This study aimed to provide scientific evidence for predicting quality markers(Q-markers) of Elephantopus scaber by establishing UPLC fingerprint of E. scaber from different geographical origins and determining the content of 13 major components, as well as conducting in vitro anti-cancer activity investigation of the main components. The chromatographic column used was Waters CORTECS UPLC C_(18)(2.1 mm×150 mm, 1.6 μm), and the mobile phase consisted of acetonitrile and 0.1% formic acid solution(gradient elution). The column temperature was set at 30 ℃, and the flow rate was 0.2 mL·min~(-1). The injection volume was 1 μL, and the detection wavelength was 240 nm. The UPLC fingerprint of E. scaber was fitted using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 edition) to determine common peaks, evaluate similarity, identify and determine the content of major components. The CCK-8 assay was used to explore the inhibitory effect of the main components on the proliferation of lung cancer cells. The results showed that in the established UPLC fingerprint of E. scaber, 35 common peaks were identified. Thirteen major components, including neochlorogenic acid(peak 1), chlorogenic acid(peak 2), cryptochlorogenic acid(peak 3), caffeic acid(peak 4), schaftoside(peak 6), galuteolin(peak 9), isochlorogenic acid B(peak 10), isochlorogenic acid A(peak 12), isochlorogenic acid C(peak 18), deoxyelephantopin(peak 28), isodeoxyelephantopin(peak 29), isoscabertopin(peak 31), and scabertopin(peak 32) were identified and quantified, and a quantitative analysis method was established. The results of the in vitro anti-cancer activity study showed that deoxyelephantopin, isodeoxyelephantopin, isoscabertopin, and scabertopin in E. scaber exhibited inhibition rates of lung cancer cell proliferation exceeding 80% at a concentration of 10 μmol·L~(-1), higher than the positive drug paclitaxel. These results indicate that the fingerprint of E. scaber is highly characteristic, and the quantitative analysis method is accurate and stable, providing references for the research on quality standards of E. scaber. Four sesquiterpene lactones in E. scaber show significant anti-cancer activity and can serve as Q-markers for E. scaber.
Humans ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Asteraceae/chemistry* ; Lung Neoplasms/drug therapy*

Abstract: To report the diagnosis, treatment and outcome of 4 patients with hematological diseases complicated with Aeromonas hydrophila bloodstream infection in the Second Affiliated Hospital of Kunming Medical University, further clarify the importance of blood culture and deepen the clinical understanding of the disease. Four patients with hematological diseases complicated with Aeromonas hydrophila bloodstream infection treated in the Second Affiliated Hospital of Kunming Medical University from 2017 to 2021 were recruited as the study objects. The clinical manifestations, blood culture collection, detection time of Aeromonas hydrophila, laboratory examination, treatment and prognosis of the patients were retrospectively analyzed. In this study, 4 cases were male patients with hematological diseases, who were in myelosuppression after chemotherapy. After fever, blood culture was collected and Aeromonas hydrophila was detected. The positive time of blood culture in 4 cases ranged from 4 to 11 hours. The results of antibiotic sensitivity showed that it was highly sensitive to the second, third and fourth generation cephalosporins, quinolones and carbapenems. Four patients were treated with imipenem cilastatin sodium in the early stage, and one patient recovered after active anti infection and leukocyte raising treatment. One patient did not complete chemotherapy due to a request for discharged, and the follow-up was unknown. Two patients developed rapidly into necrotizing fasciitis and died later. Hematological diseases complicated with Aeromonas hydrophila bloodstream infection are rare, but the mortality rate is high. For patients with repeated fever and considering infection, blood culture should be carried out as soon as possible to confirm the pathogen and drug sensitivity test. During clinical treatment, the treatment should be adjusted in time in combination with the patient's situation. In addition to anti-infection treatment, the patient's immunity should be improved and the development of necrotizing fasciitis should be vigilant. Keywords: Aeromonas hydrophila; hematologic diseases; leukemia; bloodstream infection; blood culture; necrotizing fasciitis

