OBJECTIVE To stu dy the effects of different processing me thods on the contents of the pharmacodynamic index components in Citrus aurantium and their antioxidant activity. METHODS According to the general principles of 2020 edition of Chinese Pharmacopoeia （volume Ⅳ） and the relevant processing methods in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province ，the samples of C. aurantium were prepared by steaming with water ，boiling with water ，stir-frying with vinegar ，stir-frying with wine ，stir-frying with bran ，processing with bran and processing with honey. The contents of moisture and ash in different products of C. aurantium were detected. The contents of naringin and neohesperidin in different products of C. aurantium were determined by high performance liquid chromatography. The antioxidant activity of different products was investigated by DPPH and ABTS + radical scavenging experiments and the total reducing power test. RESULTS The contents of moisture ，ash，naringin and neohesperidin were in line with the relevant requirements in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province . The content of naringin in descending order was as follow ： unprocessed sample ＞sample processed with honey ＞sample processed with bran ＞sample boiled with water ＞sample stir-fried with vinegar＞sample stir-fried with wine ＞sample stir-fried with bran ＞sample steamed with water. The content of neohesperidin in descending order was as follow ：unprocessed sample ＞sample boiled with water ＞sample processed with bran ＞sample processed with honey ＞sample stir-fried with vinegar ＞sample steamed with water ＞sample stir-fried with wine ＞sample stir-fried with bran. The samples after boiling with water ，processing with bran ，and stir-fried with bran had better DPPH radicals scavenging ability （IC50 were 7.49，8.37 and 10.22 mg/mL，respectively）. The samples after boiling with water ，steaming with water ，and processed with bran had better ABTS + radicals scavenging ability （IC50 were 1.76，2.03 and 2.72 mg/mL，respectively）. In addition ， compared with sample stir-fried with wine and processed with 发。E-mail：wanglu1286@163.com honey，unprocessed sample and other processed products of C.aurantium had bet ter total reducing ability. CONCLUSIONS After processing ，the contents of the main pharmacodynamic index components in C. aurantium have been reduced ，but they were also in line with the relevant requirements in 2015 edition of Processing Specifications of Traditional Chinese Medicine in Zhejiang Province . The antioxidant ability of some processed products has been enhanced.

The National Medical Products Administration has authorized sodium oligomannate for treating mild-to-moderate Alzheimer's disease.In this study,an LC-MS/MS method was developed and validated to quantitate sodium oligomannate in different biomatrices.The plasma pharmacokinetics,tissue distri-bution,and excretion of sodium oligomannate in Sprague-Dawley rats and beagle dogs were system-atically investigated.Despite its complicated structural composition,the absorption,distribution,metabolism,and excretion profiles of the oligosaccharides in sodium oligomannate of different sizes and terminal derivatives were indiscriminate.Sodium oligomannate mainly crossed the gastrointestinal epithelium through paracellular transport following oral administration,with very low oral bioavail-ability in rats(0.6％-1.6％)and dogs(4.5％-9.3％).Absorbed sodium oligomannate mainly resided in circulating body fluids in free form with minimal distribution into erythrocytes and major tissues.So-dium oligomannate could penetrate the blood-cerebrospinal fluid(CSF)barrier of rats,showing a con-stant area under the concentration-time curve ratio(CSF/plasma)of approximately 5％.The cumulative urinary excretion of sodium oligomannate was commensurate with its oral bioavailability,supporting that excretion was predominantly renal,whereas no obvious biliary secretion was observed following a single oral dose to bile duct-cannulated rats.Moreover,only 33.7％(male)and 26.3％(female)of the oral dose were recovered in the rat excreta within 96 h following a single oral administration,suggesting that the intestinal flora may have ingested a portion of unabsorbed sodium oligomannate as a nutrient.

