Acute liver injury (ALI) serves as a critical precursor and major etiological factor in the progression and ultimate manifestation of various hepatic disorders. The prevention and treatment of ALI is still a serious global challenge. Given the limited therapeutic options for ALI, exploring novel targeted therapeutic agents becomes imperative. The potential therapeutic efficacy of inhibiting RIPK2 is highlighted, as it may provide significant benefits by attenuating the MAPK pathway and NF-κB signaling. Herein, we propose a CMD-OPT model, a two-stage molecular optimization tool for the rapid discovery of RIPK2 inhibitors with optimal properties. Compound RP20, which targets the ATP binding site, demonstrated excellent kinase specificity, ideal oral pharmacokinetics, and superior therapeutic effects in a model of APAP-induced ALI, positioning RP20 as a promising preclinical candidate. This marks the first application of RIPK2 inhibitors in ALI treatment, opening a novel therapeutic pathway for clinical applications. These results highlight the efficacy of the CMD-OPT model in producing lead compounds from known active molecules, showcasing its significant potential in drug discovery.

Objective: To explore the predictor of lower airway inflammation among the index of nasal inflammation by investigating the expression and association of eosinophils (EOS) in the upper-lower airways and blood of patients with chronic rhinitis. Methods: A total of 162 patients with allergic rhinitis (AR), 117 patients with non-allergic rhinitis (NAR) and 104 controls were enrolled from June 2010 to December 2013 from General Hospital of Eastern Theater Command, People′s Liberation Army. All subjects were required detailed medical history collection and nasal resistance measurement. Skin prick test (SPT), blood total immunoglobin E (tIgE) and blood EOS, nasal lavage and induced sputum EOS, nasal provocation and bronchial provocation test (NPT, BPT), nasal and forced exhaled nitric oxide (NNO, FeNO) were performed in all patients. One-way analysis of variance was used for comparison between groups. LSD t test or Mann-Whitney U test was used for comparison between the two groups. Pearson or Spearman related parameter test was used for correlation analysis. Results: The nasal lavage EOS, NNO, induced sputum EOS, FeNO, blood EOS and tIgE were higher in the AR group than that in the NAR group (3.70[1.20, 14.23]/200 HP vs 1.40[0.20, 3.40]/200 HP, 673.50[466.80, 936.00] ppb vs 455.80[248.10, 705.60] ppb, 2.97[0.00, 10.63]% vs 1.00[0.23, 2.00]%, (49.28±26.37)ppb vs (34.07±19.11)ppb, 4.00[2.00, 7.00]% vs 2.00[1.00, 5.00]%, 208.01[61.70, 387.50] IU/ml vs 43.30[19.00, 122.00] IU/ml, F or χ² value was 11.442, 19.440, 70.727, 69.449, 47.453, 46.525, respectively, all P<0.05). But there was no significant difference in nasal resistance, NPT and BPT between the two groups. Nasal lavage EOS in AR group and NAR group was correlated with induced sputum EOS, FeNO, tIgE and blood EOS (r value of AR group was 0.448, 0.202, 0.159, 0.321, r value of NAR group was 0.442, 0.268, 0.268, 0.334, respectively, all P<0.05), but not with BPT. After adjustment for gender, age, height and weight, nasal EOS was positively correlated with sputum EOS. Multiple linear regression analysis showed that nasal EOS, blood EOS and SPT were factors affecting sputum EOS levels. The optimal threshold for nasal EOS to determine induced sputum EOS was 3.30/200 HP by (receiver operating characteristic,ROC) analysis. Conclusion The nasal EOS is correlated with multiple lower airway and systemic inflammatory markers, and is a risk factor for the induced sputum EOS, which can be used as an inflammation biomarker to predict the lower air inflammation.

