Objective　To investigate the effect of andrographolide on enterovirus 71 (EV71) and coxsackievirus A16 (CA16) in vitro. Methods Cytopathic CPE assay and thiazolyl blue tetrazolium bromide (MTT) assay were used to determine the maximum non-toxic dose of andrographolide on RD cells and the inhibitory effect of andrographolide on EV71 and CA16 infection in vitro. The effects of andrographolide on the virus VP1 gene and cellular inflammatory factors (IL-1β, IL-6, and TNF-α) at gene levels were detected by quantitative fluorescence PCR. Results MTT results showed that the maximum non-toxic dose of andrographolide on RD cells was 78.00 μg/mL. Upon infection with hand-foot-mouth disease viruses, cells treated with andrographolide solutions at concentrations of 78.00, 39.00, 19.50, 9.75, 4.88, and 2.44 μg/mL showed increased survival rates to (82.41±1.76)%, (79.54±2.91)%, (81.02±1.99)%, (71.81±2.26)%, (52.87±1.51)%, and (50.41±0.93)% for EV71 and (81.00±0.64)%, (79.72±1.38)%, (61.59±3.47)%, (53.37±0.53)%, (52.41±1.37)%, and (43.69±0.40)% for CA16, respectively, indicating a significant reduction in the cytopathic effect caused by EV71 and CA16. When infected with hand-foot-mouth viruses and treated with andrographolide solutions at concentrations of 78.00, 39.00, 19.50, and 9.75 μg/mL, the expression levels of VP1 in EV71 and CA16, along with pro-inflammatory factors IL-1β, IL-6, and TNF-α in RD cells, were significantly lower compared to the virus control group. These results indicated that after infection with hand-foot-mouth viruses, treatment with andrographolide solution significantly inhibits the expression of the VP1 gene and reduces the mRNA levels of IL-1β, IL-6, and TNF-α. Conclusions Andrographolide exhibits obvious in vitro antiviral effects against EV71 and, potentially through the inhibition of the expression of pro-inflammatory factors IL-1β, TNF-α, and IL-6, thereby exerting its antiviral effects.

In order to analyze the clinical characteristics of death cases in the oral emergency depart-ment of the stomatological hospital,and to improve the first aid technique before and in hospitals,we col-lected the clinical data of death cases in the Department of Oral Emergency in Peking University School and Hospital of Stomatology from January 2010 to January 2023 for retrospective analysis.General cha-racteristics,such as the patients'gender,age,chief complaint,maxillofacial diseases,systemic under-lying diseases,rescue situation,cause of death and seasonal distribution of death were summarized.The results showed that a total of 8 death cases(5 males and 3 females)occurred during the 13-year period,ranging in age from 40 to 86 years,with a median age of 66 years.Among the 8 patients,5 reported bleeding from oral cancer,1 reported chest tightness and dyspnea after oral cancer surgery,1 reported loss of consciousness after maxillofacial trauma,and 1 reported oral erosion and aphagia.All the 8 pa-tients had one or more underlying diseases,such as hypertension,coronary heart disease,diabetes,renal failure,and cerebral infarction,etc.and 2 of them showed dyscrasia.Among them,the cause of death in 5 cases was respiratory and circulatory failure caused by oral cancer rupture and hemorrhage or poor sur-gical wound healing and hemorrhage;1 case was uremia and hyperkalemia leading to circulatory failure;1 case was asphyxia caused by swelling of oral floor tissue after maxillofacial trauma;and the other case was acute myocardial infarction caused circulatory failure after oral cancer surgery.According to the vital state at the time of treatment,6 patients had loss of consciousness,respiratory and cardiac arrest before hospital,and 2 patients suffered from loss of consciousness,respiratory and cardiac arrest during treat-ment.All the patients received cardiopulmonary resuscitation and some advanced life support measures,and the average rescue time was 46 min.Due to the low incidence of death in the oral emergency depart-ment,medical personel have little experience in first aid.First aid training and drills and assessment should be organized regularly.First aid facilities should be always available and regularly maintained by special personnel,such as electrocardiogram(ECG)monitor,defibrillator,simple breathing apparatus,oxygen supply system,negative pressure suction system,endotracheal intubation and tracheotomy equip-ment.The death cases mainly occurred in the elderly patients with oral cancer bleeding and systemic un-derlying diseases.Education of emergency awareness for the elderly patients with oral cancer after surgery should be enhanced.Medical staff should strengthen first aid awareness and skills.

Objective: In this study, phage display technology was used to construct the human anti-Zika virus(ZIKV), phage antibody library and to obtain and express the monoclonal antibody. The aim was to master the preparation and expression of human phage antibody library screening method for highly specific antibodies. Methods: The whole blood samples of Zika patients were collected and the lymphocytes were isolated. The RT-PCR method was used to amplify the antibody light chain and heavy chain Fab gene from lymphocyte Ig mRNA. The pComb3H system was used to construct the gene with genetic diversity Preparation of human anti-ZIKV phage antibody library. The purified antibody library was screened by using the purified ZIKV and the obtained ZIKV E protein antigen. Results: The monoclonal antibody Fab fragment gene was successfully obtained for the ZIKV E protein antigen. The gene can be efficiently expressed in Escherichia coli. Conclusions According to the sequence analysis, this study showed that the monoclonal antibody was a new human genetically engineered antibody against ZIKV, which laid the foundation for the early diagnosis of ZIKV, and obtain a specific monoclonal antibody to ZIKV for human treatment of ZIKV infection.

Objective: To develop a micro-neutralization test for determination of neutralizing antibody against ZIKA virus (ZIKV) in human sera and to verify the acute and convalescent serum samples of 10 ZIKA virus-infected cases diagnosed by nucleic acid detection and/or virus isolation. Methods: ZIKV isolated from ZIKA cases was used to determine micro-neutralization antibody. The virus solution was prepared by infecting BHK21, VERO and VERO-E6 cell lines and viral titer was tested; 100 TCID₅₀ viral solution and 4 times diluted sera which were inactivated at 56 ℃ for 30 min were neutralized, then added the cell suspension and incubated in 5% CO₂ incubator at 37 ℃ for 7 d. The CPE was observed every day. Results: The sensitivity of BHK21, VERO and VERO-E6 was different after infection with ZIKA virus. VERO cell line was the most sensitive and showed typical CPE. VERO cell line was used to establish a micro-neutralization test for determination of neutralizing antibody against ZIKA virus in sera. Conclusions The neutralizing antibody test for zika virus in sera is a special and usefulmethod to diagnose human infection of ZIKV and to conduct population based epidemiological investigation.

Objective To determine the association of high IL-1β levels with the migration of lung cancer H460 cells with acquired resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Methods The resistant cells were referred to as H460-TR in this study. IL-1β levels in H460-TR cells and the parent H460 (H460-WT) cells were measured through RT-PCR, Western blot, and enzyme-linked immunosorbent assay. The migration capacity of the cells was determined using the migration transwell assay. The extent of migration and the activation of phosphatidyl-inositol 3-kinase/serine-threonine kinase (PI3K/Akt) were detected in H460 cells treated with or without human recombinant IL-1 or IL-1R antagonist. Results Migration capacity of H460-TR cells in the conditioned medium and its IL-1β level were higher than those of H460-WT cells . The migration capacity and Akt activation were consistent with the IL-1β level in lung cancer H460 cells. Conclusions Significantly elevated IL-1β expression in cancer cells is associated with the high migration capacity of H460-TR cells, and Akt activation. Akt signaling as the downstream pathway of IL-1β and IL-1βmay function as a therapeutic target for metastatic lung cancer.