education.

Objective:To investigate the predictive values of serum hypersensitive C-reactive protein (hs-CRP) and lipoprotein-associated phospholipase A 2 (Lp-PLA 2) for early neurological deterioration (END) and parenchymal hematoma (PH)-type 2 hemorrhagic transformation (HT) after intravenous thrombolysis in patients with acute ischemic stroke. Methods:Patients with acute ischemic stroke treated with intravenous thrombolysis in the Department of Neurology, Nanjing First Hospital, Nanjing Medical University from January 2018 to January 2021 were enrolled retrospectively. END was defined as an increase of ≥4 in the National Institutes of Health Stroke Scale (NIHSS) score at 24 h after thrombolysis compared with the baseline. PH-2 type HT was defined as parenchymal hematoma with obvious space occupying effect or hemorrhage at the distant site of infarct. Multivariate logistic regression analysis was used to determine the independent influencing factors of END and PH-2 type HT. The receiver operating characteristic (ROC) curve was used to evaluate the predictive value of hs-CRP and Lp-PLA 2 levels for END and PH-2 type HT. Results:A total of 804 patients with acute ischemic stroke treated with intravenous thrombolysis were included, of which 63 (7.8%) developed END within 24 h after intravenous thrombolysis; 41 (5.1%) developed HT, of which 38 were PH-2 type HT. Univariate analysis showed that the levels of serum hs-CRP and Lp-PLA 2 in the END group were significantly higher than those in the non-END group (all P<0.05), and the levels of serum hs-CRP and Lp-PLA 2 in the PH-2 HT group were significantly higher than those in the non-PH-2 HT group ( P<0.05). Multivariate logistic analysis showed that hs-CRP (odds ratio [ OR] 1.017, 95% confidence interval [ CI] 1.001-1.034; P=0.043) and Lp-PLA 2 ( OR 1.002, 95% CI 1.000-1.003; P=0.020) were the independent risk factors for END after intravenous thrombolysis. In addition, hs-CRP ( OR 1.019, 95% CI 1.002-1.036; P=0.027) and Lp-PLA 2 ( OR 1.002, 95% CI 1.000-1.003; P=0.018) were also the independent risk factors for PH-2 HT after intravenous thrombolysis. The ROC curve analysis showed that the areas under the curve of hs-CRP and Lp-PLA 2 for predicting END were 0.675 (95% CI 0.609-0.741; P<0.001) and 0.606 (95% CI 0.528-0.683; P=0.005) respectively, and the areas under the curve for predicting PH-2 HT were 0.641 (95% CI 0.545-0.737; P=0.003) and 0.600 (95% CI 0.500-0.699; P= 0.051) respectively. Conclusion:Higher baseline serum hs-CRP and Lp-PLA 2 are the independent predictors of END and PH-2 type HT after intravenous thrombolysis in patients with acute ischemic stroke.

Dopamine is the precursor of a variety of natural antioxidant compounds. In the body, dopamine acts as a neurotransmitter that regulates a variety of physiological functions of the central nervous system. Thus, dopamine is used for the clinical treatment of various types of shock. Dopamine could be produced by engineered microbes, but with low efficiency. In this study, DOPA decarboxylase gene from Sus scrofa (Ssddc) was cloned into plasmids with different copy numbers, and transformed into a previously developed L-DOPA producing strain Escherichia coli T004. The resulted strain was capable of producing dopamine from glucose directly. To further improve the production of dopamine, a sequence-based homology alignment mining (SHAM) strategy was applied to screen more efficient DOPA decarboxylases, and five DOPA decarboxylase genes were selected from 100 candidates. In shake-flask fermentation, the DOPA decarboxylase gene from Homo sapiens (Hsddc) showed the highest dopamine production (3.33 g/L), while the DOPA decarboxylase gene from Drosophila Melanogaster (Dmddc) showed the least residual L-DOPA concentration (0.02 g/L). In 5 L fed-batch fermentations, production of dopamine by the two engineered strains reached 13.3 g/L and 16.2 g/L, respectively. The residual concentrations of L-DOPA were 0.45 g/L and 0.23 g/L, respectively. Finally, the Ssddc and Dmddc genes were integrated into the genome of E. coli T004 to obtain genetically stable dopamine-producing strains. In 5 L fed-batch fermentation, 17.7 g/L of dopamine was produced, which records the highest titer reported to date.
Animals ; Dopa Decarboxylase/genetics* ; Dopamine/biosynthesis* ; Drosophila melanogaster/genetics* ; Escherichia coli/metabolism* ; Humans ; Metabolic Engineering

