OBJECTIVE: Based on metabonomics technology of high-performance liquid chromatography-mass spectrometry (HPLC-MS/MS) and hydrogen nuclear magnetic resonance spectroscopy (¹H NMR), the pharmacokinetic characteristics and therapeutic mechanism of Rhei Radix et Rhizoma (RhRR, Dahuang in Chinese), Eupolyphaga Steleophaga (EuS, Tubiechong in Chinese) combined with RhRR acting on acute liver injury were explored. METHODS: Models of acute liver injury were established, and the pharmacokinetic methods of five components of RhRR-EuS in rats were found by HPLC-MS/MS. The liver tissues of different groups of mice were analyzed by ¹H NMR spectroscopy combined with multivariate statistical analysis to investigate the metabolomics of RhRR-EuS and RhRR. RESULTS: Pharmacokinetic results showed there were different levels of bimodal phenomenon in different groups, and the absorption of free anthraquinone in RhRR increased after compatibility with EuS. In addition, the pathological state of acute liver injury in rats can selectively promote the absorption of emodin, chrysophanol, physcion and aloe emodin. Through 15 differential metabolites in the liver tissue of acute liver injury mice, it was revealed that RhRR-EuS and RhRR could protect the liver injury by regulating the metabolism of glutamine and glutamic acid, alanine, aspartic acid and glutamic acid, and phosphoinositide. However, the regulation of RhRR was weaker than that of RhRR-EuS. CONCLUSION For the first time, we studied the pharmacokinetics and metabolomics differences of RhRR-EuS and RhRR in rats and mice with acute liver injury, in order to provide theoretical reference for clinical treatment of liver disease by DHZCP.

Objective:To evaluate the role of Toll-like receptor 4 (TLR4)/nuclear transcription factor κB (NF-κB) signaling pathway in long-term cognitive impairment induced by multiple exposures to sevoflurane anesthesia in neonatal rats.Methods:Seventy-five SPF healthy newborn Sprague-Dawley rats of either sex, aged 6 days, weighing 12-20 g, were divided into 3 groups ( n=25 each) using a random number table method: control group (group C), multiple exposures to sevoflurane anesthesia group (group S) and TLR4 inhibitor plus multiple exposures to sevoflurane anesthesia group (group I+ S). The rats in group S and group I inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. TLR4 inhibitor TAK-242 10 mg/kg was intraperitoneally injected before each exposure to sevoflurane in group I, and the equal volume of normal saline was given instead in the other two groups. The spontaneous activity was evaluated by open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. After the behavioral test, the blood samples from the abdominal aorta were collected, and then the rats were sacrificed under deep anesthesia to isolate the hippocampal tissues for measurement of the levels of S100β and neuron-specific enolase (NSE) in serum and hippocampal interleukin-1β (IL-1β), IL-6 and tumor necrosis factor α (TNF-α) (by enzyme-linked immunosorbent assay), expression of TLR4, NF-κB p65 and phosphorylated NF-κB p65 (p-NF-κB p65) (by Western blot) and for microscopic examination of the pathological changes of hippocampal CA1 region after HE staining. Results:Compared with group C, the escape latency was significantly prolonged, the number of crossing the original platform was reduced, the TLR4 expression was up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was increased, the levels of serum S100β protein and NSE and hippocampal IL-1β, IL-6 and TNF-α were increased ( P<0.05), and the pathological changes in the hippocampal CA1 region were aggravated in group S. Compared with group S, the escape latency was significantly shortened, the number of crossing the original platform was increased, TLR4 expression was down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was decreased, the levels of S100β and NSE in serum and hippocampal IL-1β, IL-6 and TNF-α were decreased ( P<0.05), and the pathological changes in hippocampal CA1 area were significantly attenuated in group P. Conclusions:The mechanism by which multiple exposures to sevoflurane anesthesia induces long-term cognitive impairment is related to activation of TLR4/NF-κB signaling pathway and increase in hippocampal inflammatory responses in neonatal rats.

