Objective:To explore the effect of intragastric administration of diphenoxylate on inducing epilepsy in mice, as well as the changes in neurobehavioral and hippocampal neurons in mice.Methods:Forty C57 male mice aged 2-3 weeks were selected and divided into control group and diphenoxylate group using random number table method, with 20 mice in each group. The mice in the diphenoxylate group were given diphenoxylate (200 mg/kg) by gavage once a day for 14 consecutive days, while the control group mice were given an equal volume of 0.9% sodium chloride solution every day. After each gavage, the seizure status of mice within 2 hours was observed and the mice were graded based on the Racine score. Open field test, elevated cross test, and Morris water maze test were used to observe the neurobehavioral activities of mouse.A digital electroencephalogram machine was used to monitor the epileptic seizures of mice induced by diphenoxylate.Nissl staining was used to observe the damage of hippocampal neurons in mice.SPSS 25.0 software was used for statistical analysis of the data, and independent sample t-test was used for pairwise comparison. Results:The Racine grading results showed that the mice in diphenoxylate group exhibited grade 2 and 3 seizures at 1 hour after gavage. The EEG monitoring results showed that compared with before gavage, the frequency and amplitude of brain waves of mice in diphenoxylate group increased.In the open field test, the residence time in the central region of mice in diphenoxylate group was significantly lower than that of the control group ((12.21±3.37)s, (17.05±4.34)s, t=3.29, P<0.01). In the elevated cross test, the residence time in the open arm of mice in diphenoxylate group was significantly lower than that of the control group ((17.36±5.41)s, (26.70±9.06)s, t=3.31, P<0.01). In the Morris water maze test, the residence time in the platform quadrant of diphenoxylate group was significantly lower than that of the control group((22.08±6.76)s, (27.64±4.60)s, t=2.54, P<0.05). The residence time and the number of stays in the platform area of diphenoxylate group were both significantly lower than those of the control group(both P<0.05). Nissl staining showed that the number of hippocampal neurons in the CA3 region of mice treated with diphenoxylate was significantly lower than that in the control group((135.67±4.59), (140.67±2.73), P<0.05). Conclusion:Excessive diphenoxylate can induce seizures in mice, and the mice exhibit increased anxiety-like behavior and decreased spatial learning and memory abilities.

Objective:To preliminarily investigate the effect of circIGF2BP3 on autophagy in photoaged dermal fibroblasts.Methods:Human dermal fibroblasts were isolated from circumcised foreskin tissues from 6 children in the Department of Urological Surgery, Third Affiliated Hospital of Sun Yat-sen University. An ultraviolet A (UVA) -induced photoaged human dermal fibroblast model (UVA radiation group) was established by repeated UVA radiation at a dose of 10 J/cm 2 for 14 consecutive days, and human dermal fibroblasts receiving no treatment served as control group. The photoaged cell model was verified by β-galactosidase staining, Western blot analysis for determining P21 protein expression, and cell counting kit-8 (CCK8) assay for evaluating cell viability. Moreover, Western blot analysis was performed to determine the protein expression of autophagy-related proteins P62, microtubule-associated protein 1 light chain 3 (LC3) -Ⅰand LC3-Ⅱ in photoaged human dermal fibroblasts, and real-time quantitative RCR (qRT-PCR) to verify the differential expression of circIGF2BP3 between photoaged and normal human dermal fibroblasts. Furthermore, circIGF2BP3 was biologically annotated. Some cultured primary human dermal fibroblasts were divided into 4 groups: empty vector group transfected with an empty vector, UVA + empty vector group transfected with an empty vector followed by repeated UVA radiation, circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector, UVA + circIGF2BP3 group transfected with a circIGF2BP3-overexpressing lentiviral vector followed by repeated UVA radiation. Western blot analysis was performed to determine the expression of autophagy-related proteins. Statistical analysis was carried out by using t test, one-way analysis of variance and least significant difference- t test. Results:Compared with the control group, the UVA radiation group showed significantly increased proportions of β-galactosidase-positive cells (61.33% ± 5.78% vs. 6.37% ± 0.32%, t = 9.49, P < 0.01) and P21 expression (1.25 ± 0.03 vs. 1.00 ± 0.05, t = 4.26, P < 0.05), but significantly decreased cell viability (74.33% ± 3.48% vs. 100%, t = 7.38, P < 0.01). Moreover, the P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly higher in the UVA radiation group than in the control group (both P < 0.05). The relative expression of circIGF2BP3 was 0.72 ± 0.04 in the photoaged human dermal fibroblasts, which was significantly lower than that in the normal human dermal fibroblasts (1.00 ± 0.03, t = 5.46, P < 0.01). The P62 expression and LC3-Ⅱ/Ⅰ ratio were significantly lower in the circIGF2BP3 group (0.60 ± 0.01, 0.71 ± 0.01, respectively) than in the empty vector group (1.00 ± 0.02, 1.00 ± 0.01, t = 16.25, 2.75, P < 0.01, < 0.05, respectively), and lower in the UVA + circIGF2BP3 group (1.05 ± 0.02, 2.04 ± 0.05, respectively) than in the UVA + empty vector group (1.31 ± 0.02, 2.72 ± 0.14, t = 10.493, 6.472, respectively, both P < 0.01) . Conclusion:circIGF2BP3 can regulate autophagy in UVA-induced photoaged dermal fibroblasts, which provides a new potential therapeutic target for the prevention and treatment of skin photoaging.