Objective:To investigate the correlation between fluid-attenuated inversion recovery (FLAIR) vascular hyperintensities (FVHs) and outcomes after endovascular mechanical thrombectomy (EMT) in patients with anterior circulation large vessel occlusive stroke.Methods:Using " Nanjing First Hospital Stroke Database" , consecutive patients with anterior circulation large vessel occlusive stroke received EMT treatment from June 2015 to December 2018 were enrolled retrospectively. Before EMT treatment, the distal FVH grade and the American Society of Intervention and Therapeutic Neuroradiology/Society of Interventional Radiology (ASITN/SIR) collateral circulation grade were evaluated. The modified Rankin Scale was used to evaluate the functional outcome of patients at 3 months after onset, and 0-2 was defined as a good outcome. Spearman correlation analysis was used to analyze the correlation between the distal FVH grade and the ASITN/SIR collateral circulation grade. Multivariate logistic regression analysis was used to identify the independent predictors of the outcomes. Results:A total of 117 patients with acute anterior circulation large vessel occlusive stroke were enrolled, aged 70.74±12.50 years, 72 (61.5%) were male. The baseline National Institutes of Health Stroke Scale (NIHSS) score was 13.73±4.91. Seventy-four patients (63.2%) had a good outcome and 43 (36.8%) had a poor outcome. The distal FVH grade was grade 0 in 8 cases (6.84%), grade 1 in 34 cases (29.06%), and grade 2 in 75 cases (64.10%). Compared with the distal FVH low-grade group (grade 0-1), the high-grade group (grade 2) had a higher ASITN/SIR collateral circulation grade ( P<0.001) and lower baseline National Institutes of Health Stroke Scale (NIHSS) score ( P=0.026). Spearman correlation analysis showed that the distal FVH grade was significantly positively correlated with the ASITN/SIR collateral circulation grade ( r=0.620, P<0.001). Multivariate logistic regression analysis showed that the high distal FVH grade (odds ratio [ OR] 0.336, 95% confidence interval [ CI] 0.128-0.879; P=0.026) was independently associated with the good outcomes, while the higher baseline NIHSS score ( OR 1.036, 95% CI 0.988-1.229; P=0.048) and symptomatic cerebral hemorrhage ( OR 5.597, 95% CI 1.052-29.761; P=0.043) were independently associated with the poor outcomes. Conclusion:The distal FVHs can reflect the state of collateral circulation. The high grade of distal FVHs is associated with the good outcomes after EMT in patients with anterior circulation large vessel occlusive stroke.

Objective To investigate the predictive value of serum lipoprotein (a) (Lp [a]) levels for short-term functional outcomes in elderly patients with acute ischemic stroke and type 2 diabetes mellitus.Methods Elderly patients with acute ischemic stroke and type 2 diabetes mellitus admitted to Nanjing First Hospital from June 2016 to December 2016 were enrolled retrospectively.The modified Rankin scale was used to assess the outcomes at 90 d after onset;0-2 was defined as good outcome and ＞2 were defined as poor outcome.Multivariate logistic regression analysis was used to determine independent risk factors for poor outcomes,and the receiver operator characteristic (ROC) curve analysis was used to evaluate the predictive value of serum Lp(a) levels for poor outcomes.Results A total of 231 patients were enrolled,with an average age of 69.7 years and males accounting for 65.4％.The median serum Lp(a) was 166 mg/L (interquartile range 78-331 mg/L).At 90 d after onset,140 patients (60.6％) had good outcomes and 91 (39.4％) had poor outcomes.After adjustment for other confounding variables,multivariate logistic regression analysis showed that elevated serum Lp(a) (referenced to the lowest quartile,the 3rd quartile:odds ratio[OR]2.899,95％ confidence interval[CI] 1.154-7.285,P =0.024;the 4th quartile:OR 3.334,95％ CI 1.329-8.361,P =0.010),the baseline National Institute of Health Stroke Scale score (OR 1.224,95％ CI 1.143-1.310;P＜ 0.001),and complicated with coronary heart disease (OR 2.504,95％ CI 1.196-5.243;P =0.015) were the independent risk factors for poor outcomes.ROC curve analysis showed that the area under the curve of serum Lp(a) level in predicting the poor outcome was 0.775 (95％ CI 0.696-0.854;P=0.04).The optimal cut-off value was 119 mg/L,the sensitivity was 71.15％,the specificity was 75.90％,the positive predictive value was 38.52％,and the negative predictive value was 61.48％.Conclusion Serum Lp(a) level has certain predictive value for the short-term poor outcomes in elderly patients with acute ischemic stroke and type 2 diabetes mellitus.

Objective: To investigate the correlations of serum uric acid with outcomes and symptomatic intracranial hemorrhage (sICH) after intravascular mechanical thrombectomy bridged with intravenous thrombolysis in patients with ischemic stroke. Methods: From January 2015 to January 2019, patients with acute ischemic stroke admitted to Nanjing First Hospital, Nanjing Medical University and underwent intravascular mechanical thrombectomy bridged with intravenous thrombolysis were analyzed retrospectively. Demographic characteristics, vascular risk factors, laboratory findings, National Institutes of Health Stroke Scale (NIHSS) scores, onset to treatment time, and whether sICH occurred were recorded. The modified Rankin Scale was used to evaluate the outcomes at 90 d after onset, and 0 to 2 was defined as good outcome. Multivariate logistic regression models were used to determine the independent factors for outcomes and sICH. Results: A total of 144 patients were included, 54 (37.5%) had a good outcome, 90 (62.5%) had a poor outcome (including 28 deaths), and 29 (20.1%) had sICH. Serum uric acid was significantly higher in the good outcome group than in the poor outcome group (P<0.05). Serum uric acid was significantly higher in the non-sICH group than in the sICH group (P<0.05). Multivariate logistic regression analysis showed that higher serum uric acid were the independent protective factors of good outcome (odds ratio 0.82, 95% confidence interval 0.66-0.93; P<0.001) and sICH (odds ratio 0.97, 95% confidence interval 0.93-0.99; P=0.004). Conclusion High serum uric acid level is independently associated with good outcome after intravascular mechanical thrombectomybridged with intravenous thrombolysis in patients with acute ischemic stroke.