To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.
Agrobacterium ; Chromatography, Thin Layer ; Culture Media ; Dianthus ; growth & development ; Plant Growth Regulators ; Plant Leaves ; Plant Roots ; growth & development ; Plants, Genetically Modified ; Rhizobium ; Tissue Culture Techniques ; Transformation, Genetic

This study seeks to explore the early signs of cognitive impairment in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). According to polysomnography, twenty patients diagnosed with OSAHS and twenty normal controls underwent event-related potential (ERP) examination including mismatch negativity (MMN) and P300. Compared with normal controls, OSAHS patients showed significantly prolonged latency of MMN and P300 at Cz. After controlling age and body mass index (BMI), MMN latency positively correlated with apnea hypopnea index (AHI), oxygen reduction index, stage N1 sleep and arousal index, while MMN latency negatively correlated with stage N3 sleep and mean blood oxygen saturation; and P300 latency positively related to AHI and oxygen reduction index; no relationships were found among MMN latency, MMN amplitude, P300 latency and P300 amplitude. These results suggest that the brain function of automatic processing and controlled processing aere impaired in OSAHS patients, and these dysfunction are correlated with nocturnal repeatedly hypoxemia and sleep structure disturbance.
Case-Control Studies ; Cognition Disorders ; complications ; physiopathology ; Event-Related Potentials, P300 ; Humans ; Hypoxia ; physiopathology ; Oximetry ; Polysomnography ; Sleep Apnea, Obstructive ; complications ; physiopathology ; Sleep Stages

Objective:To analyze the clinical efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in patients with advanced NSCLC.Methods:We collected 23 cases of NSCLC advanced patients, who were treated in the affiliated Cancer Hospital of Guangzhou Medical University from January 2015 to March 2020. And these 23 cases of patients with first-generation EGFR-TKIs resistance were treated with the third-generation EGFR-TKI drugs. We analyzed their clinicopathological characteristics, studied their therapeutic effects, and followed up their progression-free survival (PFS).Results:It is showed that 16 of 23 cases (69.56%) were got local progression and 7 of 23 cases (31.43%) were found with systemic progression. Briefly, the median PFS of the 23 patients was 17.5 months. A total of 7 cases occurred rashes after taking EGFR-TKI, and 3 cases got abnormal liver function. Fortunately, they were all improved after symptomatic treatments. Additionally, no bone marrow suppression (granulocytes, neutropenia, thrombocytopenia, anemia) and digestive tract reactions (nausea, vomiting, diarrhea) were occurred in 23 cases of NSCLC patients. The mental and physical improvement of EGFR-TKI in the third generation of 19 patients was more obvious than that in the first generation of EGFR-TKI. Among them, 15 cases showed more obvious lesion shrinkage after third-generation EGFR-TKI treatment. 4 patients with GGO had cleaner disappearance than that of the first-generation EGFR-TKI.Conclusions:Compared with traditional chemotherapy, the first-generation EGFR-TKI resistance treatment with three-generation EGFR-TKI treatment has better efficacy with reduced toxic and side effects, and significantly improved the life quality of advanced NSCLC patients.

Objective To explore the relationship between hypoxia-inducible factor-1α (HIF-1α) expression and apoptosis in early brain injury models after subarachnoid hemorrhage (SAH).Methods Fifty-five adult male Sprague-Dawley rats were randomly assigned to five groups:sham-operated group,SAH 6 h,SAH 12 h,SAH 24 h and SAH 72 h groups (n=1 1).SAH in the later four groups was induced by modified monofilament puncture method.The rats were killed by cervical dislocation.HIF-1α expression was assessed by immunofluorescence staining.TUNEL was adopted to detect brain apoptotic cells.Immunofluorescence double staining was used to identify cell types with positive HIF-1α expression.Pearson correlation analysis was employed to analyze the relationship between HIF-1 expression percentage and TUNEL positive rate.Results As compared with those in the sham-operated group,the HIF-1 expression percentage and TUNEL positive rate in the four SAH groups was significantly higher (P＜0.05).Immunofluorescence double staining showed that neuron-specific nuclear protein staining cells were coincided with most HIF-1 positive cells,while only a few HIF-1α positive cells were coincided with glial fibrillary acidic protein staining cells.A significant positive correlation was noted between HIF-1 α expression percentage and TUNEL positive rate following SAH (r=0.737,P=0.001).Conclusion HIF-1α high expression after SAH early promotes neuronal cell apoptosis,indicating HIF-1 a might participate in the pathological progression of early brain injury after SAH.

