Objective To establish a rapid and sensitive liquid chromatography-tandem mass spectrometry(LC-MS/MS)analysis method for determination of unsymmetrical dimethylhydrazine(UDMH)contents in rat plasma and investigate the toxicokinetic characteristics of UDMH in rats.Methods Twenty-two SD rats were divided into the intravenous injection of 10 mg/kg dose group(4 females)and intragastric administration groups(low,medium and high dose,with 6 rats in each group,half males and half females).The rats were given 10mg/kg by intravenous administration and 10 mg/kg,30 mg/kg,and 90 mg/kg single dose of UDMH by gavage.Blood samples were collected from the orbital venous plexus at 0 hour before administration and at different time points after administration.The plasma samples were extracted with protein precipitation and derivatization before being analyzed using the LC-MS/MS method.Separation was carried out on a ZORBAX column(4.6mm×75mm,3.5 μm),with a mobile phase composed of 0.3％acetonitrile/formic acid at a flow rate of 1 mL/min.Propranolol was used as the internal standard.An electrospray ionization(Turbo Ionspray)source was applied and the mass spectrometer was operated in a positive MRM mode.Quantitative analysis showed that the ionization source unsymmetrical dimethylhydrazine and propranolol was at m/z:192.0→148.1,m/z:260.2→116.1,respectively.The toxicokinetic parameters were analyzed with the DAS 2.1 software.Results Quantification of UDMH exhibited a good linearity within the concentration range of 50-50000 ng/mL,with a linear correlation coefficient greater than 0.9900 and a lower limit of quantification of 50 ng/mL.The average recovery rate of UDMH was 98.1％,compared with 100.5％for the internal standard propranolol hydrochloride.The inter batch precision of standard curve samples ranged from 0.7％to 6.3％,and the relative error was between-7.1％and 6.2％.The inter batch and intra batch precision of quality control samples ranged from1.8％to 19.8％,and the relative error from-9.8％to 0.2％.The main pharmacokinetic parameters of UDMH in rats exposed to 10 mg/kg,30 mg/kg,and 90 mg/kg gavage were UC(0-t):(7624.99±2569.31),(34284.04±6657.15),(84720.88±22354.80)μg/L·h,t1/2:(0.07±0.15),(2.24±1.45),(3.04±0.90)h,Tmax:(0.75±0.27),(0.51±0.29),(0.29±0.10)h,Cmax:(4454.14±1329.45),(19442.45±9121.07),(32334.35±9882.41)μg/L,F:(77.34±26.06)％,(115.92±22.51)％,(95.48±25.19)％.Conclusion The LC-MS/MS method is highly accurate and specific,and is suitable for the toxicokinetic study of UDMH in rats.Single gavage administration of UDMH results in absorption and elimination saturation at a high dose.This study provides data for toxicological studies related to UDMH.

In order to meet the market demand for the fast and accurate genetic detection of African swine fever virus(ASFV),a new method for EIS-qPCR detection was established with a rapid,high sensitivity,and pollution prevention.Primers and probes for a duplex qPCR were designed based on the conserved region of ASFV virus p72 gene and the endogenous internal standard(EIS)cytb gene sequence in pigs.An anti-contamination system was established with uracil DNA N-gly-cosylase enzyme in the reaction system.The results showed that the method can finish the rapid qPCR detection of ASFV within 30 min with a minimal detection limit of 4.12 copies/μL.Moreo-ver,the method only detected the ASFV p72 gene,and no amplifications of classical swine fever vi-rus(CSFV),pseudorabies virus(PRV),porcine parvo virus(PPV),porcine reproductive and re-spiratory syndrome virus(PRRSV)and porcine circovirus type 2(PCV2)were observed.Repeti-tive results showed a coefficient of variation below 2％.With strong anti-pollution capacity,the method can effectively eliminate false-positive amplification caused by low-dose aerosol pollution.Detection results of 146 clinical samples showed a 100％consistence with the results of the com-mercial ASFV detection kit.Compared with similar technologies,the EIS-qPCR established in this study was faster,sensitive,and suitable for the rapid diagnosis of ASFV infection in the early stage,which provided the tool for the monitoring and precise prevention and control of African swine fever.