Objective:To evaluate the role of nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in edaravone-induced attenuation of long-term cognitive impairment caused by long-time sedation with propofol in the neonatal rats.Methods:Eighty SPF healthy newborn Sprague-Dawley rats of both sexes, aged 7 days, weighing 15-20 g, were divided into 4 groups ( n=20 each) using a random number table method: control group (group C), propofol group (group P), edaravone+ propofol group (group EP) and Nrf2 inhibitor ML385+ edaravone+ propofol group (group MEP). Propofol 75 mg/kg was intraperitoneally injected once a day for 7 consecutive days in P group, EP group and MEP group, respectively, while the equal volume of medium/long chain fat emulsion injection was intraperitoneally injected in C group. Edaravone 3 mg/kg was intraperitoneally injected at 30 min before each propofol injection in EP and MEP groups, and ML385 15 mg/kg was intraperitoneally injected simultaneously in group MEP. The spontaneous activity was evaluated by the open field test on day 29 after birth, and the cognitive function was assessed by Morris water maze test on days 30-34 after birth. The rats were sacrificed after the end of water maze test, and brains were removed and hippocampal tissues were obtained for determination of reactive oxygen species (ROS) levels (by flow cytometry), superoxide dismutase (SOD) and malondialdehyde (MDA) levels (by enzyme-linked immunosorbent assay) and expression of Nrf2 and HO-1 (by Western blot) and for microscopic examination of the pathological changes in the hippocampal CA1 area (using HE staining). Results:There was no significant difference in the speed, distance and time of stay at the center of the open field among the four groups ( P>0.05). Compared with C group, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was reduced, the levels of MDA and ROS were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the pathological injury was observed in the hippocampal CA1 region in group P. Compared with P group, the escape latency was significantly shortened, the number of crossing the original platform quadrant was increased, the levels of MDA and ROS in the hippocampus were decreased, the activity of SOD was increased, the expression of Nrf2 and HO-1 was up-regulated ( P<0.05), and the pathological injury in the hippocampal CA1 region was significantly alleviated in EP group. Compared with EP group, the escape latency was significantly prolonged, the number of crossing the original platform quadrant was reduced, the levels of MDA and ROS were increased, the activity of SOD was decreased, the expression of Nrf2 and HO-1 was down-regulated ( P<0.05), and the pathological injury was aggravated in the hippocampal CA1 region in MEP group. Conclusions:The mechanism by which edaravone attenuates long-term cognitive impairment caused by long-time sedation with propofol is related to activation of Nrf2/HO-1 signaling pathway and inhibition of oxidative stress in the neonatal rats.

Objective: To establish a metabonomics research technique based on the combination of

Lung nodules are the main manifestation of early lung cancer. So accurate detection of lung nodules is of great significance for early diagnosis and treatment of lung cancer. However, the rapid and accurate detection of pulmonary nodules is a challenging task due to the complex background, large detection range of pulmonary computed tomography (CT) images and the different sizes and shapes of pulmonary nodules. Therefore, this paper proposes a multi-scale feature fusion algorithm for the automatic detection of pulmonary nodules to achieve accurate detection of pulmonary nodules. Firstly, a three-layer modular lung nodule detection model was designed on the deep convolutional network (VGG16) for large-scale image recognition. The first-tier module of the network is used to extract the features of pulmonary nodules in CT images and roughly estimate the location of pulmonary nodules. Then the second-tier module of the network is used to fuse multi-scale image features to further enhance the details of pulmonary nodules. The third-tier module of the network was fused to analyze the features of the first-tier and the second-tier module of the network, and the candidate box of pulmonary nodules in multi-scale was obtained. Finally, the candidate box of pulmonary nodules under multi-scale was analyzed with the method of non-maximum suppression, and the final location of pulmonary nodules was obtained. The algorithm is validated by the data of pulmonary nodules on LIDC-IDRI common data set. The average detection accuracy is 90.9%.

Objective: To analyze the molecular epidemiology, genetic variations and evolution of enterovirus 71 (EV71) strains isolated in Jiangsu Province from 2009 to 2018. Methods: Statistical methods were used to analyze the data about epidemiological characteristics and results of pathogen detection in cases with EV71 infection in Jiangsu Province from 2009 to 2018. The complete VP1 sequences of 80 EV71 strains were amplified and sequenced for analysis of diversity and phylogenesis. Results: A total of 41 858 enterovirus-positive hand, foot and mouth disease cases were reported in Jiangsu Province from 2009 to 2018. EV71 was the predominant pathogen, accounting for 36.52%, and responsible for most of the severe cases. However, the percentage of EV71 among all pathogens gradually decreased over time. EV71 infection reached the peak in April to June and mainly occurred in children aged six months to five years old with higher incidence in males than in females. In terms of regional distribution, EV71 infections were characterized by area clustering in Jiangsu Province, mainly detected in Nanjing, Suzhou, Wuxi and Lianyungang. The 80 EV71 isolates belonged to C4a genotype. Nucleotide differences between them and three vaccine strains (H07, FY23 and FY7VP5)were 0.6%-5.5%, 0.8%-5.7% and 1.9%-6.9% and amino acid difference were 0-1.4%, 0.3%-2.0% and 0.3%-2.0%, respectively. Amino acid mutations in the epitopes of the 80 EV71 strains did not marked by years or regions. Conclusions EV71 strains showed obvious epidemiological characteristics in time, population and regional distribution in Jiangsu Province from 2009 to 2018.All of the 80 EV71 isolates belonged to C4a subgenotype. The nucleotide sequences between them and the vaccine strains varied greatly, but the homology of amino acids was relatively high, indicating the existence of some synonymous mutations and no risk of antigenic drift. This study would provide reference for EV71 vaccination in Jiangsu Province.