Objective: The investigate the effects and mechanism of exosomes derived from human umbilical vein endothelial cells (HUVECs) on wound healing in diabetes rabbits. Methods: The experimental research methods were used. The primary vascular endothelial cells (VECs) and human skin fibroblasts (HSFs) were extracted from skin tissue around ulcer by surgical excision of two patients with diabetic ulcer (the male aged 49 years and the female aged 58 years) admitted to Xiangya Third Hospital of Central South University in June 2019. The cells were successfully identified through morphological observation and flow cytometry. The HUVEC exosomes were extracted by ultracentrifugation and identified successfully by morphological observation, particle size detection, and Western blotting detection. Twenty female 3-month-old New Zealand rabbits were taken to create one type 2 diabetic full-thickness skin defect wound respectively on both sides of the back. The wounds were divided into exosomes group and phosphate buffer solution (PBS) group and treated accordingly, with 20 wounds in each group, the time of complete tissue coverage of wound was recorded. On PID 14, hematoxylin-eosin staining or Masson staining was performed to observe angiogenesis or collagen fiber hyperplasia (n=20). The VECs and HSFs were co-cultured with HUVEC exosomes for 24 h to observe the uptake of HUVEC exosomes by the two kinds of cells. The VECs and HSFs were divided to exosome group treated with HUVEC exosomes and PBS group treated with PBS to detect the cell proliferation on 4 d of culture with cell count kit 8, to detect and calculate the cell migration rate at 24 and 48 h after scratch by scratch test, to detect the cell migration number at 24 h of culture with Transwell test, and to detect the mRNA expressions of nuclear factor-erythroid 2-related factor 2 (NRF2) and transcription activating factor 3 (ATF3) by real time fluorescence quantitative reverse transcription polymerase chain reaction. Besides, the number of vascular branches and vascular length were observed in the tube forming experiment after 12 h of culture of VECs (n=3). The VECs and HSFs were taken and divided into PBS group and exosome group treated as before, and NRF2 interference group, ATF3 interference group, and no-load interference group with corresponding gene interference. The proliferation and migration of the two kinds of cells, and angiogenesis of VECs were detected as before (n=3). Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, independent sample t test, and least significant difference test. Results: The time of complete tissue coverage of wound in exosome group was (17.9±1.9) d, which was significantly shorter than (25.2±2.3) d in PBS group (t=4.54, P<0.05). On PID14, the vascular density of wound in PBS group was significantly lower than that in exosome group (t=10.12, P<0.01), and the collagen fiber hyperplasia was less than that in exosome group. After 24 h of culture, HUVEC exosomes were successfully absorbed by VECs and HSFs. The proliferative activity of HSFs and VECs in exosome group was significantly higher than that in PBS group after 4 d of culture (with t values of 54.73 and 7.05, respectively, P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs (with t values of 3.42 and 11.87, respectively, P<0.05 or P<0.01) and VECs (with t values of 21.42 and 5.49, respectively, P<0.05 or P<0.01) in exosome group were significantly higher than those in PBS group. After 24 h of culture, the migration numbers of VECs and HSFs in exosome group were significantly higher than those in PBS group (with t values of 12.31 and 16.78, respectively, P<0.01). After 12 h of culture, the mRNA expressions of NRF2 in HSFs and VECs in exosome group were significantly higher than those in PBS group (with t values of 7.52 and 5.78, respectively, P<0.05 or P<0.01), and the mRNA expressions of ATF3 were significantly lower than those in PBS group (with t values of 13.44 and 8.99, respectively, P<0.01). After 12 h of culture, the number of vascular branches of VECs in exosome group was significantly more than that in PBS group (t=17.60, P<0.01), and the vascular length was significantly longer than that in PBS group (t=77.30, P<0.01). After 4 d of culture, the proliferation activity of HSFs and VECs in NRF2 interference group was significantly lower than that in PBS group and exosome group (P<0.05 or P<0.01); the proliferation activity of HSFs and VECs in ATF3 interference group was significantly higher than that in PBS group (P<0.05 or P<0.01) and significantly lower than that in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in ATF3 interference group were significantly higher than those in PBS group (P<0.05 or P<0.01) and significantly lower than those in exosome group (P<0.05 or P<0.01). At 24 and 48 h after scratch, the migration rates of HSFs and VECs in NRF2 interference group were significantly lower than those in PBS group and exosome group (P<0.05 or P<0.01). After 24 h of culture, the migration numbers of VECs and HSFs in ATF3 interference group were significantly more than those in PBS group (P<0.05) and significantly less than those in exosome group (P<0.05 or P<0.01); the migration numbers of VECs and HSFs in NRF2 interference group were significantly less than those in PBS group and exosome group (P<0.01). After 12 h of culture, the vascular length and number of branches of VECs in NRF2 interference group were significantly decreased compared with those in PBS group and exosome group (P<0.01); the vascular length and number of branches of VECs in ATF3 interference group were significantly increased compared with those in PBS group (P<0.01) and were significantly decreased compared with those in exosome group (P<0.01). Conclusions: HUVEC exosomes can promote the wound healing of diabetic rabbits by promoting the proliferation and migration of VECs and HSFs, and NRF2 and ATF3 are obviously affected by exosomes in this process, which are the possible targets of exosome action.
Animals ; Female ; Humans ; Male ; Rabbits ; Collagen/metabolism* ; Diabetes Mellitus ; Exosomes/metabolism* ; Human Umbilical Vein Endothelial Cells ; Hyperplasia/metabolism* ; NF-E2-Related Factor 2/metabolism* ; RNA, Messenger/metabolism* ; Ulcer ; Wound Healing ; Middle Aged