Objective:To evaluate the effect of multimodal non-drug treatment during induction of general anesthesia on emergence delirium (ED) in the preschool children.Methods:A total of 210 pediatric patients of either sex, aged 3-6 yr, of American Society of Anesthesiologists Physical Status classification Ⅰor Ⅱ, undergoing elective operation for snoring with expected operation time <2 h, were involved in this study. The patients were divided into 2 groups according to the parity of the numbers randomly generated by the computer: multimodal non-drug treatment group (group N, n=102) and control group (group C, n=108). In group N, multiple modes of non-drug intervention (including parents′ company, carrying favorite toys, watching favorite video programs with portable multimedia devices, etc) were used during anesthesia induction, and the children left their parents and entered the operating room after completion of general anesthesia. The children directly entered the operating room with the medical staff for anesthesia induction (without parents′ company and other intervention measures) in group C. The patients were endotracheally intubated and received combined intravenous-inhalational anesthesia and general anesthesia in both groups. The anxiety was evaluated by modified Yale preoperative anxiety scale (m-YPAS) score at 24 h before operation (T 0) and immediately before induction of general anesthesia (T 1). The Pediatric Anesthesia Emergence Delirium scale score (PAED score, ED was defined as PAED score > 12), FLACC scale score and Ramsay Sedation Scale score were recorded when orientation recovered after admission to postanesthesia care unit (T 2) and at 6, 24 and 72 h after operation. The patients were divided into ED group and non-ED group (NED group) according to the occurrence of ED, and the risk factors of which P values were less than 0.05 would enter the multivariate logistic regression analysis to stratify ED-related risk factors and construct the prediction model. The accuracy of the prediction model was evaluated by the receiver operating characteristic curve. Results:Compared with group C, the m-YPAS at T 1 and PAED score and incidence of ED at T 2 were significantly decreased ( P<0.05), and no significant change was found in FLACC score and Ramsay sedation score at all time points in group N ( P>0.05). Age, m-YPAS score at T 1, multimodal non-drug treatment during anesthesia induction, FLACC score at T 2 and Ramsay sedation score at T 2 were the risk factors for ED ( P<0.05). The area under the receiver operating characteristic curve was 0.944, the 95% confidence interval was 0.914-0.974, with a Yonden index of 0.779, sensitivity of 94.9%, specificity of 83%, and the cutoff value of 0.14. Conclusions:Multimodal non-drug treatment during induction of general anesthesia can effectively reduce the development of ED in the preschool children.

Objective:To explore the influence of selective fetoscopic laser occlusion of chorioangiopagous vessels (FLOC) versus Solomon surgery in the management of twin-to-twin transfusion syndrome (TTTS) on procedure-related complications, fetal survival rate, and residual anastomoses.Methods:A total of 59 pregnant women with TTTS who underwent FLOC in the Third Affiliated Hospital of Zhengzhou University from May 2018 to March 2020 were retrospectively enrolled and divided into Solomon ( n=33) and selective FLOC groups ( n=26) based on the FLOC operation method. Placentae of 25 pregnant women (15 in the Solomon group, 10 in the selective FLOC group) with both survival twins were perfused to observe the type and diameter of the residual anastomoses. Fetal survival rate, procedure-related complications, and the type and diameter of residual anastomoses were analyzed and compared between the two groups using two independent samples t-test, Mann-Whitney U test, χ2 test and Fisher's exact test. Results:(1) The operating time of Solomon was shorter than that of selective FLOC [74 min (60-90 min) vs 95 min (81-123 min), Z=2.906, P=0.004]. But no statistically significant differences in the gestational week at operation, time of pregnancy end, and the interval between operation and pregnancy end was observed between the two groups (all P>0.05). (2) There was no statistically significant difference in the survival rate of both twins, one fetus, at least one fetus, and the incidence of postoperative twin anemia-polycythemia sequence (TAPS) and recurrent TTTS in the Solomon group and selective FLOC group [64%(21/33) vs 50%(13/26), χ2=1.107; 15%(5/33) vs 35%(9/26), χ2=3.044; 79%(26/33) vs 85%(22/26), χ2=0.326; 3%(1/33) vs 12%(3/26), χ2=1.368; 0% (0/33) vs 4%(1/26), χ2=1.118; all P>0.05]. (3) There was no statistically significant difference in the number of placentae with residual anastomoses or the number of artery-to-vein, vein-to-artery, artery-to-artery, and vein-to-vein anastomoses between the two groups (7/15 vs 6/10, 2/8 vs 4/15, 3/8 vs 4/15, 2/8 vs 5/15, 1/8 vs 2/15; Fisher's exact test, all P>0.05), but the diameter of the residual anastomoses in the Solomon group was smaller than that in the selective FLOC group [(0.8±0.3) and (2.2±0.7) mm, t=0.764, P=0.034]. (4) Among the four pregnant women developed TAPS after operation, one patient had two residual artery-to-vein anastomoses in the placenta with diameter of 0.54 mm and 0.43 mm, respectively; one patient had one artery-to-vein anastomosis with a diameter of 0.64 mm; one had one artery-to-artery and one vein-to-vein anastomosis with diameter of 1.56 mm and 1.89 mm, respectively. Conclusions:Compared with selective FLOC, Solomon surgery can reduce the vessel diameter of residual anastomoses in women with TTTS, but does not reduce postoperative complications, nor improve the pregnancy outcomes.