Objective:To investigate the protective effects of bovine lactoferrin (bLF) supplementation on intestinal mucosal tissue and its influence on of inflammatory factors in the premature rats model of necrotizing enterocolitis(NEC), and to provide the theoretical basis for prevention of NEC by bLF supplementation.Methods:Premature SD rats were randomly divided into 4 groups, 25 cases in each group.Control group: oral feeding; model group : oral feeding with lipopolysaccharides(LPS) gavage + hypoxic stimulation; high dose bLF intervention group: daily bLF (7 g/L) + oral feeding with LPS gavage + hypoxic stimulation; low dose bLF intervention group: daily bLF (2 g/L) + oral feeding with LPS gavage + hypoxic stimulation.Histopathological analysis was performed by HE staining.The expression levels of interleukin-1β(IL-1β)and interleukin-6(IL-6)in intestinal mucosa were detected by enzyme linked immunosorbent assay (ELISA).Results:(1) Morphological observation: the intestinal wall of model group was thin, and there were different degrees of pneumoconiosis and effusion in intestinal cavity.Under the microscopy, it could be observed that the intestinal tissue necrosis was serious, the intestinal villi fell off, glands arranged disorderly, epithelial edema was significant, the lamina propria and submucosa had severely edema and were separated, and there were a large number of inflammatory cells infiltrated.The above-mentioned manifestations were alleviated in the high-dose and low-dose bLF intervention groups, and no significant abnormalities were found in the control group.(2) The expression of IL-1β and IL-6 in intestinal tissue: the tissue concentration of IL-1β and IL-6 in the model group rats [(380.89±20.25) ng/L, (485.12±31.44) ng/L]were significantly higher than those in the control group[(270.69±45.58) ng/L, (212.62±89.46) ng/L]( q =9.785, 14.030, all P<0.01). The expression of IL-1β and IL-6 in mucosal tissue of ileum was significantly inhibited in hypoxic and LPS-stimulated rats fed with bLF(IL-1β: q=9.105, 8.761, all P<0.01; IL-6: q=8.175, 8.996, all P<0.01). There was no significant difference in the expression of IL-1β and IL-6 between high dose bLF(7 g/L) and low dose bLF (2 g/L) inter vention groups (IL-1β: q=-0.084, P>0.05; IL-6: q=-1.140, P>0.05). Conclusion:Enteral bLF supplementation can alleviate the damage of intestinal tissue in NEC model of premature SD rats, inhibit the expression of IL-1β and IL-6 inflammatory factors in intestinal tissue, and have a protective effect on intestinal tissue.

Objective To detect ASXL1 gene variants among patients with myelodysplastic syndrome (MDS) and explore their correlation with variants of other genes and clinical features of patients.Methods For 149 patients with MDS,genomic DNA was amplified by PCR and subject to direct sequencing to identify variants of ASXL1,U2AF1,SF3B1,DNMT3A,TET2,IDH1/2,NPM1,FLT3-ITD and C-KIT genes.Results ASXL1 variants were found among 37 patients (24.8％).Other commonly mutated genes included U2AF1 (22.8％),TET2 (11.4％),DNMT3A (9.4％),NPM1 (8.1％) and SF3B1 (6.0％).The frequency of concurrent U2AF1 and TET2 variants among patients with ASXL1 variants was slightly higher than that of wild-type patients.No significant difference was found in median age,MDS subtype,karyotype,peripheral leukocytes,hemoglobin,platelet levels,and bone marrow blast counts between the ASXL1-variant and the wild-type groups (P＞0.05).Twenty-nine patients harboring ASXL1 variants were followed up,37.9％ progressed to acute myeloid leukemia (AML).The rate of transformation in ASXL1-variant group was significantly higher than the wild-type group (37.9 ％ vs.14.1％,P＜0.01).Conclusion ASXL1showed a high frequency of variant among MDS patients,which was frequently accompanied with U2AF1 and TET2 variants.Compared with the wild type group,patients with ASXL1 variants were more likely to progress to AML.

Objective To investigate the efficacy and application of bronchoalveolar lavage in chil-dren with severe pneumonia undergoing mechanical ventilation. Methods Using a prospective randomized controlled clinical study, 202 children with severe pneumonia received mechanical ventilation in Hunan Children′s Hospital from January 2016 to January 2018 were selected as the subjects. According to the digital method,all cases were divided into treatment group (101 cases) and control group (101 cases) randomly. The patients in the control group were given conventional treatment ( anti-infection and symptomatic thera-py) . The treatment group was treated with bronchoalveolar lavage on the basis of conventional treatment. The basic situation,the respiratory function before and after the treatment,the inflammation index,the curative effect and the prognosis of two groups were analyzed. Results There were no significant differences between the two groups in gender,age,course pre-admission,pediatric critical illness score,respiratory function and in-flammation index ( P>0. 05 ) . The respiratory function indexes of the treatment group were obviously im-proved 2 hours after the treatment and the PaO2 ,PaO2/FiO2 and SaO2 were significantly higher than those of the control group[PaO2:(82. 4 ± 6. 4) mmHg(1 mmHg=0. 133 kPa) vs. (74. 0 ± 5. 5) mmHg, PaO2/FiO2:(360. 2 ± 21. 3) mmHg vs. (332. 6 ± 23. 5) mmHg,SaO2:(94. 9 ± 8. 2)％ vs. (88. 6 ± 10. 3)％], while the PaCO2 were significantly lower than the control group [ ( 37. 3 ± 10. 3 ) mmHg vs. ( 45. 8 ± 5. 5 ) mmHg],and the differences were statistically significant (P<0. 05). Five days after treatment,the WBC, PCT and CRP of treatment group were significantly lower than those in the control group[WBC:(8. 5 ± 2. 4) × 109/L vs. (11. 7 ± 3. 5) × 109/L,PCT:(1. 2 ± 0. 7) μg/L vs. (2. 3 ± 0. 9) μg/L,CRP:(9. 1 ± 3. 2) mg/L vs. (16. 5 ± 4. 7) mg/L,P<0. 05,respectively]. The total effective rate in the treatment group was significantly higher than that in the control group[93. 1％(94/101)vs. 81. 2％(82/101)]. Mechanical venti-lation duration and PICU stay in treatment group were significantly shorter than those in the control group [(148. 5 ±30. 6)h vs. (159. 6 ±47. 3)h,(220. 8 ±49. 7)h vs. (330. 7 ±94. 6)h]. The positive rate of patho-genic bacteria was significantly higher than that in the control group [79. 2％(80/101)vs. 62. 4％(63/101), P<0. 05],but there was no significant difference in the 28 days mortality of the two groups[5. 0％(5/101) vs. 5. 9％(6/101),P>0. 05]. Conclusion The bronchoalveolar lavage can improve the respiratory func-tion,reduce the inflammatory reaction,shorten mechanical ventilation duration and PICU stay in children with severe pneumonia undergoing mechanical ventilation obviously. It is worth popularizing in the PICU because of the improvement of curative effect in these children.