Objective: To study the mechanism of vocal mucosal barrier damage mediated by NF-κB and NF-κB-regulated signaling pathway via probing the expression of inflammatory factors and essential proteins for node of NF-κB signaling pathway. Methods: The patients suffering from vocal leukoplakia accompanied with larygopharyngeal reflux(LPR) were treated with oral administration of proton pump inhibitor(PPI). Mucosal specimens of vocal cord were collected from all patients before PPI treatment. And the mucosal specimens of vocal cord were collected from the patients with suspected recurrence at 8 weeks after PPI treatment. HE staining was used to observe the histopathological changes of the mucosa. ELISA was utilized to detect the levels of inflammatory factors including tumor necrosis factor(TNF)-α, interleukin(IL)-1 and IL-6. Western blot was used to detect the expression of p-p65, p-IKK and p-IκB. Immunofluorescence method was adopted to detect the entrance of p65 to cell nucleus.Data was analyzed by SPSS 23.0 software. Results: In PPI untreated group, the expressions of IL-1β, TNF-α and IL-6 in the specimens of 8 weeks after operation were not different significantly from those obtained during operation.But in the PPI-treated group, the expressions were down-regulated.The expression of p-p65 in the middle and high grade heterogenous hyperplasia group was higher than that of low level heterogenous hyperplasia group.The difference of p65 and p-p65 expression between 8 weeks after surgery and surgery in PPI-untreated group was statistically insignificant (P>0.05). The difference of p65 expression between PPI-treated group and PPI pre-treatment group was statistically insignificant (P>0.05). The expression of p-p65 in the PPI-treated group was lower than that of the PPI pre-treatment group (P<0.05). The expressions of IL-1β, TNF-α and IL-6 were positively related with that of NF-κB-p65. Immun of luorescence method revealed the entrance of p65 to cell nucleus in PPI pre-treatment group, which meant that NF-κB was activated. In the PPI-treated group, few activated p65 could be observed in the cell nucleu. Conclusion The possible mechanism of vocal mucosal barrier damage in vocal leukoplakia accompanied with LPR maybe the vocal mucosal inflammation mediated by NF-κB and NF-κB-regulated signaling pathway activated with refluxed materials.

[Abstract ] Objective Clinically, the necessity of acid suppression treatment in vocal leukoplakia is still controversial .This paper aims to investigate the roles of LPR in the pathogenesis and pathological process of vocal leukoplakia , and to clear the signifi-cance of acid suppression in the treatment of this disease through observing the influence of laryngopharyngeal reflux ( LPR) on the symptoms of postoperative vocal leukoplakia . Methods We collected 97 cases underwent vocal leukoplakia surgery from June 2013 to December 2015 in the Department of Otorhinolarygology Head and Neck Surgery , Nanjing General Hospital of Nanjing Military Re-gion.According to the results of ambulatory 24-hour double probe ( simultaneous esophageal and pharyngeal ) pH monitoring ( pH-me-try), the patients with vocal leukoplakia were divided into LPR group (n=26, laryngopharyngeal reflux) and non-LPR group(n=71, non-laryngopharyngeal reflux).Meanwhile, the patients in LPR group were then randomly divided into acid-suppressing group(n=13, oral esomeprazole ) and non-acid-suppressing ( n=13, oral placebo ) . All patients received evaluation and compared by electrolaryngendo-scope, voice handicap index-10 ( VHI-10), reflux symptom index ( RSI) and reflux finding score ( RFS) before operation and 8 weeks after operation, and observe the effect of laryngopharyngeal reflux and acid suppression on the symptoms of postoperative vocal leukoplakia . Results RSI and RFS after operation were significantly lower than before operation in LPR group[(13.6±5.8) vs (18.2±6.2), (9.2±2.4) vs (10.6±2.8), P<0.05].The difference of RSI and RFS between before and after operation in LPR group was higher than the non-LPR group[(5.8±1.4) vs (2.3±0.8), (1.2±0.6) vs (0.5±0.2), P<0.05].The difference of RSI and RFS between before and after operation in acid-suppressing group was higher than the non-acid-suppressing group[(6.6±1.2) vs (0.8±0.6), (2.6±0.6) vs (0.5±0.3), P<0.05].VHI-10 after operation was significantly lower than before operation in acid-suppressing group[(12.6±3.6) vs (15.2±4.2), P<0.05] Conclusion Standard PPI administration to patients suffering from vocal leukoplakia accompanied with LPR can alleviate the symptoms of LPR and improve hoarseness .

Objective Allergic rhinitis (AR) is studied extensively while nonallergic rhinitis (NAR) insufficiently in the recent years .The aim of this study is to describe the inflammation characteristics of different types of NAR . Methods Using the skin prick test , we investigated the characteristics , nasal cytokine levels , serum cytokine levels , and the proportion of peripheral blood Treg cells in 12 cases of AR, 10 cases of NAR with eosinophilia (EOS) syndrome (NARES), 12 cases of NAR without ES (NAR), and 11 control adults . Results The NARES patients had a signifi-cantly higher level of IFN-γ(28.89 [10.97-127.07] pg/mL) than the control (8.98 [7.88-14.90] pg/mL) and the NAR patients (7.92 [7.67-45.85] pg/mL) ( P<0.05) but a lower level of nasal IL-10 than the control ([3.97±0.68] vs [4.80±1.32] pg/mL, P<0.05) .The contents of nasal IL-4, serum IL-4, nasal IL-17 and ser-um IL-17 were all markedly higher in the AR and NARES groups than in the control (P<0.05).The proportion of CD4+CD25+FOXP3+Treg cells in the CD4+T cells in the peripheral blood was (4.5±1.3)%in the AR group and (4.0±1.8) %in the NARES group, both significantly lower than (6.5±1.0) %in the control group (P<0.05) and (6.5±1.0) %in the NAR group (P<0.05). Conclusion NAR was classified according to the EOS level into NARES and NAR without EOS, which had different mechanisms and manifestations of inflammation and similar inflammatory manifestations of Th 2 and Th17.The classification of rhinitis by the level of EOS has more practical significance .

