Objective To analyze the platelet activating factor acetylhydrolase ( PAF-AH) activity of a gene product encoded by LA2144 gene of Leptospira interrogans ( L. interrogans) , to investigate the ex-pression and secretion of LA2144 protein in various cell cultures and to further understand its function in in-ducing internal hemorrhage in an animal model. Methods The DNA sample containing LA2144 gene was extracted from L. interrogans serogroup Icterohaemorrhagiae serovar Lai strain Lai and used as the template for gene cloning by PCR. The LA2144 gene without the signal sequence coding region was amplified by PCR and inserted into a prokaryotic expression construct for the protein expression. The expressed recombinant protein, rLep-PAF-AH, was purified by Ni-NTA affinity chromatography. Spectrophotometry was used to measure the hydrolytic activity, hydrolytic efficiency, Km and Kcat values of the rLep-PAF-AH protein in hydrolyzing PAF substrate. Real-time fluorescent quantitative RT-PCR ( qRT-PCR) and Western blot assay were performed to measure the expression of LA2144 gene at mRNA and protein levels in human umbilical vein endothelial cells (HUVEC), human monocytes (THP-1) and murine macrophages (J774A. 1) with L. interrogans strain Lai infection, respectively. Each syrian hamster was intravenously injected with 100 μg of LPS-free rLep-PAF-AH for two times. Hemorrhage in the lungs, livers and kidneys were observed in three days after the injection. Results The constructed prokaryotic expression system for LA2144 gene of L. inter-rogans strain Lai could highly express the rLep-PAF-AH upon the induction of IPTG. The purified rLep-PAF-AH showed high purity with a single protein band in gel as indicated by SDS-PAGE. The efficiency of 5 μg of rLep-PAF-AH in hydrolyzing PAF substrate was 26. 6 U/L with a Km value of 82. 79 μmol/L and a Kcat value of 0. 24 S-1 . The expression of Lep-PAF-AH at mRNA level in HUVEC, THP-1 and J774A. 1 cells were significantly elevated after co-culture with L. interrogans strain Lai for 1 or 2 hours (P<0. 05). A large amount of Lep-PAF-AH were detected in the supernatants from co-cultures of L. interrogans strain Lai with the three cell lines, but not from the culture of the spirochete in EMJH medium. The signs of hemor-rhage were observed in the lung of hamsters injected with rLep-PAF-AH, but not in tissue samples from liver and kidney. Conclusion The LA2144 gene product was characterized by a stronger PAF-AH activity. The expression of LA2144 gene at mRNA and protein levels in various cell lines were enhanced during L. interro-gans infection. Moreover, the rLep-PAF-AH could induce the pulmonary hemorrhage in hamsters. This stud-y indicated that the protein encoded by LA2144 gene was an important virulence factor causing hemorrhage in hosts during L. interrogans infection.

Objective To investigate the current status of secondary prevention of cerebral infarction in Longquanyi District of Chengdu. Methods Prospective Investigation on the secondary prevention was conducted in 94 pa-tients with cerebral infarction at 3, 12 and 24 months following stroke. Results The percentages of patients receiving anti-platelet therapy, antihypertensive treatment, antidiabetic drug treatment and statin therapy were 96.8%, 92.1%, 91.7%and 63.8%at hospital discharge, 79.8%, 88.9%, 91.7%and 55.3%at 3 months, 76.1%, 75.0%and 41.4%, 63.8%at 12 months and 33.0%, 71.4%;58.3%and 22.3%at 24 months, respectively. None of stroke patients with atrial fibrillation re-ceived anticoagulation therapy. Conclusion There is a large gap between the current practice of secondary prevention and guideline in patients with cerebral infarction in Longquanyi District of Chengdu.

Objective:To explore the risk factors analysis and validation of anemia after triple-negative breast cancer (TNBC) and risk prediction model, to provide reference for reducing the incidence of anemia after TNBC-NAC.Methods:Retrospectively, 316 female TNBC-NAC patients in the breast department of our hospital from January 2016 to July 2021 were selected as the study subjects (modeling set), and the condition of anemia after the last chemotherapy was set as the observation group. In addition, 98 breast cancer patients in 2022 were selected for validation. Multivariate Logistic regression was used to analyze the effect of Fukangning capsule on TNBC-NAC anemia. We established the risk nomogram prediction model and calibration curve of anemia after chemotherapy by using R software and conducted internal and external verification. Hosmer-Lemeshow test was used to evaluate the prediction deviation between the risk prediction value of the nomogram model and the actual observed value, and the ROC curve was used to evaluate the prediction effect of the model.Results:A total of 75 (23.73%) among 316 patients developed anemia. The results of multivariate Logistic regression analysis showed that menopause [ OR (95% CI)=26.739 (5.063-141.227)], RBC [ OR (95% CI)=0.168 (0.098-0.286)], Hb level [ OR (95% CI)=0.952 (0.929-0.976)], and pathological stage Ⅲ[ OR (95% CI)=4.182 (1.759-9.946)] were independent risk factors for anemia after TNBC-NAC ( P<0.05). The nomogram prediction model established based on the above factors. Hosmer-Lemeshow test results showed that the difference between the risk prediction value and the actual observed value is ( χ2=3.68, P=0.885). The correction curve approaches to the ideal curve and the average absolute error was 0.012. The area under the ROC curve was 0.945, 95% CI was 0.918-0.972, sensitivity was 0.921 and specificity was 0.853, suggesting that the model has good differentiation and calibration degree. The external validation results showed that the nomogram predicted anemia with sensitivity of 88.7%, spectificity of 85.45% and accuracy of 86.73%. Conclusion:Menopausal status, RBC, Hb level before chemotherapy and pathological stage are independent risk factors for anemia after taking TNBC-NAC. The nomogram prediction model based on the above indicators had good discrimination and calibration, and accurately predicted the possibility of anemia after TNBC-NAC.