Objective To establish and evaluate a gas chromatography-tandem mass spectrometry(GC-MS/MS)method for the detection of six common drugs(methamphetamine,meperidine,caffeine,codeine,cocaine and ketamine)in blood,and to improve the determination basis of results.Methods The above six drugs were added into the blank blood,and GC-MS/MS was used for detection after ether extraction.The collection,quantification and confirmation were carried out under the mode of multi-reaction monitoring(MRM).The qualitative results of the above six drugs were evaluated based on the maximum allowable deviation of the retention time and relative ion abundance ratio in the qualitative results of GC-MS/MS.Results There was a good linear relationship between the six common drugs,among which ketamine and caffeine had the lowest detection limit(0.01 μg/mL),methamphetamine had the highest detection limit(0.5 μg/mL).The retention time(RT)and relative retention time(RRT)of the target substance were stable under the six supplemental levels,and the absolute deviation(ΔRTabsolute)of RT was within±0.025 min.The absolute deviation of RRT(ΔRRTabsolute)was within±0.004.The relative ion abundance ratio absolute deviation(ΔIabsolute)is±20%,and the relative ion abundance ratio relative deviation(ΔIrelative)is±50%.Conclusion This study clarified the reference range for qualitative determination of six common drugs in blood matrix detected by GC-MS/MS,and effectively supplemented the qualitative determination indicators of existing instrumental analysis methods.

Objective To prepare the biodegradable polylactic acid glycolic acid (PLGA) copolymer.encapsulated vascular endothelial growth factor(VEGF) loaded fluorescent controlled release sub-microspheres,to understand the efficiency of microspheres loading and releasing VEGF and to observe in vitro microspheres degradation.Methods VEGF-loading PLGA sub-microspheres were prepared by the two-phase solvent evaporation method,the in vitro degradation of fluorescent microspheres was observed by the laser confocal scanning electron microscopy.The drug loading efficiency and drug release curve were observed by ELISA.Results The VEGF loading PLGA fluorescent microspheres were successfully prepared by using the two-phase solvent evaporation method.The microspheres morphology was normal by using the scanning electron microscope and laser confocal microscope.The particle size was 0.5-1.0 μm with the laser particle size analyzer.The distribution was homogeneous.The VEGF loading rate and encapsulation rate detected by the quantitative ELISA were 3.91％ and 51.42 ％ respectively.The fluorescent microscope observed their slow degradation.The VEGF gentle release was detected by the quantitative ELISA,which showing linear zero order release trend.Conclusion The drug loading efficiency of VEGF-loading PLGA microspheres with 0.5-1.0 μm diameter is higher with linear zero order release,which can be directly observed by fluorescent light.

Objective To evaluate the effect of partial neuromuscular blockade (NMB) on the efficacy and safety of nerve monitoring during microvascular decompression (MVD) of facial nerve.Methods Seventy American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes,aged 39-78 yr,weighing 44-84 kg,scheduled for elective MVD,were divided into 2 groups (n=35 each) using a random number table method:control group and partial NMB group.Anesthesia was induced by intravenous injection of 3-fold ED95 cisatracurium.In control group,muscle relaxants were not used after intubation.In partial NMB group,cisatracurium was continuously infused intravenously to maintain partial NMB,and the T1/Tc ratio was maintained at 20％-40％.Intraoperative neuroelectrophysiological monitoring was performed using lateral spread response (LSR).The success rates of LSR monitoring,occurrence of body movement,requirement for anesthetics and cardiovascular agents were recorded during operation,and the patients were followed up on day 7 after surgery,and the therapeutic efficacy and occurrence of neurological complications were recorded.Results Compared with control group,the incidence of intraoperative body movement was significantly decreased,the intraoperative consumption of propofol and remifentanil was decreased,and the intraoperative requirement for vasopressors was decreased in partial NMB group (P＜0.05).There was no significant difference in the success rate of LSR monitoring,therapeutic efficacy and incidence of neurological complications between two groups (P＞0.05).Conclusion Partial NMB (T1/Tc=20％-40％) can be effectively used for MVD monitored by LSR,decrease the occurrence of the body movement,and raise the perioperative safety in patients.