Objective: To carry out mutation analysis for patients with myelodysplastic syndromes (MDS) and a normal karyotype. Methods: Targeted capture and next-generation sequencing (NGS) was carried out using a customized 49-gene panel. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutations were detected by PCR and Sanger sequencing. Results: Sixty two patients (80.5%) were found to harbor at least one mutation. Each patient has carried 2.21 mutations in average. Coexistence of ≥ 3 mutations was common (43.7%). The most commonly mutated genes were RUNX1 (23.4%, 18/77), ASXL1 (18.2%, 14/77), NPM1 (15.6%, 12/77), U2AF1 (15.6%, 12/77), DNMT3A (11.7%, 9/77). Patients with SF3B1 mutations were significantly older than those with ASXL1 mutations (P=0.023). Mutations of the DNMT3A gene were significantly associated with the blood platelet level compared with BCOR mutations (P=0.02). No significant difference was found in the number and rate of mutations between those under or above 60-year-old. Among 67 patients with clinical follow-up, 20 (29.8%) has transformed to acute myeloid leukemia, and the time of transformation has ranged from 1 to 44 months, with a average of 5.3 months. RUNX1, U2AF1 and FLT3 mutations are associated with leukemic transformation. Conclusion Coexistence of ≥ 3 mutations are frequent among patients with normal-karyotype MDS. Certain mutations are associated with age and leukemic transformation.

OBJECTIVE: To carry out mutation analysis for patients with myelodysplastic syndromes (MDS) and a normal karyotype. METHODS: Targeted capture and next-generation sequencing (NGS) was carried out using a customized 49-gene panel. FLT3 internal tandem duplication (FLT3-ITD), CALR, NPM1 and CEBPA mutations were detected by PCR and Sanger sequencing. RESULTS: Sixty-two patients (80.5%) were found to harbor at least one mutation. Each patient has carried 2.21 mutations in average. Coexistence of ≥ 3 mutations was common (43.7%). The most commonly mutated genes were RUNX1 (23.4%, 18/77), ASXL1 (18.2%, 14/77), NPM1 (15.6%, 12/77), U2AF1 (15.6%, 12/77), DNMT3A (11.7%, 9/77). Patients with SF3B1 mutations were significantly older than those with ASXL1 mutations (P=0.023). Mutations of the DNMT3A gene were significantly associated with the blood platelet level compared with BCOR mutations (P=0.02). No significant difference was found in the number and rate of mutations between those under or above 60-year-old. Among 67 patients with clinical follow-up, 20 (29.8%) has transformed to acute myeloid leukemia, and the time of transformation has ranged from 1 to 44 months, with a average of 5.3 months. RUNX1, U2AF1 and FLT3 mutations are associated with leukemic transformation. CONCLUSION Coexistence of ≥ 3 mutations are frequent among patients with normal-karyotype MDS. Certain mutations are associated with age and leukemic transformation.
Age Factors ; DNA Mutational Analysis ; Humans ; Karyotype ; Leukemia, Myeloid, Acute ; genetics ; Middle Aged ; Mutation ; Myelodysplastic Syndromes ; genetics ; Prognosis