[摘 要] 目的：探索南蛇藤提取物齐墩果烷型五环三萜（28-hydroxy-3-oxoolean-12-en-2-oic acid）协同miR-451对人胃癌AGS细胞增殖、迁移的影响及其可能的分子机制。方法：用miR-451过表达慢病毒感染AGS细胞，并用盐酸多西环素（DOX）10或100 ng/ml诱导24 h，构建过表达miR-451的细胞AGS/miR-451+。采用10、20、40、80、160 μmol/L的齐墩果烷型五环三萜处理AGS/miR-451+细胞，MTT法、划痕实验分别检测细胞增殖和迁移能力的变化，WB法检测细胞中mTOR通路及凋亡相关蛋白表达水平的变化。结果：成功构建过表达miR-451的AGS/miR-451+细胞。与未加药对照组相比，齐墩果烷型五环三萜处理后AGS/miR-451+细胞的增殖抑制率均呈时间和浓度依赖性升高（P<0.05或P<0.01），细胞迁移率均显著降低（P<0.05或P<0.01）。齐墩果烷型五环三萜处理组细胞中，mTOR 信号通路相关蛋白的表达量均有所降低（P<0.05或P<0.01）；凋亡相关蛋白中，Bcl2的表达量下降，BAX、caspase-3、caspase-1及细胞色素c的表达量升高（P<0.05或P<0.01）。结论：齐墩果烷型五环三萜能够协同miR-451抑制人胃癌AGS细胞的增殖与迁移，其机制可能与影响凋亡和mTOR信号通路相关蛋白的表达有关。

BACKGROUND: The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown. METHODS: The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies. RESULTS: We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression. CONCLUSIONS Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.

BACKGROUND: The cardiomyocyte apoptosis is considered as one of major contributions to cardiac remodeling after myocardial infarction (MI). Numerous studies find that circular RNAs (circRNAs) play pivotal roles in a variety of biological functions. However, the role of circ_0068655 in MI and human induced pluripotent stem-derived cardiomyocytes (HCMs) remains unknown. METHODS: The expression of circ_0068655, miR-498, and PRKC apoptosis WT1 regulator (PAWR) in human MI heart tissues and hypoxia subjected HCMs was evaluated with qRT-PCR and Western blot. The effects of circ_0068655 on hypoxia-induced apoptotic death and cell migration in HCMs were evaluated with qRT-PCR, cell viability, cell death ELISA (POD), and Caspase-3 activity assay, and Trans-well assay, respectively. Furthermore, luciferase assay, qRT-PCR, biotin-labeled miRNA pulldown assay, and Western blot were employed in the functional studies. RESULTS: We found that the expression of circ_0068655 and PAWR was enhanced in MI patients and hypoxia subjected HCMs; by contrast, the expression of miR-498 decreased. Inhibited expression of circ_0068655 in HMCs counteracted hypoxia-induced apoptotic death and impaired cell migration, in sharp contrast to circ_0068655 knockdown. We identified that circ_0068655 sponged an endogenous miR-498 to sequester and inhibit its activity, leading to the increased PAWR expression. CONCLUSIONS Our findings reveal that the expression of circ_0068655 can promote cardiomyocyte apoptosis through the modulation of miR-498-PAWR axis in vitro, which highlights the diagnostic and therapeutic value of circ_0068655 in patients with MI.

Objective To investigate the distribution and drug resistance to pathogenic bacterial pathogen in children with community acquired pneumonia (CAP),so as to provide recommendations for clinical rational use of anti-biotics. Methods A retrospective analysis was made on the distribution and drug resistance to bacteria in CAP chil-dren admitted to Department of Respiration,Shanghai Children′s Medical Center from January 2014 to December 2015. Results There were 463 patients with positive sputum culture,and a total of 496 strains of pathogens were found. There were 273 Galanz negative bacteria,195 Galanz positive bacteria and 28 other rare bacteria,accounted for 55. 04％,39. 31％ and 5. 65％ of the total bacteria,respectively. The main pathogens were Streptococcus pneumoniae, Staphylococcus aureus,Haemophilus influenzae,Klebsiella pneumoniae and Escherichia coli. The highest detection rate of bacteria in 1-12 months children with CAP was Staphylococcus aureus,Klebsiella pneumoniae and Escherichia coli;in > 12 months children with CAP,the highest detection rate of bacteria was Streptococcus pneumoniae,Haemophilus influenzae and Staphylococcus aureus. Both of Streptococcus pneumoniae and Staphylococcus aureus had a high resis-tance to Erythromycin,Clindamycin and Oxacillin. There were 11. 00％ Streptococcus pneumoniae and 94. 74％ taphy-lococcus aureus resistant to Penicillin,while they were not resistant to Vancomycin. Escherichia coli and Klebsiella pneumoniae both showed a high resistance to ampicillin,the second and third generation cephalosporins. Haemophilus influenzae were highly resistant to Compound sulfamethoxazole and Ampicillin. Galanz negative bacteria had the lowest resistance to Piperacillin/ Tazobactam and Amikacin. Conclusions The main pathogens of CAP in children were G -bacteria. There were some differences among the isolates at different ages of CAP. Their resistance to very common anti-biotics was very high in children.