Objective    To summarize the experience of treating adult recurrent pectus excavatum without plate turnover. Methods    Twenty-seven patients with recurrent pectus excavatum treated by thoracoscopy-assisted placement without plate turnover from 2010 to 2019 in our hospital were enrolled. There were 23 males and 4 females with the age of 3-29 (12.81±7.79) years at the first operation, and 18-29 (21.74±3.56) years at this operation. Incision of 2-3 cm at bilateral axillary midline of the deepest point of pectus excavatum was made, and an auxiliary incision under xiphoid process was adopted according to the intraoperative situation. Results    All patients underwent thoracoscopy-assisted correction of pectus excavatum without bar turnover, and subxiphoid incision was performed in 11 patients. Twenty-five patients had one bar placed, and two patients required two bars. The operation time was 28-45 (33.00±6.44) min. Postoperative Haller index (2.95±0.40) was improved compared with preoperation (4.63±1.03). The postoperative hospital stay was 4-6 (4.00±0.32) day. All patients were followed up for 1-8 years. Complications included poor wound healing in 1 patient, and steel wire fracture and displacement in 1 patient. There was no plate rotation or bar displacement. Fourteen patients removed the bar 29-84 (40.36±13.93) months after the placement. Haller index was improved to 2.43-3.61 (2.86±0.35) during removal of steel plate. Untill June 2020, there was no recurrence of pectus excavatum. Conclusion    The treatment of adult recurrent pectus excavatum without plate turnover is satisfactory, and the protection of intercostal muscle and firm fixation is the key to ensure the success of operation and long-term effects.