Objective To investigate the prevalence of pulmonary fungal infection and drug resist-ance of the pathogenic fungi among children with severe diseases in pediatric intensive care unit(PICU). Methods From July 2013 to June 2017,the complete clinical data,results of fungal culture and drug sensi-tivity of bronchoalveolar lavage fluid in 112 critically ill children with pulmonary fungus infection of PICU hospitalization in our hospital were collected. Samples of peripheral venous blood were collected meantime, including blood routine examination,C reactive protein,G test and GM test. Results One hundred and twen-ty-six fungi were isolated from sputum samples in 112 critically ill children. Severe pulmonary infection (30. 36%,34/112) was the most common form of the primary diseases, the next were severe sepsis (16. 07%,18/112)and severe malnutrition(15. 18%,17/112). Classified based on age difference,the first one was ＜1 year old (43. 75%,49/112),the second one was 1 to 3 years old(29. 46%,33/112). The fun-gal strains were predominantly Candida albicans (61. 90%) and Candida tropicalis (16. 67%),among the infectious cases 14 were diagnosed as mixed infection. Two cases of 3 cryptococcal infectious children were HIV infection,another one was malignant tumor,Cryptococcus was cultured in both sputum,pleural effusion and cerebrospinal fluid. The drug resistance rate of fluconazol in 126 strains of fungi was 12. 70%,the rate of itraconazole was 7. 14%. Generally,the fungi cultured were with very low resistance to 5-fluorocytosine,vori-conazole and amphotericin B. However,the strains of Aspergillus fumigates,Candida kruse,Candida parapsi-losis and Cryptococcus were highly resistant to fluconazol and itraconazole,but with very low resistance to 5-fluorocytosine and amphotericin B. Conclusion Candida albicans is the main pathogenic fungus of pulmo-nary fungal infection among children in PICU,and we could choose voriconazole and amphotericin B as treat-ment of critically ill children with pulmonaty fungal infection.

Objectives To determine the risk factors of ventilator-associated pneumonia (VAP) in the pediatric intensive care unit and to explore effective strategies to reduce the morbidity of VAP. Methods A retrospective analysis was conducted on 455 children admitted into the PICU of Hunan Children's Hospital from June 2014 to June 2017. The 455 children were divided into VAP group (n=43) and non-VAP group (n=412). The incidence of VAP was identified and risk factors were compared using the logistic regression analysis via SPSS 19.0 software.Results There were 311 males and 144 females with a median age of 11 months old (29 days to 9 years and 4 months). The incidence of VAP was 9.45% (43/455). Congenital laryngeal and trachea malformation with pulmonary infection was the first reason for the occurrence of VAP (23.3%), followed by congenital heart diseases with pulmonary infection (18.6%). Via univariate analysis, types of endotracheal intubation (χ2=45.33, P<0.001), duration of mechanical ventilation (Z=1.21, P=0.034), re-intubation (χ2=20.22, P=0.004), early usage of antibiotics (χ2=4.98, P=0.026),and methods of nutritional support(χ2=10.15,P=0.006)were identified as risk factors of VAP in the pediatric intensive care unit patients (P<0.05). Based on the multivariate logistic regression analysis, the followings were all independent predictor for VAP:types of endotracheal intubation(OR=1.87,95%CI:1.48~9.75),duration of mechanical ventilation(OR=1.14, 95%CI:1.08~2.35), re-intubation (OR=3.42, 95%CI:1.26~5.57), early usage of antibiotics (OR=4.55, 95%CI:2.21~8.77). Conclusions Many risk factors were found related with the occurrence of VAP. A comprehensive analysis of the host factors and iatrogenic factors should be conducted. Rational use of antibiotics and daily assessment of extubation might help reduce the incidence of VAP.

Partial least square (PLS) combining with Raman spectroscopy was applied to develop predictive models for plasma paclitaxel concentration detection. In this experiment, 312 samples were scanned by Raman spectroscopy. High performance liquid chromatography (HPLC) was applied to determine the paclitaxel concentration in 312 rat plasma samples. Monte Carlo partial least square (MCPLS) method was successfully performed to identify the outliers and the numbers of calibration set. Based on the values of degree of approach ( ), moving window partial least square (MWPLS) was used to choose the suitable preprocessing method, optimum wavelength variables and the number of latent variables. The correlation coefficients between reference values and predictive values in both calibration set ( ) and validation set ( ) of optimum PLS model were 0.933 1 and 0.926 4, respectively. Furthermore, an independent verification test was performed on the prediction model. The results showed that the correlation error of the 20 validation samples was 9.36%±2.03%, which confirmed the well predictive ability of established PLS quantitative analysis model.