Objective Nonallergic rhinitis with eosinophilia syndrome (NARES) is associated with such diseases as bronchial asthma , nasal polyps , and aspirin intolerance , but there is a lack of studies on its inflammatory conditions .The aim of this study is to describe the inflammation characteristics of NARES . Methods This study included 101 cases of allergic rhinitis ( AR) , 39 cases of NARES, and 162 adult controls .We analyzed the inflammation char-acteristics of the patients using skin prick test ( SPT) , nasal douche , fractional exhaled nitric oxide ( FeNO ) measurement , serum eosino-phil (EOS) counting, serum total IgE (tIgE) determination, induced sputum detection, and nasal and bronchial provocation tests . Results Compared with the controls , the NARES and AR groups showed significant increases in the positive rate of nasal provocation (32.1%vs 69.2%and 75.2%, P<0.05), positive rate of bronchial provocation (1.2%vs 10.3%and 14.9%, P<0.05), and FeNO level ([15.70 ±5.20] ppb vs [37.25 ±22.95] and [39.00 ±24.29] ppb, P<0.05), as well as in the serum EOS and tIgE levels, EOS count, and ratio of EOS in the induced sputum (P<0.05).However, the level of serum tIgE was significantly lower in the NARES than in the AR group (53.3 [23.3-186.0] kU/L vs 197.0 [62.6-391.0] kU/L, P<0.05). Conclusion NARES is a syndrome with nasal, lower airway, and systematic inflammation, similar to AR in inflammatory intensity.Therefore, for patients with NARES, attention should be paid not only to upper airway but also to lower airway and systemic inflammation .

Objective To accumulate operating experience and background data for housing mice in individually ventilated cages (IVC).Methods 5 weeks old Balb /c male mice(n =80) were allocated to 8 groups(n =10), which then housed in 5 or 10 per cage in 3 IVC systems(30,50 and 70 air changes /h, respectively) and one open-top cages (OTC) shelf for 8 weeks.Body weight was assessed at the initial date and every week .By the end of the experiment, necropsy was done and organs were separated and weighed .Excelland SPSS software statistics was made to draw the growth curve, and comparative analysis of body weight and organ coefficients was performed between the groups .Results 1.The growth curves of 5-mice per cage were better than that of 10-mice per cage.2.In the IVC groups, the curves trend and fluency of 50 air changes /h were more similar to that of 5-mice housed OTC group.3.The previously mentioned differences were statistically not significant (P >0.05).4.In the liver coefficients, there was a statistically significant difference between the 10-mice housed OTC group and 5-mice housed IVC group with 30 air changes /h(P <0.05), there wasn`t any other significant statistically difference with the organ coefficients between groups (P >0.05).Conclusions Based on the results of this study, the air change frequency on 50 times per hour and keeping 5 Balb/c mice per cage is recommended as the best condition for mouse housing in IVC .

Objective To investigate the frequency of micresatellite instability (MSI) and loss of heterozygosity (LOH) and select sensitive loci for studying microsatellite DNA imbalance in thymic squamous cell carcinoma. Methods 5 microsat-ellitc polymorphism markers and extrated DNA were selected from 9 specmiens of paired thymie squamous cell carcinoma/nor-real tissues. MSI and LOH in the specmiens of thymic carcinoma and relevant pericancerou tissues were detected by polymerase chain reaction (PCR) followed by 6% polyacrylamide gel electrophoresis(PAGE) with silver staining. Results MSI or LOH was detected in 9 thymic carcinoma tissues. The frequency of MSI or LOH was 66.7% (6/9) at loci of D6S1708, 33.3% (3/9) at TP53, 33.3% (3/9)at DM, 33.3% (3/9)at D11S988 and 0% (0/9)at D8S136, LOH at D6S1708 (5/6) was a common genetic alteration. DI1S988 had only LOH alteration. Conclusion D6S1708, TP53, DM, and D11S988 are sensi-tive loci for studying microsatellite DNA imbalance in thymic squamous cell carcinoma. Microsatellite DNA imbalance may play a certain role in occurrence and development of thymic squamous cell carcinoma, and the relationship between MSi or LOH.The linicopathological features of thymic squamous cell carcinoma needs further investigation.