Influenza C virus is an important respiratory pathogen not only infecting people, but also pigs, dogs, and other animals. Polymerase is central to the replication of influenza C virus and is an important target for studying the mechanism of viral replication. However, there is no commercial monoclonal antibody (MAb) targeting influenza C virus polymerase, which hampers the development of relevant research to some extent. In order to prepare MAb targeting the polymerase basic protein 2 (PB2) of influenza C virus, influenza C virus RNA-dependent RNA polymerase (RdRp, consists of PB1, PB2 and P3) was co-immunoprecipitated with Flag-tagged human acidic nuclear phosphoprotein 32A (huANP32A-Flag) from 293T cells based on the interaction between huANP32A and influenza virus RdRp. The purified RdRp was used as antigen to immunize BALB/c mice. Six positive hybridoma cell lines (7B11-5, 8A4-5, 13D9-6, 8D4-1, 8D4-3, 9F9-4) that stably secrete and recognize PB2 MAb were screened by indirect ELISA and Western blotting. The subtypes of MAb 7B11-5, 8A4-5, 8D4-1 and 8D4-3 antibody were identified as IgG1, the subtypes of MAb 13D9-6 and 9F9-4 were IgG2a and IgG3, respectively. All the light chains of the MAbs were κ chain. A hybridoma cell line 8D4-1 with high titer was further selected to prepare ascites. The titer of mouse ascites antibody was determined to be 1:64 000. Western blotting results showed that the MAb 8D4-1 had a specific immune response with ICV PB2; laser confocal assay showed that the prepared MAb 8D4-1 accurately detected the subcellular localization of PB2 subunits. Moreover, ICV RdRp was highly enriched by ANP32A. The high specific of the prepared PB2 MAb 8D4-1 may facilitate the polymerase detection, structural analysis and mechanism study of influenza C virus.
Animals ; Antibodies, Monoclonal/metabolism* ; Ascites ; Humans ; Influenzavirus C/metabolism* ; Mice ; Nuclear Proteins/metabolism* ; RNA-Binding Proteins ; RNA-Dependent RNA Polymerase/genetics* ; Viral Proteins/metabolism* ; Virus Replication

Objective:To investigate the expression of myeloid-derived suppressor cells (MDSCs) and cytokines in peripheral blood of patients with ankylosing spondylitis (AS).Methods:From June 2018 to June 2019, 50 AS patients (AS group) and 50 healthy people (control group) in Lishui People's Hospital were selected in this study.The number of PMN-MDSC and monocyte MDSC(M-MDSC) and the expression of cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, γ-IFN and IL-17) in the peripheral blood were detected by flow cytometry.The differences of MDSCs expression frequency in each group and its relationship with cytokine expression were analyzed.Results:The PMN-MDSC, M-MDSC and IL-6 in peripheral blood of the AS group were (0.22±0.08)%, (1.48±0.32)% and (20.74±5.14)ng/L, respectively, which were significantly higher than those of the control group[(0.06±0.02)%, (0.43±0.11)% and (2.52±0.53)ng/L], and the differences were statistically significant ( t=7.933, 6.310, 10.553, all P<0.05). Conclusion:IL-6 may be one of the reasons to promote the infiltration and recruitment of MDSCs, and the increase of MDSCs may be related to the occurrence and development of immune tolerance after ankylitis infection.

Objective Introduce the experience of open window thoracostomy in the treatment of bronchopleural fistula after pulmonary resection.To explore which patients are currently suitable for open window thoracostomy , how to deal with them after open window thoracostomy, and how to treat patients without window drainage.Methods In 2017, the thoracic surgery department of Shanghai Pulmonary Hospital completed 13,341 thoracic surgeries, including 10 cases of open window thoracos-tomy, and patients with BPF after other pulmonary resection were treated with conservative thoracic closed drainage .Thoracic closed drainage therapy is often accompanied by thoracic irrigation.From January 2004 to December 2017, 21 cases of chronic refractory abscess treated with autologous musculocutaneous flap implantation after pulmonary resection and open window drain-age were summarized.The treatment of chronic refractory abscess after 14 years of diagnosis was divided into three stages.The first stage is opening the abscess cavity stage, namely opening the window drainage.The second stage is elimination of abscess cavity and closure of bronchial pleural fistula.The third stage is autologous musculocutaneous flap transplantation or displace-ment to fill the abscess cavity stage.Results Compared with before open window, the 10 patients with open window thoracos-tomy showed obvious improvement in thoracic and pulmonary infection, without perioperative death.Other patients with BPF af-ter pulmonary resection without open window thoracostomy died in 2 of conservative thoracic closed drainage .From January 2004 to December 2017, 19 patients(19/21) were successfully treated with autologous musculocutaneous flap implantation af-ter pulmonary resection and open window thoracostomy, without recurrence of empyema and necrosis of skin flap, and 2 cases (2/21) were not cured due to large bronchial fistula, and local recurrence of empyema, without perioperative death.Conclu-sion Most patients with BPF after pulmonary resection are treated with closed thoracic drainage , especially those with lower lo-bectomy and with pleural irrigation.Most patients can be cured.If patients with upper lobe, middle and upper lobectomy or pneumonectomy, accompanied by BPF, chest infection and poor drainage, it is easy to develop intrapulmonary infection sprea-ding.We should do open window thoracostomy as soon as possible.The removal of the residual cavity by filling musculocutane-ous flap after open window thoracostomy is a great improvement compared with the transthoracic reconstruction .