Objective To discuss the method and clinical effect of bilobed DIEP flap for repair of degloving injuries of the hand.Methods From June, 2013 to January, 2015, the bilobed DIEP flap were designed to repair 6 cases of degloving injuries of hand, of which the small leaf flap repaired separately defect of thumb, larger leaf flap repair 2nd-5th finger and palm and dorsum.The bilobed DIEP flap design were based upon anatomic study on deep inferior epigastric artery, superior epigastric artery, intercostal artery and its anastomosis.Results The largest area of main flap was 35 cm × 12 cm, and the deputy leaves of flap was 10 cm× 7 cm.All of the 6 flaps survived.One of them experienced distal end necrosis of epidermis.But it was healed by dressing changes.All of the 6 cases were followed up for average of 12 months (range, 6-20 months).All flaps were seen with good appearance, fully recovered protectve sensation and the active motion of the thumb was recovery.Moreover, the donor sites were closed directly and the appearance were satisfactory.Conclusion The bilobed DIEP flap is a satisfying choice in repairing degloving injuries of the hand in that it has reliable blood supply, can obtain the larger area of skin and leaves small injuries in targeted area.

Objective To discuss the clinical application and evaluate the effect of repairing finger injuries using the perforator flap in forearm cubital fossa.Methods From July,2012 to December,2013,8 cases of finger injuries with totaled defect area of 2.5 cm × 4.0 cm-5.5 cm × 7.0 cm were reviewed.Among them,6 cases had phalangeal fracture,7 cases had neurovascular injury and 4 cases combined with tendon injuries.Cubital fossa flaps based on the inferior cubital perforator of radial artery were transplanted to repair the defects.Results All of the 8 flaps survived.One of them experienced distal end necrosis of epidermis and 1 cm long wound dehiscence.But it was healed by dressing change.All of the 8 cases were followed up for an average of 11 months (range,6-20 months).Sensory quality of （S） + was present in all of the flaps with two-point discrimination ranging between 7.5 mm and 9.8 mm with an average of 8.5 mm.All flaps were seen with good appearance,texture and colour.Moreover,function and appearance in donor sites were satisfactory.Conclusion The perforator flap in forearm cubital fossa is a satisfying choice in repairing small skin defects in fingers and other paas in that it is fixed,easy to dissect and leaves small injuries in targeted area.

Objective To identify the mechanisms underlying xCT deficiency by high-resolution proteomic analysis of differential protein expression in Sut and wild melanocytes .Methods The proteins,extracted from Sut and wild melano-cytes,were analyzed by two dimensional electrophoresis and PDQuest software .Subsequent MALDI-TOF mass spectrometry analysis was carried out .The protein identification was based on peptide mass fingerprint combined with the pI and the rela -tive molecular mass ( Mr) .Sequence coverage was performed with the Peptldent software on the NCBnlm website .Also,the autophagy marker protein LC3-Ⅱ, and the autophagic cell death marker protein Beclin 1 were detected by Western blotting . Results and Conclusion Twenty apparently upregulated or downregulated proteins were identified .Strikingly, important modifications in regulators of trafficking and organization of vesicles and autophagy ( Anxa3; Hist 1h2bk, NDRG1, and CaM) and in regulators of invasion and metastasis of carcinoma (S100A-4;S100A-6 and vimentin)are likely to account for dysfunctions in cell viability and cell-extracellular adhesion .These results indicate that the proteins regulating autophagy , vesicles trafficking and MAP kinase related pathways are activated and play a role in xCT-deficiency .