Objective To investigate a feasibility of using dose constraint template (DCT) to increase conformity index (CI) of planning target volume (PTV) and improve intensity modulated radiation therapy (IMRT) planning efficiency for early stage nasopharyngeal carcinoma. Methods Ten patients with pathological diagnosed and treated by IMRT were selected for this study. Target volumes were delineated with Corvus 6.3 of treatment planning system, two dose limiting regions(DLR) around PIN were added by extending from PIN,each DLR was 1 cm thick. We created three plans:Plan0,Planl and Plan2. PianO was without DLR and DCT, Planl without DLR but with DCT, Plan2 with both condition;but to compare dose distribution in PLTV and normal tissue using three plans. Results Three plans could fill equal request of dose distribution in PLTV and normal tissue, and their difference was not statistical significant. CI of Plan2 was increased and planning time was decreased significantly compared with Piano and Planl. Conclusloa Usage of DCT together with DLR can increase CI of PTV and improve IMRT planning efficiency for early stage nasopharyngeal carcinoma, planning time is shortened significantly.

BACKGROUND: Previous studies have demonstrated that neural stem cells play potential therapeutic effects on the repair of spinal cord injury. However, the time for acquiring the best allotransplantation effects remains unclear.OBJECTIVE: This study was designed to observe the repairing effects of allotransplantation of embryonic neural stem cells on the motor function of rat two posterior limbs after spinal cord injury and investigate the time effectiveness of the allotransplantation.DESIGN: A controlled observational experiment.SETTING: Laboratory of Molecular Biology, Dalian Medical University; Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology, Dalian, Liaoning Province, China.METHODS: This study was performed at the Laboratory of Molecular Biology, Dalian Medical University & Laboratory of Biomedicine, School of Environmental and Biological Science and Technology, Dalian University of Technology between July and August 2003. One albino rat of gestational 14-16 days was sacrificed for harvesting embryonic rat brain cells. Embryonic rat cerebral cortex and subcortical periventricular brain tissue were taken for in vitro culture of rat embryonic neural stem cells. An additional 30 adult Sprague Dawley rats were randomly divided into 3 groups with 10 rats in each group: control, early allotransplantation and delayed allotransplantation groups. All 30 rats were subjected to spinal cord transection injury, leading to rat paralysis of both lower extremities. Embryonic rat neural stem cells were transplanted into the rats in the early and delayed transplantation groups at 3 days and 3 weeks after injury, respectively. Following allotransplantation, motor function of rat two lower extremities was followed. At 4 weeks after allotransplantation of neural stem cells, rat spinal cord was harvested from transplanted region for immunohistochemistry in order to observe and compare the morphological change of rat spinal cord tissue among the 3 groups. The following protocol was performed in accordance with ethical guidelines stated in Guide for the use and care of laboratory animals, approved by the Committee on the Care and Use of Laboratory Animals of the Institute of Laboratory Animal Resources Commission on Life Scineces, National Research Council, China (1985).MAIN OUTCOME MEASURES: Motor functional recovery of rat two lower extremities after neural stem cell transplantation. Histomorphological change of rat spinal cord at 4 weeks after neural stem cell transplantation.RESULTS: Thirty rats were included in the final analysis. In the early and delayed transplantation groups, the motor function of rat two lower extremities was noticeably improved, in particular in the early transplantation group. In the two experimental groups, muscular strength of paralyzed rat two lower extremities began to recover 5 or 6 days after transplantation of neural stem cells. Two or three weeks later, all rats in the two experimental groups could crawl and four weeks later, two extremities could move actively (approximately approaching to score 3 prescribed as follows). In the control group, no recovery of paralyzed extremities was found. At 4 weeks after transplantation, in the early transplantation group, proliferative tissue could be visible in the spinal cord transplantation region. Through the use of microscope, a considerable number of new cells were found that presented with neuronal and glial cell-positive staining. In the control group, a cavity between two broken ends could be visible. Meanwhile, necrosis and vacuolar degeneration, and other symptoms in the stump of spinal cord were observed with a microscope. In the delayed transplantation group, the histomorphological change of spinal cord region was between the other two groups. No typical histomorphological change was found. A number of new cells were apparent with a microscope, but the number was less compared with the early transplantation group.CONCLUSION: Allotransplantation of embryonic neural stem cells promotes the recovery of rat motor function after spinal cord transection. Early transplantation acquires better therapeutic effects.