Objective:To analyze the chemical constituents in Microctis Folium by ultra performance liquid chromatography-quadrupole-time-of-flight high resolution mass spectrometry （UPLC-Q-TOF-MS/MS）. Method:Waters CORTECS UPLC C₁₈ column （2.1 mm×150 mm， 1.6 μm） was used for chromatographic separation with the mobile phase of methanol （A） -0.1% formic acid solution （B） for gradient elution （0-4 min， 14%-30%A； 4-16 min， 30%-58%A； 16-25 min， 58%-78%A； 25-25.1 min， 78%-98%A； 25.1-29 min， 98%A）， the flow rate was 0.25 mL· min^-1， the injection volume was 1 μL. The electrospray ionization （ESI） was adopted for determining the chromatographic effluent under positive and negative ion modes， the main chromatographic peaks were assigned and distinguished by Q-TOF， and the scanning range was m/z 100-1 500. Result:A total of 31 chemical constituents in Microctis Folium were identified by confirmation of reference substances， literature comparison and high resolution mass spectrometry data analysis. The chemical constituent cluster was composed of 28 flavonoids （9 flavone C-glycosides， 10 flavonols and their glycosides， 8 proanthocyanidins， 1 xanthone） and 3 organic acids （caffeic acid， p-coumaric acid， ferulic acid）. Conclusion:UPLC-Q-TOF-MS/MS technique provides a simple， rapid and accurate method for the identification of chemical constituents in Microctis Folium. Flavone C-glycosides， flavonol oxyglycosides and proanthocyanidins are the main chemical constituents. The 7 proanthocyanidins are reported for the first time in this herb. In conclusion， the chemical profile of Microctis Folium is characterized and the findings are meaningful for the in-depth quality assessment and material basis clarification of Microctis Folium.

When this paper was first published the following ethical statement was omitted in error: All animal experimental protocols were approved by the Animal Ethics Committee of Guangdong Provincial Engineering Technology Institute of Traditional Chinese Medicine (Guangzhou, Guangdong, China, Approval NO: 048483). Further, all methods were performed in accordance with the relevant guidelines and regulations. NIH mice were purchased from the Guangdong Medical Laboratory Animal Center (Guangzhou, Guangdong, China, Certificate NO.44007200031795). The authors would like to apologise for any inconvenience caused.

OBJECTIVE: To investigate the risk factors for cow's milk protein allergy (CMPA) among infants through a multicenter clinical study. METHODS: A total of 1 829 infants, aged 1-12 months, who attended the outpatient service of the pediatric department in six hospitals in Shenzhen, China from June 2016 to May 2017 were enrolled as subjects. A questionnaire survey was performed to screen out suspected cases of CMPA. Food avoidance and oral food challenge tests were used to make a confirmed diagnosis of CMPA CMPA. A multivariate logistic regression analysis was used to investigate the risk factors for CMPA. RESULTS: Among the 1 829 infants, 82 (4.48%) were diagnosed with CMPA. The multivariate logistic regression analysis showed that maternal food allergy (OR=4.91, 95%CI: 2.24-10.76, P<0.05), antibiotic exposure during pregnancy (OR=3.18, 95%CI: 1.32-7.65, P<0.05), and the introduction of complementary food at an age of <4 months (OR=3.55, 95%CI: 1.52-8.27, P<0.05) were risk factors for CMPA, while exclusive breastfeeding (OR=0.21, 95%CI: 0.08-0.58, P<0.05) and the introduction of complementary food at an age of >6 months (OR=0.38, 95%CI: 0.17-0.86, P<0.05) were protective factors. CONCLUSIONS The introduction of complementary food at an age of <4 months, maternal food allergy, and antibiotic exposure during pregnancy are risk factors for CMPA in infants.
Animals ; Cattle ; China ; Female ; Humans ; Infant ; Milk Hypersensitivity ; Milk Proteins ; Pregnancy ; Risk Factors ; Surveys and Questionnaires

Objective To introduce the application experience of a new steel bar used in minimally invasive surgery for pectus carinatum.Methods From January to October 2018, Cardiothoracic Surgery Department of Shanghai Xinhua Hospital performed a minimally invasive surgery for 25 cases of patients with pectus carinatum used a new type of steel bar.All 25 pa-tients were male, aged 10 -17 years, with an average age of(13.80 ±1.66)years.The application experience of the new bar in pectus carinatum minimally invasive surgery was summarized .Results All operations were successfully completed .The op-eration time was 35-100 min, averaged(73.44 ±17.49)min, postoperative hospital stay was 3 -6 days, averaged(3.68 ± 0.85)days.Postoperative complications included 5 cases of pneumothorax(the lung compression was about 2％ -10％, not necessary for surgical intervention).One case occured wound healing delay 1 month after operation, and healed after no surger-cal treatment.The other patients recovered smoothly.Conclusion The new steel bar is convenient to use, greatly reduces the difficulty of the pectus carinatum surgery procedure , also reduced surgical trauma and complications , has a good application prospect.