Objective To summarize the experience of integrative palliative care at Peking Union Medi-cal College Hospital and provide a reference for promoting the integrative palliative care model.Methods Twenty cases receiving integrative palliative care at Peking Union Medical College Hospital were collected.The clinical characteristics,reasons for initiating integrative palliative care,the process of integrative palliative care,and feedback from these cases were summarized.Results Insomnia(11 cases,55％)and pain(9 cases,45％)were the most common symptoms requiring control in the 20 cases.The integrative palliative care team assisted in medical decision-making for 17 cases(85％),prepared end-of-life for 9 cases(45％),assisted in the transfer for 3 cases(15％),and provided comfort care for all the 20 cases(100％).Conclusions The integrative palliative care model can help alleviate suffering in end-of-life patients and provide support to patients'families and the original medical teams.This model is worth further promotion within class A tertiary hospitals.

Pharmacokinetics (PK) is the study of the absorption, distribution, metabolism, and excretion (ADME) processes of a drug. Understanding PK properties is essential for drug development and precision medication. In this review we provided an overview of recent research on PK with focus on the following aspects: (1) an update on drug-metabolizing enzymes and transporters in the determination of PK, as well as advances in xenobiotic receptors and noncoding RNAs (ncRNAs) in the modulation of PK, providing new understanding of the transcriptional and posttranscriptional regulatory mechanisms that result in inter-individual variations in pharmacotherapy; (2) current status and trends in assessing drug-drug interactions, especially interactions between drugs and herbs, between drugs and therapeutic biologics, and microbiota-mediated interactions; (3) advances in understanding the effects of diseases on PK, particularly changes in metabolizing enzymes and transporters with disease progression; (4) trends in mathematical modeling including physiologically-based PK modeling and novel animal models such as CRISPR/Cas9-based animal models for DMPK studies; (5) emerging non-classical xenobiotic metabolic pathways and the involvement of novel metabolic enzymes, especially non-P450s. Existing challenges and perspectives on future directions are discussed, and may stimulate the development of new research models, technologies, and strategies towards the development of better drugs and improved clinical practice.

This study aimed to obtain the first national estimate of the prevalence of autism spectrum disorder (ASD) in Chinese children. We targeted the population of 6 to 12-year-old children for this prevalence study by multistage convenient cluster sampling. The Modified Chinese Autism Spectrum Rating Scale was used for the screening process. Of the target population of 142,086 children, 88.5% (n = 125,806) participated in the study. A total of 363 children were confirmed as having ASD. The observed ASD prevalence rate was 0.29% (95% CI: 0.26%-0.32%) for the overall population. After adjustment for response rates, the estimated number of ASD cases was 867 in the target population sample, thereby achieving an estimated prevalence of 0.70% (95% CI: 0.64%-0.74%). The prevalence was significantly higher in boys than in girls (0.95%; 95% CI: 0.87%-1.02% versus 0.30%; 95% CI: 0.26%-0.34%; P < 0.001). Of the 363 confirmed ASD cases, 43.3% were newly diagnosed, and most of those (90.4%) were attending regular schools, and 68.8% of the children with ASD had at least one neuropsychiatric comorbidity. Our findings provide reliable data on the estimated ASD prevalence and comorbidities in Chinese children.