OBJECTIVETo observe the regulatory effects of acupoint electric stimulation on the analgesic substances and the relevant indices of nerve-immunity-endocrine system in the patients undergoing general anesthesia anorectal operation.

METHODSOne hundred and fifty-six patients undergoing hemorrhoids and anal fistula operation were randomized into three groups, 146 cases were included in the analysis. In the No.1 group (48 cases), the conventional intravenous general anesthesia was applied. In the No.2 group (50 cases), besides the conventional intravenous general anesthesia, the acupoint transcutaneous electric stimulation was combined at Neiguan (PC 6), Shenmen (HT 7), Shangliao (BL 31) and Ciliao (BL 32). operation in the No.2 and No.3 groups were lower apparently than that in the No.1 group (<0.05,<0.01).

CONCLUSIONSDuring the general anesthesia anorectal operation, the acupoint transcutaneous electric stimulation achieves analgesic anesthesia through effectively promoting the release of body analgesic substance and reducing the stress level in the operation. With the comprehensive acupoint selection as Neiguan (PC 6) and Shenmen (HT 7) and the local acupoints, the therapeutic effects are better in comparison with the simple selection of local acupoints. In the No.3 group (48 cases), besides the conventional intravenous general anesthesia, the acupoint transcutaneous electric stimulation was combined at Shangliao (BL 31) and Ciliao (BL 32). The electric stimulation was maintained till the end of operation. The patients' saliva was collected 0.5 h before operation and 1 h after operation separately. The indices that reflect the body pain regulation and nerve-immune-endocrine secretion were detected, such as opiophin protein (OPI), secretory immunoglobulin A (SIgA), saliva amylase (sAA), cortisol (Cor) and tumor necrosis factor α (TNF-α). The pain degree was observed 1 h after operation.

RESULTSIn the No.2 group, OPI after operation was higher than that before operation (<0.05). The difference value of OPI in the No.2 group was higher apparently than that in the No.1 group and the No.3 group (both<0.05). SIgA after operation was higher than that before operation in the No.1 group (<0.05). The difference values of SIgA, sAA, Cor in the No.2 group were lower apparently than those in the No.1 group (<0.05,<0.01). TNF-αbefore and after operation and its difference value among the groups were not significant statistically (all>0.05). The pain degrees in 1 h after.

Objective To assess the efficacy of oral treatment and parenteral treatment in community acquired pneumonia( CAP) children by meta-analysis method. Methods Searches were made in MEDLINE、EMBASE and Cochrane Central Register of Controlled Trials ( CENTRAL ) from the establishment of the data base till September 2016. All randomized controlled trials about oral and parenteral treatment in community ac-quired pneumonia children were eligible. Review Manager 5. 3 was used to analyze the studies enroued in this meta-analysis. Results 4582 literatures were reviewed. Seven(n=5030)eligible trials were used for meta-a-nalysis. The treatment failure between community acquired pneumonia children treated with oral treatment and parenteral treatment was found no significant difference(OR =0. 82, 95% CI =0. 63-1. 08,P <0. 05). The treatment failure of oral treatment group was found to be significantly higher than parenteral treatment group in CAP children under 1 year of age(OR=2. 25,95%CI=1. 61-3. 14,P<0. 01). The treatment failure of children who had used antibiotics before included in the study was found to be significantly higher than those who had not used(OR=1. 94,95%CI=1. 50-2. 50,P<0. 01). The death rate of oral treatment group was found to be signif-icantly lower than the parenteral treatment group(OR=0. 31,95%CI=0. 11-0. 85,P=0. 02). There was no sig-nificant difference of relapse rate (OR=1. 28,95%CI=0. 34-4. 82) and loss to follow-up rate(OR=1. 08,95%CI=0. 77-1. 51) between the two group. Conclusion Oral treatment is as effective as parenteral treatment in CAP children. The death rate of oral treatment group is significantly lower than the parenteral treatment group.