To share the clinical experience of thoracoscopic unidirectional posterolateral basal segmentectomy via inferior pulmonary ligament. Methods    All the patients were in the healthy lateral position, with endoscopy holes in the 8th intercostal space of the middle axillary line and 2-3 cm operation holes in the 5th intercostal space of the front axillary line. Anatomical segmentectomy of the posterolateral basal vein, bronchus and artery was performed through the inferior pulmonary ligament upward in turn. The clinical data of this group were analyzed retrospectively. Results    From December 2015 to October 2018, 32 patients underwent thoracoscopic unidirectional posterolateral basal segmentectomy, including 8 males and 24 females, aged 13-71 (52.6±13.7) years. All patients successfully completed the operation, including 9 patients of left lower pulmonary posterolateral basal segmentectomy, 23 patients of right lower pulmonary posterolateral basal segmentectomy. The operation time was 80-295 (133.4 ±40.5) minutes, intraoperative bleeding volume was 20-300 (52.6±33.8) mL, drainage time was 2-14 (4.2±2.3) days, hospitalization time was 4-15 (6.9 ±2.4) days. No death occurred during hospitalization. Postoperative complications included atelectasis in 1 patient and persistent pulmonary leakage over 3 days (4 or 6 days respectively) in 2 patients , chylothorax in 1 patient. All of them recovered smoothly after non-operative treatments. Postoperative pathology showed that 29 patients of primary adenocarcinoma or atypical adenomatoid hyperplasia, including 5 patients of adenocarcinoma in situ, 9 patients of micro-invasive adenocarcinoma, 12 patients of invasive adenocarcinoma, 3 patients of atypical adenomatoid hyperplasia. One patient was of intestinal metastatic adenocarcinoma, 1 patient of inflammatory lesion and 1 patient of bronchiectasis. 3-21(9.6±4.6) lymph nodes were resected in the patients with primary pulmonary malignant tumors. And no metastasis was found. Conclusion    The operation of thoracoscopic unidirectional posterolateral basal segmentectomy via inferior pulmonary ligament is easy. There is no need to open intersegmental tissue. It can protect lung tissue better. The operative method is worthy of clinical promotion.

BACKGROUNDNucleolin (NCL) is the most abundant RNA-binding protein in the cell nucleolus and plays an important role in chromatin stability, ribosome assembly, ribosomal RNA maturation, ribosomal DNA transcription, nucleocytoplasmic transport, and regulation of RNA stability and translation efficiency. In addition to its anti-apoptotic properties, the underlying mechanisms associated with NCL-related roles in different cellular processes remain unclear. In this study, the effect of NCL on microRNA (miRNA) expression was evaluated by generating transgenic mice with myocardial overexpression of NCL and by analyzing microarrays of mature and precursor miRNAs from mice.

METHODSUsing microinjection of alpha-MyHc clone 26-NCL plasmids, we generated transgenic mice with myocardial overexpression of NCL firstly, and then mature and precursor miRNAs expression profiles were analyzed in NCL transgenic mice (n = 3) and wild-type (WT) mice (n = 3) by miRNA microarrays. Statistical Package for the Social Sciences version 16.0 software (SPSS, Inc., Chicago, IL, USA) was used to perform Student's t-test, and statistical significance was determined at P < 0.05.

RESULTSSeveral miRNAs were found to be differentially expressed, of which 11 were upregulated and 4 were downregulated in transgenic mice with myocardial overexpression of NCL compared to those in WT mice. Several differentially expressed miRNAs were subsequently confirmed and quantified by real-time quantitative reverse transcription-polymerase chain reaction. Bioinformatics analysis was used for the prediction of miRNA targets. Furthermore, in vitro experiments showed that NCL regulated miR-21 expression following hydrogen peroxide preconditioning.

CONCLUSIONSMyocardial-protection mechanisms exerted by NCL might be mediated by the miRNAs identified in this study.