Non-arterial Inflammatory ischemic optic neuropathy(NAION)is a common ocular disease in middle and old ages.Symptoms of optic nerve dysfunction in NAION are caused by cerebral ischemia,which leads to optic atrophy.Recent studies have focused on the early interventions targeting NAION′s risk factors to protect the optic nerve and retinal ganglion cells.This article reviews recent progress in studies on the etiology and risk factors of NAION and potential therapies that may help to preserve optic nerve function in NAION.

Objective To summarize experience in the treatment of chronic refractory empyema with autologous myocutaneous flap implantation.Methods From January 2004 to December 2017,26 patients had been treated with autologous myocutaneous flap implantation in Shanghai Pulmonary Hospital for chronic refractory empyema.Among them,24 were men and 2 were women.The mediam age was 50.1 years(14-74 years).21 of them had medical histories of lung resection because of basic diseases(most of which accepted surgeries in other hospitals).Complications appeared after surgeries.15 of them had bronchopleural fistula while windowing,which could not be cured by conservative treatments such as drainage.Then we performed open-window thoracostomy and long-time dressing.6 of 21 had experienced pneumonectomy.Other 5 patients did not have primary operational histories.They experienced dressing by windowing because of chronic refractory empyema after the in effective conservative treatments like drainage without pulmonary re-expansion.Results No respiratory complications occurred in these patients.The catheters were successfully removed within 5 days and the patients were discharged within 3-6 weeks after the operations.The median follow-up period was 9 months.24 cases were successful with no recurrence of empyema or flap necrosis,the other 2 cases underwent recurrence of empyema.Conclusion The application of autologous myocutaneous flaps for the treatment of chronic refractory empyema is an effective and continuously improving method.

This paper was aimed to study the albiflorin activity of Tang-Bi-Kang (TBK) granules in protecting Schwann cells (SCs) of rat's sciatic nerve.The establishment of SCs oxidative stress model was the condition of 150 mmol×L-1 Dglucose with different concentrations.And the incubation time was 48 h.The experiment groups were the high-dose,middle-dose and low-dose (100 μM,20 μM,4 μM) albiflorin group,the model group,the vitamin C (100 μM) group,and the normal group.Flow cytometry was used to detect the content of ROS in SCs.Fluorescence microscope was used to observe the condition of ROS fluorescence in SCs.And CCK-8 was used to detect the cell activity.The results showed that by using CCK-8 to detect cell proliferation,after 48 h,there was a significant difference between the model group and the normal group (P＜0.01);the albiflorin group compared with the model group (P＜0.01).It indicated that albiflorin can promote the proliferation of SCs.Detecting the ROS fluorescence content,it showed that compared with the model group (Glu 150 mM),the 100 μM,20 μM,and 4 μM albiflorin group,it was P＜0.01 for each group.It showed that albiflorin could relieve the ROS in SCs and alleviate oxidative stress.It was concluded that albiflorin can increase the proliferation of SCs and improve the state of oxidative stress with the protection of SCs.

Through comprehensively characterizing components in blood after oral administration of Tang-Bi-Kang (TBK) granules by UPLC-ESI-MSn,this study was aimed to explain the pharmaceutical material basis of TBK initially.UPLC-LTQ-Orbitrap was used under both positive and negative ion modes of electrospray ionization.The blank serum and rat serum after oral administration of TBK were analyzed.Components in rat serum were identified and characterized based on ion fragment information,evenelectron law,nitrogen rule and so on.Reference data was used to establish the UPLC-ESI-MSn method.The results showed that after oral administration of TBK granules,15 components were detected in the serum,of which 13 components were taken as the prototype to blood and 2 metabolites.It was concluded that constituents of TBK granules in rat serum were generated from compatibility of all herbal medicines.In rat serum,most of the components had been absorbed by rat's metabolism;a few were absorbed as the prototype.This research provided references for pharmacodynamic material basis and metabolism of TBL granules in vivo.