Objective: Subthreshold attention-deficit/hyperactivity disorder (ADHD) has been suggested to be a “morbid condition” which also needs medical attention. Methods: The present study recruited 89 children with subthreshold ADHD (sADHD), 115 children with diagnosed ADHD (cADHD), and 79 healthy controls (HC) to explore the clinical manifestation, executive functions (EFs) of sADHD, and the caregiver strain. The clinical manifestation was evaluated through clinical interviews and parent-reports. Executive functions were assessed both experimentally and ecologically. Caregiver strain was measured by a parent-reported questionnaire. Results: For the clinical manifestation, both sADHD and cADHD indicated impairments when compared with HC. The comorbidities and the scaled symptoms indicated that the externalizing behaviors were relatively less serious in sADHD than cADHD, whereas the internalizing behaviors between two groups were comparable. For ecological EFs, sADHD scored between cADHD and HC in inhibition and working memory. For experimental EFs, sADHD was comparable to cADHD in inhibition, shifting, and was worse than cADHD in verbal working memory. For the caregiver strain, all scores of sADHD were between that in cADHD and that in HC. Conclusion Our present findings supported the suggestion of subthreshold ADHD as “morbid condition,” which should be treated with caution in clinical practice, especially for the internalizing behaviors and some key components of EFs.

Objective To investigate the relationship between thyroid function and brain volume in patients with Alzheimer's disease(AD).Methods A total of 64 AD patients(AD group)and 36 patients with mild cognitive impairment(MCI group)admitted in our department from January 2020 to March 2022 were enrolled in this study.Another 19 healthy individuals who had no cogni-tive impairment or psychiatric disorders were enrolled and served as normal control group.Their levels of free triiodothyronine(FT3),free thyroxine(FT4)and thyroid stimulating hormone(TSH)were detected by electrochemical luminescence assay.Results FT3 level was significantly decreased in the MCI group and AD group than the normal control group[2.50(2.28,2.60)ng/L and 2.07(1.97,2.30)ng/L vs 2.76(2.55,2.93)ng/L,P＜0.05],and the decrease in the AD group was more obvious than that in the MCI group.The volumes of the midbrain,pons,medulla oblon-gata,hippocampus,amygdala and temporal lobe were significantly smaller in the AD group than the MCI group(P＜0.05,P＜0.01).Spearman correlation analysis and multivariate linear regres-sion analysis showed that in the AD patients,FT3 and FT4 levels were positively while TSH level was negatively correlated with the volumes of both right and left hippocampus and amygdala(P＜0.05,P＜0.01),and TSH level was also negatively correlated with the left temporal lobe volume(P＜0.05).Conclusion Thyroid dysfunction is associated with reduced brain volume in AD patietns,and may contribute to the progression of AD cognitive dysfunction and brain atrophy.

Patients with coronary heart disease have recurrent cardiovascular events after systemic treatment is often one of the leading factors increasing the death and hospitalization rates, thereby adding to the economic burden of society. According to foreign and domestic reports, poor compliance of cardiac rehabilitation in patients with coronary heart disease is a common problem at present. Therefore, in the rehabilitation of patients with coronary heart disease, Improving cardiac rehabilitation compliance is important in regaining health and cost saving during coronary heart disease recovery treatment. This study summarized the status and influence factors of cardiac rehabilitation compliance among foreign and domestic coronary heart disease patients and the interventions recommended for improving cardiac rehabilitation compliance.

OBJECTIVE To establish HPLC characteristic chro matogram of Jianpi yifei biyan prescription standard decoction ， to select the quality control index components and determine their contents. METHODS HPLC method combined with Similarity Evaluation System of TCM Chromatographic Fingerprint （2004 edition）were used to establish the characteristic chromatogram of 10 batches of Jianpi yifei biyan prescription standard decoction ；the similarity evaluation and common peaks identification were also carried out. Using common peak area of characteristic chromatogram as variables ，SPSS 26.0 software and SIMCA 14.1 software were used to perfor m cluster analysis （CA），principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）；differential components with variable important i n pro jection（VIP）value greater than 1.5 were screened；the contents of cimifugin and differential components were determined by the same method. RESULTS A total of 24 common characteristic peaks were identified ， and the similarities of 10 batches of samples were higher than 0.960；eight characteristic peaks were identified by comparison with reference substance. CA and PCA results revealed that the samples were classified into 3 categories.OPLS-DA analysis showed that 3 components with VIP value greater than 1.5， which were prim-O-glucosylcimifugin （peak 2），calycosin 7-O-β-D-glucopyranoside （peak 4） and 5-O-methylvisammioside （peak 6） in descending order. The linear ranges of prim- O- glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside were 0.010 7-0.213 0，0.007 8- 0.156 0，0.008 0-0.160 0，0.009 8-0.195 0 μg（r＞0.999），respectively. RSD values of precision ，repeatability and stability tests （24 h） were all less than 2％. Average recoveries were 105.98％（RSD＝1.75％，n＝6），98.06％（RSD＝3.87％，n＝6），96.38％（RSD＝ 4.03％ ，n＝6） and 104.17％（RSD＝1.27％ ，n＝6）. The contents of the above 4 components in 10 batches of samples were 12.12-18.87，3.86-6.40，3.10-4.27 and 11.17-15.79 μ g/mL，respectively. CONCLUSIONS The established HPLC characteristic chromatographic method is stable and feasible ，it can be used for the quality control of Jianpi yifei biyan prescription standard decoction. Prim- O-glucosylcimifugin，calycosin 7-O-β-D-glucopyranoside，cimifugin and 5-O-methylvisammioside can be used as the index components for quality control of the standard decoction.